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Homology and homoplasy of swimming behaviors and neural circuits in Nudipleura molluscs


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How neural circuit evolution relates to behavioral evolution is not well understood. Here the relationship between neural circuits and behavior is explored with respect to the swimming behaviors of the Nudipleura (Mollusca, Gastropoda, Opithobranchia). Nudipleura is a diverse monophyletic clade of sea slugs among which only a small percentage of species can swim. Swimming falls into a limited number of categories, the most prevalent of which are rhythmic left-right body flexions (LR) and rhythmic dorsal-ventral body flexions (DV). The phylogenetic distribution of these behaviors suggests a high degree of homoplasy. The central pattern generator (CPG) underlying DV swimming has been well characterized in Tritonia diomedea and in Pleurobranchaea californica. The CPG for LR swimming has been elucidated in Melibe leonina and Dendronotus iris, which are more closely related. The CPGs for the categorically distinct DV and LR swimming behaviors consist of nonoverlapping sets of homologous identified neurons, whereas the categorically similar behaviors share some homologous identified neurons, although the exact composition of neurons and synapses in the neural circuits differ. The roles played by homologous identified neurons in categorically distinct behaviors differ. However, homologous identified neurons also play different roles even in the swim CPGs of the two LR swimming species. Individual neurons can be multifunctional within a species. Some of those functions are shared across species, whereas others are not. The pattern of use and reuse of homologous neurons in various forms of swimming and other behaviors further demonstrates that the composition of neural circuits influences the evolution of behaviors.
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Homology and homoplasy of swimming behaviors and
neural circuits in the Nudipleura (Mollusca,
Gastropoda, Opisthobranchia)
James M. Newcomb
, Akira Sakurai
, Joshua L. Lillvis
, Charuni A. Gunaratne
, and Paul S. Katz
Department of Biology, New England College, Henniker, NH 03242; and
Neuroscience Institute, Georgia State University, Atlanta, GA 30302
Edited by John C. Avise, University of California, Irvine, CA, and approved April 23, 2012 (received for review February 29, 2012)
How neural circuit evolution relates to behavioral evolution is not
well understood. Here the relationship between neural circuits
and behavior is explored with respect to the swimming behaviors
of the Nudipleura (Mollusca, Gastropoda, Opithobranchia). Nudi-
pleura is a diverse monophyletic clade of sea slugs among which
only a small percentage of species can swim. Swimming falls into
a limited number of categories, the most prevalent of which are
rhythmic leftright body exions (LR) and rhythmic dorsalventral
body exions (DV). The phylogenetic distribution of these behav-
iors suggests a high degree of homoplasy. The central pattern
generator (CPG) underlying DV swimming has been well charac-
terized in Tritonia diomedea and in Pleurobranchaea californica.
The CPG for LR swimming has been elucidated in Melibe leonina
and Dendronotus iris, which are more closely related. The CPGs for
the categorically distinct DV and LR swimming behaviors consist of
nonoverlapping sets of homologous identied neurons, whereas
the categorically similar behaviors share some homologous iden-
tied neurons, although the exact composition of neurons and
synapses in the neural circuits differ. The roles played by homolo-
gous identied neurons in categorically distinct behaviors differ.
However, homologous identied neurons also play different roles
even in the swim CPGs of the two LR swimming species. Individual
neurons can be multifunctional within a species. Some of those
functions are shared across species, whereas others are not.
The pattern of use and reuse of homologous neurons in various
forms of swimming and other behaviors further demonstrates
that the composition of neural circuits inuences the evolution
of behaviors.
rhythmic movement
species differences
ehavior and neural mechanisms can be considered to rep-
resent two different levels of biological organization (14).
Nevertheless, the evolution of behavior and the evolution of
neural circuits underlying behavior are intertwined. For example,
it has been suggested that the properties of neural circuits affect
the evolvability of behavior; the evolution of particular behaviors
could be constrained or promoted by the organization of neural
circuits (59). Darwin and the early ethologists recognized that
behaviors, like anatomical features, are heritable characters that
are amenable to a phylogenetic approach (1013). The use of
behavioral traits to determine phylogenies has been validated
several times (1417), and the historical debates about homology
and homoplasy of behavior have been thoroughly reviewed (24,
15, 17, 18). Examining the neural bases for independently
evolved (i.e., homoplastic) behaviors within a clade could pro-
vide insight into fundamental aspects of neural circuit organi-
zation. However, it is difcult enough to determine the neural
basis for behavior in one species. Doing this in several species
with quantiable behaviors is even more challenging.
Studies of the neural bases of swimming behaviors in the
Nudipleura (Mollusca, Gastropoda, Opisthobranchia) offer such
a possibility. These sea slugs exhibit well differentiated catego-
ries of swimming behaviors, and their nervous systems have large
individually identiable neurons, allowing the neural circuitry
underlying the swimming behaviors to be determined with
cellular precision.
Here we will summarize what is known about the phylogeny of
Nudipleura, their swimming behaviors, and the neural circuits
underlying swimming. We will also provide data comparing the
roles of homologous neurons. We nd that neural circuits un-
derlying the behaviors of the same category are composed of
overlapping sets of neurons even if they most likely evolved in-
dependently. In contrast, neural circuits underlying categorically
distinct behaviors use nonoverlapping sets of neurons. Further-
more, homologous neurons can have different functions in dif-
ferent behaviors and even in similar behaviors.
Phylogeny of Nudipleura
The Nudipleura form a monophyletic clade within Opistho-
branchia (Gastropoda) that contains two sister clades: Pleuro-
branchomorpha and Nudibranchia (1921) (Fig. 1). Molecular
evidence suggests that the two sister groups separated approxi-
mately 125 Mya (21). Nudibranchia (or, informally, nudi-
branchs), which are shell-less and have a slug-shaped appearance
with naked gills, were traditionally classied as their own or-
der. The most recently agreed upon taxonomic classication
system for nudibranchs uses unranked clades instead of orders,
suborders, and superfamilies (22). There are at least 2,000 to
3,000 identied nudibranch species (23). Studies that used
morphological and molecular data support the monophyly of
Nudibranchia (1921, 2426).
Within Nudibranchia, there are two monophyletic clades (19):
Euctenidiacea (Anthobranchia) (27, 28) and Cladobranchia (25).
Euctenidiacea includes Doridacea, which is larger than Clado-
branchia, subdividing into 25 families (28). Within Cladobranchia,
Bornellidae forms a sister group to the other subclades (25).
Aeolidida is a monophyletic clade with Lomanotidae as a sister
group (25). What was traditionally called Dendronotida forms
a paraphyletic grouping. A recent study was unable to include
the nudibranch Melibe in Cladobranchia because of a 12-bp de-
letion in its genome (25). However, its natural afnity with Tethys
in terms of shared derived characteristics strongly suggests that it
belongs in Cladobranchia, as we have indicated in Fig. 1. There
are several additional unresolved relations in Nudibranchia, most
This paper results from the Arthur M. Sackler Colloquium of the National Academy of
Sciences, In the Light of Evolution VI: Brain and Behavior, held January 1921, 2012, at
the Arnold and Mabel Beckman Center of the National Academies of Sciences and Engi-
neering in Irvine, CA. The complete program and audio les of most presentations are
available on the NAS Web site at
Author contributions: J.M.N., A.S., J.L.L., C.A.G., and P.S.K. designed research; J.M.N., A.S.,
J.L.L., and C.A.G. performed research; J.M.N., A.S., J.L.L., C.A.G., an d P.S.K. analyzed data;
and J.M.N., A.S., J.L.L., C.A.G., and P.S.K. wrote the paper.
The authors declare no conict of interest.
This article is a PNAS Direct Submission.
To whom correspondence should be addressed. E-mail:
This article contains supporting information online at
1073/pnas.1201877109/-/DCSupplemental. PNAS Early Edition
notably in Dendronotida and Doridacea. Consideration of loco-
motor behavior and neural circuits may help resolve these relations.
Categories of Locomotor Behavior
Crawling is the primary form of locomotion for all Nudipleura
(2931). The majority of species crawl via mucociliary locomo-
tion; cilia on the bottom of the foot beat and propel the animal
over a surface of secreted mucus. The speed of crawling is af-
fected by efferent serotonergic and peptidergic neurons that
control the ciliary beat frequency (3032). Some species also use
muscular crawling, which relies on waves of contraction or ex-
tension and contraction of the foot. Crawling is a trait shared
with most Opisthobranchia and is therefore plesiomorphic to the
Nudipleura. Only three nudibranch species do not crawl because
they are truly pelagic: Phylliroë atlantica, Phylliroë bucephala, and
Cephalopyge trematoides (33). This is also true for gastropods in
general; there are 40,000 marine gastropod species but only
approximately 150 are pelagic (33).
In addition to crawling, a limited number of benthic species
can also swim (34). We classify swimming in the Nudipleura into
seven general categories: (i) leftright exion (LR), (ii) dorsal
ventral exion (DV), (iii) leftright undulation (LU), (iv ) dorsal
ventral undulation (DU), (v) asymmetric undulation (AU), (vi)
breaststroke (BS), and (vii) apping (F) (Table S1).
LR swimming is characterized by the attening of the body in
the sagittal plane and repeated leftright bending near the
midpoint of the body axis with the head and tail coming together
laterally (Fig. 2A). This movement propels the animal through
the water. Some animals, such as Melibe leonina, exhibit foot-rst
directionality, presumably because the dorsal cerata create drag.
Other animals, such as Tambja eliora, proceed headrst, with the
tail lagging slightly, causing the body to take on an S form (34).
Animals in the genus Plocamopherus typically have a dorsal crest
at the posterior end of the body that may act as a paddle and
cause the head to proceed the tail (35).
Plocamopherus ceylonicus (35, 36) and Plocamopherus maderae
(37) swim with LR exions when dislodged from a substrate or
disturbed in some way. Tambja appears to use LR swimming as
an escape response; contact with the predacious nudibranch
Roboastra will elicit swimming in Tambja (34, 38). LR swim-
ming in
Melibe and Dendronotus iris can be initiated in response
to loss of contact with the substrate or in response to t he touch
of a pr edatory sea star (39, 40). Melibe may also swim seasonally
to disperse (41). The exio n cycle period for Melibe and Den-
dronotus is approximately 3 s, and swim bouts can la st many
minutes (39, 40 ).
As its name suggests, Bornella anguilla swims with an eel-like
movement caused by waves of muscular contraction (42).
Therefore, unlike other members of its genus, it is classied as an
LU swimmer. LU swimming, which otherwise is found mostly in
pelagic species, may be a further renement of LR swimming.
DV swimming involves the animal attening its body in the
horizontal plane and repeatedly bending such that the tail and
head meet in alternation above and below the midpoint of the
Dirona -NS
Archidoris - NS
Hypselodoris - NS
Cadlina - NS
Pleurobranchaea - DV
Tomthompsonia - ?
Burthella - NS?
Pleurobranchus - AU
Bathyberthella - NS?
Euselenops - F
Berthellina - NS?
Hexabranchus - DV/DU
Sebadoris - DV/DU
Trapania - LR
Dendronotus -LR
Lomanotus -LR
Melibe -LR
Tethys -LR
Aeolidella - BS
Hermissenda - LR
Pteraeolidia - LR
Tritonia - DV
Marionia - DV
Tochuina -NS
Scyllaea -LR
Notobryon - LR
Phyllir -LU
Cephalopyge -LU
Cumanotus - BS
Flabellina cynara - BS
Flabellina iodinea - LR
Flabellina telja - LR
Flabellina trophina - NS
Aphelodoris - DV
Discodoris - DV
Diaulula - NS
Triopha fulgarans - LR
Plocamopherus - LR
Triopha catalinae - NS
Tamja - LR
Nembrotha - LR
Bornella -LR
Armina - NS
Family GenusClade
Fig. 1. An abbreviated phylogeny of the Nudipleura with reference to their
behavior. Only the genera of the speci es listed in Table S1 are shown here
unless species differences exist within the genus. The phylogenic relation-
ships are based on refs. 1921, 25, 26, 28. The references for the behavior are
listed in Table S1. Note that this gure represents all the known swimming
species and only a tiny fraction of the more than 2,000 species that are not
capable of swimming or for which there are no published reports of swim-
ming. LR, left-right exion; NS, nonswimmer; DV, dorsal-ventral exion; LU,
left-right undulation; BS, breast stroke; DU, dorsal-ventral undulation; AU,
asymmetric undulation; F, apping.
Le-Right Flexion
Dorsal-Ventral Flexion
Fig. 2. Two examples of swimming behaviors. (A) LR swimming exhibited
by M. leonina. The ventral side of the animal is shown with the mouth at the
top of the image. During swimming, the foot is narrowed to a strip and the
animal rhythmically exes its body leftward and rightward, bending at
a point midway along the body axis. (B) DV swimming exhibited by T. dio-
medea. The animal starts on the substrate, shown at the bottom with its
head to the right. It launches with a ventral exion, where the head and tail
meet under the foot. Then, it exes so that the head and tail meet above the
dorsal body surface. The foot is attened and expanded to the width of the
body. A, anterior; P, posterior.
| Newcomb et al.
body (Fig. 2B). Tritonia diomedea and Pleurobranchaea californica
are two examples of DV swimmers that have been extensively
studied (4346). Swim bouts for Tritonia and Pleurobranchaea last
less than 1 min and are triggered by contact with a predatory sea
star or in the laboratory by high salt solutions or electric shock
(47). The exion cycle period under natural conditions is 5 to 10 s
in Tritonia (48) and 3 to 6 s in Pleurobranchaea (49).
DU swimming, like DV swimming, involves movement in
dorsal and ventral directions, but here there are progressive
symmetric waves of body wall or mantle muscular contraction.
The Spanish Dancer, Hexabranchus sanguineus, and other
members of that genus are famous for their amboyant swim-
ming behavior (34, 50, 51). Hexabranchus swimming differs in
several ways from the DV swimming of Tritonia and Pleuro-
branchaea; in addition to the symmetrical undulation of the
lateral fringes of the mantle, it has a shorter exion cycle period
(24 s), swim bouts occur spontaneously, and swimming can last
for long periods of time.
F swimming is similar to DV swimming in that the movement
is bilaterally symmetric and dorsalventral in orientation, but
instead of the head and tail meeting, the lateral edges of the
mantle or foot rise and fall. F swimming is much more common
in Opisthobranchia outside of the Nudipleura, such as Clione
limacina (52) and many species of Aplysia (53, 54).
AU and BS are less common forms of locomotion. AU is
characteristic of Pleurobranchus membranaceus (55) in which the
animal swims upside down using its mantle as a passive keel
while producing alternating muscular waves along its foot. BS
involves the use of appendages including cerata and tentacles to
stroke the water in a manner similar to a human swimmers
movements. Only four nudibranch species have been described
as exhibiting this type of behavior (Table S1).
Phylogenetic Distribution of Swimming Behaviors
As noted earlier, we have been unable to nd reports of swim-
ming by 97% to 98% of nudibranch species and approximately
half the major subfamilies in the Pleurobranchomorpha clade.
However, this does not mean they are not capable of swimming.
Some species swim only as a high threshold escape response.
Still, it is highly probable that the vast majority of the Nudipleura
cannot and do not swim. This discussion is limited to species for
which the type of swimming has been reported or for which
swimming has been explicitly tested and shown not to occur.
LR swimming is by far the most prevalent of the six modes of
swimming exhibited by nudibranchs: of the 60 nudibranch species
documented to swim in the scientic literature, 40 species use LR
or LU (Table S1). These 40 species are phylogenetically dispa-
rate, encompassing species in Doridacea and Cladobranchia (Fig.
1). Within the latter, there are LR swimmers in Aeolidoidea and
Dendronotoidea. In Doridacea, all but one of the LR swimmers is
in the family Polyceridae. There are no LR swimmers in the
Pleurobranchomorpha or, to our knowledge, in any other Opis-
thobranch clade. This suggests that LR swimming is a derived
characteristic of the nudibranch clade.
Unlike LR swimming, DV swimming is found in Nudibranchia
and in Pleurobranchomorpha (Fig. 1). DV swimming is not pre-
sent outside of Nudipleura and is therefore likely to be a synapo-
morphy of this clade. However, it is not widely displayed wi thin
Scenario 1 Scenario 2
Scenario 3 –revised phylogeny
Scenario 4
Pleurobranchidae - DV
Doridae - DV
Hexabranchidae - DV/DU
Discodoridae - DV/DU
Goniodorididae - LR
Bornellidae - LR
Dendronodae - LR
Lomanodae - LR
Aeolida - LR
Tethydae - LR
Polyceridae - LR
Scyllaeidae - LR
Tritoniidae - DV
Pleurobranchidae - DV
Doridae - DV
Hexabranchidae - DV/DU
Discodoridae - DV/DU
Goniodorididae - LR
Bornellidae - LR
Dendronodae - LR
Lomanodae - LR
Aeolida - LR
Tethydae - LR
Polyceridae - LR
Scyllaeidae - LR
Tritoniidae - DV
Pleurobranchidae - DV
Doridae - DV
Hexabranchidae - DV/DU
Discodoridae - DV/DU
Goniodorididae - DV*
Bornellidae - LR
Dendronodae - LR
Lomanodae - LR
Aeolida - LR
Tethydae - LR
Polyceridae - LR
Scyllaeidae - LR
Tritoniidae - DV
Pleurobranchidae - DV
Doridae - DV
Hexabranchidae - DV/DU
Discodoridae - DV/DU
Goniodorididae - DV*
Bornellidae - LR
Dendronodae - LR
Lomanodae - LR
Aeolida - LR
Tethydae - LR
Polyceridae - LR
Scyllaeidae - LR
Tritoniidae - DV
Fig. 3. Possible evolutionary scenarios explaining the phylogenetic distribution of swimming behaviors. Just the families of the DV and LR swimming animals
are shown. (A) In scenario 1, DV swimming is a synapomorphy of the Nudipleura that was lost and replaced six times by LR swimming. (B) In scenario 2, LR
swimming is a synapo morphy of the Nudibranchi a. DV swimming then reappears four times in different nudibranch lineages. (C) For scenario 3, the phy-
logenetic tree of Dendronotida is altered to group LR swimmers together. Goniodorididae (asterisk), which includes T. velox, is switched from LR to DV (as
discussed in the text). This reduces the number of transitions to LR from six in scenario 1 to four. (D) Scenario 4 is similar to scenario 2, with Goniodoridid ae
(asterisk) switched to DV. This represents the most parsimonious explanation if DV swimming is ancestral, with just three transitions from the basal DV state.
Newcomb et al. PNAS Early Edition
Nudibranchia, appearing in just one family of Dendronotida
(Tritoniidae) and in three families of Doridacea (Discodorididae,
Dorididae, and Hexi branchid ae). Discodori didae an d Hexi-
branchidae also exhibit dorsal ventral undulations (i.e., DU).
Evolution of Swimming Behaviors
There are a number of possible scenarios that could account for
the phylogenetic distribution of swimming behaviors among the
Nudipleura. Considering the extreme rarity of swimming, it is
possible, maybe even likely, that swimming evolved on multiple
occasions from nonswimming species. The repeated gain of a
function such as rhythmic movement could suggest that there is
a predisposition toward these behaviors. The repeated appear-
ance of LR and DV swimming may simply indicate that these
two basic movements are the most likely to occur in a slug-sha-
ped body with few appendages. When appendages such as
moveable cerata are present, they have been repeatedly used for
BS swimming. In the absence of such appendages, the only
means of swimming are with LR-like or DV-like movements.
Given the presence of swimming across the phylogeny, it is
possible that, rather than evolving independently many times
from nonswimmers, swimming behaviors were repeatedly lost.
Although this may lead to more transformations, it may be easier
to lose a character than to gain one, as has been seen in other
systems (5660).
For the moment, we will only consider the possible evolu-
tionary scenarios that include transformations from one swim-
ming state to another and ignore nonswimmers. It is generally
the case that members of the same genus and often the same
family exhibit the same form of swimming (Table S1), allowing
us to group them together (Fig. 3). Here we will consider po-
tential scenarios involving just the evolution of DV and LR
swimming. It is possible that the ancestral species was able to
swim by using DV and LR movements. However, this seems
unlikely because there are no extant species that exhibit both
these behaviors. It is also unlikely that the ancestral state was LR
swimming because of its absence in Pleurobranchomorpha.
Consider scenario 1 (Fig. 3A) in which DV swimming arose once
at the base of the Nudipleura and LR swimming evolved in-
dependently several times. In this scenario, DV swimming behav-
iors in Pleurobranchomorpha, Doridacea, and Cladobranchia are
homologous because they are shared by a common ancestor. Sce-
nario 1 would also suggest that LR swimming evolved indepen-
dently as many as seven times. Because of the unresolved branches
in the phylogeny, there may be fewer switches in phenotype than
this. In scenario 2 (Fig. 3B), LR swimming evolved once in the
Nudibranchia and DV swimming reevolved independently as many
as four times. Again, the number of homoplastic events could be
lower if the bifurcations in the phylogeny were better resolved.
The phylogenetic distribution of the swimming behavior
suggests a resolution to the Dendronotida phylogen y, with
Tritoniidae branching off separately from the LR swimmers.
This would reduce the number of homoplastic events in Cla-
dobranchia according to scenario 1 from ve to three (scenario
3; Fig. 3C).
The phylogenetic distribution of the behavior also calls into
question the accuracy of a report about the behavior of Trapania
velox. Outside of the family Polyceridae, T. velox (family:
Goniodorididae) is the only doridacean reported to swim with
leftright exions. Farmer (34) categorized T. velox as an LR
swimmer based on a previous report by Cockerell (61), who
described T. velox as being, very active when swimming with an
undulating motion on the surface of the water. However, there
is no indication as to the plane of movement. Farmer (34)
reported working with this rare species and being unsuccessful at
making it swim, and was thus unable to provide any additional
information. We were unable to nd any other reports of its
behavior. If T. velox is reclassied as a DV swimmer, it would
further decrease the number of homoplastic events in scenario 1
from seven to four (Fig. 3C). Thus, examining the phylogenetic
distribution of behavior makes a prediction about the behavior of
this rare species.
Redening T. velox as a DV swimmer also suggests a fourth
scenario (Fig. 3D), whereby LR swimming arose independently
in Cladobranchia and Polyceridae. This would also involve
reevolution of DV swimming in Tritoniidae. Scenario 4 would
therefore be the most parsimonious explanation for the phylo-
genetic distribution of swimming behaviors if one does not take
into account the hundreds of nonswimming species.
Neural Circuits Underlying Swimming
With our potential scenarios about the homology and homoplasy
of swimming behaviors, it is now of interest to compare the
neural mechanisms for these behaviors. The neural activity that
underlies rhythmic DV and LR movements originates from
central pattern generator (CPG) circuits (62). These swim CPGs
are composed of neurons whose anatomical and physiological
properties allow them to be individually identiable from animal
to animal within a species. The same sets of characteristics can
be used to identify homologous neurons in other species (63).
This allows the composition of neural circuits and the roles of
homologous neurons to be compared across species. The neural
circuits underlying swimming have been determined in two DV
swimmers [T. diomedea (44) and P. californica (45, 64)] and two
LR swimmers [M. leonina (40, 65) and D. iris (40)]. We can now
begin to compare neural circuits underlying behaviors of animals
to address phylogenetic and functional hypotheses.
5 sec
10 mV
5 sec
Tritonia diomedea
Pleurobranchaea californica
Fig. 4. The neural circuits and swim motor patters for the DV swimmers
Tritonia and Pleurobranchaea.(A) The Tritonia swim CPG consists of three
neuron types: DSI, C2, and VSI. (B) Simultaneous intracellular microelectrode
recordings show that two contralateral DSIs re bursts of action potentials in
phase with each other and slightly ahead of the two C2s. VSI (not recorded
here) res action potentials in the interburst interval. The motor pattern is
initiated by electrical stimulation of a body wall nerve (stim). (C) The Pleu-
robranchaea swim CPG contains ve types of neurons (64). The As neurons
are homologues of the DSIs. A1 is homologous to C2. A10 is strongly elec-
trically coupled to A1 and, for simplicity, is shown together with it. A3 is not
found in Tritonia. The Ivs neuron has not been found, but has been postu-
lated to exist based on recordings of inhibitory postsynaptic potentials in
other neurons. (D) Simultaneous intracellular recordings from an A3, As, and
A1. The As neuron leads the A1 neuron just as DSI leads C2. The swim motor
pattern is initiated by electrical stimulation of a body wall nerve (stim). In A
and C, the small lled circles represent inhibitory synapses, the triangles are
excitatory synapses and combinations are mixed inhibition and excitation.
The resistor symbol represents electrical synapses. B and D are previously
unpublished recordings.
| Newcomb et al.
DV Swim CPGs. The neural basis for DV swimming was rst
studied in T. diomedea (43, 6669). The swim CPG consists of
just three neuron types (Fig. 4A). On each side of the brain,
there are three dorsal swim interneurons (DSIs), one ventral
swim interneuron (VSI), and one cerebral interneuron 2 (C2),
for a total of 10 neurons (44, 47). The DSIs initiate the dorsal
exion cycle in which C2 participates. C2 then excites VSI, which
inhibits DSI and C2 and elicits the ventral phase of the move-
ment. As would be expected for a DV swimmer, the contralateral
counterparts for each neuron re in relative synchrony (Fig. 4B).
The neurons comprising the CPG for DV swimming in P.
californica include DSI and C2 homologues called As and A1,
respectively (49, 64). The connectivity and activity of these
homologues is similar in both species (Fig. 4 C and D). The
homologue of the Tritonia VSI has not been identied in Pleu-
robranchaea, although there is synaptic input to As and A1
during the ventral phase of the motor pattern that may arise
from such a neuron (i.e., Ivs neuron) (64). Alternatively, ventral
phase synaptic input may arise from a neuron that is not ho-
mologous to VSI, but serves a similar role.
There are also Pleurobranchaea swim CPG neurons (A3 and
A10) that have not been identied in Tritonia. Despite more than
40 y of electrophysiological study concentrated in the area where
the A3 and A10 somata would be, no neurons with equivalent
synaptic connectivity or activity have been found in Tritonia.
Thus, either these neurons do not exist in Tritonia or they cannot
be recognized with electrophysiological criteria.
With the information available about the swim CPGs in Tri-
tonia and Pleurobranchaea, we can currently say that some ho-
mologous neurons are used for similar functions in distantly
related species. This result is compatible with any of the phylo-
genetic scenarios (Fig. 3). If DV swimming is homologous (sce-
narios 1 or 3; Fig. 3 A and C), the similarities in the DV swim
CPGs in Tritonia and Pleurobranchaea could be a result of their
homology and the potential differences in the swim CPGs could
represent divergence of the circuit architecture. The differences
in the swim CPGs may just as readily reect independent
evolutionary paths (scenarios 2 or 4; Fig. 3 B and D), which
might suggest a predisposition to use certain neurons to produce
these behaviors.
LR Swim CPGs. The LR swim CPG was rst described in M. leo-
nina (65, 70). The published circuit consists of a pair of bi-
laterally represented neurons: swim interneuron 1 (Si1) and swim
interneuron 2 (Si2; Fig. 5A). Based on their anatomy and neu-
rochemistry, these neurons are not homologous to any of the
Tritonia or Pleurobranchaea swim CPG neurons.
In the Melibe swim CPG, each neuron reciprocally inhibits the
two contralateral counterparts (Fig. 5B). There is also strong
electrical coupling between the ipsilateral Si1 and Si2, causing
them to re in phase with each other and 180° out of phase with
the contralateral pair (Fig. 5C). This bursting pattern drives the
leftright alternations of the swimming behavior (71).
Homologues of the Melibe Si1 and Si2 were identied in D. iris
based on anatomical, neurochemical, and electrophysiological
features (40). However, there are important differences in the
neural circuit formed by these neurons (Fig. 5D). Although the
contralateral Si2 neurons reciprocally inhibit each other, Si1
does not inhibit or receive inhibition from either contralateral
neuron. Instead, Si1 exhibits strong electrical coupling to its
contralateral counterpart (Fig. 5E). During a swim motor pat-
tern, the contralateral Si2 neurons re bursts of action potentials
in alternation, but the Si1 pair re irregularly (Fig. 5F). Thus,
whereas both Si1 and Si2 are members of the LR swim CPG in
Melibe, only Si2 is in Dendronotus.
If LR swimming in Melibe and Dendronotus is homologous, as
would be expected from scenarios 2, 3, or 4 (Fig. 3 BD), this
would be an example in which the neural mechanisms diverged
while the behavior stayed the same. However, it could be the
case that the differences in neural mechanism reect a different
evolutionary origin for LR swimming in Melibe and Dendronotus
as in scenario 1 (Fig. 3A ).
Melibe leonina
Dendronotus iris
2 sec
2 sec
1 sec
2 nA
2 nA
2 sec
2 nA
2 nA
L-Si1 R-Si1
Fig. 5. Neural circuitry and swim motor pattern for the LR swimmers Melibe and Dendronotus.(A)IntheMelibe swim CPG (65), there are two bilaterally
represented neurons Si1 and Si2 that are mutually inhibitory across the midline and exhibit strong electrical coupling ipsilaterally (as indicated by thicker
resistor symbol). (B) Depolarization of one Si1 by injecting 2nA of current into it hyperpolarizes the contralateral counterpart. (C) The Melibe swim motor
pattern consists of ipsilateral synchrony and alternation with the contralateral side. (D)InDendronotus, the inhibitory connections to and from Si1 are absent,
and the electrical coupling between the contralateral Si1 pair dominates (40). (E) Depolarization of an Si1 with 2nA current injection depolarizes the con-
tralateral counterpart. (F)IntheDendronotus swim motor pattern, the left and right Si2 re alternating bursts of action potentials, but the Si1s re
irregularly. In A and D, the shaded boxes represent the functional CPGs. All recordings are previously unpublished.
Newcomb et al. PNAS Early Edition
Functions of DV Swim CPG Neurons in Other Species
DSI and C2 homologues can be recognized by using neuroan-
atomical and neurochemical criteria, allowing them to be
identied in species that are no t DV swimmers (Table 1). The
DSIs are serotonergic (72, 73) and have a charac teristic axon
projection pattern (67). They have been identied in 10 differ-
ent genera, including two opisthobranchs outside of the Nudi-
pleura (74). Elec trophysiological traits of the DSI homologues
show little correlation with the type of behavior prod uced by
the species (74). C2 has been identied based on peptide im-
munoreactivity and characteristic morphology in ve genera
within the Nudipleura (75). Thus, these DV swim CPG neu-
rons are present regardless of the animals mode of locomot ion.
This suggests that the swimming CPGs were built upon pre-
viousl y existing neural circuits, coopting existing neurons for
new functions.
The DV swim CPG neurons are not members of the LR swim
CPGs. The DSI and C2 homologs in Melibe are not rhythmically
active in phase with the motor pattern (Fig. 6A), nor are the DSI
homologues rhythmically active during the Dendronotus swim
motor pattern (Fig. 6B). Thus, categorically distinct behaviors are
produced by CPGs containing nonoverlapping sets of neurons.
It was shown that the DSI homologues in Melibe do have an
effect on the production of the swim motor pattern; they can
initiate a motor pattern in a quiescent preparation and hyper-
polarization can temporarily halt an ongoing motor pattern (85).
In contrast to Tritonia, in which the DSIs are an integral part of
the DV swim CPG, in Melibe, they act as extrinsic modulators.
Thus, the functions of homologous neurons differ in species with
different behaviors.
The DSIs are not dedicated to one function even within
a species. In Pleurobranchaea, the DSI homologues synapse onto
serotonergic neurons that increase ciliary beating and thereby
increase the speed of crawling (86). In Tritonia, DSI accelerates
crawling through synapses onto the efferent peptidergic pedal
neuron Pd5, which in turn increases cilia beat frequency (87).
DSI homologues in the nonswimming Tochuina tetraquetra and
Triopha catalinae also monosynaptically excite homologues of
Pd5 and presumably increasing the speed of crawling (74). In
Hermissenda, which produces LR exions, the DSI homologues
do not increase ciliary beating, but instead excite motor neurons
that cause contraction of the anterior foot (77). In the more
distantly related opisthobranch, Aplysia californica, DSI homo-
logues also initiate muscular crawling (78). Whereas, in the pe-
lagic opisthobranch, C. limacina, the DSI homologues increase
the frequency of parapodial wing apping and excite motor
neurons that innervate the wings (83, 88). Thus, the DSI
homologues share common functions in controlling the foot and/
or locomotion.
The C2 and DSI homologues have additional roles outside of
locomotion. In Pleurobranchaea, the C2 homologue (A1) sup-
presses feeding through its connections to feeding-related
interneurons (49). In contrast, the DSI homologues (As) have
the opposite effect by exciting a number of feeding interneurons
(86). This is a shared function with other opisthobranchs such as
A. californica, in which the DSI homologues (CC9-10) help excite
one of the same feeding interneurons as in Pleurobranchaea
, the
metacerebral cell (78). Thus, individual neurons are multifunc-
tional. Some functions are shared across species, whereas other
functions are particular to some species.
A phylogenetic analysis of the neural basis for swimming in the
Nudipleura has revealed several interesting aspects about the
evolution of behavior. First, the basic building blocks of neural
circuits, namely the neurons, are shared across diverse species.
For example, DSI homologues are found across Opistho-
branchia. Second, neurons, which are multifunctional within
a species, appear to take on additional functions over the course
of evolution. For instance, the DSI homologues are involved in
several behaviors in various species, including generating DV
swimming or enhancing other types of locomotion such as en-
hancing LR swimming or wing apping. They also accelerate
crawling and promote feeding. It is reasonable to expect that
highly interconnected interneurons would not be dedicated to
a single function, but would dynamically interact with many
neurons involved in a variety of different behaviors.
This comparative analysis has also revealed that species with
categorically similar behaviors such as the two DV swimmers,
Tritonia and Pleurobranchaea, or the two LR swimmers, Melibe
and Dendronotus, have overlapping sets of neurons in the swim
CPG circuits. In contrast, the CPGs underlying categorically
distinct behaviors consist of nonoverlapping sets of neurons.
However, even in species that exhibit similar behaviors such as
Melibe and Dendronotus, the CPG circuits can differ in neuronal
and synaptic composition. Thus, although behavior itself is not
a predictor of its underlying neural mechanism, it is a good
rst approximation.
We do not understand why the circuits in Melibe and Den-
dronotus differ. There could be functional reasons; perhaps Si1,
Table 1. Homologous neurons identied in different species with different behaviors
OpisthobranchiaDV swimmers LR swimmers Nonswimmers
DSI Tritonia (76) Melibe (74) Armina (74) Aplysia (7881)
Pleurobranchaea (64) Dendronotus (74) Triopha (74) Clione (82, 83)
Hermissenda (77) Tochina (74)
C2 Tritonia (76, 84) Melibe (75)
Pleurobranchaea (49) Hermissenda (75)
Flabellina (75)
5 sec
Melibe leonina
5 sec
Dendronotus iris
Fig. 6. Homologues of the Tritonia DV swim CPG neurons are not rhyth-
mically active during LR swim motor patterns. (A)InMelibe, the C2 and DSI
homologues do not display any rhythmic bursting in phase with the swim
motor pattern reected in the alternating ring pattern of the left and right
Si. (B)InDendronotus, a contralateral pair of DSI homologues exhibit syn-
chronous irregular spiking that shows no relation to the ongoing LR swim
motor pattern displayed by two contralateral pedal motor neurons (L-Pd and
R-Pd). All recordings are previously unpublished.
| Newcomb et al.
which is not rhythmically active in Dendronotus, has an addi-
tional function that is incompatible with swimming in that spe-
cies. There may also be phylogenetic reasons; perhaps Melibe
and Dendronotus independently evolved swim CPGs and came
up with different circuit organizations. Whatever the reason, the
results show that analogous behaviors can be generated by cir-
cuits with different circuit architectures. Recent work in inver-
tebrates has shown that there can be variability in neural circuits
that is not reected in the performance of the behavior even
across individuals within a species (89, 90).
There is a great degree of behavioral homoplasy. Although
scenario 4 (Fig. 3D) may be the most parsimonious explanation
for the phylogenetic distribution of the swimming behaviors, it
should be kept in mind that only approximately 2% to 3% of
nudibranch species have been reported to swim. Therefore, there
is probably even more behavioral homoplasy than any of the
scenarios in Fig. 3 indicate. It is conceivable that swimming arose
independently in each family where it is found, 16 times in all
(Fig. 1 and Table S1).
Given that Tritonia and Pleurobranchaea are very distantly re-
lated within the Nudipleura clade, it is even more likely that they
independently evolved DV swim CPGs. If so, the incorporation of
DSI and C2 homologues into such a circuit represents parallel
evolution, whereby homologous structures independently came to
have similar functions (9194). This has been suggested for other
systems as well. For example, homologous brain nuclei appear to
be involved in vocal learning in lineages of birds that evolved song
independently (95, 96). Similarly, interaural coincidence detec-
tion circuits arose independently in the brainstem nuclei of birds
and mammals (97). Finally, the appearance of similar cortical
areas are correlates with the independent evolution of precision
hand control in primates (98), suggesting that constraints in cor-
tical organization led to the evolution of similar neural mecha-
nisms underlying dexterity (99).
If homologous neurons are repeatedly i ncorporated into
neural circuits for analogous behaviors, it suggests that these
neurons may be part of a more readily achievable state for
swimming. Thus, the ne rvous system m ay affect the evo lvability
of behavior because some congurations of exist ing neurons
could be more robust than others. The concept of evolvability
rst arose from genet ics (100, 101), but has since been appli ed
to nervous systems (5, 79). Exploring the aspects of neural
organ ization that lead to repeated evoluti on of particular
behaviors will point to the factors that are most important for
behavioral output.
ACKNOWLEDGMENTS. We thank Arianna Tamvacakis for feedback on the
manuscript. This work was supported by National Science Foundation
Integrative Organismal Systems Grants 0814411, 1120950, and 1011476.
1. Striedter GF, Northcutt RG (1991) Biological hierarchies and the concept of homol-
ogy. Brain Behav Evol 38:177189.
2. Rendall D, Di Fiore A (2007) Homoplasy, homology, and the perceived special status
of behavior in evolution. J Hum Evol 52:504521.
3. Lauder GV (1994) Homology, form, and function. Homology: The Hierachical Basis of
Comparative Biology, ed Hall BK (Academic, San Diego), pp 151196.
4. Lauder GV (1986) Homology, analogy, and the evolution of behavior. Evolution of
Animal Behavior, eds Nitecki MH, Kitchell JA (Oxford Univ Press, New York), pp 940.
5. Katz PS (2011) Neural mechanisms underlying the evolvability of behaviour. Philos
Trans R Soc Lond B Biol Sci 366:20862099.
6. Carlson BA, et al. (2011) Brain evolution triggers increased diversication of electric
shes. Science 332:583586.
7. Bendesky A, Bargmann CI (2011) Genetic contributions to behavioural diversity at
the gene-environment interface. Nat Rev Genet 12:809820.
8. Airey DC, Castillo-Juarez H, Casella G, Pollak EJ, DeVoogd TJ (2000) Variation in the
volume of zebra nch song control nuclei is heritable: developmental and evolu-
tionary implications. Proc Biol Sci 267:20992104.
9. Yamamoto K, Vernier P (2011) The evolution of dopamine systems in chordates.
Front Neuroanat 5:21.
10. Darwin C (1876) The Origin of Species by Natural Selection, or the Preservation of
Favoured Races in the Struggle for Life (John Murray, London).
11. Heinroth O (1911) Beiträge zur Biologie, namentlich Ethologie und Psychologie der
Anatiden. Verhanalung des V Internationalen Ornithologen Kongresses, Berlin, 1910
12. Whitman CO (1899) Animal Behavior. Biological Lectures of the Marine Biological
Laboratory (Woods Hole Marine Biological Laboratory, Woods Hole, MA).
13. Lorenz K (1981) The Foundations of Ethology (Springer-Verlag, New York).
14. De Queiroz A, Wimberger PH (1993) The usefulness of behavior for phylogeny es-
timation: Levels of homoplasy in behavioral and morphological characters. Evolution
15. Wenzel JW (1992) Behavioral homology and phylogeny. Annu Rev Ecol Syst 23:
16. Stuart AE, Hunter FF, Currie DC (2002) Using behavioural characters in phylogeny
reconstruction. Ethol Ecol Evol 14:129139.
17. Proctor HC (1996) Measures of homoplasy. Homoplasy: The Recurrence of Similarity
in Evolution, eds Sanderson MJ, Hufford L (Academic, San Diego), pp 131149.
18. Foster SA, Cresko WA, Johnson KP, Tlusty MU, Willmott HE (1996) Homoplasy: The
Recurrence of Similarity in Evolution
, eds Sanderson MJ, Hufford L (Academic, San
Diego), pp 245269.
19. Waegele H, Willan RC (2000) Phylogeny of the nudibranchia. Zool J Linn Soc 130:
20. Wollscheid-Lengeling E, Boore J, Brown W, Waegele H (2001) The phylogeny of
Nudibranchia (Opisthobranchia, Gastropoda, Mollusca) reconstructed by three mo-
lecular markers. Org Divers Evol 1:241256.
21. Göbbeler K, Klussmann-Kolb A (2010) Out of Antarctica?new insights into the
phylogeny and biogeography of the Pleurobranchomorpha (Mollusca, Gastropoda).
Mol Phylogenet Evol 55:9961007.
22. Bouchet P, et al. (2005) Classication and nomenclator of gastropod families. Mal-
acologia 47:1368.
23. Behrens DW (2005) Nudibranch Behavior (New World Publications, Jacksonville, FL).
24. Dinapoli A, Klussmann-Kolb A (2010) The long way to diversityphylogeny and
evolution of the Heterobranchia (Mollusca: Gastropoda). Mol Phylogenet Evol 55:
25. Pola M, Gosliner TM (2010) The rst molecular phylogeny of cladobranchian opis-
thobranchs (Mollusca, Gastropoda, Nudibranchia). Mol Phylogenet Evol 56:931941.
26. Vonnemann V, Schrodl M, Klussmann-Kolb A, Wagele H (2005) Reconstruction of the
phylogeny of the Opisthobranchia (Mollusca: Gastropoda) by means of 18S and 28S
rRNA gene sequences. J Molluscan Stud 71:113125.
27. Valdes A (2003) A phylogenetic analysis and systematic revision of the cryptobranch
dorids (Mollusca, Nudibranchia, Anthobranchia). Zool. J. Linn. Soc. Lond. 136:
28. Thollesson M (1999) Phylogenetic analysis of dorid nudibranchs (Gastropoda: Dor-
idacea) using the mitochondrial 16S rRNA gene. J Molluscan Stud 65:335353.
29. Chase R (2002) Behavior and Its Neural Control in Gastropod Molluscs (Oxford Univ
Press, New York).
30. Audesirk G, McCaman RE, Willows AOD (1979) The role of serotonin in the control of
pedal ciliary activity by identied neurons in Tritonia diomedia. Comp Biochem
Physiol 62C:8791.
31. Audesirk G (1978) Central neuronal control of cilia in Tritonia diamedia. Nature 272:
32. Willows AOD, Pavlova GA, Phillips NE (1997) Modulation of ciliary beat frequency by
neuropeptides from identied molluscan neurons. J Exp Biol 200:1433
33. Lalli CM, Gilmer RW (1989) Pelagic Snails (Stanford Univ Press, Stanford, CA).
34. Farmer WM (1970) Swimming gastropods (Opisthobranchia and Prosobranchia).
Veliger 13:7389.
35. Rudman WB, Darvell BW (1990) Opisthobranch molluscs of Hong Kong: Part 1. Go-
niodorididae, Onchidorididae, Triophidae, Gymnodorididae, Chromodorididae (Nu-
dibranchia). Asian Marine Biology 7:3179.
36. Marshall JG, Willan RC (1999) Nudibranchs of Heron Island, Great Barrier Reef - a
survey of the Opisthobranchia (sea slugs) of Heron and Wistari Reefs (Backhuys,
Leiden, The Netherlands).
37. Lowe RT (1842) Description of a new dorsibranchiate gasteropod discovered at
Madeira. Proc Zool Soc Lond 10:5153.
38. Pola M, Cervera JL, Gosliner TM (2006) Description of two new phanerobranch
nembrothid species (Nudibranchia: Polyceridae: Doridacea). J Mar Biol Assoc U K 86:
39. Lawrence KA, Watson WH, 3rd (2002) Swimming behavior of the nudibranch Melibe
leonina. Biol Bull 203:144151.
40. Sakurai A, Newcomb JM, Lillvis JL, Katz PS (2011) Different roles for homologous
interneurons in species exhibiting similar rhythmic behaviors. Curr Biol 21:
41. Mills CE (1994) Reproduction and Development of Marine Invertebrates,eds
Wilson, Jr WH, Stricker SA, Shinn GL (Johns Hopkins Univ Press, Baltimore), pp
3133 19.
42. Johnson S (1984) A new Indo-West pacic species of the dendronotacean nudi-
branch Bornella (Mollusca: Opisthobranchia) with anguilliform swimming behav-
iour. Micronesica 19:1726.
43. Willows AO (1967) Behavioral acts elicited by stimulation of single, identiable brain
cells. Science 157:570574.
44. Katz PS (2009) Tritonia swim network. Scholarpedia 4:3638.
Newcomb et al. PNAS Early Edition
45. Gillette R, Jing J (2001) The role of the escape swim motor network in the organi-
zation of behavioral hierarchy and arousal in Pleurobranchaea. Am Zool 41:983992.
46. Davis WJ, Mpitsos GJ (1971) Behavioral choice and habituation in the marine mollusk
Pleurobranchaea californica MacFarland (Gastropoda, Opisthobranchia). Z Vgl
Physiol 75:207232.
47. Katz PS (2010) Handbook of Microcircuits, eds Shepherd G, Grillner S (Oxford Univ
Press, New York), pp 443449.
48. Hume RI, Getting PA, Del Beccaro MA (1982) Motor organization of Tritonia
swimming. I. Quantitative analysis of swim behavior and exion neuron ring pat-
terns. J Neurophysiol 47:6074.
49. Jing J, Gillette R (1995) Neuronal elements that mediate escape swimming and
suppress feeding behavior in the predatory sea slug Pleurobranchaea. J Neuro-
physiol 74:19001910.
50. Edmunds M (1968) On the swimming and defensive response of Hexabranchus
marginatus (Mollusca, Nudibranchia). J. Linnean Soc 47:425429.
51. Gohar HAF, Soliman GN (1963) The biology and development of Hexabranchus
sanguineus (Rüpp. and Leuck.) (Gastropoda, Nudibranchiata). Publ Mar Biol Sta
Ghardaqa (Red Sea) 12:219247.
52. Arshavsky YuI, et al. (1986) Control of locomotion in marine mollusc Clione limacina.
VI. Activity of isolated neurons of pedal ganglia. Exp Brain Res 63:106112.
53. Donovan DA, Pennings SC, Carefoot TH (2006) Swimming in the sea hare Aplysia
brasiliana: Cost of transport, parapodial morphometry, and swimming behavior. J
Exp Mar Biol Ecol 328:7686.
54. Bebbington A, Hughes GM (1973) Locomotion in Aplysia (Gastropoda, Opistho-
branchia). J Molluscan Stud 40:399405.
55. Thompson TE, Slinn DJ (1959) On the biology of the opisthobranch Pleurobranchus
membranaceus. J Mar Biol Assoc U K 38:507524.
56. Moczek AP, Cruickshank TE, Shelby A (2006) When ontogeny reveals what phylog-
eny hides: Gain and loss of horns during development and evolution of horned
beetles. Evolution 60:23292341.
57. Duboué ER, Keene AC, Borowsky RL (2011) Evolutionary convergence on sleep loss in
cavesh populations. Curr Biol
58. Harshman J, et al. (2008) Phylogenomic evidence for multiple losses of ight in ratite
birds. Proc Natl Acad Sci USA 105:1346213467.
59. Wiens JJ, Kuczynski CA, Duellman WE, Reeder TW (2007) Loss and re-evolution of
complex life cycles in marsupial frogs: Does ancestral trait reconstruction mislead?
Evolution 61:18861899.
60. Whiting MF, Bradler S, Maxwell T (2003) Loss and recovery of wings in stick insects.
Nature 421:264267.
61. Cockerell TDA (1901) Three new nudibranchs from California. J Malacol 8:8587.
62. Delcomyn F (1980) Neural basis of rhythmic behavior in animals. Science 210:
63. Croll RP (1987) Identied neurons and cellular homologies. Nervous Systems in In-
vertebrates, ed Ali MA (Plenum, New York), pp 4159.
64. Jing J, Gillette R (1999) Central pattern generator for escape swimming in the no-
taspid sea slug Pleurobranchaea californica. J Neurophysiol 81:654667.
65. Thompson S, Watson WH, 3rd (2005) Central pattern generator for swimming in
Melibe. J Exp Biol 208:13471361.
66. Dorsett DA, Willows AOD, Hoyle G (1969) Centrally generated nerve impulse se-
quences determining swimming behavior in Tritonia. Nature 224:711712.
67. Getting PA, Lennard PR, Hume RI (1980) Central pattern generator mediating
swimming in Tritonia. I. Identication and synaptic interactions. J Neurophysiol 44:
68. Getting PA (1981) Mechanisms of pattern generation underlying swimming in Tri-
tonia. I. Neuronal network formed by monosynaptic connections. J Neurophysiol 46:
69. Getting PA (1983) Neural control of swimming in Tritonia. Symposia of the Society
for Experimental Biology, No. 37, Neural Origin of Rhythmic Movements,, eds
Roberts A, Roberts BL (Cambridge Univ Press, New York), pp 89128.
70. Watson WH, Lawrence KA, Newcomb JM (2001) Neuroethology of Melibe leonina
swimming behavior. Am Zool 41:10261035.
71. Watson WH, 3rd, Newcomb JM, Thompson S (2002) Neural correlates of swimming
behavior in Melibe leonina. Biol Bull 203:152
72. Katz PS, Getting PA, Frost WN (1994) Dynamic neuromodulation of synaptic strength
intrinsic to a central pattern generator circuit. Nature 367:729731.
73. McClellan AD, Brown GD, Getting PA (1994) Modulation of swimming in Tritonia:
Excitatory and inhibitory effects of serotonin. J Comp Physiol A Neuroethol Sens
Neural Behav Physiol 174:257266.
74. Newcomb JM, Katz PS (2007) Homologues of serotonergic central pattern generator
neurons in related nudibranch molluscs with divergent behaviors. J Comp Physiol A
Neuroethol Sens Neural Behav Physiol 193:425443.
75. Lillvis JL, Gunaratne CA, Katz PS (2012) Neurochemical and neuroanatomical iden-
tication of central pattern generator neuron homologues in Nudipleura molluscs.
PLoS ONE 7:e31737.
76. Getting PA (1977) Neuronal organization of escape swimming in Tritonia. J Comp
Physiol A Neuroethol Sens Neural Behav Physiol 121:325342.
77. Tian LM, Kawai R, Crow T (2006) Serotonin-immunoreactive CPT interneurons in
Hermissenda: Identication of sensory input and motor projections. J Neurophysiol
78. Jing J, Vilim FS, Cropper EC, Weiss KR (2008) Neural analog of arousal: Persistent
conditional activation of a feeding modulator by serotonergic initiators of loco-
motion. J Neurosci 28:1234912361.
79. Xin YP, Koester J, Jing J, Weiss KR, Kupfermann I (2001) Cerebral-abdominal inter-
ganglionic coordinating neurons in Aplysia. J Neurophysiol 85:174186.
80. Wright WG, Jones K, Sharp P, Maynard B (1995) Widespread anatomical projections
of the serotonergic modulatory neuron, CB1, in Aplysia. Invert Neurosci 1:173183.
81. Mackey SL, Kandel ER, Hawkins RD (1989) Identied serotonergic neurons LCB1 and
RCB1 in the cerebral ganglia of Aplysia produce presynaptic facilitation of siphon
sensory neurons. J Neurosci 9:42274235.
82. Panchin YV, Popova LB, Deliagina TG, Orlovsky GN, Arshavsky YI (1995) Control of
locomotion in marine mollusk Clione limacina. VIII. Cerebropedal neurons. J Neu-
rophysiol 73:19121923.
83. Satterlie RA, Norekian TP (1995) Serotonergic modulation of swimming speed in the
pteropod mollusc Clione limacina. III. Cerebral neurons. J Exp Biol 198:917930.
84. Taghert PH, Willows AOD (1978) Control of a xed action pattern by single, central
neurons in the marine mollusk, Tritonia diomedea.
J Comp Physiol 123:253259.
85. Newcomb JM, Katz PS (2009) Different functions for homologous serotonergic in-
terneurons and serotonin in species-specic rhythmic behaviours. Proc Biol Sci 276:
86. Jing J, Gillette R (2000) Escape swim network interneurons have diverse roles in
behavioral switching and putative arousal in Pleurobranchaea. J Neurophysiol 83:
87. Popescu IR, Frost WN (2002) Highly dissimilar behaviors mediated by a multifunc-
tional network in the marine mollusk Tritonia diomedea. J Neurosci 22:19851993.
88. Arshavsky YI, Deliagina TG, Orlovsky GN, Panchin YV, Popova LB (1992) Interneur-
ones mediating the escape reaction of the marine mollusc Clione limacina. J Exp Biol
89. Roffman RC, Norris BJ, Calabrese RL (2011) Animal-to-animal variability of connec-
tion strength in the leech heartbeat central pattern generator. J Neurophysiol 107:
90. Goaillard JM, Taylor AL, Schulz DJ, Marder E (2009) Functional consequences of
animal-to-animal variation in circuit parameters. Nat Neurosci 12:14241430.
91. Hoekstra HE, Price T (2004) Evolution. Parallel evolution is in the genes. Science 303:
92. Scotland RW (2011) What is parallelism? Evol Dev 13:214227.
93. Wake DB, Wake MH, Specht CD (2011) Homoplasy: From detecting pattern to de-
termining process and mechanism of evolution. Science 331:10321035.
94. Sanderson MJ, Hufford L (1996) Homoplasy: The Recurrence of Similarity in Evolu-
tion (Academic, San Diego).
95. Hara E, Rivas MV, Ward JM, Okanoya K, Jarvis ED (2012) Convergent differential
regulation of parvalbumin in the brains of vocal learners. PLoS ONE 7:e29457.
96. Feenders G, et al. (2008) Molecular mapping of movement-associated areas in the
avian brain: A motor theory for vocal learning origin. PLoS ONE 3:e1768.
97. Schnupp JW, Carr CE (2009) On hearing with more than one ear: Lessons from
evolution. Nat Neurosci 12:692697.
98. Padberg J, et al. (2007) Parallel evolution of cortical areas involved in skilled hand
use. J Neurosci 27:1010610115.
99. Krubitzer L (2009) In search of a unifying theory of complex brain evolution. Ann N Y
Acad Sci 1156:4467.
100. Masel J, Trotter MV (2010) Robustness and evolvability. Trends Genet 26:406
101. Kirschner M, Gerhart J (1998) Evolvability. Proc Natl Acad Sci USA 95:84208427.
| Newcomb et al.
... Animals belonging to the larger clade of sea slugs called Heterobranchia exhibit a wide variety of swimming behaviors that differ in mode of propulsion, directionality, and function (see reviews : Farmer, 1970;Willows, 2001). The neural basis of rhythmic swimming has been studied in depth in species belonging to the clade Nudipleura (Fig. 1), which includes Pleurobranchaea californica and the nudibranchs: Tritonia diomedea, Melibe leonina, Dendronotus iris, and Hermissenda crassicornis (Newcomb et al., 2012). These studies were facilitated by the simple swimming behaviors and the large, identifiable neurons that constitute relatively uncomplicated circuits. ...
... Dendronotus, Scyllaea, and Melibe form a clade of LR swimming species. The phylogeny is based on RNA sequencing data (Goodheart et al., 2015) and morphological traits (Newcomb et al., 2012). PRINTED ...
... There are more than 2,000 different species in the monophyletic Nudipleura clade. Of those, fewer than 70 species have been observed to swim (Newcomb et al., 2012). The distribution of swimming in the phylogeny suggests that nonswimming (NS) is the ancestral behavioral state. ...
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This article compares the neural basis for swimming in sea slugs belonging to the Nudipleura clade of mollusks. There are two primary forms of swimming. One, dorsal/ventral (DV) body flexions, is typified by Tritonia diomedea and Pleurobranchaea californica. Although Tritonia and Pleurobranchaea evolved DV swimming independently, there are at least two homologous neurons in the central pattern generators (CPGs) underlying DV swimming in these species. Furthermore, both species have serotonergic neuromodulation of synaptic strength intrinsic to their CPGs. The other form of swimming is with alternating left/right (LR) body flexions. Melibe and Dendronotus belong to a clade of species that all swim with LR body flexions. Although the swimming behavior is homologous, their swim CPGs differ in both cellular composition and in the details of the neural mechanisms. Thus, similar behaviors have independently evolved through parallel use of homologous neurons, and homologous behaviors can be produced by different neural mechanisms.
... Neural networks composed of homologous neurons can be compared across species (Newcomb et al. 2012). Three serotonergic neurons, called variously DSI, As, or CeSP (, ...
... In Tritonia diomedea and Pleurobranchaea californica, the DSIs are members of a central pattern generator (CPG) network that underlies a swimming behavior consisting of alternating dorsal and ventral (DV) flexions (Getting 1989a;Jing and Gillette 1999;Katz 2009). Phylogenetic analysis suggests that although the neurons are homologous, the CPGs and swimming behaviors arose independently and are thus homoplastic (Newcomb et al. 2012). In species lacking the behavior and CPG, the neurons play other roles such as modulating a different CPG (Newcomb and Katz 2009). ...
... Unlike Tritonia and Pleurobranchaea, these species swim using alternating left-right (LR) whole body flexions. The neurons in the Tritonia and Pleurobranchaea DV swim CPGs are not homologous to those in the LR swim CPGs (Newcomb et al. 2012). Thus, the neural circuits underlying the two forms of swimming evolved using non-overlapping sets of identified neurons. ...
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New genetic and molecular methodologies are making it ever more feasible to explore the diversity of motor systems beyond the traditional few model organisms. This chapter reviews the phylogeny and evolution of motor systems, drawing on examples from a diverse array of invertebrate and vertebrate taxa representing a wide variety of motor systems. Phylogenetic analysis allows people to disentangle evolutionary history from current function. Evolutionary studies thus tell us not only about evolution but also about fundamental patterns and processes. As with phylogenetic trees, homology and homoplasy are always hypotheses about evolutionary paths that change as more information about the phylogeny and trait become available. Homology and homoplasy can be considered at any level of biological organization: genes, proteins, neurons, neural structures, or behaviour. In some species individual neurons can be identified across animals based on morphological, neurochemical, or developmental criteria. Understanding how behaviors and neural networks evolved provides insights into how they function and why.
... In many invertebrates, individual neurons can be identified from animal to animal within a species, allowing the neural circuits to be determined with cellular precision. Moreover, homologous neurons can be identified across species, permitting comparative analyses of CPG circuits and the rhythmic behaviours that they produce[4,5]. Thus CPGs provide extraordinary opportunities to study the evolution of behaviour and neural circuits. ...
... In nudibranch molluscs, homologous neurons differ in function in species exhibiting different behaviours. Most nudibranchs do not swim, but those species that do, generally use one of two modes, alternating dorsal –ventral (DV) whole body flexions or rhythmic left–right (LR) flexions[4]. Neurons that are part of the DV swim CPG have homologues in species that produce LR swimming[28 –30], but these homologous neurons are not part of the LR swim CPG. ...
... Neurons that are part of the DV swim CPG have homologues in species that produce LR swimming[28 –30], but these homologous neurons are not part of the LR swim CPG. Instead, the two types of CPG are composed of non-overlapping sets of neurons[4]. Nonetheless, homologues of DV swim CPG neurons can have a neuromodulatory effect on the LR swim CPG[31]. ...
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Comparisons of rhythmic movements and the central pattern generators (CPGs) that control them uncover principles about the evolution of behaviour and neural circuits. Over the course of evolutionary history, gradual evolution of behaviours and their neural circuitry within any lineage of animals has been a predominant occurrence. Small changes in gene regulation can lead to divergence of circuit organization and corresponding changes in behaviour. However, some behavioural divergence has resulted from large-scale rewiring of the neural network. Divergence of CPG circuits has also occurred without a corresponding change in behaviour. When analogous rhythmic behaviours have evolved independently, it has generally been with different neural mechanisms. Repeated evolution of particular rhythmic behaviours has occurred within some lineages due to parallel evolution or latent CPGs. Particular motor pattern generating mechanisms have also evolved independently in separate lineages. The evolution of CPGs and rhythmic behaviours shows that although most behaviours and neural circuits are highly conserved, the nature of the behaviour does not dictate the neural mechanism and that the presence of homologous neural components does not determine the behaviour. This suggests that although behaviour is generated by neural circuits, natural selection can act separately on these two levels of biological organization. © 2015 The Author(s) Published by the Royal Society. All rights reserved.
... Dendro not us bilaterally synchronous bursts of action potentials [73]. In addition, a peptidergic neuron, which has been identified in many other nudipleura species [51], has also been recruited into the escape swim circuit of both species. ...
... The evolution of behaviour and of neural circuits underlying behaviour is intertwined. Studying the neural elements implicated in the evolvability of behaviour among different species has the classical difficulty of distinguishing between homology and homoplasy (but seeNewcomb et al., 2012), a problem that is not present when studying interpopulation differences within a single species. It has been argued that by using interpopulation comparisons, microevolutionary processes can be explicitly investigated, because more populations are likely to be found in the environments that actually shaped their brains (Gonda et al., 2011). ...
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Interpopulation comparisons in species that show behavioural variations associated with particular ecological disparities offer good opportunities for assessing how environmental factors may foster specific functional adaptations in the brain. Yet, studies on the neural substrate that can account for interpopulation behavioural adaptations are scarce. Predation is one of the strongest driving forces for behavioural evolvability and, consequently, for shaping structural and functional brain adaptations. We analysed the escape response of crabs Neohelice granulata from two isolated populations exposed to different risks of avian predation. Individuals from the high-risk area proved to be more reactive to visual danger stimuli (VDS) than those from an area where predators are rare. Control experiments indicate that the response difference was specific for impending visual threats. Subsequently, we analysed the response to VDS of a group of giant brain neurons that are thought to play a main role in the visually guided escape response of the crab. Neurons from animals of the population with the stronger escape response were more responsive to VDS than neurons from animals of the less reactive population. Our results suggest a robust linkage between the pressure imposed by the predation risk, the response of identified neurons and the behavioural outcome.
... Zalypsis, currently in phase II clinical trials for the treatment of various cancers; made from a chemical isolated from Jorunna funebris)789. Strong phylogenetic hypotheses of Cladobranchia will be useful in understanding the evolution of these chemical defences and the evolution of other character traits, such as the ability of many cladobranch species to sequester nematocysts [6] and swimming behaviours [10]. To date, there have been only two large-scale phylogenies published specifically on Cladobranchia [11,12], the first of which used the three genes most commonly used in nudibranch phylogenetics (mitochondrial 16S rRNA and cytochrome oxidase I (COI), and nuclear histone H3) [11]. ...
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Cladobranchia (Gastropoda: Nudibranchia) is a diverse (approx. 1000 species) but understudied group of sea slug molluscs. In order to fully comprehend the diversity of nudibranchs and the evolution of character traits within Cladobranchia, a solid understanding of evolutionary relationships is necessary. To date, only two direct attempts have been made to understand the evolutionary relationships within Cladobranchia, neither of which resulted in well-supported phylogenetic hypotheses. In addition to these studies, several others have addressed some of the relationships within this clade while investigating the evolutionary history of more inclusive groups (Nudibranchia and Euthyneura). However, all of the resulting phylogenetic hypotheses contain conflicting topologies within Cladobranchia. In this study, we address some of these long-standing issues regarding the evolutionary history of Cladobranchia using RNA-Seq data (transcriptomes). We sequenced 16 transcriptomes and combined these with four transcriptomes from the NCBI Sequence Read Archive. Transcript assembly using Trinity and orthology determination using HAMSTR yielded 839 orthologous groups for analysis. These data provide a well-supported and almost fully resolved phylogenetic hypothesis for Cladobranchia. Our results support the monophyly of Cladobranchia and the sub-clade Aeolidida, but reject the monophyly of Dendronotida.
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The wide variety of animal behaviours that can be observed today arose through the evolution of their underlying neural circuits. Advances in understanding the mechanisms through which neural circuits change over evolutionary timescales have lagged behind our knowledge of circuit function and development. This is particularly true for central neural circuits, which are experimentally less accessible than peripheral circuit elements. However, recent technological developments — including cross-species genetic modifications, connectomics and transcriptomics — have facilitated comparative neuroscience studies with a mechanistic outlook. These advances enable knowledge from two classically separate disciplines — neuroscience and evolutionary biology — to merge, accelerating our understanding of the principles of neural circuit evolution. Here we synthesize progress on this topic, focusing on three aspects of neural circuits that change over evolutionary time: synaptic connectivity, neuromodulation and neurons. By drawing examples from a wide variety of animal phyla, we reveal emerging principles of neural circuit evolution. Understanding how brain circuits have been altered by evolution can provide insight into their development and function. Prieto-Godino and colleagues provide an overview of our current understanding of the principles of central circuit evolution, drawing on numerous examples from across the animal kingdom.
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A fundamental question in comparative neuroethology is the extent to which synaptic wiring determines behavior versus the extent to which it is constrained by phylogeny. We investigated this by examining the connectivity and activity of homologous neurons in different species. Melibe leonina and Dendronotus iris (Mollusca, Gastropoda, Nudibranchia) have homologous neurons and exhibit homologous swimming behaviors consisting of alternating left-right (LR) whole body flexions. Yet, a homologous interneuron (Si1) differs between the two species in its participation in the swim motor pattern (SMP) and synaptic connectivity. Here we examine Si1 homologs in two additional nudibranchs: Flabellina iodinea, which evolved LR swimming independently of Melibe and Dendronotus, and Tritonia diomedea, which swims with dorsal-ventral (DV) body flexions. In Flabellina, the contralateral Si1s exhibit alternating rhythmic bursting activity during the SMP and are members of the swim central pattern generator (CPG), as in Melibe. The Si1 homologs in Tritonia do not burst rhythmically during the DV SMP, but are inhibited and receive bilaterally synchronous synaptic input. In both Flabellina and Tritonia, the Si1 homologs exhibit reciprocal inhibition as in Melibe. However, in Flabellina the inhibition is polysynaptic, whereas in Tritonia it is monosynaptic, as in Melibe. In all species, the contralateral Si1s are electrically coupled. These results suggest that Flabellina and Melibe convergently evolved a swim CPG that contains Si1; however, they differ in monosynaptic connections. Connectivity is more similar between Tritonia and Melibe, which exhibit different swimming behaviors. Thus, connectivity between homologous neurons varies independently of both behavior and phylogeny.
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The discovery of shared behavioral processes across phyla is a significant step in the establishment of a comparative study of behavior. We use immobility as an origin and reference for the measurement of fly locomotor behavior; speed, walking direction and trunk orientation as the degrees of freedom shaping this behavior; and cocaine as the parameter inducing progressive transitions in and out of immobility. We characterize and quantify the generative rules that shape Drosophila locomotor behavior, bringing about a gradual buildup of kinematic degrees of freedom during the transition from immobility to normal behavior, and the opposite narrowing down into immobility. Transitions into immobility unfold via sequential enhancement and then elimination of translation, curvature and finally rotation. Transitions out of immobility unfold by progressive addition of these degrees of freedom in the opposite order. The same generative rules have been found in vertebrate locomotor behavior in several contexts (pharmacological manipulations, ontogeny, social interactions) involving transitions in-and-out of immobility. Recent claims for deep homology between arthropod central complex and vertebrate basal ganglia provide an opportunity to examine whether the rules we report also share common descent. Our approach prompts the discovery of behavioral homologies, contributing to the elusive problem of behavioral evolution.
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Gastropods comprise the second largest class in the Animal Kingdom with 60,000 to 80,000 living species occupying ecological niches covering the globe. Anatomy, behaviour and development vary significantly between the five clades of Patellogastropoda, Vetigastropoda,Neritimorpha, Caenogastropoda and Heterobranchia. Generally, the central nervous system consists of paired cerebral, buccal, pleural and pedal ganglia and five ganglia of thevisceral loop, but the ganglia demonstrate vary degrees of asymmetry (chiastoneury, euthyneury) and fusion due to the combined processes of centralizationand torsion, which is a 180o rotation of the posterior portion of the body that occurs early in larval development. Giant, identifiable neurons with characteristic locations, axonal morphology and physiological properties have led to the adoption of some heterobranchs as ‘model organisms’ for investigation of motor and centralpattern generation activity, molecular basis of learning and memory, and single cell transcriptomes. Gastropods also possess extensive peripheral nervous systems containing axons efferent and afferentto the central ganglia and also large numbers of peripheral neurons located within different organs of the body. Most of the classical, small molecule neurotransmitters identified in vertebrates are also found in the central and peripheral neuronsof gastropods together with numerous neuropeptides. The first neural elements (cells of the apical organ, posterior pioneerneurons, peripheralsensory neurons)and also many central neurons appear during trochophore-veliger larval stages, although many more neurons are added during metamorphosis and postlarval development.Gastropods thus provide a unique diversity of form, function and development of the nervous systems offering the opportunity to investigate adaptive evolution of the nervous system at levels of analysis ranging from behaviour to its molecular underpinnings.
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Studies on homologies of identified neurons offer the promise for an understanding of the evolution of gross neural structures and behaviors in terms of the evolution of single nerve cells. Strong cases now exist in the literature for cellular homologies and evidence is available that permits an initial evaluation of which specific features of nerve cells appear to be most conserved through evolution and which features appear to be plastic and therefore permit adaptive variations in the morphology of the nervous system and in its behavioral manifestations. However, due to the relatively small number of putative cellular homologies which have been studied to date, generalizations may be of questionable accuracy. Much more information is necessary in the form of more examples of identifiable cells with known functions. Such examples will possibly allow better insights into how nerve cells adapt to pressures for changes in function. New techniques must also be employed which allow for the sampling of different types of cells than have usually been identified and homologized in the past. Finally, broader phyletic surveys of such neurons are also necessary to test the generality of hypotheses on the conservation and plasticity of neuronal features through evolution.
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A 400 bp region of the mitochondrial 16S rRNA gene was sequenced in 24 dorid nudibranch species. In addition a dendronotid nudibranch and a notaspidean were sequenced for use as outgroup. The sequences were characterized with respect to spatial variation and secondary structure. The data were analysed using parsimony, exploring the effects of alignment and weighting on the optimal hypothesis, and a distance analysis was also performed. The optimal hypotheses differed between the analyses, although the clades with substantial bootstrap proportions were usually present in all cases. These clades are in general congruent with existing classifications, and correspond to relationships within family level taxa. The relationships between these clades were inconclusive and the monophyletic status of higher taxa of particular interest (e.g., Eudoridoidea, Anadoridoidea) could neither be corroborated nor dismissed.
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Neural mechanisms underlying the escape reaction have been extensively studied for over 20 years in many species of animals belonging to several different phyla (for a review, see Eaton, 1984). Among molluscs, the escape reaction has been most thoroughly studied in Tritonia. In this animal a strong mechanical or chemical stimulus applied to the posterior part of the body evokes swimming (for a review, see Getting, 1983; Getting and Dekin, 1985). Electrophysiological experiments on the isolated nervous system revealed that neurones constituting the central pattern generator (CPG) for swimming were strongly depolarized in response to sensory stimulation and that this depolarization caused rhythmic activity in the CPG. However, attempts to identify command neurones mediating the escape reaction in Tritonia have failed (Getting, 1983). This paper deals with the escape reaction in another gastropod mollusc, Clione limacina (subclass Opistobranchia, order Pterapoda). We have been able to identify some of the interneurones involved in this animal's escape reaction. Clione swims using rhythmic movements of two wings, which are controlled by the
Abstract How ecological, developmental and genetic mechanisms interact in the genesis and subsequent diversification of morphological novelties is unknown for the vast majority of traits and organisms. Here we explore the ecological, developmental, and genetic underpinnings of a class of traits that is both novel and highly diverse: beetle horns. Specifically, we focus on the origin and diversification of a particular horn type, those protruding from the pronotum, in the genus Onthophagus, a particularly speciose and morphologically diverse genus of horned beetles. We begin by documenting immature development of nine Onthophagus species and show that all of these species express pronotal horns in a developmentally transient fashion in at least one or both sexes. Similar to species that retain their horns to adulthood, transient horns grow during late larval development and are clearly visible in pupae. However, unlike species that express horns as adults, transient horns are resorbed during pupal development. In a large number of species this mechanisms allows fully horned pupae to molt into entirely hornless adults. Consequently, far more Onthophagus species appear to possess the ability to develop pronotal horns than is indicated by their adult phenotypes. We use our data to expand a recent phylogeny of the genus Onthophagus to explore how the widespread existence of developmentally transient horns alters our understanding of the origin and dynamics of morphological innovation and diversification in this genus. We find that including transient horn development into the phylogeny dramatically reduces the number of independent origins required to explain extant diversity patters and suggest that pronotal horns may have originated only a few times, or possibly only once, during early Onthophagus evolution. We then propose a new and previously undescribed function for pronotal horns during immature development. We provide histological as well as experimental data that illustrate that pronotal horns are crucial for successful ecdysis of the larval head capsule during the larval-to-pupal molt, and that this molting function appears to be unique to the genus Onthophagus and absent in the other scarabaeine genera. We discuss how this additional function may help explain the existence and maintenance of developmentally transient horns, and how at least some horn types of adult beetles may have evolved as exaptations from pupal structures originally evolved to perform an unrelated function.
It is widely believed that behavior is more evolutionarily labile and/or more difficult to characterize than morphology, and thus that behavioral characters are not as useful as morphological characters for estimating phylogenetic relationships. To examine the relative utility of behavior and morphology for estimating phylogeny, we compared levels of homoplasy for morphological and behavioral characters that have been used in systematic studies. In an analysis of 22 data sets that contained both morphological and behavioral characters we found no significant difference between mean consistency indices (CIs, which measure homoplasy) within data sets for the two types of characters. In a second analysis we compared overall CIs for 8 data sets comprised entirely of behavioral characters with overall CIs for 32 morphological data sets and found no significant difference between the two types of data sets. For both analyses, 95% confidence limits on the difference between the two types of characters indicate that, even if given the benefit of the doubt, morphological characters could not have substantially higher mean CIs than behavioral characters. These results do not support the idea that behavioral characters are less useful than morphological characters for the estimation of phylogeny.