Article

Stability of inbred mouse strains in behavior and brain size between laboratories and across decades

Department of Psychology, University of Alberta, Edmonton, AB, Canada T6G 2E9.
Proceedings of the National Academy of Sciences (Impact Factor: 9.67). 11/2006; 103(44):16364-9. DOI: 10.1073/pnas.0605342103
Source: PubMed

ABSTRACT

If we conduct the same experiment in two laboratories or repeat a classical study many years later, will we obtain the same results? Recent research with mice in neural and behavioral genetics yielded different results in different laboratories for certain phenotypes, and these findings suggested to some researchers that behavior may be too unstable for fine-scale genetic analysis. Here we expand the range of data on this question to additional laboratories and phenotypes, and, for the first time in this field, we formally compare recent data with experiments conducted 30-50 years ago. For ethanol preference and locomotor activity, strain differences have been highly stable over a period of 40-50 years, and most strain correlations are in the range of r = 0.85-0.98, as high as or higher than for brain weight. For anxiety-related behavior on the elevated plus maze, on the other hand, strain means often differ dramatically across laboratories or even when the same laboratory is moved to another site within a university. When a wide range of phenotypes is considered, no inbred strain appears to be exceptionally stable or labile across laboratories in any general sense, and there is no tendency to observe higher correlations among studies done more recently. Phenotypic drift over decades for most of the behaviors examined appears to be minimal.

Full-text

Available from: Alexander A Bachmanov, Nov 25, 2014
Stability of inbred mouse strain differences in
behavior and brain size between laboratories
and across decades
Douglas Wahlsten*
, Alexander Bachmanov
, Deborah A. Finn
§
, and John C. Crabbe
§
*Department of Psychology, University of Alberta, Edmonton, AB, Canada T6G 2E9;
Monell Chemical Senses Center, Philadelphia, PA 19104-3308;
and
§
Department of Behavioral Neuroscience, Oregon Health and Science University, and Portland Alcohol Research Center and Veterans Affairs Hospital,
Portland, OR 97239-3098
Edited by Joseph S. Takahashi, Northwestern University, Evanston, IL, and approved September 7, 2006 (received for review June 26, 2006)
If we conduct the same experiment in two laboratories or repeat
a classical study many years later, will we obtain the same results?
Recent research with mice in neural and behavioral genetics
yielded different results in different laboratories for certain phe-
notypes, and these findings suggested to some researchers that
behavior may be too unstable for fine-scale genetic analysis. Here
we expand the range of data on this question to additional
laboratories and phenotypes, and, for the first time in this field, we
formally compare recent data with experiments conducted 30–50
years ago. For ethanol preference and locomotor activity, strain
differences have been highly stable over a period of 40 –50 years,
and most strain correlations are in the range of r 0.85–0.98, as
high as or higher than for brain weight. For anxiety-related
behavior on the elevated plus maze, on the other hand, strain
means often differ dramatically across laboratories or even when
the same laboratory is moved to another site within a university.
When a wide range of phenotypes is considered, no inbred strain
appears to be exceptionally stable or labile across laboratories in
any general sense, and there is no tendency to observe higher
correlations among studies done more recently. Phenotypic drift
over decades for most of the behaviors examined appears to be
minimal.
agonistic behavior anxiety ethanol preference gene– environment
interaction locomotor activity
I
f we conduct the same experiment in two laboratories or repeat
a classical study many years later, will we obtain the same
results? Recent research with mice in neural and behavioral
genetics yielded somewhat different results in different labora-
tories for certain phenotypes (1–4), and these findings evoked
what Pfaff (5) ter med an ‘‘old stereotype,’’ i.e., that behavior may
be too unstable for fine-scale genetic analysis. Here we expand
the range of dat a on this question to additional laboratories and
phenot ypes, and, for the first time in this field, we formally
c ompare recent data with experiments c onducted 30–50 years
ago. We find that several kinds of behavior are genetically quite
st able.
It is now c ommon practice to backcross targeted mut ations in
mice onto the same st andard, inbred strain background in
dif ferent laboratories. If the background genot ype on which
mut ations are placed does not behave c onsistently across labo-
ratories, our understanding of the effects of gene manipulations
on behaviors may be compromised. Comprehensive information
on a wide range of inbred strain characteristics is now being
c ompiled in the Mouse Phenome Database (6) in a manner that
facilit ates comparisons among dif ferent phenotypes studied in
dif ferent laboratories. Many investigators are now using gene
ex pression information for brain tissue of recombinant inbred
strains from the Gene Net work (www.genenetwork.org) to com-
pare phenotypes across strains (7). What constitutes a high
bet ween-trait strain correlation depends critically on the mag-
n itude of the strain correlation for the same trait measured in
dif ferent laboratories or at different times.
To repeat a genetic experiment, the genotype of the animals
must be replicable. Brother-by-sister mating can achieve genetic
purit y in an inbred strain after 60 generations (8). Derivation of
st andard strains of mice commenced with the DBA strain in
1909, BALB in 1913, and C57BL in 1921 (9). By 1950, many
strains maint ained by The Jackson Laboratory (Bar Harbor,
ME) had undergone 40 generations of full sibling mating.
Today, when two laboratories obtain samples of the same strain,
they can be confident that the genotypes are almost identical, but
genetic cont amination can occur when rigorous inbreeding is not
practiced (10–12). Consequently, in this analysis we compare
only data reported for the same substrains from one supplier, the
The Jackson Laboratory (laboratory code J), in almost all cases.
Only simultaneous study of identical tests given to identical
mouse strains can evaluate replicabilit y in the strict sense, and
any differences in results must arise from the inevitable differ-
ence between the environments in the research facilities external
to the test situation itself. Comparisons of reasonably similar
tests given to nearly identical strains in different years in
dif ferent laboratories, however, evaluate the robustness of be-
havioral test results to methodological differences, including
small genetic differences, that exist bet ween most published
reports. Details of the test situation usually dif fer to some extent
bet ween laboratories (13), and the field is principally concerned
with this category.
We selected for historical comparisons studies based on
previously published reports of tests given to inbred strains from
The Jackson Laboratory. First, they had to provide data on at
least five of the same substrains examined by at least one of our
four laboratories. Sec ond, they had to provide sufficient infor-
mation in the published article that we c ould determine the
sample size, mean score, and standard deviation for each strain,
so that an unweighted-means ANOVA could be perfor med to
c ompare studies (14). Third, they had to measure essentially the
same behavior on a scale that allowed meaningful comparisons.
Results
Brain Weight. Unlike behavior, weight of a dead and fixed brain
does not fluctuate f rom moment to moment or day to day, and
Author contributions: D.W., A.B., D.A.F., and J.C.C. designed research; D.W., A.B., D.A.F.,
and J.C.C. performed research; D.W. analyzed data; and D.W., A.B., D.A.F., and J.C.C. wrote
the paper.
The authors declare no conflict of interest.
This article is a PNAS direct submission.
Freely available online through the PNAS open access option.
Data deposition: The data reported in this paper have been deposited in the Mouse
Phenome Database, www.jax.orgphenome (accession no. MPD108).
To whom correspondence should be sent at the present address: Great Lakes Institute,
University of Windsor, Windsor, ON, Canada N9B 3P4. E-mail: wahlsten@uwindsor.ca.
© 2006 by The National Academy of Sciences of the USA
16364–16369
PNAS
October 31, 2006
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it is measured on the same scale in grams in all studies. Fig. 1A
c ompares data collected in the Edmonton laboratory with
strain means in four other data sets. ANOVAs comparing all
possible pairs of dat a sets and det ails of methods for measuring
brains in each study are provided in Tables 1 and 2 and
Suppor ting Tex t, which are published as supporting informa-
tion on the PNAS web site. The Edmonton and Portland strain
means were very similar indeed (r 0.98), and average weights
across all strains were almost identical at the t wo sites (0.452
and 0.456 g, respectively) (15). Methods of fixation were very
similar in the Williams (16) study, and the c orrelation of
Edmonton data w ith their strain means was very high (r
0.88). The t wo older studies (17, 18) both weighed unfixed
brains, and fresh brain weights are ex pected to be slightly
larger than fixed brain weights, but there is no reason to expect
major strain-dependent dif ferences in shrinkage (19).The sim-
ilarit y bet ween the Edmonton data and the Roderick et al.
means collected 30 years earlier on the same strains (17) was
very high (r 0.84), whereas the Storer (18) and Edmonton
dat a were not ably less similar (r 0.54).
There were some discrepancies between older and more
recent data. The clearly greater brain size of C57BL6J than AJ
in our study and the Williams study was not seen in the Roderick
et al. (17) and Storer (18) studies. On the other hand, the tight
clustering of means for strains AKRJ, C57BL6J, and SMJ was
seen in all four studies spann ing a range of 35 years. Strain
SWRJ brains were considerably heavier (by 46 mg) than strain
SJLJ in the Storer data, in contrast w ith the other three studies.
Results from the ANOVAs (Table 1) reveal that the strain-by-
laboratory interaction was in all but one case significant, yet it
was subst antial by our criteria (interaction effect size more than
half the strain main effect size; see Methods) only for the
c omparisons of the Storer data (18) with brain weights from the
Wahlsten laboratory (15).
Fig. 1. Correlations between means of inbred strains observed in different laboratories. The dashed line indicates identical results for two laboratories, and
the gray line is the best fit of data from the y axis to data on the x axis, plotted for the actual range of data. When this line is above the dashed line, the means
scores in laboratory Y were generally greater than for the corresponding strains tested in laboratory X. Numbers 1–21 correspond to strains shown at the bottom
of the figure, with the most common strains shown as colored numerals. Effect sizes from the ANOVAs are shown for the strain main effect and strain-by-
laboratory interaction. The significance (P) of the interaction effect is also indicated. NS denotes an interaction not significant at P 0.05. (A) Brain weight
measured in the Edmonton laboratory versus four other laboratories. Data for Edmonton and Portland were collected in 2002 and published in ref. 15. (B)
Preference for 10% ethanol solution versus tap water. Nine additional strains not studied in the D.W. and J.C.C. laboratories were included in this setof
comparisons (22, BALBcJ; 23, BUBBnJ; 24, C57BLKSJ; 25, CBAJ; 26, CEJ; 27, ILnJ; 28, LPJ; 29, RIIISJ; 30, SEAGnJ). (C) Locomotor activity in Edmonton versus
four other laboratories, scaled to centimeters per minute.
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GENETICS
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Ethanol Preference. The remarkable preference of the C57BL6J
mouse strain for a 10% ethanol solution versus tap water and the
dist aste of DBA 2J for ethanol was originally discovered in 1959
(20), and the techn ique of measuring ethanol preference was
thoroughly ex plored by Fuller (21). In 1966 Rodgers (22) used
the two-bottle preference test to document a wide range of
preference scores for 15–19 inbred strains, and another large
survey of 15 strains was reported 27 years later by Belk nap et al.
(23). The older data are compared here with t wo new inbred
strain surveys done by A.B. (28 strains) and D.A.F. (22 strains)
(Table 3, which is published as supporting infor mation on the
PNAS web site). We present data f rom each study where a
t wo-bottle preference test of 10% ethanol versus tap water was
done with bottle position changed systematically over a period of
at least 4 days. Methods of testing preference are described in
det ail in Table 4, which is published as supporting information
on the PNAS web site. The results of D.A.F. could not be
c ompared with Belknap et al. (23) because the two studies
included only two identical substrains (C57BL6J and DBA2J).
The strain c orrelations among the four studies done 30 or more
years apart are impressive indeed (Fig. 1B and Table 1) and for
three data sets versus Rodgers (22) are slightly greater than the
strain correlations seen for brain weight in independent studies.
A considerable number of other studies with fewer inbred strains
has been published over the years, and, to our knowledge, the
high ethanol preference of C57BL6J versus the pronounced
aversion to ethanol of DBA2J has been confirmed in every
report. Thus, this behavioral difference is highly robust with
respect to laboratory environments and the fine details of
methods used to assess preference.
A lthough strain correlations are generally very high, the
ANOVAs (Table 1) point to several significant strain-by-
laboratory interactions. These t wo indicators of replicability are
not in c onflict, because the interaction effects are invariably
much smaller that the massive main effects of strain. Neverthe-
less, closer inspection of strain distributions (Fig. 1B) reveals
some noteworthy differences for certain pairs of strains. If an
investigator works with only a few inbred strains and happens to
include one or two that are especially sensitive to conditions of
rearing andor testing, these kinds of interaction effects could
under mine replicability.
Locomotor Activity. Depending on the test conditions, locomotion
is used to index complex traits such as general health, ex plor-
atory drive, novelty seeking, anxiet y, and the potential future
preference for drugs of abuse. The first large survey of 15 inbred
strains in 1953 by Thompson (24) involved a simple count by a
human of the lines crossed by the mouse in a box divided into 5
5-inch squares. Fifteen years later, Southwick and Clark (25)
used the same size enclosure but c ounted entries into 6 6-inch
squares. Fifty years after Thompson, Liu and Gershenfeld (26)
used a computer to monitor beam breaks of photocells spaced
2.5 cm apart, whereas the J.C.C. and D.W. laboratories used
video tracking at 25 frames per second. Further details of the test
in all laboratories are specified in Table 5, which is published as
supporting information on the PNAS web site. Results are
c ompared in Fig. 1C and Table 1. To aid comparisons among
studies, we converted all dat a to a common measurement scale,
centimeters traveled per minute.
Fig. 1C and Table 1 indicate that results of studies done 40–50
years apart are remarkably similar. Every laboratory found that
AJ was the least active and C57BL6J was among the most
active strains, and the same result has been reported in virtually
every other published study involving fewer strains. In only one
c omparison was the correlation not ably lower and the interac-
tion term really large relative to the strain main effect: Liu and
Gershenfeld (26) versus Thompson (24). Thompson (24) found
large dif ferences between C3HHeJ and AJ and between
BALBcJ and C57BL6J, whereas Liu and Gershenfeld (26)
observed only minor differences between these strain pairs. This
was not a consequence of the great time span bet ween the
studies, because the Thompson data (24) were highly correlated
with results of Southwick and Clark (25) as well as our own very
recent data.
Elevated Plus Maze. Anxiety or anxiety-like behavior, as assessed
by the elevated plus maze (27, 28), is very sensitive to environ-
ment al variation. In our previous study of eight mouse strains on
the elevated plus maze in three laboratories, average anxiety-like
behavior levels were much lower in Edmonton than Portland for
almost all strains (1), even though we used physically identical
Fig. 2. Percent time spent exploring the two open arms of the elevated plus
maze. Lines, effect sizes, and strain numbers in A and B have the same
meanings as in Fig. 1. (A) For data in the Edmonton and Portland laboratories,
the value for the wild-derived inbred strain PERAEi (number 15 in yellow
circle) in Portland was considerably greater than in Edmonton, but other
means were generally very similar for the two laboratories. (B) There were
remarkable discrepancies for the strains BALBcByJ (no. 3), C3HHeJ (no. 5),
and C57BL6J (no. 6) in the Edmonton laboratory versus the study of Trullas
and Skolnick (30). (C) Four of the same strains were tested in 1998 (1) and again
in 2002 as part of the Mouse Phenome Project using the same apparatus and
protocols as in 1998, but the actual laboratories had moved to new locations
in each university, resulting in substantially different data, especially in Edm-
onton, where every strain was above 30% (blue line) in 1998. (D) Four inbred
strains were tested recently in eight laboratories, indicated in the figure by the
surname of the first author; complete references (30–35) are provided in the
text. The substrains were not identical across all laboratories shown in D.
Dashed lines show that the profile of strain means was very similar for three
studies [Ducottet and Belzung (32), Griebel et al. (33), and Khrapova et al.
(34)], but remarkable differences between other laboratories are evident.
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Page 3
apparatus and test protoc ols. Five years later we ran the same
test in Edmonton and Portland using the same mazes and
protoc ols from the previous study, expanding the sample to 21
inbred strains. This time, our two laboratories obt ained very
similar results (Fig. 2A). ANOVA indicated very large differ-
ences among strains, no significant laboratory difference, and a
very small strain-by-laboratory interaction (Table 1).
Four of the strains in the current study were also tested in our
earlier study (1). As shown in Fig. 2C, open ar m exploration
increased markedly for strain BALBcByJ in Portland, whereas
it decreased considerably for strains AJ, BALBcByJ, and
DBA2J in Edmonton. The apparatus and protocols were the
same, but laboratory environments changed considerably at both
sites. The colony and testing rooms in Edmonton were moved to
another wing of the same building, into an area where several
other mammalian species were housed in a bustling, centralized
an imal facility, whereas the testing laboratory in Portland was
moved from a large facility where many mouse studies were
being done at the same time to a more isolated location in
another building where the mice for this study were housed in the
same room with the test apparatus.
The elevated plus maze test was originally devised for use with
rats and later was adapted for use with mice (29), then applied
to several inbred strains by Trullas and Skoln ick (30). Our
physical maze and procedures and those used by Trullas and
Sk olnick (30) were almost the same (Table 6, which is published
as supporting infor mation on the PNAS web site), because we
found their version to be serviceable and therefore copied it. The
results of our current study and the Tr ullas and Skolnick (30)
study were drastically different for strains C3HHeJ, BALB
cByJ, and C57BL6J (Fig. 2B), and the strain-by-laboratory
interaction was large relative to the main effect of strain.
Studies with 6–11 inbred strains on the elevated plus maze
have been reported from several laboratories (31–35), but
unfortunately the number of specific substrains in common with
our experiment or the study of Trullas and Skolnick (30) was four
or fewer in every inst ance. By treating substrains from different
suppliers as effectively the same strain, it was possible to
c ompute correlations involving three to six strains in common.
Mean scores for four strains that were examined in all eight
laboratories are shown in Fig. 2D. Three studies yielded strain
means that were positively correlated for four to six strains in
c ommon (32–34), whereas the results of Yilmazer-Hanke et al .
(35) were negatively correlated with each of these three studies
but positively correlated with the data from Edmonton and
Portland.
Many factors can influence mouse anxiety (36). Consequently,
the experimenter needs to choose test parameters very carefully.
At the same time, the great similarity of apparatus and proce-
dures in the synchron ized Edmonton and Portland experiments
and the independent test by Trullas and Skolnick (30) did not
yield similar data (Fig. 2B). Furthermore, we could detect no
major differences or similarities in methods, as described in
Table 6, that c ould account for the patterns of strain correlations
among the eight studies (Fig. 2D). This suggests that variations
in the local laboratory environments had import ant influences
on the manifest ation of mouse anxiet y.
Other Behaviors. Agonistic behavior of male mice appears to be
strongly influenced by rearing and testing conditions (37). Two
early studies found opposite rank orderings of two strains (38,
39), an interaction that Ginsburg (40) later attributed to forceps
handling of young mice before weaning in one of the laborato-
ries. Maxson et al. (41) demonstrated Y chromosome ef fects on
agon istic behavior and their interaction with genetic background
(41), but the consomic line differences vanished when the strains
were moved into a specific pathogen-f ree laboratory, a gene
environment interaction that was eventually traced to the acid-
ified water (37). Selective breeding is very effective in creating
large line differences if the males are reared in isolation, but the
line difference largely disappears when they are instead reared
in groups (42–44). Recent inbred strain surveys (45–48) used
very different test methods andor different substrains, and they
dif fered substantially in many details from the 1968 multiple-
strain comparison by Southwick and Clark (25). The available
dat a did not warrant a formal statistical comparison bet ween
studies, and we could not evaluate the stability of sc ores over
decades.
A number of multiple strain surveys of behavior have been
done recently and are reported in the Mouse Phenome Database
(www.jax.orgphenome). At the present time, however, the
Mouse Phenome Database includes few instances where the
same phenotype was examined by dif ferent laboratories. We
have compared data on the accelerating rotarod and water
escape learning in two laboratories, but our refined versions were
c onsiderably different from previous t asks. In the future, it will
be informative to assess replicability by carefully copying the
apparatus and protocols of earlier studies.
Discussion
It is evident from these comparisons among studies that inbred
strain differences for ethanol preference and locomotor activity
are highly replicable across laboratories and even decades,
whereas strain differences in elevated plus maze exploration are
strongly dependent on local conditions. Data on agon istic be-
havior suggest that it is also very sensitive to pretesting envi-
ronment, but stability over decades cannot yet be fairly judged.
Thus, there is no basis for view ing mouse behavior as inherently
unst able or unsuitable for genetic analysis in any general sense.
Some of the classical strain differences in behav ior that were
disc overed 40–50 years ago are highly robust and still evident
today. Other kinds of tests are more labile. These findings do not
imply that tests of anxiety and agonistic interactions are bad
tests. On the contrary, the tests are very sensitive to genuine
individual and strain dif ferences in the constr ucts they are
designed to assess, but the processes underlying those constructs
are also very sensitive to environmental conditions.
The data do not indicate that any one strain is especially stable
or labile across a wide range of behavioral phenotypes. Certain
extreme strain differences on certain tasks, such as C57BL6J
versus DBA2J on ethanol preference and C57BL6J versus AJ
on motor activity, are highly replicable over laboratories and
decades, but those strains show considerable variation for other
phenot ypes such as anxiety-related and agonistic behaviors.
C57BL6J quickly develops obesity and diabetes on a high-fat
diet (49) and as a neonate is easily primed by loud noise so that
it later suffers audigen ic seizures (50). C57BL6J serves as a
good choice when backcrossing a new mutation onto a standard
strain background, not because it is markedly stable phenotyp-
ically, but because it breeds reasonably well and is so widely used
as a de facto standard in different laboratories.
Comparisons of current and classical data suggest that minor
genetic changes over several decades have been of little conse-
quence for mouse behavioral testing and, ipso facto, demonstrate
the robustness of carefully conducted behav ioral assays. It
appears that substrain dif ferentiation is generally not a major
threat to replicabilit y of behavioral data over several decades,
provided the substrains are formed af ter the root strain has
already been inbred for many generations. A study of 1,638
single-nucleotide poly morphisms observed that differences
among several C57BL6 substrains represented residual het-
eroz ygosity at the time substrains were separated as well as new
mut ations (51). Allelic differences at 12 of 342 microsatellite loci
were noted between the C57BL6J and C57BL6NTac sub-
strains that were separated in 1951, 150 generations ago (52).
A C57BL6J mouse from The Jackson Laboratory today will not
Wahlsten et al. PNAS
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GENETICS
Page 4
be identical nucleotide-for-nucleotide to the same strain 50 years
ago. It will nevertheless be far more similar to its own ancestor
in 1950 than to substrains separated early in the history of a
major strain, for example C57BL10, C57L, and C58 (51), or to
another substrain separated around 1950.
A ll of the phenot ypes examined here rank as highly complex
traits. Quantitative trait locus analyses have indicated influences
of multiple loci for brain weight (16) and for the behavioral traits
studied (53). A minor change in one or two of many pertinent
genes should not shift the mean score of a specific strain
appreciably when it is tested several decades later. There does
not seem to be any tendency in the dat a reviewed here for strains
to differ more substantially when the studies were conducted
many years apart. For example, the ethanol preference data of
A.B. were more similar to those of Rodgers (22) from 40 years
earlier than those of Belknap et al. (23) just 10 years earlier.
A lthough the Thompson (24) activit y dat a were not highly
c orrelated with the data from Liu and Gershenfeld (26), they
were very similar to more recent data from the Edmonton
laboratory.
The importance of many fine det ails of husbandry and testing
is recognized on the basis of carefully c ontrolled experiments
c onducted within a single laboratory. Phytoestrogens in the diet
(54), cage enrichment (4), cage position in the colony room (55),
size of the drinking spout orifice (56), shipping before testing
(57), and even the specific experimenter administering the test
(4, 58) have been found to af fect laboratory mouse physiology
and behavior. Nevertheless, just as a single gene usually ac counts
for little variance on its own, it is highly unlikely that a multi-
dimensional laboratory difference in behavioral dat a can be
traced to a single environmental variant. A small environmental
ef fect might be evident within a carefully controlled ex periment,
whereas the same effect might not be audible amidst the
cac ophony of multiple laboratory-specific parameters in rearing
and testing.
The sample of phenotypes in this systematic comparison of
recent and classical data sets is not sufficient to warrant strong
c onclusions about what kinds of behavior should be most and
least robust across laboratories. Very tentatively, we suggest that
things more closely associated with sensory input and motor
output will tend to be less affected by minor variants in the
laboratory environment, whereas behaviors related to emotional
and social processes will be more labile. A similar classification
regarding degree of genetic influence has been proposed on
theoretical grounds by Lipp (59).
Methods
Animals and Laboratories. Data on brain weight, open field activ-
it y, and elevated plus maze were collected simultaneously in the
J.C.C. laboratory in Portland and the D.W. laboratory in Edm-
onton, whereas data on ethanol preference were collected in the
A.B. and D.A.F. laboratories as separate studies done w ithin a
few months of each other. We all obtained the animals from The
Jackson Laboratory at 46 weeks of age. The D.W. and J.C.C.
laboratories evaluated the same 21 inbred strains, c onsisting of
priorit y lists A and B of the Mouse Phenome Project (129S1
SvImJ, AJ, A KRJ, BALBcByJ, C3HHeJ, C57BL6J,
C57LJ, C58J, CASTEiJ, DBA2J, FVBNJ, MOLFEiJ,
NODLtJ, NZBBlNJ, PER AEiJ, PLJ, SJLJ, SMJ,
SPRETEiJ, and SWRJ) plus the strain BTBR T tfJ from
list D (www.jax.orgphenome), which has an interesting but
viable loss of forebrain commissures (15). The A.B. laboratory
studied 28 strains, and the D.A.F. laboratory studied 22 strains
(Table 3), 10 of which were common to at least t wo studies but
were in addition to those studied by the J.C.C. and D.W.
laboratories (129P3J, BALBcJ, BUBBnJ, C57BLKsJ,
CBAJ, CEJ, ILnJ, LPJ, RIIISJ, and SEAGnJ). No at-
tempt was made to equate the housing conditions in the four
laboratories, but conditions were nevertheless quite similar, as
described in Table 7, which is published as supporting informa-
tion on the PNAS web site.
Data Analysis. There currently is no st andard in the field for what
size a strain correlation denotes a bona fide replication of results.
Instead, we rely strongly on close inspection of the data by those
having extensive experience with a particular kind of test. Strictly
speak ing, replication of results in two laboratories requires a
large strain difference but no strain-by-laboratory interaction
ef fect. The magn itude of a strain difference can be ex pressed as
partial
2
, the proportion of variance attributable to the differ-
ences among strain means when laboratory differences are
removed f rom the equation. The partial
2
can also be estimated
for an interaction effect (1). If
2
for an interaction effect is
c onsiderably less than
2
for a strain main effect, then we
c onsider the interaction to be relatively small, even if its statis-
tical sign ificance (P value) is beyond reproach. Likewise, to be
c onsidered large or substantial in a strain-by-laboratories study,
the interaction ef fect size should be at least half the strain main
ef fect size when dat a are compared across two laboratories.
We thank Naomi Yoneyama, Andrea Wetzel, Maria Theodorides, Sue
Burkhart-Kasch, Janet D. Dorow, Jason R. Sibert, Jason P. Schlumbohm,
Charlotte D. Wenger, Chia-Hua Yu, Pamela Metten, Brandie Moisan,
Sean F. Cooper, Tera Mosher, Tim Frigon, and Elizabeth Munn for
assistance in collecting the data. This work was supported in part by
National Institutes of Health grants to D.W. (Grant AA12714), A.B.
(Grant AA11028), J.C.C. (Grant AA10270 and Integrative Neuro-
science Initiative on Alc oholism Consortium Grant AA13519), and
D.A.F. (Integrative Neuroscience Initiative on Alcoholism Consortium
Grants AA134785, AA10760, AA12439, and AA13478); Department of
Veterans Affairs grants (to J.C.C. and D.A.F.); and Natural Sciences and
Engineering Research Council Grant 45825 (to D.W.).
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GENETICS
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  • Source
    • "Housing laboratory animals (defined as intended for research or teaching use, regardless of housing type [Association for Assessment and Accreditation of Laboratory Animal Care International, 2015a]) in simple and uniform conditions is done to limit environmental variability and increase the internal validity of studies, as well as facilitate sample collection, treatments, and husbandry procedures. Recently, it has been demonstrated that highly controlled and standardized laboratory environments for rodent models are generating reduced repeatability and reproducibility in research outcomes [Branchi et al., 2011; Crabbe et al., 1999; Paylor, 2009; Richter et al., 2009 Richter et al., , 2010 Richter et al., , 2011 Schumann et al., 2014; Wahlsten et al., 2006; Wurbel, 2000]. For example, rodent tests comparing results from four repeated experiments using subjects in highly controlled cages (same aged subjects and one enrichment item) versus subjects in less controlled cages (varying aged subjects and enrichment items) show that the highly controlled condition had low within experiment variation, but significant between experiment differences, resulting in low repeatability; the latter condition, however , had greater within experiment variation and low between experiment differences, resulting in greater repeatability in research results [Richter et al., 2009 [Richter et al., , 2010 [Richter et al., , 2011. "
    [Show abstract] [Hide abstract] ABSTRACT: Macaque species, specifically rhesus (Macaca mulatta), are the most common nonhuman primates (NHPs) used in biomedical research due to their suitability as a model of high priority diseases (e.g., HIV, obesity, cognitive aging), cost effective breeding and housing compared to most other NHPs, and close evolutionary relationship to humans. With this close evolutionary relationship, however, is a shared adaptation for a socially stimulating environment, without which both their welfare and suitability as a research model are compromised. While outdoor social group housing provides the best approximation of a social environment that matches the macaque behavioral biology in the wild, this is not always possible at all facilities, where animals may be housed indoors in small groups, in pairs, or alone. Further, animals may experience many housing changes in their lifetime depending on project needs, changes in social status, management needs, or health concerns. Here, we review the evidence for the physiological and health effects of social housing changes and the potential impacts on research outcomes for studies using macaques, particularly rhesus. We situate our review in the context of increasing regulatory pressure for research facilities to both house NHPs socially and mitigate trauma from social aggression. To meet these regulatory requirements and further refine the macaque model for research, significant advances must be made in our understanding and management of rhesus macaque social housing, particularly pair-housing since it is the most common social housing configuration for macaques while on research projects. Because most NHPs are adapted for sociality, a social context is likely important for improving repeatability, reproducibility, and external validity of primate biomedical research. Am. J. Primatol. © 2016 Wiley Periodicals, Inc.
    Full-text · Article · Feb 2016 · American Journal of Primatology
  • Source
    • "Such predictive data suggest that genes like H2-d1, H2-k1, C1qb, Cx3cl1, and Polr3b may be involved in the behavioral processes that are related to and inherently important for social activities of animals. Another interesting finding was that many of these immune genes are associated with the brain morphology (Table 2) (MPD #108) [39]. For example, Cx3cl1 is positively associated with the length of the corpus callosum (CC) (r = 0.89, P = 0.003,Table 2). "
    [Show abstract] [Hide abstract] ABSTRACT: Social deficit is one of the core symptoms of neuropsychiatric diseases, in which immune genes play an important role. Although a few immune genes have been shown to regulate social and emotional behaviors, how immune gene network(s) may jointly regulate sociability has not been investigated so far. To decipher the potential immune-mediated mechanisms underlying social behavior, we first studied the brain microarray data of eight inbred mouse strains with known variations in social behavior and retrieved the differentially expressed immune genes. We then made a protein-protein interaction analysis of them to find the major networks and explored the potential association of these genes with the behavior and brain morphology in the mouse phenome database. To validate the expression and function of the candidate immune genes, we selected the C57BL/6 J and DBA/2 J strains among the eight inbred strains, compared their social behaviors in resident-intruder and 3-chambered social tests and the mRNA levels of these genes, and analyzed the correlations of these genes with the social behaviors. A group of immune genes were differentially expressed in the brains of these mouse strains. The representative C57BL/6 J and DBA/2 J strains displayed significant differences in social behaviors, DBA/2 J mice being less active in social dominance and social interaction than C57BL/6 J mice. The mRNA levels of H2-d1 in the prefrontal cortex, hippocampus, and hypothalamus and C1qb in the hippocampus of the DBA/2 J strain were significantly down-regulated as compared to those in the C57BL/6 J strain. In contrast, Polr3b in the hippocampus and Tnfsf13b in the prefrontal cortex of the DBA/2 J strain were up-regulated. Furthermore, C1qb, Cx3cl1, H2-d1, H2-k1, Polr3b, and Tnfsf13b were predicted to be associated with various behavioral and brain morphological features across the eight inbred strains. Importantly, the C1qb mRNA level was confirmed to be significantly correlated with the sociability in DBA/2 J but not in C57BL/6 J mice. Our study provided evidence on the association of immune gene network(s) with the brain development and behavior in animals and revealed neurobiological functions of novel brain immune genes that may contribute to social deficiency in animal models of neuropsychiatric disorders.
    Full-text · Article · Apr 2015 · Journal of Neuroinflammation
  • Source
    • "under study (Sena et al., 2010). Although less well studied, there is no reason to assume that issues regarding quality of research are different in psychopharmacology than in other fields of CNS research (Wahlsten et al., 2006; Button et al., 2013; Groenink et al., 2014). 2.2.5. "
    [Show abstract] [Hide abstract] ABSTRACT: Psychopharmacology has had some bad publicity lately. Frankly, there have been some major problems along the way in developing new effective drugs for psychiatric disorders. After a prolonged period of high investments but low success rates, big pharmaceutical companies seem to retract their activities in the psychopharmacology field. Yet, the burden of mental disorders is likely to keep on growing in the next decades. In this position paper, we focus on drug development for depression and anxiety disorders, to narrow the scope of the assay. We describe the current situation of the psychopharmacology field, and analyse some of the methods and paradigms that have brought us here, but which should perhaps change to bring us even further. In addition, some of the factors contributing to the current stagnation in psychopharmacology are discussed. Finally, we suggest a number of changes that could lead to a more rational strategy for central nervous system drug development and which may circumvent some of the pitfalls leading to "me too" approaches. Central to the suggested changes, is the notion that mental disorders do not lead to several symptoms, but a network of causally related symptoms convolutes into a mental disorder. We call upon academia to put these changes in the early phases of drug development into effect. Copyright © 2015. Published by Elsevier B.V.
    Full-text · Article · Mar 2015 · European journal of pharmacology
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