Cell Excitability Necessary for Male Mating Behavior
in Caenorhabditis elegans Is Coordinated
by Interactions Between Big Current
and Ether-A-Go-Go Family K+Channels
Brigitte LeBoeuf and L. Rene Garcia1
Howard Hughes Medical Institute, Department of Biology, Texas A&M University, College Station, Texas 77843
ABSTRACT Variations in K+channel composition allow for differences in cell excitability and, at an organismal level, provide flexibility
to behavioral regulation. When the function of a K+channel is disrupted, the remaining K+channels might incompletely compensate,
manifesting as abnormal organismal behavior. In this study, we explored how different K+channels interact to regulate the neuro-
muscular circuitry used by Caenorhabditis elegans males to protract their copulatory spicules from their tail and insert them into the
hermaphrodite’s vulva during mating. We determined that the big current K+channel (BK)/SLO-1 genetically interacts with ether-a-go-
go (EAG)/EGL-2 and EAG-related gene/UNC-103 K+channels to control spicule protraction. Through rescue experiments, we show
that specific slo-1 isoforms affect spicule protraction. Gene expression studies show that slo-1 and egl-2 expression can be upregulated
in a calcium/calmodulin-dependent protein kinase II-dependent manner to compensate for the loss of unc-103 and conversely, unc-
103 can partially compensate for the loss of SLO-1 function. In conclusion, an interaction between BK and EAG family K+channels
produces the muscle excitability levels that regulate the timing of spicule protraction and the success of male mating behavior.
through cross talk by the various molecular signaling compo-
nents. This signaling is essential for fine-tuning cell excitabil-
ity levels to generate appropriate responses and is partially
accomplished by a variety of K+channels. Organisms have
a large number of K+channels that have multiple isoforms
and ancillary subunits (Salkoff et al. 2006; Torres et al. 2007;
Fodor and Aldrich 2009). In excitable cells, a few K+channel
types are responsible for the maintenance of depolarization;
the rest are proposed to be modifiers, fine-tuning the partic-
ular cell or cell type for its specific function (Bargmann 1998;
Santi et al. 2003).
The importance of these modifiers is highlighted by the
multitude of human disorders that exist as a result of
defective K+channels. These disorders include epilepsy,
which can be caused by dysfunction of a variety of channels
HE neuronal and muscular networks that execute be-
havioral responses to stimuli are ultimately maintained
(Schmitt et al. 2000; Du et al. 2005), long Q-T syndrome,
one form of which can be caused by mutations in the HERG
gene and leads to cardiac arrhythmias and sudden death
(Sanguinetti et al. 1995), hypertension caused by kidney
malfunction (Grimm and Sansom 2010), and erectile dys-
function (Werner et al. 2005).
The K+channels, which fine-tune cell excitability, need to
work together. When there is a defective or absent channel,
the remaining K+channels might compensate in an incom-
plete manner. However, the structure and function of the
compensating channels likely differ from the conventional
one, resulting in imperfect regulation of cell excitability. De-
termining how defective K+channels change the dynamics
of the remaining K+channels will provide insights into how
the integration of their functions maintains appropriate ex-
To address how the regulation of cell excitability and
behavior is coordinated by fine-tuning K+channels, we used
Caenorhabditis elegans male mating as a model system. The
stereotyped steps of this behavior include contact response,
backward locomotion and vulval scanning, turning, vulva
sensing, repetitive spicule insertion attempts, complete spicule
Copyright © 2012 by the Genetics Society of America
Manuscript received September 7, 2011; accepted for publication December 3, 2011
1Corresponding author: Department of Biology, 3258 Texas A&M University, College
Station, TX 77843. E-mail: email@example.com
Genetics, Vol. 190, 1025–1041March 2012
penetration, and sperm transfer. Males utilize a pair of copu-
latory spicules to breach the hermaphrodite vulva. The spi-
cules must be maintained inside the male tail prior to
mating and during mating until vulva penetration (Liu and
Sternberg 1995). We have previously identified two K+chan-
nels, ether-a-go-go (EAG) and EAG-related gene (ERG), that
inhibit protraction until the vulva has been breached (Garcia
and Sternberg 2003; LeBoeuf et al. 2007). However, these
two K+channels do not account for all the regulation neces-
sary to coordinate spicule protraction. In this article we iden-
tify the big current (BK) SLO1 K+channel as an additional
regulator of male sex muscle excitability. Since these three K+
channels function in the mating circuit to regulate behavior,
we explored their effects on each other and elucidated how
their interplay maintains the cell excitability that allows
a male to successfully sire progeny.
Materials and Methods
Strains and culture methods
The following strains were used in this study: unc-103(n1213)
(Park and Horvitz 1986), unc-103(sy557) (Garcia and Stern-
berg 2003), and pha-1(e2123) (Schnabel and Schnabel 1990)
on LGIII; unc-43(sy574) (LeBoeuf et al. 2007) on LGIV; him-5
(e1490) (Hodgkin et al. 1979), slo-1(js379) (Wang et al.
2001), slo-1(rg432) (this work), egl-2(rg4) (LeBoeuf et al.
2007), and egl-2(n693) (Reiner et al. 1995) on LGV. C. elegans
is a naturally hermaphroditic species with a low incidence of
males; to generate a larger percentage of males, strains in this
study contained the him-5(e1490) allele. Animals were main-
tained on NGM plates seeded with Escherichia coli strain OP50
at 20? (Brenner 1974).
Protraction constitutive assay
A total of 20–30 virgin L4 males were isolated on NGM plates
with OP50. The males were allowed to develop into adults
overnight and scored as positive for spicule protraction if at
least one spicule partially extended from the cloaca. P values
were determined using GraphPad Prism (version 4.03).
Identification of the slo-1(rg432) mutation
We sequenced all of slo-1, including the promoter region,
from the genomic DNA of an unc-103(n1213); egl-2(rg4)
strain with a lower-than-normal instance of the protraction
constitutive (Prc) phenotype. We discovered that rg432
changes the sequence TGACATTTATATTATCATTTT to TGA-
CATTTATTTTATCATTTT in the intronic region between exons
11 and 12 (exons numbered according to Wang et al. 2001).
pBK1 contains the slo-1 promoter and isoform slo-1(A2;B0;
C1;D0) tagged with GFP and was provided by Michael No-
net (Washington University, St. Louis). To express slo-1(A2;
B0;C1;D0):GFP in specific tissues, we removed the slo-1 pro-
moter from pBK1 and added the XbaI restriction site in front
of the cDNA via single-site mutagenesis with primers
FpBK1XbaI and pBK1r to create plasmid pBL161. pBL161
was then cut with XbaI, blunt-ended, and Gateway vector
conversion reading frame cassette (RfC) A (Invitrogen,
Carlsbad, CA) was ligated into the site to create plasmid
pBL176. The tissue-specific constructs were created using
LR clonase (Invitrogen) to recombine the pBL176 destina-
tion vector with the vectors containing the tissue-specific
promoters. pBL176 was recombined with pLR35 (Paex-3,
pan-neuronal) (LeBoeuf et al. 2007), pLR21 (Punc-103E,
sex muscles) (Reiner et al. 2006), and pLR92 (Pacr-8,
body-wall muscles) (LeBoeuf et al. 2007) to create plasmids
pBL179, pBL180, and pBL181, respectively.
We cloned slo-1(A2;B0;C0;D0) into plasmid pSX322YFP
to generate pBL185cc. slo-1(A2;B0;C0;D0) was cloned from
PCR-amplified cDNA using primers Fslo1cDNASphI and
Rslo1cDNAXbaI. The cDNA was generated from poly(A)
RNA using Qiagen’s (Valencia, CA) Oligotex mRNA Mini
kit and Invitrogen’s SuperScript First-Strand Synthesis sys-
tem for RT–PCR. Both the plasmid and PCR product were
cut with SphI and XbaI and ligated together to generate
pBL185cc. In this manner, we cloned nine different slo-1
full-length cDNA isoforms, and we discovered a new exon
located between exons 2 and 3. We cut pBL185cc with NheI
and ligated Gateway RfC A (Invitrogen) in front of slo-1(A2;
B0;C0;D0) to create plasmid pBL192. pBL192 contained
point mutations in the slo-1 cDNA that changed amino acids.
To correct the slo-1 sequence, we cut pBL192 and a plasmid
containing slo-1(A2;B0;C0;D0) provided by Brandon Johnson
and Miriam Goodman (Stanford University, Palo Alto, CA)
with BseRI. The short fragment from digested slo-1(A2;B0;
C0;D0) was ligated to the long fragment from digested
pBL192 to create pBL224. We recombined a PCR fragment
of the slo-1 promoter generated with primers Fslo1p and
Slo1pr from N2 genomic DNA with pDG15 using BP clonase
(Invitrogen) to generate plasmid pBL153 (the same promoter
region reported in Wang et al. 2001). We performed an LR
reaction between pBL153 and pBL224 to generate plasmid
A plasmid containing a Gateway RfC, slo-1(A1;B0;C0;
D0), and GFP was created by cutting pBL176 and a plasmid
containing slo-1(A1;B0;C0;D0) provided by Miriam Good-
man with StuI and HpaI. The long restriction enzyme prod-
uct from cutting pBL176 was ligated to the short restriction
enzyme product from slo-1(A1;B0;C0;D0) to generate
pBL198. pBL198 was recombined with pLR21 and pBL153
via LR clonase to create plasmids pBL204 and pBL198,
Plasmids containing mutated unc-103 genomic DNAwere
generated using single-site mutagenesis on plasmid pLR67,
which contains unc-103 genomic DNA plus Gateway RfC C.1
(Invitrogen) (Reiner et al. 2006). The primers psy557A and
phosrevsy557A were used to create pLR74 containing the
sy557A mutation, and the primers psy557B and phosrev-
sy557B were used to create pLR75 containing the sy557B
B. LeBoeuf and L. R. Garcia
mutation. pLR74 was recombined with pLR21 and pLR28
(Punc-103F, sex neurons) (Reiner et al. 2006) using LR clo-
nase to create plasmids pBL35 and pBL36, respectively.
pLR75 was recombined with pLR21 and pLR28 using LR
clonase to create plasmids pBL34 and pBL37, respectively.
pTG44 containing the unc-103E promoter driving egl-2
cDNA was constructed as previously described (LeBoeuf
et al. 2007). We added the n693gf mutation by performing
single-site mutagenesis on pTG44 with primers fegl2n698gf
and regl2698gf to create plasmid pBL111 (LeBoeuf et al.
pBL160 was created via an LR reaction between pBL153
(Pslo-1) and pLR186 (Invitrogen Gateway RfC C.1:DsRed1-
E5) (LeBoeuf et al. 2011).
Plasmids containing the construct of interest were injected
into 1-day-old adult hermaphrodites of the appropriate
strain following standard procedures (Mello et al. 1991).
Either 50 ng/ml pBL66 or 100 ng/ml pBX1 were used as
a transgenic marker (Granato et al. 1994; LeBoeuf et al.
2007). pUC18 was used as carrier DNA to complete each
injection mixture to a final concentration of 200 ng/ml. For
each plasmid injected, concentrations were as follows: 20
ng/ml was used for pBL34, pBL35, pBL36, pBL37, pBK1,
pBL180, pBL179, pBL181, and pBL199 and 50 ng/ml was
used for pBL111, pBL160, pBL226, and pBL204. Once stable
transgenic lines were obtained, males from two or more in-
dependent transgenic lines were scored for the Prc pheno-
type following the procedure previously described.
To obtain a stable expression line to measure fluorescent
levels in males carrying Pslo-1:DsRed1-E5, the transgene
was integrated using trimethylpsoralen following standard
procedures (Anderson 1995) to create transgenic line rgIs2.
Image acquisition and quantification
Males carrying the rgIs2 transgene were isolated at mid-L4.
Images were either taken immediately, or the males were
allowed to develop into adults on NGM plates containing E.
coli OP50 for 1 or 2 days. Males were immobilized on 10%
Noble agar (dissolved in water) pads with Polybead poly-
styrene 0.1-mm microspheres (Polysciences, Warrington,
PA) and covered with a cover slip. Images were taken using
an Olympus BX51 microscope with a ·40 objective. The
image signal was split into two channels using the Dual View
Simultaneous Imaging system by Photometrics (Tucson, AR).
Pictures were taken by the Hamamatsu EM-CCD digital cam-
era ImagEM using HC Image (version 126.96.36.199) (Bridgewater,
NJ). All images were taken with the gain, sensitivity, and
exposure time held constant. The one exception is at the
2-day time point, when the red channel was overexposed at
the standard settings. One image was taken at the standard
settings and a second image was taken when the gain was
lowered so the red channel was no longer overexposed. Data
from both images were used to obtain the mean gray level for
the red channel (see below).
Images were analyzed using Hamamatsu Simple PCI
(version 188.8.131.52). A region of interest (ROI) was placed over
the male tail in both the red and green channels and in the
background of both the red and green channels to get the
background fluorescent level. The ROI was held to a constant
size using the “ROI clone” tool. The mean gray level was
recorded for each ROI, and the background fluorescence
was subtracted. For 2 day-old adult males, the mean gray
level was determined for the green channel under standard
settings as well as both the green and red channel with the
gain lowered. Since the green channel was not overexposed
in either image, a ratio between the high gain and low gain
image was obtained. This ratio was then combined with the
low gain red mean gray level to approximate the standard
gain red mean gray level. The analyzed images were not
modified in any way. Statistical analysis was performed using
GraphPad Prism (version 4.03).
Quantitative real-time PCR
Each strain of worms was cultivated on six 100-mm NGM
plates containing E. coli OP50. Worms were harvested be-
fore they starved by washing the plates with M9 buffer.
Worms were collected at 1000 rpm and the M9 and E. coli
were removed. The worms were washed with an additional
1 ml of M9 to remove additional E. coli. For experiments
where hermaphrodites were segregated from males, worms
were first placed on a 20-mM Nitex nylon filter to remove
larva, and then on a 35-mM Nitex nylon filter to separate
males and hermaphrodites (Sefar Filtration, Depew, NY).
Worms were divided into four 1.5-ml microcentrifuge tubes
and 250 ml Tri Reagent (Sigma-Aldrich, St. Louis) and 0.5
mm zirconium oxide beads (Next Advance, Cambridge, MA)
were added. The Bullet Blender (Next Advance) set for
3 min at speed 8 was used to break open the worms. The
debris, Tri Reagent, and beads were spun down for 1 min at
top speed in a microcentrifuge. The Tri Reagent and RNA
were moved to a fresh RNase-free 0.5-microcentrifuge tube.
Ten microliters of glycogen and 50 ml of chloroform were
added, and the mixture was shaken by hand and centrifuged
for 10 min at 12,000 rpm to separate the layers. The aque-
ous layer was transferred to a fresh 0.5-RNase-free micro-
centrifuge tube. Two ethanol precipitations were performed
to remove any salts that might interfere with the ability to
separate poly(A) RNA from total RNA. The quality of the
total RNA was determined using a 2% agarose gel and the
quantity was measured using a spectrophotometer. Poly(A)
RNA was obtained using Ambion’s Poly(A) Purist kit (Ap-
plied Biosystems, Austin, TX). cDNA was made using the
SuperScript First-Strand Synthesis system for RT–PCR (Invi-
trogen) with 200 ng of starting poly(A) RNA from each
sample and replicated at least one time. Reference genes
were determined as done in Vandesompele et al. (2002)
and Hoogewijs et al. (2008). Thirteen reference genes
were considered: act-1, gpd-3, F23B2.13, rrn-1.1, cdc-42,
Overlapping K+Channels in Mating
Kapiloff, M. S., J. M. Mathis, C. A. Nelson, C. R. Lin, and M. G.
Rosenfeld, 1991 Calcium/calmodulin-dependent protein ki-
nase mediates a pathway for transcriptional regulation. Proc.
Natl. Acad. Sci. USA 88: 3710–3714.
Keren, H., M. Donyo, D. Zeevi, C. Maayan, T. Pupko et al.,
2010 Phosphatidylserine increases IKBKAP levels in familial
dysautonomia cells. PLoS ONE 5: e15884.
Kim, H., J. T. Pierce-Shimomura, H. J. Oh, B. E. Johnson, M. B.
Goodman et al., 2009The dystrophin complex controls BK
channel localization and muscle activity in Caenorhabditis ele-
gans. PLoS Genet. 5: e1000780.
Kim, J. M., R. Beyer, M. Morales, S. Chen, L. Q. Liu et al.,
2010 Expression of BK-type calcium-activated potassium chan-
nel splice variants during chick cochlear development. J. Comp.
Neurol. 518: 2554–2569.
Lamba, J. K., Y. S. Lin, E. G. Schuetz, and K. E. Thummel,
2002 Genetic contribution to variable human CYP3A-medi-
ated metabolism. Adv. Drug Deliv. Rev. 54: 1271–1294.
LeBoeuf, B., T. R. Gruninger, and L. R. Garcia, 2007
vation attenuates seizures through CaMKII and EAG K+ Chan-
nels. PLoS Genet. 3: e156.
LeBoeuf, B., X. Guo, and L. R. Garcia, 2011
sient starvation persist through direct interactions between
CaMKII and ether-a-go-go K+ channels in C. elegans males.
Neuroscience 175: 1–17.
Lee, J., and C. F. Wu, 2010Orchestration of stepwise synaptic
growth by K+ and Ca2+ channels in Drosophila. J. Neurosci.
Lee, U. S., and J. Cui, 2010 BK channel activation: structural and
functional insights. Trends Neurosci. 33: 415–423.
Li, H., W. Li, A. K. Gupta, P. J. Mohler, M. E. Anderson et al.,
2010 Calmodulin kinase II is required for angiotensin II-medi-
ated vascular smooth muscle hypertrophy. Am. J. Physiol. Heart
Circ. Physiol. 298: H688–H698.
Lints, R., and S. W. Emmons, 1999
neurotransmitter identity among Caenorhabditis elegans ray sen-
sory neurons by a TGFbeta family signaling pathway and a Hox
gene. Development 126: 5819–5831.
Little, G. H., Y. Bai, T. Williams, and C. Poizat, 2007
calcium/calmodulin-dependent protein kinase IIdelta preferen-
tially transmits signals to histone deacetylase 4 in cardiac cells.
J. Biol. Chem. 282: 7219–7231.
Liu, J., M. Asuncion-Chin, P. Liu, and A. M. Dopico, 2006
kinase II phosphorylation of slo Thr107 regulates activity and
ethanol responses of BK channels. Nat. Neurosci. 9: 41–49.
Liu, K. S., and P. W. Sternberg, 1995
mating behavior in Caenorhabditis elegans. Neuron 14: 79–89.
Liu, Q., B. Chen, Q. Ge, and Z. W. Wang, 2007a
+/calmodulin-dependent protein kinase II modulates neuro-
transmitter release by activating BK channels at Caenorhabditis
elegans neuromuscular junction. J. Neurosci. 27: 10404–10413.
Liu, Y., B. LeBoeuf, and L. R. Garcia, 2007b
muscarinic acetylcholine receptors enhance nicotinic acetylcho-
line receptor signaling in Caenorhabditis elegans mating behav-
ior. J. Neurosci. 27: 1411–1421.
Liu, Y., B. LeBeouf, X. Guo, P. A. Correa, D. G. Gualberto et al.,
2011 A cholinergic-regulated circuit coordinates the main-
tenance and bi-stable states of a sensory-motor behavior dur-
ing Caenorhabditis elegans male copulation. PLoS Genet. 7:
Livak, K. J., and T. D. Schmittgen, 2001
expression data using real-time quantitative PCR and the 2
(-Delta Delta C(T)). Method. Methods 25: 402–408.
Longman, D., R. H. Plasterk, I. L. Johnstone, and J. F. Caceres,
2007 Mechanistic insights and identification of two novel fac-
tors in the C. elegans NMD pathway. Genes Dev. 21: 1075–1085.
The effects of tran-
Patterning of dopaminergic
Sensory regulation of male
Analysis of relative gene
MacDonald, S. H., P. Ruth, H. G. Knaus, and M. J. Shipston,
2006 Increased large conductance calcium-activated potas-
sium (BK) channel expression accompanied by STREX variant
downregulation in the developing mouse CNS. BMC Dev. Biol.
Maquat, L. E., 2004Nonsense-mediated mRNA decay: splicing,
translation and mRNP dynamics. Nat. Rev. Mol. Cell Biol. 5:
McKim, K. S., K. Peters, and A. M. Rose, 1993
required for meiotic chromosome pairing in Caenorhabditis ele-
gans. Genetics 134: 749–768.
Mello, C. C., J. M. Kramer, D. Stinchcomb, and V. Ambros,
1991 Efficient gene transfer in C.elegans: extrachromosomal
maintenance and integration of transforming sequences. EMBO
J. 10: 3959–3970.
Mongan, N. P., A. K. Jones, G. R. Smith, M. S. Sansom, and D. B.
Sattelle, 2002 Novel alpha7-like nicotinic acetylcholine recep-
tor subunits in the nematode Caenorhabditis elegans. Protein Sci.
Oruganti, S. R., S. Edin, C. Grundstrom, and T. Grundstrom,
2011 CaMKII targets Bcl10 in T-cell receptor induced activa-
tion of NF-kB. Mol. Immunol. 48: 1448–1460.
Park, E. C., and H. R. Horvitz, 1986
effects on the behavior and morphology of the nematode Cae-
norhabditis elegans. Genetics 113: 821–852.
Ramirez, M. T., X. L. Zhao, H. Schulman, and J. H. Brown,
1997The nuclear deltaB isoform of Ca2+/calmodulin-depen-
dent protein kinase II regulates atrial natriuretic factor gene
expression in ventricular myocytes. J. Biol. Chem. 272:
Reiner, D. J., D. Weinshenker, and J. H. Thomas, 1995
dominant mutations affecting muscle excitation in Caenorhabdi-
tis elegans. Genetics 141: 961–976.
Reiner, D. J., E. M. Newton, H. Tian, and J. H. Thomas,
1999 Diverse behavioural defects caused by mutations in Cae-
norhabditis elegans unc-43 CaM kinase II. Nature 402: 199–203.
Reiner, D. J., D. Weinshenker, H. Tian, J. H. Thomas, K. Nishiwaki
et al., 2006 Behavioral genetics of Caenorhabditis elegans unc-
103-encoded erg-like K(+) channel. J. Neurogenet. 20: 41–66.
Ronkainen, J. J., S. L. Hanninen, T. Korhonen, J. T. Koivumaki, R.
Skoumal et al., 2011 Ca2+-calmodulin-dependent protein ki-
nase II represses cardiac transcription of the L-type calcium
channel 1C-subunit gene (Cacna1c) by DREAM translocation.
J. Physiol. 589: 2669–2686.
Salkoff, L., A. Butler, G. Ferreira, C. Santi, and A. Wei, 2006
conductance potassium channels of the SLO family. Nat. Rev.
Neurosci. 7: 921–931.
Sanguinetti, M. C., C. Jiang, M. E. Curran, and M. T. Keating,
1995 A mechanistic link between an inherited and an acquired
cardiac arrhythmia: HERG encodes the IKr potassium channel.
Cell 81: 299–307.
Santi, C. M., A. Yuan, G. Fawcett, Z. W. Wang, A. Butler et al.,
2003Dissection of K+ currents in Caenorhabditis elegans mus-
cle cells by genetics and RNA interference. Proc. Natl. Acad. Sci.
USA 100: 14391–14396.
Schmitt, N., M. Schwarz, A. Peretz, I. Abitbol, B. Attali et al.,
2000A recessive C-terminal Jervell and Lange-Nielsen muta-
tion of the KCNQ1 channel impairs subunit assembly. EMBO J.
Schnabel, H., and R. Schnabel, 1990
ation gene, pha-1, from Caenorhabditis elegans. Science 250:
Sulston, J., M. Dew, and S. Brenner, 1975
in the nematode Caenorhabditis elegans. J. Comp. Neurol. 163:
Two types of sites
Mutations with dominant
An organ-specific differenti-
B. LeBoeuf and L. R. Garcia
Sulston, J. E., D. G. Albertson, and J. N. Thomas, 1980
norhabditis elegans male: postembryonic development of non-
gonadal structures. Dev. Biol. 78: 542–576.
Sulston, J. E., E. Schierenberg, J. G. White, and J. N. Thomson,
1983 The embryonic cell lineage of the nematode Caenorhab-
ditis elegans. Dev. Biol. 100: 64–119.
Sze, J. Y., M. Victor, C. Loer, Y. Shi, and G. Ruvkun, 2000
and metabolic signalling defects in a Caenorhabditis elegans
serotonin-synthesis mutant. Nature 403: 560–564.
Terskikh, A., A. Fradkov, G. Ermakova, A. Zaraisky, P. Tan et al.,
“Fluorescent timer”: protein that changes color with
time. Science 290: 1585–1588.
Torres, Y. P., F. J. Morera, I. Carvacho, and R. Latorre, 2007
marriage of convenience: beta-subunits and voltage-dependent
K+ channels. J. Biol. Chem. 282: 24485–24489.
Tseng-Crank, J., C. D. Foster, J. D. Krause, R. Mertz, N. Godinot
et al., 1994 Cloning, expression, and distribution of function-
ally distinct Ca(2+)-activated K+ channel isoforms from human
brain. Neuron 13: 1315–1330.
Vandesompele, J., K. De Preter, F. Pattyn, B. Poppe, N. Van Roy
et al., 2002Accurate normalization of real-time quantitative
RT-PCR data by geometric averaging of multiple internal control
genes. Genome Biol. 3: RESEARCH0034.
Wang, Z. W., O. Saifee, M. L. Nonet, and L. Salkoff, 2001
potassium channels control quantal content of neurotransmitter
release at the C. elegans neuromuscular junction. Neuron 32:
Watanabe, Y., S. Kato, Y. Adachi, and K. Nakashima, 2000
shift, nonsense and non amino acid altering mutations in SOD1
in familial ALS: report of a Japanese pedigree and literature
review. Amyotroph. Lateral Scler. Other Motor Neuron Disord.
Weinshenker, D., G. Garriga, and J. H. Thomas, 1995
pharmacological analysis of neurotransmitters controlling egg
laying in C. elegans. J. Neurosci. 15: 6975–6985.
Weinshenker, D., A. Wei, L. Salkoff, and J. H. Thomas, 1999
of an ether-a-go-go-like K(+) channel by imipramine rescues egl-
2 excitation defects in Caenorhabditis elegans. J. Neurosci. 19:
Werner, M. E., P. Zvara, A. L. Meredith, R. W. Aldrich, and M. T.
Nelson, 2005Erectile dysfunction in mice lacking the large-
conductance calcium-activated potassium (BK) channel. J. Phys-
iol. 567: 545–556.
Xie, J., and D. P. McCobb, 1998
potassium channels by stress hormones. Science 280: 443–446.
Yuan, P., M. D. Leonetti, A. R. Pico, Y. Hsiung, and R. MacKinnon,
2010Structure of the human BK channel Ca2+-activation ap-
paratus at 3.0 Å resolution. Science 329: 182–186.
Zhang, T., S. Miyamoto, and J. H. Brown, 2004
calcium and calcium/calmodulin-dependent protein kinase II:
friends or foes? Recent Prog. Horm. Res. 59: 141–168.
Control of alternative splicing of
Communicating editor: R. Anholt
Overlapping K+Channels in Mating