Chemical Senses Advance Access originally published online on June 6, 2006
Chemical Senses 2006 31(7):613-619; doi:10.1093/chemse/bjj066
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Attenuation of the Production of Inositol 1,4,5-Trisphosphate in the Mouse Vomeronasal Organ by Antibodies Against the 
q/11 Subfamily of G-Proteins
Department of Biological Sciences, Alabama State University, Montgomery, AL 36101-0271, USA
Correspondence to be sent to: Kennedy S. Wekesa, Department of Biological Sciences, Alabama State University, Montgomery, AL 36101-0271, USA. e-mail: kwekesa{at}alasu.edu
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and methodsResultsDiscussionAcknowledgementsReferences |
|---|
The social and reproductive behaviors of most mammals are modulated by pheromones, which are perceived by the vomeronasal organ (VNO). Vomeronasal transduction in vertebrates is activated through G-protein–coupled receptors, which in turn leads to the generation of inositol 1,4,5-trisphosphate (IP3) and diacylglycerol (DAG) by the activity of phospholipase C. DAG has been shown to gate the transient receptor potential channel 2, whereas IP3 may play a role in stimulating the release of calcium from the endoplasmic reticulum store. To investigate the role of the alpha subunits of Gq/11 in the transduction process, microvillar membranes from female mice VNO were preincubated with a selective C-terminal peptide antibody against G
q/11 and then stimulated with adult male urine. Incubation of VNO membranes with antibodies against Gq/11 blocked the production of IP3 in a dose-dependent manner. We were also able to impair the production of IP3 when we stimulated with 2-heptanone or 2,5-dimethylpyrazine in the presence of antibodies against the alpha subunit of Gq/11. 2-Heptanone is a known pheromone that has been linked to VIR receptors. Thus, our observations indicate that the alpha subunits of Gq/11 play a role in pheromonal signaling in the VNO.
Key words: 2,5-dimethylpyrazine, G-proteins, 2-heptanone, IP3, pheromones, vomeronasal
|
Introduction |
|---|
|
TopAbstractIntroductionMaterials and methodsResultsDiscussionAcknowledgementsReferences |
|---|
The social and reproductive behaviors of most mammals are modulated by pheromones, which are chemical signals among conspecifics. The perception of pheromones is mainly mediated by the vomeronasal organ (VNO) which is located at the base of the nasal septum. The VNO contains a lumen that communicates via a duct with the oral or nasal cavity. Therefore, chemical stimuli in urine and glandular secretions when inhaled can act upon the dendritic microvilli of bipolar chemosensory neurons in the VNO. These chemosensory neurons express two classes of putative pheromone receptor gene families, V1R and V2R, segregated in zones (Halpern et al., 1995
; Berghard and Buck, 1996; Jia and Halpern, 1996; Wekesa and Anholt, 1999; Dulac, 2000; Pantages and Dulac, 2000). The V1R family of putative pheromone receptors is coexpressed with the G-protein Gi2 on the apical zone of the vomeronasal neuroepithelium that projects to the anterior aspect of the accessory olfactory bulb. The V2R family of receptors are coexpressed with the G-protein Go in the basal zone that projects to the posterior aspect of the accessory olfactory bulb (Halpern et al., 1995; Berghard and Buck, 1996; Jia and Halpern, 1996; Wekesa and Anholt, 1999; Dulac, 2000; Pantages and Dulac, 2000). V1R receptors are structurally similar to olfactory receptors although evolutionarily unrelated, and there are estimated to be about 150 different types (Herrada and Dulac, 1997; Matsunami and Buck, 1997; Ryba and Tirindelli, 1997; Rodriguez et al., 2002). V2Rs on the other hand have a long extracellular N-terminal region believed to be involved in ligand binding and are similar to metabotropic glutamate receptors. There are estimated to be 100 V2Rs in rodents, arrayed into several subfamilies (Herrada and Dulac, 1997; Matsunami and Buck, 1997; Ryba and Tirindelli, 1997).
Electrophysiological and calcium-imaging studies have shown that the V1R class of receptors respond to volatile chemicals including 2-heptanone and 2,5-dimethylpyrazine (Leinders-Zufall et al., 2000; Boschat et al., 2002), whereas the V2R receptors respond to larger nonvolatile compounds such as MHC class I peptides (Krieger et al., 1999; Leinders-Zufall et al., 2004). 2-Heptanone found in both male and female urine extends the estrous cycle in female mice, whereas 2,5-dimethylpyrazine found only in female urine acts to delay puberty in females (Novotny et al., 1985; Jemiolo et al., 1989; Novotny, 2003). MHC class I peptides are involved in social recognition signals that convey information about genetic individuality (Leinders-Zufall et al., 2004).
The transduction cascade in the VNO of vertebrates is distinct and is driven by phospholipase-C (PLC)–induced production of diacylglycerol (DAG) and inositol 1,4,5-trisphosphate (IP3), and the subsequent increase in intracellular calcium concentration (Luo et al., 1994; Inamura et al., 1997a,b; Wekesa and Anholt, 1997; Holy et al., 2000; Inamura and Kashiwayanagi, 2000; Leinders-Zufall et al., 2000; Cinelli et al., 2002; Wekesa et al., 2003). DAG gates the transient receptor potential channel 2 (TRPC2), whereas IP3 may a play a role in stimulating the release of calcium from the endoplasmic reticulum store (Lucas et al., 2003).
Several G-protein subunits have been identified in the vomeronasal neurons, including Gi2, Go, Gq/11, ß
2, G2, and G8 (Berghard et al., 1996; Jia and Halpern, 1996; Tirindelli and Ryba, 1996; Runnenburger et al., 2002; Wekesa et al., 2003), but the G-protein alpha subunits that may be involved in pheromonal signaling have yet to be clearly shown. In order to determine which of the G-protein alpha subunits plays a role in the activation of PLC, we used antibodies prepared against the C-terminus. This approach is based on the evidence that the C-terminal region of the G-protein alpha subunits is involved in receptor coupling (Birnbaumer et al., 1990), and therefore, antibodies that bind selectively to this region can disrupt alpha subunit activity. These antibodies uncouple G-proteins from their receptors in situ and have previously been used to selectively block activation of transducin, Gs, Gi2, Go, and Gq (Cerione et al., 1988; Mackenzie et al., 1988; McFadzean et al., 1989; Simonds et al., 1989; McClue and Milligan, 1990).
Here we show that antibodies against Gq/11 can block the production of IP3 in both VIR and V2R receptors in the female VNO when stimulated by whole adult male urine or isolated urinary compounds such as 2-heptanone or 2,5-dimethylpyrazine.
|
Materials and methods |
|---|
|
TopAbstractIntroductionMaterials and methodsResultsDiscussionAcknowledgementsReferences |
|---|
Animals
CD-1 mice were originally obtained from Charles River Laboratories (Kingston, NY) and maintained in a breeding colony in the Department of Biological Sciences at Alabama State University. Animals were housed in Institutional Animal Care and Use Committee inspected and approved facilities and cared for according to the National Institutes of Health (NIH) Guide for Care and Use of Laboratory Animals. Mice were kept in Nalgene cages 26 x 21 x 14 cm, at 25°C room temperature, and a 12:12 h light:dark cycle. Food and water were provided ad libitum.
Membrane preparations
VNOs from female mice, up to 4 weeks old, were dissected from their crevices in the nasal cavity, removed from the cartilaginous capsule, and frozen on dry ice. The tissues were then minced with a razor blade and subjected to sonication for 2–5 min in ice-cold phosphate-buffered saline (PBS). The resulting suspension was layered on a 45% (w/w) sucrose cushion and centrifuged at 4°C for 30 min at 40,000 rpm in a Beckman SW55Ti rotor. The membrane fraction on top of the sucrose was collected and centrifuged as before for 15 min to pellet the membranes. The membranes were resuspended in 100 µl of ice-cold PBS. Protein was then determined according to the method of Lowry et al. (1951), using bovine serum albumin as standard. The procedure used for the preparation of microvillar membranes is modeled after well-established methods (Anholt, 1995; Wekesa and Anholt, 1999; Wekesa et al., 2003). These preparations have been previously characterized (Anholt et al., 1986; Anholt 1995; Wekesa and Anholt, 1997) and are sufficiently enriched in chemosensory membranes. Membranes from the liver were prepared by homogenizing the tissue in PBS with a Teflon homogenizer. Membranes were collected by centrifugation, washed once, and suspended in PBS.
Second messenger assays
The chemicals 2-heptanone, 2,5-dimethylpyrazine, and angiotensin II (AngII) were purchased from Aldrich Chemical Co. (Milwaukee, WI). For IP3 assays, reactions were incubated for 1 min at 37°C in 25 mM Tris-acetate buffer pH 7.2, 5 mM Mg-acetate, 1 mM dithiothreitol, 0.5 mM adenosine triphosphate, 0.1 mM CaCl2, 0.1 mg/ml bovine serum albumin, 10 µM guanosine triphosphate, and 20 µg VNO membrane proteins. Reactions were terminated by the addition of 1 M trichloroacetic acid. IP3 was measured with a kit from Perkin Elmer, Inc. (Boston, MA) according to the manufacturer's instructions and is based on displacement of [3H] IP3 from a specific IP3 binding protein. Differences between experimental and control animals were analyzed by analysis of variance.
Antibodies
The antibodies against alpha subunits of G-proteins were purchased from Calbiochem (La Jolla, CA). The C-terminus of the G alpha subunits is a defined receptor contact site, and the overexpression of peptides corresponding to this region competitively blocks interaction between G-protein–coupled receptors (GPCRs) and the targeted G-proteins (Gilchrist et al., 2001). Consistent with this, the last five C-terminal amino acids of Gq/11 are important for maintaining receptor selectivity and signaling capacity (Gilchrist et al., 2001). The antibody against the alpha subunit of Gq/11 was raised against the last 12 amino acids on the carboxy-tail peptide (QLNLKEYNLV). The antibody against the alpha subunit of Gi2 was raised against the C-terminal peptide KNNLDCGLF.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and methodsResultsDiscussionAcknowledgementsReferences |
|---|
Inhibition of AngII-mediated stimulation of IP3 in membranes from liver
The peptide hormone AngII regulates vasoconstriction, water and salt balance, and neuromodulation and cellular growth through actions on two types of AngII receptors type 1, AT1, and type 2, AT2 (de Gasparo et al., 1995). The AT1 receptor is the principal mediator of biological actions of AngII. AT1 receptors are GPCRs and activate G-proteins through the membrane-proximal regions of the third cytoplasmic loop and C-terminus in the receptor (Sano et al., 1997; Kai et al., 1998). AT1 activation by AngII results in the generation of IP3 and DAG, via a PLC–coupled G-protein, Gq/11, to cause release of calcium from intracellular stores and activation of protein kinase C (Griendling et al., 1997). Previous experiments using antibodies against Gq/11 impaired the production of IP3 in this well-studied system (Gutowski et al., 1991). In order to demonstrate that our antibodies can indeed block Gq/11, we used the AngII system in the liver. Incubation of liver membranes with AngII resulted in a robust increase of IP3 (P < 0.05; Figure 1), whereas liver membranes that had been preincubated with 100-fold dilution of antibodies against Gq/11 did not show an increase in IP3. This experiment confirmed that our antibodies were able to disrupt the activity of the alpha subunit of Gq/11.
|
Dose-dependent inhibition of the production of IP3 in female VNO membranes by antibodies against G
q/11
To study transduction pathways activated by pheromonal stimuli, we developed preparations enriched in microvillar membranes from VNOs of females. Incubation of microvillar VNO membranes from females with adult male urine results in a significant increase in the production of IP3 (P < 0.05; Figure 2). In order to determine if Gq/11 mediated the activation of PLC, we preincubated our VNO membranes in varying concentrations of antibodies against Gq/11. We observed that stimulation of VNO membranes with male urine in the presence of antibodies against Gq/11 impaired the production of IP3 in a dose-dependent manner.
|
Female VNO response to male urine is not impaired by antibodies against G
i2
In order to determine if the alpha subunits of Gi2 have the same effects as Gq/11, we preincubated our VNO membranes with antibodies against Gi2. Incubation of Gi2 antibodies in the presence of male urine did not impair the production of IP3 (Figure 3).
|
Female VNO response to urinary chemicals is impaired by antibodies against G
q/11
The stimulation of microvillar membranes from VNOs of female mice with 2-heptanone and 2,5-dimethylpyrazine resulted in an increase in the production of IP3 as compared to the control (P < 0.05; Figure 4). This effect was blocked by preincubation of the VNO membranes with antibodies against Gq/11.
|
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and methodsResultsDiscussionAcknowledgementsReferences |
|---|
Understanding the signal transduction cascade in the VNO is an important step in understanding how this system encodes pheromonal information which is important for regulating reproductive behaviors and physiology. There is a lot of information on the molecular entities (receptors and G-proteins) that may be involved in pheromonal signaling but how these different molecules actually interact is not clear. Several studies using immunohistochemical techniques or in situ hybridization have shown that there are three G-proteins on the microvillar surface of the VNO, G
i2, Go, and Gq/11 (Wekesa and Anholt, 1997; Berghard and Buck, 1996). Roles for activation of phosphoinositide hydrolysis have been reported for Gi2 (Ohta et al., 1985) and for Go (Moriarty et al., 1990; Blitzer et al., 1993).
Gi2 has been implicated in inhibition of adenylyl cyclase (Watkins et al., 1992; Wong et al., 1992; Taussig et al., 1993), potassium channel activation (Yatani et al., 1988; Kobayashi et al., 1990), and modulation of calcium channels (Linder et al., 1990). The principal functions of Go appear to be inhibition of neuronal calcium channels (Linder et al., 1990; Kleuss et al., 1991) and activation of potassium channels (Van Dongen et al., 1988; Kobayashi et al., 1990). In contrast, Gq and G11 are specialized for only one signal transduction function, activation of PLC (Taylor et al., 1991; Lee et al., 1992).
In our studies, we have been able to show that incubation of VNO membranes with antibodies against Gq/11 was able to impair the production of IP3 when stimulated with 2-heptanone and 2,5-dimethylpyrazine (Figure 4). Although the response to these two compounds was higher than basal levels, it was significantly lower than stimulation with whole urine (Figure 4). This would suggest that these two compounds stimulate a subset of the VNO neurons, whereas urine which contains a mixture of different pheromones would stimulate either all or a higher subset of pheromone receptors. Previous experiments using electrophysiological and biochemical techniques have shown that 2-heptanone and 2,5-dimethylpyrazine stimulates V1R receptors (Boschat et al., 2002; Thompson et al., 2004). 2-Heptanone induces aggression in males and puberty in females (Novotny et al., 1985, 1999), and the receptors that are responsible for this response are the VIRb2 receptors (Boschat et al., 2002). These data would therefore suggest that even though V1R receptors are coupled to Gi2, the alpha subunit of Gq/11 plays a role in the transduction cascade.
Although there is a significant amount of data showing the receptors (V1Rs, V2Rs) and G-protein subunits (Go, Gi2, Gq/11, Gß) that may be involved in the pheromonal signaling, there are almost no studies showing the interactions of these proteins in the VNO. Studies using membrane preparations and electrophysiology have clearly shown that PLC plays a major role in the transduction cascade (Krieger et al., 1999; Holy et al., 2000; Lucas et al., 2003; Wekesa et al., 2003). Furthermore, blocking the G-protein alpha subunits of Go and Gi2 by adenosine diphosphate ribosylation using pertussis toxins does not block PLC-mediated production of IP3 (Wekesa et al., 2003). Although the previous study showed that the alpha subunits of Go and Gi2 do not play a role in the transduction cascade, it did not conclusively show that blocking the alpha subunit of Gq/11 impairs the activation of PLC and therefore impair the production of IP3. In this current study, we show that by using antibodies against the C-terminus of Gq/11, we can block the production of IP3 in female VNOs that are stimulated with adult male urine or specific pheromones such as 2-heptanone and 2,5-dimethylpyrazine (Figures 2 and 3).
Several questions regarding signal transduction in the VNO are still unanswered, such as the role of IP3 and the role of calcium once it has entered the cell. Studies by Liman (2003) show that VNO microvilli have a calcium-activated cation channel which could be opened by calcium ions that either enter the cell through the TRPC2 channel or from the endoplasmic reticulum. Opening of this channel would enhance the receptor potential and therefore amplify the signal. Several studies show that IP3 levels increase in the VNO membrane preparations during stimulation with urinary pheromones (Kroner et al., 1996; Wekesa and Anholt, 1997; Sasaki et al., 1999; Wekesa et al., 2003). Furthermore, injection of IP3 into rat VNO neurons induces inward current responses under whole-cell voltage clamp conditions (Inamura et al., 1997b). Ruthenium red, an IP3 receptor inhibitor, reduces action potential firing induced by urine in rat VNO (Inamura et al., 1997a). Also immunohistochemical studies by Brann et al. (2002) show the presence of IP3 receptors throughout the sensory neuroepithelium of the rat VNO. All these studies support the notion that IP3 plays a role in pheromonal signaling.
In summary, we conclude that the production of IP3 in the VNO by urine, 2 heptanone, and 2,5-dimethylpyrazine is mediated by the alpha subunit of Gq/11.
|
Acknowledgements |
|---|
|
TopAbstractIntroductionMaterials and methodsResultsDiscussionAcknowledgementsReferences |
|---|
This work was supported by grants from the NIH (GM08219 and P20 MD000547) to K.S.W. We would like to thank Dr Ronald McMillon and two anonymous reviewers for critical comments on the manuscript.
|
References |
|---|
|
TopAbstractIntroductionMaterials and methodsResultsDiscussionAcknowledgementsReferences |
|---|
Anholt RRH. (1995) Preparation of olfactory cilia. In Spielman AL and Brand JG (Eds.). Experimental Cell Biology of Taste and Olfaction, Current Techniques and Protocols (CRC Press, New York) pp. 163–168.
Anholt RRH, Aebi U, Snyder SH. (1986) A partially purified preparation of isolated chemosensory cilia from olfactory epithelium of the bullfrog Rana catesbina. J. Neurosci. 6:1962–1969.[Abstract]
Berghard A and Buck LB. (1996) Sensory transduction in vomeronasal neurons: evidence for Go, Gi2, and adenylyl cyclase II as major components of a pheromone signaling cascade. J. Neurosci. 16:909–918.
Berghard A, Buck LB, Liman ER. (1996) Evidence for distinct signaling mechanisms in two mammalian olfactory sense organs. Proc. Natl. Acad. Sci. USA 93:2365–2369.
Birnbaumer L, Abramowitz J, Brown AM. (1990) Receptor-effector coupling by G proteins. Biochim. Biophys. Acta 1031:163–224.[Medline]
Blitzer RD, Omri G, De Vivo M, Carty DJ, Premont RT, Codina J, Birnbaumer L, Cotecchia S, Caron MG, Lefkowitz RJ, Landau EM, Iyenger R. (1993) Coupling of the expressed 1ß-adrenergic receptor to the phospholipase C pathway in Xenopus oocytes: the role of Go. J. Biol. Chem. 268:7532–7537.
Boschat C, Pelofir C, Randin O, Roppolo D, Luscher C, Broillet MC, Rodriguez I. (2002) Pheromone detection mediated by a V1r vomeronasal receptor. Nat. Neurosci. 5:1261–1262.[CrossRef][Web of Science][Medline]
Brann JH, Dennis JC, Morrison EE, Fadool DA. (2002) Type specific inositol 1,4,5-trisphosphate receptor localization in the vomeronasal organ and its interaction with a transient receptor channel TRPC2. J. Neurochem. 83:1452–1460.[CrossRef][Web of Science][Medline]
Cerione RA, Kroll S, Rajaram R, Unson C, Goldsmith P, Spiegel AM. (1988) An antibody directed against the carboxyl-terminal decapeptide of the alpha subunit of the retinal GTP-binding protein, transducin. Effects on transducin function. J. Biol. Chem. 263:9345–9352.
Cinelli AR, Wang D, Chen P, Liu W, Halpern M. (2002) Calcium transients in the garter snake vomeronasal organ. J. Neurophysiol. 87:1449–1472.
de Gasparo M, Husain A, Alexander W, Catt KJ, Chiu AT, Drew M, Goodfriend T, Harding JW, Ingami T, Timmermans PB. (1995) Proposed update of angiotensin receptor nomenclature. Hypertension 25:924–927.
Dulac C. (2000) Sensory coding of pheromone signals in mammals. Curr. Opin. Neurobiol. 10:511–518.[CrossRef][Web of Science][Medline]
Gilchrist A, Vanhauwe JF, Li A, Thomas TO, Voyno-Yasenetskaya T, Hamm HE. (2001) G minigenes expressing C-terminal peptides serve as specific inhibitors of thrombin-mediated endothelial activation. J. Biol. Chem. 276:25672–25679.
Griendling KK, Ushio-Fukai M, Lassegue B, Alexander RW. (1997) Angiotensin II signaling in vascular smooth muscle: new concepts. Hypertension 29:366–373.
Gutowski S, Smrcka A, Nowak L, Wu D, Simon M, Strenweis PC. (1991) Antibodies to the q subfamily of guanine nucleotide-binding regulatory protein subunits attenuate activation of phosphatidylinositol 4,5-biphosphate hydrolysis by hormones. J. Biol. Chem. 266:20519–20524.
Halpern M, Shapiro LS, Jia C. (1995) Differential localization of G proteins in the opossum vomeronasal system. Brain Res. 677:157–161.[CrossRef][Web of Science][Medline]
Herrada G and Dulac C. (1997) A novel family of putative pheromone receptors in mammals with a topographically organized and sexually dimorphic distribution. Cell 90:763–770.[CrossRef][Web of Science][Medline]
Holy TE, Dulac D, Mesiter M. (2000) Responses of vomeronasal neurons to natural stimuli. Science 289:1569–1572.
Inamura K and Kashiwayanagi M. (2000) Inward current responses to urinary substances in rat vomeronasal sensory neurons. Eur. J. Neurosci. 12:3529–3536.[CrossRef][Web of Science][Medline]
Inamura K, Kashiwayanagi M, Kurihara K. (1997a) Blockage of urinary responses by inhibitors of IP3-mediated pathway in rat vomeronasal sensory neurons. Neurosci. Lett. 233:129–132.[CrossRef][Web of Science][Medline]
Inamura K, Kashiwayanagi M, Kurihara K. (1997b) Inositol-1,4,5-trisphosphate induces responses in receptor neurons in rat vomeronasal sensory slices. Chem. Senses 22:93–103.
Jemiolo B, Andreolini F, Xie TM, Wiesler D, Novotny MV. (1989) Puberty-affecting synthetic analogs of urinary chemosignals in the house mouse, Mus domesticus. Physiol. Behav. 46:293–298.[CrossRef][Medline]
Jia C and Halpern M. (1996) Subclasses of vomeronasal receptor neurons: differential expression of G proteins (Gi2 and Go) and segregated projections to the accessory olfactory bulb. Brain Res. 719:117–128.[CrossRef][Web of Science][Medline]
Kai H, Alexander RW, Ushio-Fukay M, Lyons PR, Akers M, Griendling KK. (1998) G-protein binding domains of the angiotensin II ATIA receptors mapped with synthetic peptides selected from the receptor sequence. Biochem. J. 332:781–787.
Kleuss C, Hescheler J, Ewel C, Rosentahl W, Schultz G, Wittig B. (1991) Assignments of G-protein subtypes to specific receptors inducing inhibition of calcium currents. Nature 353:43–48.[CrossRef][Medline]
Kobayashi I, Shibasaki H, Takahashi K, Tohyama K, Kurachi Y, Ito H. (1990) Purification and characterization of five different subunits of guanine-nucleotide-binding proteins in bovine brain membranes: their physiological properties concerning the activities of adenylate cyclase and atrial muscarinic potassium ion channels. Eur. J. Biochem. 191:499–506.[Web of Science][Medline]
Krieger J, Schmitt A, Lobel D, Gudermann T, Schultz G, Breer H, Boekhoff I. (1999) Selective activation of G protein subtypes in the vomeronasal organ upon stimulation with urine-derived compounds. J. Biol. Chem. 274:4655–4662.
Kroner C, Breer H, Singer AG, O'Connell RJ. (1996) Pheromone-induced second messenger signaling in the hamster vomeronasal organ. Neuroreport 7:2989–2992.[Web of Science][Medline]
Lee CH, Park D, Wu D, Rhee SG, Simon MI. (1992) Members of the Gq subunit gene family activate phospholipase C ß isozymes. J. Biol. Chem. 267:16044–16047.
Leinders-Zufall T, Brennan P, Widmayer P, Chandramani PS, Pavicic AM, Jager M, Li XH, Breer H, Zufall F, Boehm T. (2004) MHC class I peptides as chemosensory signals in the vomeronasal organ. Science 306:1033–1037.
Leinders-Zufall T, Lane AP, Punche AC, Ma W, Novotny MV, Shipley MT, Zufall F. (2000) Ultrasensitive pheromone detection by mammalian vomeronasal neurons. Nature 405:792–796.[CrossRef][Medline]
Liman ER. (2003) Regulation by voltage and adenine nucleotides of Ca2+-activated cation channel from hamster vomeronasal sensory neurons. J. Physiol. 548:777–787.
Linder ME, Ewald DA, Miller RJ, Gilman AG. (1990) Purification and characterization of Go and three types of Gi after expression in Escherichia coli. J. Biol. Chem. 265:8243–8251.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. (1951) Protein measurement by Folin phenol reagent. J. Biol. Chem. 193:265–275.
Lucas P, Ukhanov K, Leinders-Zufall T, Zufall F. (2003) A diacylglycerol-gated channel in vomeronasal neuron dendrites is impaired in TRPC2 mutant mice: mechanism of pheromone transduction. Neuron 40:551–561.[CrossRef][Web of Science][Medline]
Luo Y, Lu S, Chen P, Wang D, Halpern M. (1994) Identification of chemoattractant receptors and G-proteins in the vomeronasal system of garter snakes. J. Biol. Chem. 269:16867–16877.
Mackenzie FR, Kelly ECH, Unson CG, Spiegel AM, Milligan G. (1988) Antibodies which recognize the C-terminus of the inhibitory guanine-nucleotide-binding protein (Gi) demonstrate that opioid peptides and foetal-calf serum stimulate the high-affinity GTPase activity of two separate pertussis-toxin substrates. Biochem. J. 249:653–659.[Web of Science][Medline]
Matsunami H and Buck L. (1997) A multigene family encoding a diverse array of putative pheromone receptors in mammals. Cell 90:775–784.[CrossRef][Web of Science][Medline]
McClue SJ and Milligan G. (1990) The 2 ß adrenergic receptor of undifferentiated neuroblastoma glioma hybrid NG108-15 cells, interacts directly with the guanine nucleotide binding protein, Gi2. FEBS Lett 269:430–434.[CrossRef][Web of Science][Medline]
McFadzean I, Mullaney I, Brown DA, Milligan G. (1989) Antibodies to the GTP binding protein, Go, antagonize noradrenaline-induced calcium current inhibition in NG108-15 hybrid cells. Neuron 3:177–182.[CrossRef][Web of Science][Medline]
Moriarty TM, Padrell E, Carty DJ, Omri G, Landau EM, Iyengar R. (1990) Go protein as signal transducer in the pertussis toxin-sensitive phosphatidylinositol pathway. Nature 343:79–82.[CrossRef][Medline]
Novotny M, Harvey S, Jemiolo B, Alberts J. (1985) Synthetic pheromones that promote intermale aggression in mice. Proc. Natl Acad. Sci. USA 82:2059–2061.
Novotny M, Ma W, Zidek L, Daev E. (1999) Recent biochemical insights into puberty acceleration, estrus induction, and puberty delay in the house mouse. In Johnston RE, Muller-Schwarze D, Sorenson PW (Eds.). Advances in Chemical Signals in Vertebrates (Kluwer Academic/Plenum Publishers, New York) pp. 99–116.
Novotny MV. (2003) Pheromones, binding proteins and receptor responses in rodents. Biochem. Soc. Trans. 31:117–122.[Web of Science][Medline]
Ohta H, Okajima F, Ui M. (1985) Inhibition by islet-activating protein of a chemotactic peptide-induced early breakdown of inositol phospholipids and calcium mobilization in guinea pig neutrophils. J. Biol. Chem. 260:15771–15780.
Pantages E and Dulac C. (2000) A novel family of candidate pheromone receptors in mammals. Neuron 28:835–845.[CrossRef][Web of Science][Medline]
Rodriguez I, Del Punta K, Rothman S, Ishii T, Mombaerts P. (2002) Multiple new and isolated families within the mouse superfamily of V1R vomeronasal receptors. Nat. Neurosci. 5:134–140.[CrossRef][Web of Science][Medline]
Runnenburger K, Breer H, Boekhoff I. (2002) Selective G protein beta gamma-subunit compositions mediate phospholipase C activation in the vomeronasal organ. Eur. J. Cell Biol. 81:539–547.[CrossRef][Web of Science][Medline]
Ryba NJP and Tirindelli R. (1997) A new multigene family of putative pheromone receptors. Neuron 19:371–379.[CrossRef][Web of Science][Medline]
Sano T, Ohyama K, Yamano Y, Kakagomi Y, Nakazawa S, Kikyo M, Shirai H, Blank JS, Exton JH, Ingami T. (1997) A domain for G protein coupling in carboxyl-terminal tail of rat angiotensin II receptor type IA. J. Biol. Chem. 272:23631–23636.
Sasaki K, Okamoto K, Inamura K, Tokumitsu Y, Kashiwayanagi M. (1999) Inositol-1,4,5-trisphosphate accumulation induced by urinary pheromones in female rat vomeronasal epithelium. Brain Res 823:161–168.[CrossRef][Web of Science][Medline]
Simonds WF, Goldsmith PK, Woodard CJ, Unson CG, Spiegel AM. (1989) Gi2 mediates 2-adrenergic inhibition of adenylyl cyclase in platelet membranes: in situ identification with G C-terminal antibodies. Proc. Natl Acad. Sci. USA 86:7809–7813.
Taussig R, Iniguez-Lluhi JA, Gilman AG. (1993) Inhibition of adenylyl cyclase by Gi. Science 261:218–221.
Taylor SJ, Chae HZ, Rhee SG, Exton JH. (1991) Activation of the ß1 isozyme of phospholipase C by the subunits of the Gq class of G proteins. Nature 350:516–518.[CrossRef][Medline]
Thompson RN, Robertson BK, Napier A, Wekesa KS. (2004) Sex-specific responses to urinary chemicals by the mouse vomeronasal organ. Chem. Senses 29:749–754.
Tirindelli R and Ryba NJ. (1996) The G-protein gamma-subunit G gamma 8 is expressed in the developing axons of olfactory and vomeronasal neurons. Eur. J. Neurosci. 8:2388–2398.[CrossRef][Web of Science][Medline]
Van Dongen AMJ, Codina J, Olate J, Mattera R, Joho R, Birnbaumer L, Brown AM. (1988) Newly identified brain potassium channels gated by the guanine nucleotide binding protein Go. Science 242:1433–1437.
Watkins DC, Johnson GL, Malbon CC. (1992) Regulation of the differentiation of teratocarcinoma cells into primitive ectoderm by Gi2. Science 258:1373–1375.
Wekesa KS and Anholt RRH. (1997) Pheromone regulated production of inositol-(1,4,5)-trisphosphate in the mammalian vomeronasal organ. Endocrinology 138:3497–3504.
Wekesa KS and Anholt RRH. (1999) Differential expression of G proteins in the mouse olfactory system. Brain Res 837:117–126.[CrossRef][Web of Science][Medline]
Wekesa KS, Miller S, Napier A. (2003) Involvement of Gq/11 in signal transduction in the mammalian vomeronasal organ. J. Exp. Biol. 206:827–832.
Wong YH, Conklin BR, Bourne HR. (1992) Gi2-mediated hormonal inhibition of cyclic AMP accumulation. Science 255:339–342.
Yatani A, Hamm H, Coding J, Mazzoni MR, Birnbaumer L, Brown AM. (1988) A monoclonal antibody to the alpha subunit of Gk blocks muscarinic activation of atrial K+ channels. Science 241:828–831.
Accepted 15 May 2006
CiteULike 
Connotea 
Del.icio.us What's this?
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||