Disruption of the Type III Adenylyl Cyclase Gene Leads to Peripheral and Behavioral Anosmia in Transgenic Mice

Wong, Scott T.; Trinh, Kien; Hacker, Beth M.; Chan, Guy C.‐K.; Lowe, Graeme; Gaggar, Anuj; Xia, Zhengui; Gold, Geoffrey H.; Storm, Daniel R.

doi:10.1016/s0896-6273(00)00060-x

Cited by 415 publications

(395 citation statements)

References 62 publications

(3 reference statements)

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“…ACs are known to be expressed in different types of cilia. For example, olfactory sensory cilia possess AC2, AC3, and AC4 (2,58). Motile cilia in airway epithelia have soluble AC (47).…”

Section: Discussionmentioning

confidence: 99%

Cholangiocyte primary cilia are chemosensory organelles that detect biliary nucleotides via P2Y₁₂ purinergic receptors

Masyuk

Gradilone²,

Bañales

et al. 2008

American Journal of Physiology-Gastrointestinal and Liver Physi

151

121

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Masyuk AI, Gradilone SA, Banales JM, Huang BQ, Masyuk TV, Lee S-O, Splinter PL, Stroope AJ, LaRusso NF. Cholangiocyte primary cilia are chemosensory organelles that detect biliary nucleotides via P2Y 12 purinergic receptors. Am J Physiol Gastrointest Liver Physiol 295: G725-G734, 2008. First published August 7, 2008 doi:10.1152/ajpgi.90265.2008.-Cholangiocytes, the epithelial cells lining intrahepatic bile ducts, contain primary cilia, which are mechano-and osmosensory organelles detecting changes in bile flow and osmolality and transducing them into intracellular signals. Here, we asked whether cholangiocyte cilia are chemosensory organelles by testing the expression of P2Y purinergic receptors and components of the cAMP signaling cascade in cilia and their involvement in nucleotide-induced cAMP signaling in the cells. We found that P2Y 12 purinergic receptor, adenylyl cyclases (i.e., AC4, AC6, and AC8), and protein kinase A (i.e., PKA RI-␤ and PKA RII-␣ regulatory subunits), exchange protein directly activated by cAMP (EPAC) isoform 2, and A-kinase anchoring proteins (i.e., AKAP150) are expressed in cholangiocyte cilia. ADP, an endogenous agonist of P2Y 12 receptors, perfused through the lumen of isolated rat intrahepatic bile ducts or applied to the ciliated apical surface of normal rat cholangiocytes (NRCs) in culture induced a 1.9-and 1.5-fold decrease of forskolininduced cAMP levels, respectively. In NRCs, the forskolin-induced cAMP increase was also lowered by 1.3-fold in response to ATP-␥S, a nonhydrolyzed analog of ATP but was not affected by UTP. The ADP-induced changes in cAMP levels in cholangiocytes were abolished by chloral hydrate (a reagent that removes cilia) and by P2Y 12 siRNAs, suggesting that cilia and ciliary P2Y12 are involved in nucleotide-induced cAMP signaling. In conclusion, cholangiocyte cilia are chemosensory organelles that detect biliary nucleotides through ciliary P2Y 12 receptors and transduce corresponding signals into a cAMP response. liver; ADP; adenylyl cyclases; cAMP; protein kinase A; exchange protein directly activated by cAMP; A-kinase anchoring protein 150 CHOLANGIOCYTES, the epithelial cells lining intrahepatic bile ducts (IBDs), contain primary cilia, nonmotile, solitary organelles extending from the apical plasma membrane into the ductal lumen (21,28,29). In many cell types, including cholangiocytes, primary cilia function as sensory organelles detecting multiple (i.e., mechano-, osmo-, chemo-) stimuli and transducing them into intracellular signaling (12,16,27,40,46). However, although increasing evidence suggests the ability of primary cilia to act as mechano-and osmosensors (16, 24, 28, 34 -37, 40 -42, 49, 50, 55), less data support their chemosensory functions. To function as chemosensory organelles, primary cilia should possess receptors and associated signaling cascades through which signals induced by specific ligands are transmitted into the cell. Such mechanism exists in Caenorhabditis elegans neuronal primary cilia, which express specific G protein-coupled...

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“…ACs are known to be expressed in different types of cilia. For example, olfactory sensory cilia possess AC2, AC3, and AC4 (2,58). Motile cilia in airway epithelia have soluble AC (47).…”

Section: Discussionmentioning

confidence: 99%

Cholangiocyte primary cilia are chemosensory organelles that detect biliary nucleotides via P2Y₁₂ purinergic receptors

Masyuk

Gradilone²,

Bañales

et al. 2008

American Journal of Physiology-Gastrointestinal and Liver Physi

151

121

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“…Although mice deficient in either mAC VIII or G␣ olf are fertile but rarely surviving (43,44), homozygous matings of mAC III K.O. mice were unproductive, although they showed normal mounting and anogenital sniffing behaviors (45). However, it remains to be determined if OR-mediated sperm signaling pathways uncovered in humans can be transferred to the mouse model, because fertilization barriers between species may also be reflected on the molecular level.…”

Section: Discussionmentioning

confidence: 99%

Particulate Adenylate Cyclase Plays a Key Role in Human Sperm Olfactory Receptor-mediated Chemotaxis

Spehr

Schwane

Riffell

et al. 2004

Journal of Biological Chemistry

141

131

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Human sperm chemotaxis is a critical component of the fertilization process, but the molecular basis for this behavior remains unclear. Recent evidence shows that chemotactic responses depend on activation of the sperm olfactory receptor, hOR17-4. Certain floral scents, including bourgeonal, activate hOR17-4, trigger pronounced Ca 2؉ fluxes, and evoke chemotaxis. Here, we provide evidence that hOR17-4 activation is coupled to a cAMP-mediated signaling cascade. Multidimensional protein identification technology was used to identify potential components of a G-protein-coupled cAMP transduction pathway in human sperm. These products included various membrane-associated adenylate cyclase (mAC) isoforms and the G olf -subunit. Using immunocytochemistry, specific mAC isoforms were localized to particular cell regions. Whereas mAC III occurred in the sperm head and midpiece, mAC VIII was distributed predominantly in the flagellum. In contrast, G olf was found mostly in the flagellum and midpiece. The observed spatial distribution patterns largely correspond to the spatiotemporal character of hOR17-4-induced Ca 2؉ changes. Behavioral and Ca 2؉ signaling responses of human sperm to bourgeonal were bioassayed in the presence, or absence, of the adenylate cyclase antagonist SQ22536. This specific agent inhibits particulate AC, but not soluble AC, activation. Upon incubation with SQ22536, cells ceased to exhibit Ca 2؉ signaling, chemotaxis, and hyperactivation (faster swim speed and flagellar beat rate) in response to bourgeonal. Particulate AC is therefore required for induction of hOR17-4-mediated human sperm behavior and represents a promising target for future design of contraceptive drugs.

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“…Upon ligand binding, odorant receptors activate the G protein G␣ olf , which stimulates type III adenylyl cyclase to increase intracellular levels of cAMP (4)(5)(6)(7)(8)(9). This second messenger then directly opens cAMPsensitive, cyclic nucleotide-gated (CNG) channels in the plasma membrane, resulting in the initial depolarization of the cell (3,4).…”

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confidence: 99%

Contribution of the receptor guanylyl cyclase GC-D to chemosensory function in the olfactory epithelium

Leinders‐Zufall

Cockerham

Michalakis

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

196

267

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The mammalian main olfactory epithelium (MOE) recognizes and transduces olfactory cues through a G protein-coupled, cAMPdependent signaling cascade. Additional chemosensory transduction mechanisms have been suggested but remain controversial. We show that a subset of MOE neurons expressing the orphan receptor guanylyl cyclase GC-D and the cyclic nucleotide-gated channel subunit CNGA3 employ an excitatory cGMP-dependent transduction mechanism for chemodetection. By combining gene targeting of Gucy2d, which encodes GC-D, with patch clamp recording and confocal Ca 2؉ imaging from single dendritic knobs in situ, we find that GC-D cells recognize the peptide hormones uroguanylin and guanylin as well as natural urine stimuli. These molecules stimulate an excitatory, cGMP-dependent signaling cascade that increases intracellular Ca 2؉ and action potential firing. Responses are eliminated in both Gucy2d-and Cnga3-null mice, demonstrating the essential role of GC-D and CNGA3 in the transduction of these molecules. The sensitive and selective detection of two important natriuretic peptides by the GC-D neurons suggests the possibility that these cells contribute to the maintenance of salt and water homeostasis or the detection of cues related to hunger, satiety, or thirst.O dor recognition by canonical olfactory sensory neurons (OSNs) of the main olfactory epithelium (MOE) begins when odor molecules bind to any one of many hundred G protein-coupled odorant receptors (1-4). Upon ligand binding, odorant receptors activate the G protein G␣ olf , which stimulates type III adenylyl cyclase to increase intracellular levels of cAMP (4-9). This second messenger then directly opens cAMPsensitive, cyclic nucleotide-gated (CNG) channels in the plasma membrane, resulting in the initial depolarization of the cell (3, 4). OSNs that use this cAMP-mediated transduction cascade respond to a wide variety of odors, including food odors, volatile pheromones, and peptides that bind major histocompatibility complex proteins (4, 10-12). A recently discovered second family of G protein-coupled receptors expressed in the MOE, the trace amine-associated receptors, may also function in the recognition of some odors or pheromones by coupling to the canonical cAMP pathway (13). However, there has been intense debate as to whether the cAMP cascade is the only excitatory sensory transduction mechanism in the MOE (11,14,15).A subpopulation of ciliated MOE neurons that express the orphan receptor guanylyl cyclase GC-D (16) are prime candidates to mediate cAMP-independent odor recognition. GC-D cells lack key components of the canonical OSN odor transduction cascade, including G␣ olf , type III adenylyl cyclase, the Ca 2ϩ /calmodulin-dependent phosphodiesterase PDE1C2, the cAMP-specific phosphodiesterase PDE4A, and the cAMPsensitive CNG channel subunits CNGA2 and CNGB1b (17,18). Instead, these neurons express a cGMP-specific CNG channel subunit, CNGA3, and a cGMP-stimulated phosphodiesterase, PDE2 (17,18). Although these cells were identified more than a de...

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Disruption of the Type III Adenylyl Cyclase Gene Leads to Peripheral and Behavioral Anosmia in Transgenic Mice

Cited by 415 publications

References 62 publications

Cholangiocyte primary cilia are chemosensory organelles that detect biliary nucleotides via P2Y₁₂ purinergic receptors

Cholangiocyte primary cilia are chemosensory organelles that detect biliary nucleotides via P2Y₁₂ purinergic receptors

Particulate Adenylate Cyclase Plays a Key Role in Human Sperm Olfactory Receptor-mediated Chemotaxis

Contribution of the receptor guanylyl cyclase GC-D to chemosensory function in the olfactory epithelium

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Disruption of the Type III Adenylyl Cyclase Gene Leads to Peripheral and Behavioral Anosmia in Transgenic Mice

Cited by 415 publications

References 62 publications

Cholangiocyte primary cilia are chemosensory organelles that detect biliary nucleotides via P2Y12 purinergic receptors

Cholangiocyte primary cilia are chemosensory organelles that detect biliary nucleotides via P2Y12 purinergic receptors

Particulate Adenylate Cyclase Plays a Key Role in Human Sperm Olfactory Receptor-mediated Chemotaxis

Contribution of the receptor guanylyl cyclase GC-D to chemosensory function in the olfactory epithelium

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Resources

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Cholangiocyte primary cilia are chemosensory organelles that detect biliary nucleotides via P2Y₁₂ purinergic receptors

Cholangiocyte primary cilia are chemosensory organelles that detect biliary nucleotides via P2Y₁₂ purinergic receptors