Synaptotagmin IX Regulates Ca2+-dependent Secretion in PC12 Cells

Fukuda, Mitsunori; Kowalchyk, Judith A.; Zhang, Xiaodong; Martin, Thomas F.J.; Mikoshiba, Katsuhiko

doi:10.1074/jbc.c100588200

Cited by 93 publications

(109 citation statements)

References 38 publications

Supporting

Mentioning

101

Contrasting

Order By: Relevance

“…vesicular protein that modulates exocytosis by enhancing the interaction of synaptotagmin with the SNARE protein SNAP25 (28). On the other hand, Syt IX is a synaptotagmin isoform expressed in synaptic vesicles of neuronal cells that regulates Ca 2ϩ -dependent exocytosis (26,27). The role played by Syt IX and Snapin in SNARE-dependent exocytosis suggests that their interaction with TRPV1 may modulate aspects of TRPV1 trafficking and/or delivery to the plasma membrane.…”

Section: Discussionmentioning

confidence: 99%

Regulated Exocytosis Contributes to Protein Kinase C Potentiation of Vanilloid Receptor Activity

Morenilla‐Palao

Planells-Cases

García-Sanz

et al. 2004

Journal of Biological Chemistry

386

302

View full text Add to dashboard Cite

The vanilloid receptor-1 (TRPV1) plays a key role in the perception of peripheral thermal and inflammatory pain. TRPV1 expression and channel activity are notably up-regulated by proalgesic agents. The transduction pathways involved in TRPV1 sensitization are still elusive. We have used a yeast two-hybrid screen to identify proteins that associate with the N terminus of TRPV1. We report that two vesicular proteins, Snapin and synaptotagmin IX (Syt IX), strongly interact in vitro and in vivo with the TRPV1 N-terminal domain. In primary dorsal root ganglion neurons, TRPV1 co-distributes in vesicles with Syt IX and the vesicular protein synaptobrevin. Neither Snapin nor Syt IX affected channel function, but they notably inhibited protein kinase C (PKC)-induced potentiation of TRPV1 channel activity with a potency that rivaled the blockade evoked by botulinum neurotoxin A, a potent blocker of neuronal exocytosis. Noteworthily, we found that PKC activation induced a rapid delivery of functional TRPV1 channels to the plasma membrane. Botulinum neurotoxin A blocked the TRPV1 membrane translocation induced by PKC that was activated with a phorbol ester or the metabotropic glutamate receptor mGluR5. Therefore, our results indicate that PKC signaling promotes at least in part the SNARE-dependent exocytosis of TRPV1 to the cell surface. Taken together, these findings imply that activitydependent delivery of channels to the neuronal surface may contribute to the buildup and maintenance of thermal inflammatory hyperalgesia in peripheral nociceptor terminals.TRPV1 1 is a capsaicin-, proton-and heat-sensitive, cationselective ion channel expressed in nociceptors that participates in the transduction of noxious chemical and thermal stimuli by sensory nerve endings in peripheral tissues (1-3). Heterologous expression of TRPV1 cDNA results in ionic currents that recapitulate most of the functional properties displayed by native capsaicin-and heat-activated currents in sensory neurons (1, 2). For instance, TRPV1 exhibits a time-and Ca 2ϩ -dependent desensitization, a long lasting refractory state during which the receptor does not respond to vanilloids or other stimuli (2). In addition, the channel activity of TRPV1 is remarkably upregulated by inflammatory mediators through the activation of phospholipase C and protein kinases A and C (PKA and PKC) signaling pathways (4 -13). Recent evidence shows that an increase in TRPV1 expression in peripheral nociceptors is critical for the maintenance of inflammatory hyperalgesia (14, 15). The involvement of TRPV1 in heat hypersensitivity is further underscored by the reduced sensitivity of mice lacking TRPV1 (16, 17) and by mice treated with receptor-specific antagonists (18).TRPV1 belongs to the family of transient receptor potential channels, which structurally resembles the family of voltagegated potassium or cyclic nucleotide-gated channels (19). Accordingly, these channels are presumed to be tetrameric assemblies of identical subunits (Fig. 1A), although heteromeric assemblies have be...

show abstract

Section: Discussionmentioning

confidence: 99%

Regulated Exocytosis Contributes to Protein Kinase C Potentiation of Vanilloid Receptor Activity

Morenilla‐Palao

Planells-Cases

García-Sanz

et al. 2004

Journal of Biological Chemistry

386

302

View full text Add to dashboard Cite

show abstract

“…For instance, recombinant Syt IX (or I) transiently overexpressed is often targeted to the plasma membrane of PC12 cells (supplemental Fig. 2), whereas endogenous Syt IX (or I) is clearly localized on dense-core vesicles, but not at the plasma membrane (25,64). Consistent with these observations, the present study on the PC12 cell line stably expressing Syt VII-GFP clearly demonstrated that the targeting of Syt VII molecules to specific membrane compartment(s) and the dynamics of Syt VII molecule during densecore vesicle exocytosis depend on the transfection method (transient expression versus stable expression).…”

Section: Discussionmentioning

confidence: 99%

“…Antibody-uptake Experiments-PC12 cells cultured on 35-mm glass bottom dishes were incubated for 15 min at 37°C with the anti-Syt IX-N rabbit polyclonal antibody (10 g/ml) and anti-FLAG tag mouse monoclonal antibody (1 g/ml) in either high KCl buffer (56 mM KCl, 95 mM NaCl, 2.2 mM CaCl 2 , 0.5 mM MgCl 2 , 5.6 mM glucose, and 15 mM HEPES-KOH, pH 7.4) or low KCl buffer (5.6 mM KCl, 145 mM NaCl, 2.2 mM CaCl 2 , 0.5 mM MgCl 2 , 5.6 mM glucose, and 15 mM HEPES-KOH, pH 7.4) as described previously (25,45,50). The cells were immediately washed twice with phosphate-buffered saline and then fixed with 4% paraformaldehyde as described above.…”

Section: Methodsmentioning

confidence: 99%

“…For instance, Syt I, the best characterized Syt isoform abundant on synaptic vesicles, is now widely believed to function as the major Ca 2ϩ sensor for neurotransmitter release (15)(16)(17) and regulates synaptic vesicle docking, fusion, and recycling steps via its C2 domains (18 -21). Syts I and IX are present on the dense-core vesicles in some endocrine cells and regulate their Ca 2ϩ -dependent exocytosis (13,(22)(23)(24)(25)(26)(27)(28)(29). Syt IV regulates glutamate release from astrocytes (30).…”

Section: Synaptotagmin (Syt)mentioning

confidence: 99%

“…during dense-core vesicle exocytosis and endocytosis (42,43), we next sought to determine whether Syt VII-GFP-containing dense-core vesicles are fully mature enough to undergo regulated exocytosis in response to Ca 2ϩ -stimulation. To do so, we performed an N-terminal antibody-uptake experiment as described previously (25,45,50). As anticipated, both anti-FLAG tag antibody (Fig.…”

Section: Syt VII a Putative Vesicular Ca 2ϩ Sensor For Exocytosismentioning

confidence: 99%

See 2 more Smart Citations

Synaptotagmin VII Is Targeted to Dense-core Vesicles and Regulates Their Ca2+-dependent Exocytosis in PC12 Cells

Fukuda

Kanno

Satoh

et al. 2004

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Exocytosis and Synaptic Vesicle Function

Shin

2014

Comprehensive Physiology

View full text Add to dashboard Cite

Synaptic vesicles release their vesicular contents to the extracellular space by Ca(2+)-triggered exocytosis. The Ca(2+)-triggered exocytotic process is regulated by synaptotagmin (Syt), a vesicular Ca(2+)-binding C2 domain protein. Synaptotagmin 1 (Syt1), the most studied major isoform among 16 Syt isoforms, mediates Ca(2+)-triggered synaptic vesicle exocytosis by interacting with the target membranes and SNARE/complexin complex. In synapses of the central nervous system, synaptobrevin 2, a major vesicular SNARE protein, forms a ternary SNARE complex with the plasma membrane SNARE proteins, syntaxin 1 and SNAP25. The affinities of Ca(2+)-dependent interactions between Syt1 and its targets (i.e., SNARE complexes and membranes) are well correlated with the efficacies of the corresponding exocytotic processes. Therefore, different SNARE protein isoforms and membrane lipids, which interact with Syt1 with various affinities, are capable of regulating the efficacy of Syt1-mediated exocytosis. Otoferlin, another type of vesicular C2 domain protein that binds to the membrane in a Ca(2+)-dependent manner, is also involved in the Ca(2+)-triggered synaptic vesicle exocytosis in auditory hair cells. However, the functions of otoferlin in the exocytotic process are not well understood. In addition, at least five different types of synaptic vesicle proteins such as synaptic vesicle protein 2, cysteine string protein α, rab3, synapsin, and a group of proteins containing four transmembrane regions, which includes synaptophysin, synaptogyrin, and secretory carrier membrane protein, are involved in modulating the exocytotic process by regulating the formation and trafficking of synaptic vesicles.

show abstract

Synaptotagmin IX Regulates Ca2+-dependent Secretion in PC12 Cells

Cited by 93 publications

References 38 publications

Regulated Exocytosis Contributes to Protein Kinase C Potentiation of Vanilloid Receptor Activity

Regulated Exocytosis Contributes to Protein Kinase C Potentiation of Vanilloid Receptor Activity

Synaptotagmin VII Is Targeted to Dense-core Vesicles and Regulates Their Ca2+-dependent Exocytosis in PC12 Cells

Exocytosis and Synaptic Vesicle Function

Contact Info

Product

Resources

About