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At least 19 recordsLinked to original sources
PubMed1999

The subcellular localizations of atypical synaptotagmins III and VI. Synaptotagmin III is enriched in synapses and synaptic plasma membranes but not in synaptic vesicles.

Multiple synaptotagmins are expressed in brain, but only synaptotagmins I and II have known functions in fast, synchronous Ca2+-triggered neurotransmitter release. Synaptotagmin III was proposed to regulate other aspects of synaptic vesicle exocytosis, particularly its slow component. Such a function predicts that synaptotagmin III should be an obligatory synaptic vesicle protein, as would also be anticipated from its high homology to synaptotagmins I and II. To test this hypothesis, we studied the distribution, developmental expression, and localization of synaptotagmin III and its closest homolog, synaptotagmin VI. We find that synaptotagmins III and VI are present in all brain regions in heterogeneous distributions and that their levels increase during development in parallel with synaptogenesis. Furthermore, we show by immunocytochemistry that synaptotagmin III is concentrated in synapses, as expected. Surprisingly, however, we observed that synaptotagmin III is highly enriched in synaptic plasma membranes but not in synaptic vesicles. Synaptotagmin VI was also found to be relatively excluded from synaptic vesicles. Our data suggest that synaptotagmins III and VI perform roles in neurons that are not linked to synaptic vesicle exocytosis but to other Ca2+-related nerve terminal events, indicating that the functions of synaptotagmins are more diverse than originally thought.

Animals↗
PubMed2001

Synaptotagmin 13: structure and expression of a novel synaptotagmin.

Synaptotagmins represent a family of putative vesicular trafficking proteins. With synaptotagmin 13, we have now identified a novel synaptotagmin, making this one of the largest families of trafficking proteins. Similar to synaptotagmins 3, 4, 6, 7, 9, and 11, synaptotagmin 13 is expressed at highest levels in brain but is also detectable at lower levels in non-neuronal tissues. Synaptotagmin 13 is composed of the canonical domains of synaptotagmins that include an N-terminal transmembrane region and two C-terminal cytoplasmic C2-domains (C2A- and C2B-domain) and a connecting sequence between the transmembrane region and the C2-domains. Different from most other synaptotagmins, however, synaptotagmin 13 does not have an N-terminal sequence preceding the transmembrane region, and features an unusually long connecting sequence that is proline-rich. Furthermore, the C2-domains of synaptotagmin are degenerate and lack almost all of the residues involved in Ca2+ binding, suggesting that synaptotagmin 13 is not a Ca2+-binding protein unlike most other synaptotagmins. Our data demonstrate that synaptotagmins represent a larger and more complex gene family than previously envisioned.

Amino Acid Sequence↗
PubMed1998

Inositol 1,3,4,5-tetrakisphosphate binding activities of neuronal and non-neuronal synaptotagmins. Identification of conserved amino acid substitutions that abolish inositol 1,3,4,5-tetrakisphosphate binding to synaptotagmins III, V, and X.

Synaptotagmins I and II are essential for Ca2+-regulated exocytosis of synaptic vesicles from neurons, probably serving as Ca2+ sensors. This Ca2+-sensing function is thought to be disrupted by binding of an inositol 1,3,4,5-tetrakisphosphate (IP4) to the C2B domain of synaptotagmin I or II (Fukuda, M., Moreira, J. E., Lewis, F. M. T., Sugimori, M., Niinobe, M., Mikoshiba, K., and Llinás, R. (1995) Proc. Natl. Acad. Sci. U.S.A. 92, 10708-10712). Recently, several synaptotagmin isoforms, expressed outside the nervous system, have been identified in rats and proposed to be involved in constitutive vesicle traffic. To test whether the inositol high polyphosphates also regulate constitutive vesicle traffic by binding to the non-neuronal synaptotagmins, we examined the IP4 binding properties of the recombinant C2 domains of both neuronal (III, V, X, and XI) and non-neuronal (VI-VIII and IX) synaptotagmins. The C2B domains of synaptotagmins VII-IX and XI had strong IP4 binding activity, but the C2B domain of synaptotagmin VI showed very weak IP4 binding activity. In contrast, there was no significant IP4 binding activity of the C2B domains of synaptotagmins III, V, and X or any of the C2A domains. A phylogenetic tree of the C2 domains of 11 isoforms revealed that synaptotagmins III, V, VI, and X (IP4-insensitive or very weak IP4-binding isoforms) belong to the same branch. Based on the sequence comparison between the IP4-sensitive and -insensitive isoforms, we performed site-directed mutagenesis of synaptotagmin III and identified several amino acid substitutions that abolish IP4 binding activity. Our data suggest that the inositol high polyphosphates might also regulate constitutive vesicle traffic via binding to the IP4-sensitive non-neuronal synaptotagmins.

Amino Acid Sequence↗
PubMed1994

Synaptotagmin III is a novel isoform of rat synaptotagmin expressed in endocrine and neuronal cells.

Synaptotagmin (p65), an integral membrane protein of synaptic vesicles, is thought to be involved in calcium-dependent exocytosis of synaptic vesicles. Here, we report the cloning and tissue distribution of a novel isoform of synaptotagmin, designated synaptotagmin III. The cDNA clones encoding synaptotagmin III have been isolated from a rat brain cDNA library. Rat synaptotagmin III is a protein of 588 amino acids having 40.5, 38.3, and 64.0% identity with rat synaptotagmin I, rat synaptotagmin II, and o-p65-C, a third synaptotagmin isoform of marine ray Discopyge ommata, respectively. The region of the two internal repeats homologous to the regulatory domain (C2 domain) of protein kinase C is highly conserved among synaptotagmin I, II, and III. RNA blotting studies reveal that synaptotagmin III mRNA is expressed in brain, various endocrine tissues, and hormone-secreting clonal cells. These results suggest that rat synaptotagmin III is a mammalian homolog of o-p65-C and is involved in Ca(2+)-dependent exocytosis of secretory vesicles in endocrine cells, as well as in neurons.

Amino Acid Sequence↗
PubMed1991

Synaptotagmin II. A novel differentially distributed form of synaptotagmin.

Synaptotagmin is a synaptic vesicle membrane protein with properties suggestive of a role in synaptic vesicle exocytosis (Perin, M. S., Fried, V. A., Mignery, G. A., Jahn, R., and Südhof, T. C. (1990) Nature 345, 260-263). Here, we report the structure of a novel form of synaptotagmin named synaptotagmin II that is highly homologous to the originally described synaptotagmin, now referred to as synaptotagmin I. Synaptotagmins I and II exhibit the same overall structure, containing a small intravesicular sequence that is glycosylated, a single transmembrane region, and a large carboxyl-terminal cytoplasmic sequence that includes two copies of an internal repeat homologous to the regulatory region of protein kinase C. The homology between synaptotagmins I and II is not uniformly distributed across the molecule but is highest in their carboxyl-terminal regulatory repeats (88% sequence identity) and lowest in their amino-terminal intravesicular sequences (46% sequence identity). RNA blots demonstrate complementary patterns of expression for synaptotagmins I and II, with synaptotagmin I preferentially expressed in rostral, phylogenetically younger brain regions, and synaptotagmin II predominantly expressed in caudal, phylogenetically older brain regions. With this description of two forms of synaptotagmin, all major synaptic vesicle proteins implicated in membrane traffic have now been shown to be present in several isoforms with differential distributions, suggesting that this is a general organizational principle of the mammalian brain.

Amino Acid Sequence↗
PubMed1995

Synaptotagmin I and 1B4 are identical: implications for synaptotagmin distribution in the primate brain.

We have determined that the human cDNA sequence of the previously described primate brain mRNA species 1B4 is nearly identical (99.95% similarity) to that of human Synaptotagmin I. The apparent identity of Synaptotagmin I with 1B4, whose distribution in the brain of the monkey Cynomolgous was determined previously by Northern blot and in situ hybridization (ISH) analyses, reveals the Synaptotagmin I is differentially expressed in the primate brain. Primate Synaptotagmin I mRNA is enriched in hindbrain structures relative to forebrain structures by Northern blot analysis. By ISH analysis, primate Synaptotagmin I mRNA is highly expressed in occipital cortex and lateral geniculate (visual system components) and differentially expressed across topographic cortical boundaries between inferior and superior temporal gyrus (a polymodal zone with visual, auditory and somatosensory inputs) and between areas 17 and 18 of the visual cortex (primary and secondary visual areas). Cortical expression is also enriched in layers V and VI, which contain large pyramidal projection neurons. Synaptotagmin I's greater association with large projection neurons and with some components of visual sensory transduction could reflect a requirement of these neural components for greater synaptic activity. Synaptotagmin I expression in the primate brain is also dissimilar to Synaptotagmin I expression in rodents. Thus, variation of Synaptotagmin I expression has occurred during mammalian evolution, perhaps as a consequence of the larger size and neurotransmitter requirements of primate neurons.

Base Sequence↗
PubMed2002

Synaptotagmin I-DeltaC2B. A novel synaptotagmin isoform with a single C2 domain in the bovine adrenal medulla.

Synaptotagmin I is a 65 kDa type 1 membrane glycoprotein found in secretory organelles that plays a key role in regulated exocytosis. We have characterised two forms (long and short) of synaptotagmin I that are present in the bovine adrenal medulla. The long form is a type I integral membrane protein which has two cytoplasmic C2 domains and corresponds to the previously characterised full-length synaptotagmin I isoform. The short-form synaptotagmin I-DeltaC2B has the same structure in the lumenal and transmembrane sequences, but synaptotagmin I-DeltaC2B is truncated such that it only has a single cytoplasmic C2 domain. Analysis of synaptotagmin I-DeltaC2B expression indicates that synaptotagmin I-DeltaC2B is preferentially expressed in the bovine adrenal medulla. However, it is absent from the dense core chromaffin granules. Furthermore, when expressed in the rat pheochromocytoma cell line PC12 bovine synaptotagmin I-DeltaC2B is largely absent from dense core granules and synaptic-like microvesicles. Instead, indirect immunofluorescence microscopy reveals the intracellular location of synaptotagmin I-DeltaC2B to be the plasma membrane.

Adrenal Medulla↗
PubMed1997

The first C2 domain of synaptotagmin is required for exocytosis of insulin from pancreatic beta-cells: action of synaptotagmin at low micromolar calcium.

The Ca2+- and phospholipid-binding protein synaptotagmin is involved in neuroexocytosis. Its precise role and Ca2+-affinity in vivo are unclear. We investigated its putative function in insulin secretion which is maximally stimulated by 10 microM cytosolic free Ca2+. The well-characterized synaptotagmin isoforms I and II are present in pancreatic beta-cell lines RINm5F, INS-1 and HIT-T15 as shown by Northern and Western blots. Subcellular fractionation and confocal microscopy revealed their presence mainly on insulin-containing secretory granules whereas only minor amounts were found on synaptic vesicle-like microvesicles. Antibodies or Fab-fragments directed against the Ca2+-dependent phospholipid binding site of the first C2 domain of synaptotagmin I or II inhibited Ca2+-stimulated, but not GTPgammaS-induced exocytosis from streptolysin-O-permeabilized INS-1 and HIT-T15 cells. Transient expression of wild-type synaptotagmin II did not alter exocytosis in HIT-T15 cells. However, mutations in the Ca2+-dependent phospholipid binding site of the first C2 domain (Delta180-183, D231S) again inhibited only Ca2+-, but not GTPgammaS-evoked exocytosis. In contrast, mutations in the IP4-binding sites of the second C2 domain (Delta325-341; K327,328, 332Q) did not alter exocytosis. Synaptotagmin II mutated in both C2 domains (Delta180-183/K327,328,332Q) induced greater inhibition than mutant Delta180-183, suggesting a discrete requirement for the second C2 domain. Thus, synaptotagmin isoforms regulate exocytotic events occurring at low micromolar Ca2+.

Animals↗
PubMed2000

Distinct self-oligomerization activities of synaptotagmin family. Unique calcium-dependent oligomerization properties of synaptotagmin VII.

Synaptotagmins constitute a large protein family, characterized by one transmembrane region and two C2 domains, and can be classified into several subclasses based on phylogenetic relationships and biochemical activities (Fukuda, M., Kanno, E., and Mikoshiba, K. (1999) J. Biol. Chem. 274, 31421-31427). Synaptotagmin I (Syt I), a possible Ca(2+) sensor for neurotransmitter release, showed both Ca(2+)-dependent (via the C2 domain) and -independent (via the NH(2)-terminal domain) self-oligomerization, which are thought to be important for synaptic vesicle exocytosis. However, little is known about the relationship between these two interactions and the Ca(2+)-dependent oligomerization properties of other synaptotagmin isoforms. In this study, we first examined the Ca(2+)-dependent self-oligomerization properties of synaptotagmin family by co-expression of T7- and FLAG-tagged Syts (full-length or cytoplasmic domain) in COS-7 cells. We found that Syt VII is a unique class of synaptotagmins that only showed robust Ca(2+)-dependent self-oligomerization at the cytoplasmic domain with EC(50) values of about 150 micrometer Ca(2+). In addition, Syt VII preferentially interacted with the previously described subclass of Syts (V, VI, and X) in a Ca(2+)-dependent manner. Co-expression of full-length and cytoplasmic portion of Syts VII (or II) indicate that Syt VII cytoplasmic domain oligomerizes in a Ca(2+)-dependent manner without being tethered at the NH(2)-terminal domain, whereas Ca(2+)-dependent self-oligomerization at the cytoplasmic domain of other isoforms (e.g. Syt II) occurs only when the two molecules are tethered at the NH(2)-terminal domain.

Animals↗
PubMed1995

Synaptotagmin V: a novel synaptotagmin isoform expressed in rat brain.

Regulated Ca(2+)-dependent release of transmitters from synaptic vesicles is an important characteristic of chemical neurotransmission. Synaptotagmins are abundant synaptic vesicle transmembrane proteins that probably function as Ca2+ sensors. Molecular cloning has identified four different synaptotagmin isoforms in mammals. We report here the cloning and sequencing of a novel isoform of 386 amino acids. Synaptotagmin V is 54% identical in sequence to synaptotagmin I and possesses all the domains that characterise this multigene family. It is expressed at high levels in rat brain, but not in spinal cord or a number of peripheral non-neuronal tissues.

Amino Acid Sequence↗
PubMed1999

Calcium-dependent oligomerization of synaptotagmins I and II. Synaptotagmins I and II are localized on the same synaptic vesicle and heterodimerize in the presence of calcium.

Synaptotagmins constitute a large family of membrane proteins characterized by their distinct distributions and different biochemical features. Genetic evidence suggests that members of this protein family are likely to function as calcium sensors in calcium-regulated events in neurons, although the precise molecular mechanism remains ill defined. Here we demonstrate that different synaptotagmin isoforms (Syt I, II, and IV) are present in the same synaptic vesicle population from rat brain cortex. In addition, Syt I and II co-localize on the same small synaptic vesicle (SSV), and they heterodimerize in the presence of calcium with a concentration dependence resembling that of the starting phase of SSV exocytosis (EC50 = 6 +/- 4 microM). The association between Syt I and Syt II was demonstrated by immunoprecipitation of the native proteins and the recombinant cytoplasmic domains and by using fluorescence resonance energy transfer (FRET). Although a subpopulation of SSV containing Syt I and IV can be isolated, these two isoforms do not show a calcium-dependent interaction. These results suggest that the self-association of synaptotagmins with different calcium binding features may create a variety of calcium sensors characterized by distinct calcium sensitivities. This combinatorial hypothesis predicts that the probability of a single SSV exocytic event is determined, in addition to the gating properties of the presynaptic calcium channels, by the repertoire and relative abundance of distinct synaptotagmin isoforms present on the SSV surface.

Amino Acid Sequence↗
PubMed2000

Genomic structures of synaptotagmin II protein: comparison of exon-intron organization of the synaptotagmin gene family.

Synaptotagmin (Syt) I (or II) is thought to be a Ca(2+) sensor for neurotransmitter release in the rostral (or caudal) region of the mammalian brain. In this study, we first report the genomic structures and the transcription initiation site of the mouse syt II gene. Syt II protein coding regions consist of 8 exons with a total size of about 7 kbp. Exon 2 encodes the transmembrane domain, and exons 3-5 and 6-8 encode the C2A and C2B domains, respectively, although the precise intron positions within the two C2 domains are different. The syt II gene structures are quite similar to the mouse syt VIII and human syt IX genes, but not Caenorhabditis elegans syt I, human syt VII, or the mouse syt III genes, indicating that the exon-intron patterns of the functional domain of synaptotagmins (especially the C2A domain) have not been as well conserved during evolution as among isoforms. The only conserved feature among the synaptotagmin gene family seems to have a single exon that encodes the whole transmembrane domain.

Amino Acid Sequence↗
PubMed2003

Distribution of the presynaptic calcium sensors, synaptotagmin I/II and synaptotagmin III, in the goldfish and rodent retinas.

Synaptic vesicle exocytosis is triggered by rises in calcium up to 100 microM at the site of vesicle fusion. The synaptic vesicle proteins synaptotagmin 1 and 2 (Syt I and Syt II) bind calcium at similarly high concentrations and have been proposed as the calcium sensors for fast neurotransmitter release. However, 1 microM calcium produces tonic transmitter release at photoreceptor and bipolar cell synapses in the goldfish retina, suggesting that these synapses use a higher affinity calcium sensor. Immunofluorescent staining with a panel of Syt I/II antibodies detected Syt I/II in both photoreceptor and bipolar cell terminals of the rodent retina. By contrast, no staining of either photoreceptor or protein kinase C (PKC)-labeled bipolar cell terminals was detected in the goldfish retina with any of the Syt I/II antibodies. The high affinity calcium sensor synaptotagmin 3 (Syt III) was localized to the synaptic layers of both goldfish and rodent retinas; however, while Syt III was associated with PKC-labeled bipolar cell terminals in the goldfish retina, it did not co-localize with PKC in the mouse retina. These results suggest that, unlike in their mammalian counterparts, synaptic vesicle exocytosis in goldfish photoreceptor and bipolar cell terminals utilizes a calcium sensor other than Syt I/II, possibly Syt III.

Animals↗
PubMed1999

Variations of synaptotagmin I, synaptotagmin IV, and synaptophysin mRNA levels in rat hippocampus during the estrous cycle.

Periodic changes in ovarian steroid levels during fertility cycles affect learning both in humans and in rats in parallel with electrophysiological and morphological fluctuations in selective neuronal populations. In particular, during the estrous cycle of the female rat, hippocampal CA1 region undergoes cyclic modifications in synaptic density. To investigate the molecular mechanisms involved in synaptic remodeling during the estrous cycle, we analyzed the expression of three presynaptic markers, synaptotagmin I, synaptotagmin IV, and synaptophysin, in the female adult rat brain by in situ hybridization. Relative abundance in mRNA for these three markers was quantified at four phases of the estrous cycle: diestrus, proestrus (AM and PM), and estrus. mRNA levels for syt1 exhibited cyclic variations in pyramidal neurons of the CA3 region of hippocampus during the estrous cycle, while mRNA levels for syt4 and SYN were relatively invariant in this or other regions of the hippocampus. Because CA3 pyramidal neurons make synaptic contacts in CA1, modulation of syt1 expression in CA3 may participate in the changes in synaptic density observed in CA1 during the estrous cycle. Furthermore, both syt1 and SYN mRNA varied cyclically in layer II, but not in layer III of entorhinal cortex, while syt4 remained unchanged throughout the cycle. These data suggest that regular variations in steroid hormone levels during fertility cycles may alter the properties of several networks involved in information processing and learning and memory through altered levels of presynaptic proteins.

Animals↗
PubMed2001

Three-dimensional structure of the synaptotagmin 1 C2B-domain: synaptotagmin 1 as a phospholipid binding machine.

Synaptotagmin 1 probably functions as a Ca2+ sensor in neurotransmitter release via its two C2-domains, but no common Ca2+-dependent activity that could underlie a cooperative action between them has been described. The NMR structure of the C2B-domain now reveals a beta sandwich that exhibits striking similarities and differences with the C2A-domain. Whereas the bottom face of the C2B-domain has two additional alpha helices that may be involved in specialized Ca2+-independent functions, the top face binds two Ca2+ ions and is remarkably similar to the C2A-domain. Consistent with these results, but in contrast to previous studies, we find that the C2B-domain binds phospholipids in a Ca2+-dependent manner similarly to the C2A-domain. These results suggest a novel view of synaptotagmin function whereby the two C2-domains cooperate in a common activity, Ca2+-dependent phospholipid binding, to trigger neurotransmitter release.

Amino Acid Sequence↗
PubMed2002

The C2A domain of synaptotagmin-like protein 3 (Slp3) is an atypical calcium-dependent phospholipid-binding machine: comparison with the C2A domain of synaptotagmin I.

The synaptotagmin-like protein (Slp) family consists of an N-terminal Rab27-binding domain and C-terminal tandem C2 motifs, and although it has been suggested to regulate Rab27-dependent membrane trafficking, such as Ca2+-regulated granule exocytosis in T-lymphocytes [Kuroda, Fukuda, Ariga and Mikoshiba (2002) J. Biol. Chem. 277, 9212-9218], little is known about the Ca2+-binding property of the Slp family. In this study, I demonstrated that the C2A domain of Slp3 exhibits Ca(2+)-dependent phospholipid-binding activity similar to that of the C2A domain of synaptotagmin I (Syt I) with regard to phospholipid selectivity, bivalent cation selectivity and effect of ionic strength. This finding was surprising because the C2A domains of other C-terminal-type (C-type) tandem C2 proteins require five conserved acidic residues in the putative Ca2+-binding loops 1 and 3 on the top of the beta-sandwich structure for their Ca2+-/phospholipid-binding activity, whereas the C2A domain of Slp3 contains only one conserved acidic residue in the putative Ca2+-binding loop 1. Site-directed mutagenesis and chimaeric analysis of the C2A domains of Syt I and Slp3 showed that Glu-336 and Glu-337 in the putative Ca2+-binding loop 1 and polybasic sequence (Lys-359, Lys-360 and Lys-361) in the beta-4 strand of the C2 structure are crucial for Ca2+-dependent phospholipid-binding activity of the Slp3 C2A domain, whereas the similar polybasic sequence in the C2A domain of Syt I is dispensable for Ca2+-dependent phospholipid-binding activity. These results indicate that the C2A domain of Slp3 is an atypical Ca2+-/phospholipid-binding machine, compared with other C-type tandem C2 proteins.

Amino Acid Sequence↗
PubMed2002

Vesicle-associated membrane protein-2/synaptobrevin binding to synaptotagmin I promotes O-glycosylation of synaptotagmin I.

Synaptotagmin I (Syt I), an evolutionarily conserved integral membrane protein of synaptic vesicles, is now known to regulate Ca2+-dependent neurotransmitter release. Syt I protein should undergo several post-translational modifications before maturation and subsequent functioning on synaptic vesicles (e.g. N-glycosylation and fatty acylation in vertebrate Syt I), because the apparent molecular weight of Syt I on synaptic vesicles (mature form, 65,000) was much higher than the calculated molecular weight (47,400) predicted from the cDNA sequences both in vertebrates and invertebrates. Common post-translational modification(s) of Syt I conserved across phylogeny, however, have never been elucidated. In the present study, I discovered that dithreonine residues (Thr-15 and Thr-16) at the intravesicular domain of mouse Syt I are post-translationally modified by a complex form of O-linked sugar (i.e. the addition of sialic acids) in PC12 cells and that the O-glycosylation of Syt I in COS-7 cells depends on the coexpression of vesicle-associated membrane protein-2 (VAMP-2)/synaptobrevin. I also showed that a transmembrane domain of Syt I directly interacts with isolated VAMP-2, but not VAMP-2, in the heterotrimeric SNARE (SNAP receptor) complex (vesicle SNARE, VAMP-2, and two target SNAREs, syntaxin IA and SNAP-25). Since di-Thr or di-Ser residues are often found at the intravesicular domain of invertebrate Syt I, and VAMP-dependent O-glycosylation was also observed in squid Syt expressed in COS-7 cells, I propose that VAMP-dependent O-glycosylation of Syt I is a common modification during evolution and may have important role(s) in synaptic vesicle trafficking.

Amino Acid Sequence↗
PubMed2002

Non-polarized distribution of synaptotagmin IV in neurons: evidence that synaptotagmin IV is not a synaptic vesicle protein.

Synaptotagmin IV (Syt IV) expression is regulated by neuronal development and by depolarization in the brain and in neuronal cell cultures. In cultures, immunocytochemical analysis has shown that Syt IV is localized at the Golgi and at the tips of growing neurites, but little was known about associations between Syt IV and vesicles or organelles [J. Neurochem. 74 (2000) 518]. In this study we performed an electron microscopic (EM) analysis of developing mouse neocortex to determine the exact localization of Syt IV in native mouse tissues. In neurons of layer II/III, Syt IV was found to be localized in the dendrites and axons, and at the Golgi in the cell body. Some Syt IV signals were clearly associated with vesicles and/or organelles, but EM and cell fractionation studies showed no Syt IV signals at synaptic vesicles. Detection of fluorescence protein-tagged Syt IV (Syt IV-EGFP) in hippocampal neurons also showed the presence of Syt IV-EGFP vesicles or organelles in the axons and dendrites. These results suggest that Syt IV regulates non-polarized membrane trafficking in neurons, which may be involved in synaptic plasticity or neuronal development.

Animals↗