title: Synaptotagmin VII is targeted to secretory organelles in PC12 cells, where it functions as a high-affinity calcium sensor.
abstract: Synaptotagmin (syt) I is thought to act as a Ca2+ sensor that regulates neuronal exocytosis. Fifteen additional isoforms of syt have been identified, but their functions are less well understood. Here, we used PC12 cells to test the idea that different isoforms of syt impart cells with distinct metal (i.e., Ca2+, Ba2+, and Sr2+) requirements for secretion. These cells express syt's I and IX (syt IX sometimes referred to as syt V), which have low apparent metal affinities, at much higher levels than syt VII, which we show has a relatively high apparent affinity for metals. We found that syt I and VII partially colocalize on large dense core vesicles and that upregulation of syt VII produces a concomitant increase in the divalent cation sensitivity of catecholamine release from PC12 cells. Furthermore, RNA interference-mediated knockdown of endogenous syt VII reduced the metal sensitivity of release. These data support the hypothesis that the complement of syt's expressed by a cell, in conjunction with their metal affinity, determines the divalent cation sensitivity of exocytosis.
authors.affiliation: Department of Physiology, University of Wisconsin, 1300 University Ave., SMI 129, Madison, WI 53706, USA.
authors.forename: Ping
authors.initials: P
authors.lastname: Wang
authors.forename: Michael C
authors.initials: MC
authors.lastname: Chicka
authors.forename: Akhil
authors.initials: A
authors.lastname: Bhalla
authors.forename: David A
authors.initials: DA
authors.lastname: Richards
authors.forename: Edwin R
authors.initials: ER
authors.lastname: Chapman
mesh_terms: D000818:Animals; D001464:Barium / Q000737:chemistry; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism*; D002395:Catecholamines / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005089:Exocytosis; D005982:Glutathione Transferase / Q000378:metabolism; D015151:Immunoblotting; D007150:Immunohistochemistry; D008055:Lipids / Q000737:chemistry; D008081:Liposomes / Q000737:chemistry; D008247:Lysosomes / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009474:Neurons / Q000378:metabolism; D009841:Oligonucleotides / Q000737:chemistry; D016716:PC12 Cells; D020033:Protein Isoforms; D017434:Protein Structure, Tertiary; D034622:RNA Interference; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry; D013324:Strontium / Q000737:chemistry; D014162:Transfection; D015854:Up-Regulation
chemical_list: D002395:Catecholamines; D018076:DNA, Complementary; D008055:Lipids; D008081:Liposomes; D009841:Oligonucleotides; D020033:Protein Isoforms; D011994:Recombinant Proteins; D001464:Barium; D005982:Glutathione Transferase; D002118:Calcium; D013324:Strontium