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Differential distribution of synaptotagmin-1, -4, -7, and -9 in rat adrenal chromaffin cells

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Abstract

Neurons and certain kinds of endocrine cells, such as adrenal chromaffin cells, have large dense-core vesicles (LDCVs) and synaptic vesicles or synaptic-like microvesicles (SLMVs). These secretory vesicles exhibit differences in Ca2+ sensitivity and contain diverse signaling substances. The present work was undertaken to identify the synaptotagmin (Syt) isoforms present in secretory vesicles. Fractionation analysis of lysates of the bovine adrenal medulla and immunocytochemistry in rat chromaffin cells indicated that Syt 1 was localized in LDCVs and SLMVs, whereas Syt 7 was the predominant isoform present in LDCVs. In contrast to PC12 cells and the pancreatic β cell line INS-1, Syt 9 was not immunodetected in LDCVs in rat chromaffin cells. Double-staining revealed that Syt 9-like immunoreactivity was nearly identical with fluorescent thapsigargin binding, suggesting the presence of Syt 9 in the endoplasmic reticulum (ER).The exogenous expression of Syt 1-GFP in INS-1 cells, which had a negligible level of endogenous Syt 1, resulted in an increase in the amount of Syt 9 in the ER, suggesting that Syt 9 competes with Syt 1 for trafficking from the ER to the Golgi complex. We conclude that LDCVs mainly contain Syt 7, whereas SLMVs contain Syt 1, but not Syt 7, in rat and bovine chromaffin cells.

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Acknowledgments

We thank I. Niki (Oita University, Oita, Japan) and C. B. Wollheim (University Medical Center, Geneva, Switzerland) for providing INS-1 cells. Our thanks are also due to M. Courel (University of California, San Diego, USA) for the plasmid pCMV-CgA-EGFP and to H.-H. Gerdes (University of Bergen, Bergen, Norway) for the plasmid pcDNA3-hCgB-EGFP. We are grateful to T. Hatama for technical assistance.

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Authors and Affiliations

  1. Department of Cell and Systems Physiology, University of Occupational and Environmental Health School of Medicine, Kitakyushu, 807-8555, Japan

    Hidetada Matsuoka, Keita Harada & Masumi Inoue

  2. Department of Psychiatry, University of Occupational and Environmental Health School of Medicine, Kitakyushu, 807-8555, Japan

    Jun Nakamura

  3. Department of Developmental Biology and Neurosciences, Graduate School of Life Sciences, Tohoku University, Aobayama, Aoba-ku, Sendai, Miyagi, 980-8578, Japan

    Mitsunori Fukuda

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  1. Hidetada Matsuoka
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  2. Keita Harada
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  3. Jun Nakamura
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  4. Mitsunori Fukuda
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  5. Masumi Inoue
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Correspondence to Masumi Inoue.

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This work was supported in part by MEXT KAKENHI (21026029 to M.I.) and JSPS KAKENHIs (21500360 to M.I. and 22790222 to H.M.).

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Matsuoka, H., Harada, K., Nakamura, J. et al. Differential distribution of synaptotagmin-1, -4, -7, and -9 in rat adrenal chromaffin cells. Cell Tissue Res 344, 41–50 (2011). https://doi.org/10.1007/s00441-011-1131-8

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  • DOI: https://doi.org/10.1007/s00441-011-1131-8

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