We imaged and analysed the motion of single insulin secretory granules near the plasma membrane in live pancreatic β-cells, from normal and diabetic Goto–Kakizaki (GK) rats, using total internal reflection fluorescence microscopy (TIRFM). In normal rat primary β-cells, the granules that were fusing during the first phase originate from previously docked granules, and those during the second phase originate from ‘newcomers’. In diabetic GK rat β-cells, the number of fusion events from previously docked granules were markedly reduced, and, in contrast, the fusion from newcomers was still preserved. The dynamic change in the number of docked insulin granules showed that, in GK rat β-cells, the total number of docked insulin granules was markedly decreased to 35% of the initial number after glucose stimulation. Immunohistochemistry with anti-insulin antibody observed by TIRFM showed that GK rat β-cells had a marked decline of endogenous insulin granules docked to the plasma membrane. Thus our results indicate that the decreased number of docked insulin granules accounts for the impaired insulin release during the first phase of insulin release in diabetic GK rat β-cells.
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Research Article|
June 22 2004
TIRF imaging of docking and fusion of single insulin granule motion in primary rat pancreatic β-cells: different behaviour of granule motion between normal and Goto–Kakizaki diabetic rat β-cells
Mica OHARA-IMAIZUMI;
Mica OHARA-IMAIZUMI
1Department of Biochemistry (II), Kyorin University School of Medicine, Shinkawa 6-20-2, Mitaka, Tokyo 181-8611, Japan
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Chiyono NISHIWAKI;
Chiyono NISHIWAKI
1Department of Biochemistry (II), Kyorin University School of Medicine, Shinkawa 6-20-2, Mitaka, Tokyo 181-8611, Japan
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Toshiteru KIKUTA;
Toshiteru KIKUTA
1Department of Biochemistry (II), Kyorin University School of Medicine, Shinkawa 6-20-2, Mitaka, Tokyo 181-8611, Japan
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Shintaro NAGAI;
Shintaro NAGAI
1Department of Biochemistry (II), Kyorin University School of Medicine, Shinkawa 6-20-2, Mitaka, Tokyo 181-8611, Japan
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Yoko NAKAMICHI;
Yoko NAKAMICHI
1Department of Biochemistry (II), Kyorin University School of Medicine, Shinkawa 6-20-2, Mitaka, Tokyo 181-8611, Japan
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Shinya NAGAMATSU
Shinya NAGAMATSU
1
1Department of Biochemistry (II), Kyorin University School of Medicine, Shinkawa 6-20-2, Mitaka, Tokyo 181-8611, Japan
1To whom correspondence should be addressed (e-mail shinya@kyorin-u.ac.jp).
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Publisher: Portland Press Ltd
Received:
March 16 2004
Revision Received:
May 04 2004
Accepted:
May 05 2004
Accepted Manuscript online:
May 06 2004
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2004
Biochem J (2004) 381 (1): 13–18.
Article history
Received:
March 16 2004
Revision Received:
May 04 2004
Accepted:
May 05 2004
Accepted Manuscript online:
May 06 2004
Citation
Mica OHARA-IMAIZUMI, Chiyono NISHIWAKI, Toshiteru KIKUTA, Shintaro NAGAI, Yoko NAKAMICHI, Shinya NAGAMATSU; TIRF imaging of docking and fusion of single insulin granule motion in primary rat pancreatic β-cells: different behaviour of granule motion between normal and Goto–Kakizaki diabetic rat β-cells. Biochem J 1 July 2004; 381 (1): 13–18. doi: https://doi.org/10.1042/BJ20040434
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