Rab8a is involved in membrane trafficking of Kir6.2 in the MIN6 insulinoma cell line

Keiichiro Uchida, Masatoshi Nomura, Tadashi Yamamoto, Yoshihiro Ogawa, Noriyoshi Teramoto

Research output: Contribution to journalArticle

Abstract

Although ATP-sensitive K+ (KATP) channels play an important role in the secretion of insulin by pancreatic beta cells, the mechanisms that regulate the intracellular transport of KATP channel subunit proteins (i.e., Kir6.2 and sulfonylurea receptor 1 (SUR1)) to the plasma membrane remain uncharacterized. We investigated the possibility that an interaction between KATP channel subunit proteins and Rab8a protein, a member of the RAS superfamily, may be involved in the membrane trafficking of KATP channels. Co-immunoprecipitation and immunostaining experiments using co-expression systems with fluorescent protein-tagged Kir6.2 were carried out to identify the coupling of KATP channels and Rab8a proteins in the insulin-secreting cell line, MIN6. Rab8a protein co-localized with Kir6.2 protein, a channel pore subunit (in a granular pattern), and with insulin. Knockdown of the Rab8a gene with RNA interference using small interfering RNA systems caused reductions in the amount of total KATP and plasma membrane surface KATP channels without decreasing the messenger RNA transcription of the KATP channel subunits. Rab8a gene knockdown also enhanced glucose-induced insulin secretion. These results suggest that Rab8a may be involved in membrane trafficking of KATP channels and the maintenance of normal insulin secretion in the MIN6 pancreatic beta cell line.

Original languageEnglish
Pages (from-to)877-887
Number of pages11
JournalPflugers Archiv European Journal of Physiology
Volume471
Issue number6
DOIs
Publication statusPublished - Jun 1 2019

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KATP Channels
Insulinoma
Cells
Membranes
Cell Line
Insulin
Insulin-Secreting Cells
Gene Knockdown Techniques
Protein Subunits
Cell membranes
Proteins
Genes
Sulfonylurea Receptors
Cell Membrane
Transcription
RNA Interference
Immunoprecipitation
Small Interfering RNA
Adenosine Triphosphate
Maintenance

All Science Journal Classification (ASJC) codes

  • Physiology
  • Clinical Biochemistry
  • Physiology (medical)

Cite this

Rab8a is involved in membrane trafficking of Kir6.2 in the MIN6 insulinoma cell line. / Uchida, Keiichiro; Nomura, Masatoshi; Yamamoto, Tadashi; Ogawa, Yoshihiro; Teramoto, Noriyoshi.

In: Pflugers Archiv European Journal of Physiology, Vol. 471, No. 6, 01.06.2019, p. 877-887.

Research output: Contribution to journalArticle

Uchida, Keiichiro ; Nomura, Masatoshi ; Yamamoto, Tadashi ; Ogawa, Yoshihiro ; Teramoto, Noriyoshi. / Rab8a is involved in membrane trafficking of Kir6.2 in the MIN6 insulinoma cell line. In: Pflugers Archiv European Journal of Physiology. 2019 ; Vol. 471, No. 6. pp. 877-887.
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