β-cell lines derived from transgenic Cpe(fat)/Cpe(fat) mice are defective in carboxypeptidase E and proinsulin processing

Oleg Varlamov, Lloyd D. Fricker, Hisasi Furukawa, Donald F. Steiner, Stephen H. Langley, Edward H. Leiter

Research output: Contribution to journalArticle

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Abstract

A spontaneous point mutation in the coding region of the carboxypeptidase E (CPE) gene in Cpe(fat)/Cpe(fat) mice affects proinsulin processing. Cell lines derived from the pancreatic β-cells of Cpe(fat)/C(fat) mice were generated by crossing C57BLKS/J-Cpe(fat)/+ mice with NOD mice expressing the simian virus 40 large T oncogene under the control of the rat insulin II promoter. Two cell lines, designated NIT-2 and NIT-3, were cultured from adenomatous islets obtained from F2 littermates and were compared with the NIT-1 cell line previously developed from mice with wild-type CPE. Electron microscopy of the cultured NIT-2 and -3 cells showed increased numbers of enlarged and electron-lucent granules compared with NIT- 1 cells Pro-CPE, but not the mature form of CPE, is present in NIT-2 and -3 cells, and neither pro-CPE nor CPE are secreted into the medium. Immunocytochemistry shows the pro-CPE to be localized to an endoplasmic reticulum-like structure in NIT-3 cells. Proinsulin is less extensively processed in NIT-2 and -3 cells than in NIT-1 cells indicating that the Cpe(fat) mutation affects both the endopeptidase and carboxypeptidase reactions. The secretion of insulin/proinsulin from NIT-2 and -3 cells is significantly elevated by secretagogues, indicating that CPE is not required for sorting proinsulin into the regulated pathway.

Original languageEnglish (US)
Pages (from-to)4883-4892
Number of pages10
JournalEndocrinology
Volume138
Issue number11
DOIs
StatePublished - 1997

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Carboxypeptidase H
Proinsulin
Fats
Cell Line
Insulin
Carboxypeptidases
Endopeptidases
Inbred NOD Mouse
Simian virus 40
Oncogenes
Point Mutation
Adipocytes
Endoplasmic Reticulum
Electron Microscopy
Immunohistochemistry
Electrons

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

β-cell lines derived from transgenic Cpe(fat)/Cpe(fat) mice are defective in carboxypeptidase E and proinsulin processing. / Varlamov, Oleg; Fricker, Lloyd D.; Furukawa, Hisasi; Steiner, Donald F.; Langley, Stephen H.; Leiter, Edward H.

In: Endocrinology, Vol. 138, No. 11, 1997, p. 4883-4892.

Research output: Contribution to journalArticle

Varlamov, Oleg ; Fricker, Lloyd D. ; Furukawa, Hisasi ; Steiner, Donald F. ; Langley, Stephen H. ; Leiter, Edward H. / β-cell lines derived from transgenic Cpe(fat)/Cpe(fat) mice are defective in carboxypeptidase E and proinsulin processing. In: Endocrinology. 1997 ; Vol. 138, No. 11. pp. 4883-4892.
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