Glucagon gknk kxpression is altered by amino acm limitation

G. L. Paul, A. V. Reddy, H. R. Gaskins, N. F. Shay

Research output: Contribution to journalArticle

Abstract

Recent evidence indicates that aniino acids <AA) influence gene expression in a number of mammalian sy stems. Since AA stimulate glucagon release from pancreatic cells, the purpose of this study was to determine it' individual AA influence glucagon gene expression. Mouse α-TC6 cells, uhich lack preproinsulin mRNA and secrete only glucagon, were maintained in DMHM containing 25 m M glucose. 15% horse serum, and 2.5% fetal call serum, fells were seeded at 200.000 cells/ml into either 6 cm dishes or 2-4-\vell plates. At 70% confluence, cells were incubated in serum-free maintenance media for 24 hr, then in serum-free treatment media containing 12.5 m M glucose and either 0 or 20 AA or 20 AA minus phe. met. leu. 01 trp for another 24 hr. Northern anal)sis was performed using a 516 bp mouse preproglucagon eDNA produced by RT-PCR from rat preproglucagon ei)NA primers. We found that 20 AA and minus leu enhanced glucagon mRNA abundance 3- and 5-fold, respective!}, compared to 0 AA. Similarly, immunoreactive glucagon secretion was enhanced 4- and 5-told for 20 AA and minus leu. respectively, versus U AA. A 24-hr lime course study using a nbonuclease protection assay to detect glucagon mRNA abundance m [he 0 AA. 2(1 AA. and minus leu treatments indicated that glucagon me.ssaue IcveK decreased o\er time in all groups, and lhat levels were ! .7 and 2.5-fbld greater in the 20 AA and minus leu, respectively, versus 0 AA alter 24 lit We conclude that leu may be imolved in the regulation of gtucagon mRKA leveU in α-TC6 cells.

Original languageEnglish (US)
Pages (from-to)A652
JournalFASEB Journal
Volume10
Issue number3
StatePublished - Dec 1 1996

Fingerprint

glucagon
Glucagon
Proglucagon
Serum-Free Culture Media
Gene expression
Messenger RNA
cells
Gene Expression
Glucose
gene expression
glucose
mice
Serum
Horses
Rats
Assays
reverse transcriptase polymerase chain reaction
Maintenance
secretion
horses

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

Paul, G. L., Reddy, A. V., Gaskins, H. R., & Shay, N. F. (1996). Glucagon gknk kxpression is altered by amino acm limitation. FASEB Journal, 10(3), A652.

Glucagon gknk kxpression is altered by amino acm limitation. / Paul, G. L.; Reddy, A. V.; Gaskins, H. R.; Shay, N. F.

In: FASEB Journal, Vol. 10, No. 3, 01.12.1996, p. A652.

Research output: Contribution to journalArticle

Paul, GL, Reddy, AV, Gaskins, HR & Shay, NF 1996, 'Glucagon gknk kxpression is altered by amino acm limitation', FASEB Journal, vol. 10, no. 3, pp. A652.
Paul, G. L. ; Reddy, A. V. ; Gaskins, H. R. ; Shay, N. F. / Glucagon gknk kxpression is altered by amino acm limitation. In: FASEB Journal. 1996 ; Vol. 10, No. 3. pp. A652.
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