ob gene expression and secretion of leptin following differentiation of rat preadipocytes to adipocytes in primary culture

Sharon Elizabeth Mitchell, William Rees, L. J. Hardie, Nigel Hoggard, M Tadayyon, J R Arch, P Trayhurn

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

Expression of the ob gene and production of leptin have been examined on differentiation of rat fibroblastic preadipocytes to adipocytes in primary culture. Preadipocytes were obtained from the inguinal fat pad of suckling rats, and following differentiation the cells contained lipid droplets and the mRNAs for both lipoprotein lipase and adipsin were detected by Northern blotting. ob mRNA was not, however, detected on Northern blots, but analysis by RT-PCR indicated that the ob gene was expressed, particularly after differentiation. Measurement of leptin in the culture medium by ELISA showed that the ob gene product was secreted by adipocytes from approximately 4 days after the induction of differentiation. Leptin production was sustained over a 2-week period with a peak at 8-10 days post-induction. Dexamethasone stimulated leptin production, while an inhibition was observed with the beta-adrenoceptor agonist isoprenaline. These results demonstrate that following the differentiation of fibroblastic preadipocytes to adipocytes in primary culture, leptin is secreted with the cells responding to stimuli which regulate production of the hormone.
Original languageEnglish
Pages (from-to)360-364
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume230
Issue number2
DOIs
Publication statusPublished - 13 Jan 1997

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Leptin
Cell culture
Adipocytes
Gene expression
Rats
Gene Expression
Northern Blotting
Genes
Complement Factor D
Messenger RNA
Lipoprotein Lipase
Groin
Isoproterenol
Adrenergic Receptors
Dexamethasone
Culture Media
Adipose Tissue
Cell Differentiation
Enzyme-Linked Immunosorbent Assay
Fats

Keywords

  • Adipocytes
  • Adipose Tissue
  • Animals
  • Cell Differentiation
  • Cells, Cultured
  • Complement Factor D
  • DNA Primers
  • Enzyme-Linked Immunosorbent Assay
  • Fibroblasts
  • Kinetics
  • Leptin
  • Lipoprotein Lipase
  • Male
  • Obesity
  • Oligonucleotide Probes
  • Oligonucleotides, Antisense
  • Polymerase Chain Reaction
  • Protein Biosynthesis
  • Rats
  • Rats, Inbred Strains
  • Serine Endopeptidases
  • Time Factors

Cite this

ob gene expression and secretion of leptin following differentiation of rat preadipocytes to adipocytes in primary culture. / Mitchell, Sharon Elizabeth; Rees, William; Hardie, L. J.; Hoggard, Nigel; Tadayyon, M; Arch, J R; Trayhurn, P.

In: Biochemical and Biophysical Research Communications, Vol. 230, No. 2, 13.01.1997, p. 360-364.

Research output: Contribution to journalArticle

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abstract = "Expression of the ob gene and production of leptin have been examined on differentiation of rat fibroblastic preadipocytes to adipocytes in primary culture. Preadipocytes were obtained from the inguinal fat pad of suckling rats, and following differentiation the cells contained lipid droplets and the mRNAs for both lipoprotein lipase and adipsin were detected by Northern blotting. ob mRNA was not, however, detected on Northern blots, but analysis by RT-PCR indicated that the ob gene was expressed, particularly after differentiation. Measurement of leptin in the culture medium by ELISA showed that the ob gene product was secreted by adipocytes from approximately 4 days after the induction of differentiation. Leptin production was sustained over a 2-week period with a peak at 8-10 days post-induction. Dexamethasone stimulated leptin production, while an inhibition was observed with the beta-adrenoceptor agonist isoprenaline. These results demonstrate that following the differentiation of fibroblastic preadipocytes to adipocytes in primary culture, leptin is secreted with the cells responding to stimuli which regulate production of the hormone.",
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