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Porcine granulosa cells do not express transforming growth factor-beta 2 (TGF-beta 2) messenger ribonucleic acid: molecular basis for their inability to produce TGF-beta activity comparable to that of rat granulosa cells.

Publication ,  Journal Article
Mulheron, GW; Mulheron, JG; Danielpour, D; Schomberg, DW
Published in: Endocrinology
December 1992

In contrast to rat granulosa cells (GC), GC of the pig and cow produce very low levels of transforming growth factor-beta (TGF-beta)-like activity in vitro. Because cultured rat GC predominantly express TGF-beta 2 messenger RNA (mRNA) and secrete high levels of the protein, we hypothesized that TGF-beta 2 mRNA expression by porcine GC might be absent or diminished, thus providing a molecular explanation(s) for their relatively low levels of TGF-beta production. We tested this hypothesis by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay analysis. When analyzed by RT-PCR, porcine GC RNA from 1-3 mm follicles did not yield the expected 489 base pair (bp) TGF-beta 2 product but instead generated a smaller than anticipated 240 bp species; porcine testis RNA generated both the 240 and the anticipated 489 bp products. Sequencing these species indicated that the smaller form was not a novel TGF-beta 2 splice variant, and that the 489-bp product was porcine TGF-beta 2. This is the first reported nucleotide sequence for porcine TGF-beta 2; it is 90% and 91% identical to murine and human TGF-beta 2 sequences, respectively. Further RT-PCR analysis of porcine GC RNA resulted in the identification of bp products representing TGF-beta 1 and TGF-beta 3 mRNA. Enzyme-linked immunosorbent assay analysis of porcine GC conditioned medium confirmed the presence of TGF-beta 1 at very low levels. TGF-beta 2 was undetectable. Comparable analysis of GC from the diethylstilbestrol-treated prepubertal rat demonstrated the presence of TGF-beta 1 and TGF-beta 3 mRNA by RT-PCR and very low levels of the corresponding protein products in conditioned culture medium. Collectively, these results suggest that the inability of porcine GC to express TGF-beta 2 mRNA could explain the very low levels of TGF-beta activity secreted by these cells in vitro.

Duke Scholars

Published In

Endocrinology

DOI

ISSN

0013-7227

Publication Date

December 1992

Volume

131

Issue

6

Start / End Page

2609 / 2614

Location

United States

Related Subject Headings

  • Transforming Growth Factor beta
  • Testis
  • Swine
  • Rats
  • RNA-Directed DNA Polymerase
  • RNA, Messenger
  • Polymerase Chain Reaction
  • Molecular Sequence Data
  • Male
  • Immunologic Techniques
 

Citation

APA
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Mulheron, G. W., Mulheron, J. G., Danielpour, D., & Schomberg, D. W. (1992). Porcine granulosa cells do not express transforming growth factor-beta 2 (TGF-beta 2) messenger ribonucleic acid: molecular basis for their inability to produce TGF-beta activity comparable to that of rat granulosa cells. Endocrinology, 131(6), 2609–2614. https://doi.org/10.1210/endo.131.6.1280203
Mulheron, G. W., J. G. Mulheron, D. Danielpour, and D. W. Schomberg. “Porcine granulosa cells do not express transforming growth factor-beta 2 (TGF-beta 2) messenger ribonucleic acid: molecular basis for their inability to produce TGF-beta activity comparable to that of rat granulosa cells.Endocrinology 131, no. 6 (December 1992): 2609–14. https://doi.org/10.1210/endo.131.6.1280203.
Journal cover image

Published In

Endocrinology

DOI

ISSN

0013-7227

Publication Date

December 1992

Volume

131

Issue

6

Start / End Page

2609 / 2614

Location

United States

Related Subject Headings

  • Transforming Growth Factor beta
  • Testis
  • Swine
  • Rats
  • RNA-Directed DNA Polymerase
  • RNA, Messenger
  • Polymerase Chain Reaction
  • Molecular Sequence Data
  • Male
  • Immunologic Techniques