Real-time monitoring of cAMP response element binding protein signaling in porcine granulosa cells modulated by ovarian factors

Pei Jian He, Yasunori Fujimoto, Nobuhiko Yamauchi, Masa Aki Hattori

Research output: Contribution to journalArticlepeer-review

14 Citations (Scopus)

Abstract

The present study was performed to establish a real-time monitoring of the cAMP response element binding protein (CREB) signalling using granulosa cells, and to assess the modulation of CREB activity by potential ovarian autocrine/paracrine and oocyte-derived factors. Granulosa cells were isolated from porcine follicles and cultured for 2 days, and then transfected with CRE-containing pGL3. The cells were directly stimulated or cultured with FSH, LH, forskolin, or a permeable cAMP analog, and/or IGF-I, EGF, bFGF, TGF-β2 or TNF-α, or cumulus-oocyte complex (COCs) for the real-time monitoring of CREB signaling. The activation pattern of CREB signaling consisted of three distinct phases, i.e., burst, attenuation and refractory. In contrast to FSH, LH, and forskolin, a cAMP analog induced the prolonged activation, although three distinct phases were observed at its high concentration. Of all the autocrine/paracrine factors, only IGF-I slightly induced CREB activity. On the other hand, TGF-β2 and TNF-β significantly repressed FSH-stimulated transcriptional activation of CREB by 30% (P < 0.05) and 45% (P< 0.05), respectively. Additionally, coculture with COCs caused a significant suppression of transcriptional activation of CREB signaling stimulated by FSH. These results indicate that ovarian autocrine/paracrine factors such as IGF-I, TGF-β2, TNF-α and oocyte-derived factors modulate the CREB signaling. The present study provides a new approach for direct signaling study on transcription factors under the influences of potential factors.

Original languageEnglish
Pages (from-to)177-184
Number of pages8
JournalMolecular and cellular biochemistry
Volume290
Issue number1-2
DOIs
Publication statusPublished - Oct 1 2006

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Clinical Biochemistry
  • Cell Biology

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