Gap junctions in myometrial cell cultures: Evidence for modulation by cyclic adenosine 3':5'-monophosphate

H. D. Dookwah, R. Barhoumi, T. R. Narasimhan, S. H. Safe, R. C. Burghardt

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

Abstract

Primary cultures of myometrial cells from juvenile rats, continuous cultures maintained by serial passage, and a pSV3neo-transfected myometrial cell line were established and utilized for the study of development and modulation of gap junctional intercellular communication (GJIC) in vitro. The smooth muscle origin and homogeneity of the cultures were verified by immunofluorescence staining of α-smooth muscle actin and cellular desmin. Although gap junctions were not detected in thin sections of juvenile and adult myometrial tissues by transmission electron microscopy, they were detected in cultured myometrial cells derived from juvenile and adult animals. The presence of GJIC in cultured cells was confirmed using a fluorescence recovery after photo-bleaching assay. Administration of exogenous estradiol-17β (10-7 M) resulted in an increase in GJIC in primary and passage 9 myometrial cultures, whereas pSV3neo-transfected myometrial cells were not significantly different from untreated controls. The lack of estrogen responsiveness in pSV3neo-transfected cultures correlated with lower levels of estrogen receptors than in primary cultures. Addition of 1 mM 8-bromo-cAMP resulted in rapid (within 2 min) increases in dye transfer in both control and estradiol-17β-primed primary cultures. Uncoupling of cells by treatment with 1 mM 1-octanol, followed by addition of 1 mM 8-bromo-cAMP, resulted in increased GJIC in control and estradiol-17β- primed cultures, although up-regulation of GJIC in estradiol-17β-primed cultures was much greater than in control cultures. Comparative experiments carried out on a spontaneously immortalized rat granulosa cell line (SIGC), which expresses the same connexin43 species as myometrial cells, exhibited similar responses to exogenous 8-bromo-cAMP following uncoupling of gap junctions with octanol. While the results of these investigations may not be extrapolated to myometrium in vivo, they suggest that myometrial cell culture may offer additional opportunities to explore the temporal expression and modulation of GJIC in myometrium.

Original languageEnglish (US)
Pages (from-to)397-407
Number of pages11
JournalBiology of Reproduction
Volume47
Issue number3
DOIs
StatePublished - 1992

ASJC Scopus subject areas

  • Reproductive Medicine
  • Cell Biology

Fingerprint

Dive into the research topics of 'Gap junctions in myometrial cell cultures: Evidence for modulation by cyclic adenosine 3':5'-monophosphate'. Together they form a unique fingerprint.

Cite this