Expression of granulosa cell-specific genes and induction of apoptosis in conditionally immortalized granulosa cell lines established from H-2Kb-tsA58 transgenic mice.
Granulosa cell lines have been established from H-2Kb-tsA58 transgenic mice. Using immunocytochemistry, significant amounts of insulin-like growth factor-I (IGF-I) were found in all cell lines investigated, whereas estrogen and progesterone receptor expression could be detected in only some of the lines. All cell lines showed low basal production of the gonadal steroids estradiol and progesterone. The genes for the ovarian paracrine regulators IGF-I and basic fibroblast growth factor were expressed, as well as the genes for anti-Müllerian hormone and for the P450 side-chain cleavage enzyme (P450scc). Expression of P450scc could be shown to be up-regulated in the cell lines under conditions mimicking the hormonal environment of the luteinizing granulosa cells in vivo. Inactivation of the temperature-sensitive SV40 T antigen by a shift of the cell lines to the nonpermissive temperature of 39.5 degrees C led to massive induction of apoptosis in several cell lines. These cell lines will allow a detailed study of the mechanisms regulating the expression of granulosa cell-specific functions, as well as the induction of granulosa cell apoptosis.
Walther
N
N
Institute for Hormone and Fertility Research, University of Hamburg, D-22529 Hamburg, Germany. walther@ihf.de
Einspanier
A
A
Jansen
M
M
D000818:Animals; D017209:Apoptosis / Q000235:genetics* / Q000502:physiology*; D002460:Cell Line; D004967:Estrogens / Q000096:biosynthesis; D005260:Female; D005786:Gene Expression Regulation / Q000235:genetics* / Q000502:physiology*; D006107:Granulosa Cells / Q000378:metabolism / Q000502:physiology*; D006133:Growth Substances / Q000096:biosynthesis; D007150:Immunohistochemistry; D051379:Mice; D008822:Mice, Transgenic; D011374:Progesterone / Q000096:biosynthesis; D011962:Receptors, FSH / Q000096:biosynthesis / Q000235:genetics; D011974:Receptors, LH / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013256:Steroids / Q000096:biosynthesis
D004967:Estrogens; D006133:Growth Substances; D011962:Receptors, FSH; D011974:Receptors, LH; D013256:Steroids; D011374:Progesterone