Expression of Granulosa Cell-Specific Genes and Induction of Apoptosis in Conditionally Immortalized Granulosa Cell Lines Established from H-2Kb-tsA58 Transgenic Mice

Abstract

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.

Establishment and characterization of a noradrenergic adrenal chromaffin cell line, tsAM5NE, immortalized with the temperature-sensitive SV40 T-antigen

“…In fact, it was earlier reported that, in some of tsSV40T-immortalized cell lines, the upshift to the nonpermissive temperature not only induces growth arrest but also causes apoptotic cell death Yanai and Obinata, 1994;Zheng et al, 1994;Guenal et al, 1997;Walther et al, 1999;Tavelin et al, 1999;Wade et al, 1999). In fact, it was earlier reported that, in some of tsSV40T-immortalized cell lines, the upshift to the nonpermissive temperature not only induces growth arrest but also causes apoptotic cell death Yanai and Obinata, 1994;Zheng et al, 1994;Guenal et al, 1997;Walther et al, 1999;Tavelin et al, 1999;Wade et al, 1999).…”

“…In contrast, at the non-permissive temperature of 39uC, tsAM5NE cells underwent cell death when cultured in the presence of serum alone. In fact, it was earlier reported that, in some of tsSV40T-immortalized cell lines, the upshift to the nonpermissive temperature not only induces growth arrest but also causes apoptotic cell death Yanai and Obinata, 1994;Zheng et al, 1994;Guenal et al, 1997;Walther et al, 1999;Tavelin et al, 1999;Wade et al, 1999). Because p53 has the ability to induce apoptotic cell death (Evan et al, 1998;Gottlieb et al, 1998), it has been suggested that the apoptotic death of tsSV40T-immortalized cells upon temperature upshift reflects the biological activity of p53 released from its complex with the thermolabile T-antigen (Murata et al, 2006;Yanai and Obinata, 1994;Zheng et al, 1994).…”

Neuronal differentiation elicited by glial cell line‐derived neurotrophic factor and ciliary neurotrophic factor in adrenal chromaffin cell line tsAM5D immortalized with temperature‐sensitive SV40 T‐antigen

“…In contrast, at the nonpermissive temperature, tsAM5D cells underwent cell death when cultured in the presence of serum alone. In fact, it was reported that, in some of tsSV40T‐immortalized cell lines, the upshift to the nonpermissive temperature not only induces growth arrest but also causes apoptotic cell death (Vayssiere et al,1994; Yanai and Obinata,1994; Zheng et al,1994; Guenal et al,1997; Walther et al,1999; Tavelin et al,1999; Wade et al,1999). Because p53 has the ability to induce apoptotic cell death (Evan and Littlewood,1998; Gottlieb and Oren,1998), it has been suggested that the apoptotic death of tsSV40T‐immortalized cells upon temperature upshift reflects the biological activity of p53 released from its complex with thermolabile T antigen (Yanai and Obinata,1994; Zheng et al,1994).…”