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The aim of the present work was to determine the relative transcript abundance of the oestrogen receptor alpha (ERα) and beta (ERβ) gene in long-term cultures (up to 18 days) of pig granulosa cells (GC) obtained from small (1-2 mm; SF-GC) and large (5-7 mm; LF-GC) follicles. GC were cultured in the presence of 10% foetal calf serum (FCS) and one of the following growth factors: leukaemia inhibitory factor (LIF), basic fibroblast growth factor (bFGF), or stem cell factor (SCF). The relative transcript abundance of the investigated genes was measured by real time PCR (RTPCR). In comparison with the control (medium only), LIF significantly increased (P<0.05-0.01) ERα gene expression in SF-GC after 3, 15 and 18 days of culture and after 12 days in LF-GC. The ERβ mRNA level in SF-GC was significantly decreased (P<0.01) by all of the investigated factors after 3, 6, 9 and 12 days of incubation. In conclusion, expression of both types of oestrogen receptors during prolonged culture of pig GC under different conditions suggests the role of oestrogens acting via their receptors in maintaining survivability of porcine granulosa cells in vitro.
The aim of the study was to investigate the effect of testosterone (T) and dihydrotestosterone (DHT) applied alone or in combination with antiandrogen (flutamide) on proliferation, progesterone secretion and telomerase activity (TA) of pig granulosa cells (GC) derived from small (1-3 mm; SFGC) and large (5-7 mm; LF-GC) ovarian follicles. Cells were treated with investigated factors for 48 h. 5α-DHT stimulated (P<0.05-0.01) proliferation of SF and LF granulosa cells. Flutamide applied individually and in a combination with testosterone and DHT stimulated (P<0.05-0.01) proliferation of pig GC from small and large antral follicles. Flutamide had no effect on progesterone synthesis in small as well as in large follicle GC. Antiandrogen applied individually and in a combination with testosterone and DHT enhanced (P<0.05-0.01) telomerase actitvity in SF- and LF-GC. The results of the study suggest the involvement of androgen receptor in telomerase activity regulation in pig GC and a link between telomerase and the proliferation status of GC.
White Short-Haired (WSH) and Brown Short-Haired (BSH) are two protected local goat dairy breeds in Czech Republic. A genetic characterization of both breeds is necessary to preserve them from extinction and to exploit their genetic variation. For this purpose a study was carried out on the genetic polymorphism of the CSN2 and CSN3 loci. Genomic analysis was performed by PCR method. Both breeds were characterized by the absence of CSN20 allele. The CSN3 locus was found polymorphic, with the CSN3A, CSN3B and CSN3C alleles frequency of 0.15, 0.80 and 0.05 in WSH and 0.52, 0.40 and 0.08 in BSH, respectively.
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