Steady‐state level of kit ligand mRNA in goat ovaries and the role of kit ligand in preantral follicle survival and growth in vitro |
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Authors: | Juliana JH Celestino Jamily B Bruno Isabel B Lima‐Verde Maria Helena T Matos Mércia Viviane A Saraiva Roberta N Chaves Fabricio S Martins Anderson P Almeida Rodrigo MS Cunha Laritza F Lima Khesller PO Name Claudio C Campello José Roberto V Silva Sônia N Báo José Ricardo Figueiredo |
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Institution: | 1. Faculty of Veterinary Medicine, LAMOFOPA, PPGCV, State University of Ceara, Fortaleza, CE, Brazil;2. Biotecnology Nucleus of Sobral (NUBIS), Federal University of Ceara, Sobral, CE, Brazil;3. Laboratory of Electron Microscopy, Department of Cell Biology, University of Brasilia, Brasilia, DF, Brazil |
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Abstract: | The aims of this study were to investigate steady‐state level of Kit Ligand (KL) mRNA and its effects on in vitro survival and growth of caprine preantral follicles. RT‐PCR was used to analyze caprine steady‐state level of KL mRNA in primordial, primary, and secondary follicles, and in small (1–3 mm) and large (3–6 mm) antral follicles. Furthermore, ovarian fragments were cultured for 1 or 7 days in Minimal Essential Medium (MEM+) supplemented with KL (0, 1, 10, 50, 100, or 200 ng/ml). Noncultured (control) and cultured fragments were processed for histology and transmission electron microscopy (TEM). RT‐PCR demonstrated an increase in steady‐state level of KL mRNA during the transition from primary to secondary follicles. Small antral follicles had higher steady‐state levels of KL mRNA in granulosa and theca cells than large follicles. After 7 days, only 50 ng/ml of KL had maintained the percentage of normal follicles similar to control. After 1 day, all KL concentrations reduced the percentage of primordial follicles and increased the percentage of growing follicles. KL at 10, 50, 100, or 200 ng/ml increased primary follicles, compared to MEM+ after 7 days. An increase in oocyte and follicular diameter was observed at 50 ng/ml of KL. TEM confirmed ultrastructural integrity of follicles after 7 days at 50 ng/ml of KL. In conclusion, the KL mRNAs were detected in all follicular categories. Furthermore, 50 ng/ml of KL maintained the integrity of caprine preantral follicle cultured for 7 days and stimulated primordial follicle activation and follicle growth. Mol. Reprod. Dev. 77: 231–240, 2010. © 2009 Wiley‐Liss, Inc. |
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