Steady-state level of kit ligand mRNA in goat ovaries and the role of kit ligand in preantral follicle survival and growth in vitro

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...

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Published in:	Molecular reproduction and development Vol. 77; no. 3; pp. 231 - 240
Main Authors:	Celestino, Juliana J.H., Bruno, Jamily B., Lima-Verde, Isabel B., Matos, Maria Helena T., Saraiva, Mércia Viviane A., Chaves, Roberta N., Martins, Fabricio S., Almeida, Anderson P., Cunha, Rodrigo M.S., Lima, Laritza F., Name, Khesller P.O., Campello, Claudio C., Silva, José Roberto V., Báo, Sônia N., Figueiredo, José Ricardo
Format:	Journal Article
Language:	English
Published:	Hoboken Wiley Subscription Services, Inc., A Wiley Company 01-03-2010
Subjects:	Analysis of Variance Animals Cell Survival Female Goats - metabolism Goats - physiology Oocytes - cytology Oocytes - metabolism Ovarian Follicle - cytology Ovarian Follicle - metabolism Ovarian Follicle - physiology Ovary - cytology RNA, Messenger - genetics RNA, Messenger - metabolism Statistics, Nonparametric Stem Cell Factor - genetics Stem Cell Factor - metabolism Stem Cell Factor - physiology Tissue Culture Techniques
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Summary:	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.
Bibliography:	ArticleID:MRD21138 ark:/67375/WNG-HCV82CZ6-4 istex:CF0D18041D046FE7BF349D07B8CD17AFC9813AA0 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1040-452X 1098-2795
DOI:	10.1002/mrd.21138