title: Steady-state level of kit ligand mRNA in goat ovaries and the role of kit ligand in preantral follicle survival and growth in vitro.
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.
authors.affiliation: Faculty of Veterinary Medicine, LAMOFOPA, PPGCV, State University of Ceara, Fortaleza, CE, Brazil. juliana_jales@yahoo.com.br
authors.forename: Juliana J H
authors.initials: JJ
authors.lastname: Celestino
authors.forename: Jamily B
authors.initials: JB
authors.lastname: Bruno
authors.forename: Isabel B
authors.initials: IB
authors.lastname: Lima-Verde
authors.forename: Maria Helena T
authors.initials: MH
authors.lastname: Matos
authors.forename: Mércia Viviane A
authors.initials: MV
authors.lastname: Saraiva
authors.forename: Roberta N
authors.initials: RN
authors.lastname: Chaves
authors.forename: Fabricio S
authors.initials: FS
authors.lastname: Martins
authors.forename: Anderson P
authors.initials: AP
authors.lastname: Almeida
authors.forename: Rodrigo M S
authors.initials: RM
authors.lastname: Cunha
authors.forename: Laritza F
authors.initials: LF
authors.lastname: Lima
authors.forename: Khesller P O
authors.initials: KP
authors.lastname: Name
authors.forename: Claudio C
authors.initials: CC
authors.lastname: Campello
authors.forename: José Roberto V
authors.initials: JR
authors.lastname: Silva
authors.forename: Sônia N
authors.initials: SN
authors.lastname: Báo
authors.forename: José Ricardo
authors.initials: JR
authors.lastname: Figueiredo
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D002470:Cell Survival; D005260:Female; D006041:Goats / Q000378:metabolism / Q000502:physiology*; D009865:Oocytes / Q000166:cytology / Q000378:metabolism*; D006080:Ovarian Follicle / Q000166:cytology / Q000378:metabolism* / Q000502:physiology; D010053:Ovary / Q000166:cytology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D018709:Statistics, Nonparametric; D019089:Stem Cell Factor / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D046509:Tissue Culture Techniques
chemical_list: D012333:RNA, Messenger; D019089:Stem Cell Factor