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) 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.
Celestino
Juliana J H
JJ
Faculty of Veterinary Medicine, LAMOFOPA, PPGCV, State University of Ceara, Fortaleza, CE, Brazil. juliana_jales@yahoo.com.br
Bruno
Jamily B
JB
Lima-Verde
Isabel B
IB
Matos
Maria Helena T
MH
Saraiva
Mércia Viviane A
MV
Chaves
Roberta N
RN
Martins
Fabricio S
FS
Almeida
Anderson P
AP
Cunha
Rodrigo M S
RM
Lima
Laritza F
LF
Name
Khesller P O
KP
Campello
Claudio C
CC
Silva
José Roberto V
JR
Báo
Sônia N
SN
Figueiredo
José Ricardo
JR
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
D012333:RNA, Messenger; D019089:Stem Cell Factor