The effects of insulin and follicle-simulating hormone (FSH) during in vitro development of ovarian goat preantral follicles and the relative mRNA expression for insulin and FSH receptors and cytochrome P450 aromatase in cultured follicles.
The actions of different concentrations of insulin alone or in combination with follicle-stimulating hormone (FSH) were evaluated by in vitro follicular development and mRNA expression of cytochrome P450 aromatase (CYP19A1) and as receptors for insulin (INSR) and FSH (FSHR) from isolated, cultured goat preantral follicles. Goat preantral follicles were microdissected and cultured for 18 days in the absence or presence of insulin (5 and 10 ng/ml or 10 μg/ml) alone or in combination with FSH. After 18 days, the addition of the maximum concentration of insulin to the culture medium reduced follicular survival and antrum formation rates significantly compared to the other treatments. However, when FSH was added to the culture medium, no differences between these two parameters were observed. Preantral and antral follicles from the fresh control as well as from all cultured follicles still presented a normal ultrastructural pattern. In medium supplemented with FSH, only insulin at 10 ng/ml presented oocytes with higher rates of meiosis resumption compared to control, as well as oocytes in metaphase II. Treatment with insulin (10 ng/ml) plus FSH resulted in significantly increased levels of INSR and CYP19A1 mRNA compared to that with other treatments. In conclusion, 10 ng/ml insulin associated with FSH was more efficient in promoting resumption of oocyte meiosis, maintaining survival, stimulating follicular development, and increasing expression of the INSR and CYP19A1 genes in goat preantral follicles.
Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary, State University of Ceara, Fortaleza-CE, Brazil. rncvet@gmail.com
Roberta N
RN
Chaves
Ana Beatriz G
AB
Duarte
Giovanna Q
GQ
Rodrigues
Juliana J H
JJ
Celestino
Gerlane M
GM
Silva
Claudio Afonso P
CA
Lopes
Anderson P
AP
Almeida
Mariana A M
MA
Donato
C A
CA
Peixoto
Arlindo A A
AA
Moura
Carlos H
CH
Lobo
Yann
Y
Locatelli
Pascalle
P
Mermillod
Claudio C
CC
Campello
Jose Ricardo
JR
Figueiredo
D000818:Animals; D001141:Aromatase / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D002478:Cells, Cultured; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D006041:Goats / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D059471:In Vitro Oocyte Maturation Techniques / Q000379:methods; D007328:Insulin / Q000494:pharmacology*; D006080:Ovarian Follicle / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D011972:Receptor, Insulin / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D011962:Receptors, FSH / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D015895:Relative Value Scales
D007328:Insulin; D012333:RNA, Messenger; D011962:Receptors, FSH; D005640:Follicle Stimulating Hormone; D001141:Aromatase; D011972:Receptor, Insulin