The Expression Characteristics Of Partial Gene During The Porcine Follicualr Atresia

In mammalian ovaries, a large number of follicles undergo atresia during follicular growth and development. Follicuar atresia is a multi-factors involved complex regulation process, some research has confirmed the regulation role of many factors, but the mechanism of follicular atresia is subject to further research, which has crucial guiding significance in animal reproduction control and definition the mechanism of ovulation dysfunction disease. The antral follicles in 3-5mm diameter were successfully separated and classified different stages of atresia though comprehensive methods "morphological determination and progesterone/estrogen ratio". Based on the exact classification of the follicles, we measured the changes in the expression of partial follicular atresia related gene in porcine ovary follicles during follicular atresia, analyzed the correlationship among the genes and the physiological situation of the follicles.The classification of different stages of atresia through morphological determination and progesterone/estrogen ratio(P/E). The morphological determination:Healthy follicles (H) had a homogenous and pink theca interna, visible and uniformly distributed capillaries, such a clear follicular fluid that the cumulus-oocyte complex (COC) was discernable and granulosa cell layer was integrity. Early atretic follicles (EA) was turbid, dark orange coloured, the COC was illegible and granulose cell layer was partially detachment and crinkled. Progressed atretic follicles (PA) had a non-homogenous, dark gray theca interna and few capillaries, follicular fluid was so flocculent that the COC was not discernable and granulose cell layer seriously lost and deposited. The progesterone/estrogen ratio:health follicles P4/E2<5;Early atretic follicles 5<P4/E2<10; Progressed atretic follicles P4/E2 >10.The expression changes of gene in porcine ovary follicles during follicular atresia. Total RNA was extracted from healthy, early and progressed atretic follicles, detected the changes of gene expression by Real Time RT-PCR for VEGFA,Cx43,Egr-1,CYP1A1 FasL,Foxol,Caspase-3,IL-1β,P53,Bcl-XL,Mcl-1,Bcl-2,Bax,Bak,Bid and Bim.The expression levels of VEGFA gene decreased in the porcine ovary follicles with the progress of follicular atresia, there is significant difference among healthy, early and progressed atretic follicles. The expression Cx43,Egr-1 and IL-1βis significant higher in early and progressed atretic follicles than the health follicles. CYP1A1,FasL,Foxo1,Caspase-3 and P53 gene expression increased in the porcine ovary follicles with the progress of follicular atresia, the expression of CYP1A1,FasL,Foxoland P53 is high in the progressed atretic follicles, decreased much in health follicles, there is significant difference between health and the other two groups. The expression of Caspase-3 is significant difference between progressed atretic follicles and the other two groups. Bcl-2,Mcl-1,Bid and Bim gene expression increased with the process of follicular atresia, there is there is significant difference between health and the other two groups. The expression of Bcl-XL and Bak is high in progressed atretic follicle, there is significant difference between progressed atretic follicles and the other two groups. Bax gene expression increased with the process of follicular atresia, there is no significant difference among healthy, early and progressed atretic follicles.The correlationship between the expression of gene and P4/E2 ratio. The correlation coefficient of VEGFA expression and P4/E2 ratio is -0.349, there is no significant correlation. The correlation coefficient of Cx43 expression and P4/E2 ratio is 0.510, reached significant positive correlation. The correlation coefficient of Egr-1 expression and P4/E2 ratio is 0.301, there is no significant correlation. The correlation coefficient of CYP1A1 expression and P4/E2 ratio is 0.295, there is no significant correlation. The correlation coefficient of FasL expression and P4/E2 ratio is 0.509, reached significant positive correlation. The correlation coefficient of Foxol expression and P4/E2 ratio is 0.420, reached significant positive correlation. The correlation coefficient of Caspase-3 expression and P4/E2 ratio is 0.398, showed significant positive correlation. The correlation coefficient of IL-1βexpression and P4/E2 ratio is 0.749, showed significant positive correlation. The correlation coefficient of P53 expression and P4/E2 ratio is 0.611, showed significant positive correlation. The correlation coefficient of Bcl-XL expression and P4/E2 ratio is 0.559, showed significant positive correlation. The correlation coefficient of Mcl-1 expression and P4/E2 ratio is 0.546, reached significant positive correlation. The correlation coefficient of Bcl-2 expression and P4/E2 ratio is 0.481, reached significant positive correlation. The correlation coefficient of Bax expression and P4/E2 ratio is 0.038, there is no significant correlation. The correlation coefficient of Bak expression and P4/E2 ratio is 0.478, reached significant positive correlation. The correlation coefficient of Bid expression and P4/E2 ratio is 0.619, reached significant positive correlation.The correlation coefficient of Bim expression and P4/E2 ratio is 0.632, showed significant positive correlation.