Expression Analysis of BMF Gene During the Ovaries at Different Stages of Estrus and Follicular Development in Small-tailed Han Sheep

doi:10.7668/hbnxb.20193551

Abstract

Abstract:

The aim of this experiment was to investigate the biological function of BMF gene(Bcl-2 modifying factor)in the follicular development of Small-tailed Han Sheep,and to provide the theoretical basis for further exploration of its regulatory mechanism in the future.Firstly,the synchronization estrus was implemented in Small-tailed Han Sheep.Then,we used Quantitative Real-time PCR(RT-qPCR),Western Blot(WB)and Immunofluorescence staining(IF)to detect the expression of the BMF gene and its encoded protein in the ovaries at different physiological stages;subsequently,we isolated follicles of different diameters and analyzed the differences in expression of the target gene in large,middle and small follicles.The results showed that the expression of BMF mRNA was significantly higher in the ovary at 42 h after thrombectomy than at 18 h after thrombectomy(P<0.05),and the results of its encoded protein expression of BMF were basically consistent with the mRNA.BMF protein was mainly expressed in follicular granulosa cells and follicular membrane cells.We further isolated follicles and found that the expression of both the BMF gene and its encoded protein in middle follicles was significantly higher than that in large and small follicles(P<0.05).These results suggest that BMF gene may be involved in the ovarian cyclic activity and play an important role in follicular development.

Key words: Small-tailed Han Sheep, BMF, Follicle, Ovary, Reproduction

LI Yingying, ZHANG Haokun, JIA Haofan, GUO Jingyao, YANG Xixi, WANG Zhiliang, LI Mingna, WANG Xinrong. Expression Analysis of BMF Gene During the Ovaries at Different Stages of Estrus and Follicular Development in Small-tailed Han Sheep[J]. Acta Agriculturae Boreali-Sinica, 2023, 38(2): 217-223. doi: 10.7668/hbnxb.20193551.

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Figures/Tables 6

Tab.1 Information of primer sequences

基因 Gene	引物序列(5'-3') Primer sequences (5'-3')	产物大小/bp Amplification size
BMF	F:CCAGTTCCATCGGCTTCATA	115
	R:CCCATTCCTGTTCTCATCTCC
β-actin	F:CTTCCAGCCTTCCTTCCTGG	180
	R:GCCAGGGCAGTGATCTCTTT

Fig.1 Expression of BMF mRNA in ovary of Small-tailed Han Sheep at different physiological stages The different letters means significant difference(P < 0.05). The same as Fig.2,4,5.

Fig.2 Expression of BMF protein in ovary of Small-tailed Han Sheep at different physiological stages A.BMF blotting bands in ovaries at different physiological stages;B.BMF protein expression in different physiological stages of ovary.

Fig.3 Distribution of BMF protein in ovary of Small-tailed Han Sheep at different physiological stages(200×) GC.Granulosa cells;LC.Leydig cells;TC.Theca cells.

Fig.4 Expression of BMF mRNA in follicles of different diameters

Fig.5 Expression of BMF protein in follicles of different diameters A.BMF imprinted bands in follicles of different diameters:S.Small follicles,M.Middle follicles,L.Large follicles;B.Expression of BMF protein in follicles of different diameters.

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