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. 2020 Mar;75(1):101-108.
doi: 10.22092/ari.2018.122232.1220. Epub 2020 Mar 1.

Effect of Point Mutation in the Growth Differentiation Factor 9 Gene of Oocytes on the Sterility and Fertility of Awassi Sheep

H Al-Mutar  1   1 , L Younis  2
Affiliations

Effect of Point Mutation in the Growth Differentiation Factor 9 Gene of Oocytes on the Sterility and Fertility of Awassi Sheep

H Al-Mutar et al. Arch Razi Inst. 2020 Mar.

Abstract

Growth differentiation factor 9 (GDF9) plays a critical role in ovarian follicular development and ovulation rate. The present study aimed to investigate the correlation between the single-nucleotide polymorphism (SNP) of the GDF9 gene and reproductive performance variables, such as fertility and sterility in Awassi sheep. Forty pairs of ovaries from a total of 40 slaughtered Iraqi Awassi ewes were used in this study. Twenty of the ovaries were collected from sterile ewes and the other 20 ovaries were taken from fertile ewes for genomic DNA extraction, polymerase chain reaction, and sequencing to detect GDF9 gene polymorphism. Follicles and oocytes of all the 40 ovaries were evaluated and compared with the results of genotyping. Furthermore, histopathological and microscopic evaluations were performed for 40 ovarian tissues of the two groups. The sequence analysis revealed that exon I had three SNPs, including T(114)C, G(129)R, and G(199)A. The first two SNPs were silent mutations and the last mutation was missense responsible for the substitution of glutamic acid with lysine at position 67. The current study showed a significant increase (P≤0.01) in GG, AA, CC, GA, and GG genotypes at G(129)R, G(199)A, T(114)C, G(129)R, and G(199)A loci, respectively. Moreover, the TT genotype in locus T(114)C was recorded to significantly augment (P≤0.05) in the fertile ewes. Mutant GA genotype of the G(129)R locus led to a significant (P≤0.05) increase in the percentage of follicles (4-8 mm) and oocytes number, compared to wild GG. On the other hand, a significant decrease was recorded in the mutant AA genotype in G(199)A, compared to wild GG. Differences between CC and TT genotypes at T(114)C locus were not significant. Histopathological examination revealed hypoplasia in the ovarian tissue of sterile ewes accompanied by fibrous connective tissue invasion and follicles degeneration. However, in the fertile ewes, the ovarian tissues were normal with the presence of numerous corpus albicans and degenerative corpus luteum. According to the findings of this study, the homozygote mutation in fertile ewes minimized the number of follicles and oocytes leading to sterility, while the heterozygote mutation was reported in the fertile Awassi ewes.

Keywords: Fertility; Heterozygote; Homozygote; Infertility.

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Conflict of interest statement

The authors declare that they have no conflict of interest.

Figures

Figure 1
Figure 1
PCR product of exon I GDF9 gene; M: DNA ladder 100–10000 bp; Lane 1–19: s PCR samples.
Figure 2
Figure 2
Wild-type and new variants of the exon I of GDF9 gene; (A): T(114)C, (B): G(129)R, and (C): G(199)A.
Figure 3
Figure 3
Ovarian histological sections of sterile ewes; (A) hypoplatic ovarian tissue; (B) follicular cells degeneration (H&E ×40).
Figure 4
Figure 4
Histological analysis of the ovaries of normal fertile ewes; (A) normal follicular development with the presence of many corpus albicans; (B) degenerative corpus luteum.
See this image and copyright information in PMC

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