Molecular dissection of a quantitative trait locus: a phenylalanine-to-tyrosine substitution in the transmembrane domain of the bovine growth hormone receptor is associated with a major effect on milk yield and composition.

Recent studies challenge the previous view that apoptosis within the granulosa cells of the maturing ovarian follicle is a reflection of aging and consequently a marker for poor quality of the contained oocyte. On the contrary, apoptosis within the granulosa cells is an integral part of normal development and has limited predictive capability regarding oocyte quality or the ensuing pregnancy rate in in vitro fertilization programs. This review article covers our revised understanding of the process of apoptosis within the ovarian follicle, its three phenotypes, the major signaling pathways underlying apoptosis as well as the associated mitochondrial pathways.

/pdf/granulosa-cell-apoptosis-in-the-ovarian-follicle-a-changing-2ln2rj243a.pdf

Granulosa Cell Apoptosis in the Ovarian Follicle-A Changing View.

Bone Morphogenetic Protein (BMP) signaling in animal reproductive system development and function.

The multiplex polymerase chain reaction (PCR) method is fast and accuracy for screening several polymorphism loci in a single reaction in large samples. Therefore, this study aimed to identify three novel insertion/deletion (indel) loci in goat growth hormone receptor (GHR) gene by using multiplex PCR method and the genotypes in three indel loci are identified in 918 individuals from three Chinese goat breeds, as well as to evaluate its associations with growth traits. After validating the accuracy by taking the conventional PCR method, the concordance between these two methods was 100%. Moreover, P14 locus polymorphism was significantly associated with six growth traits of Hainan black goats. Briefly, an economic multiplex PCR method is presented to simultaneously and accurately detect three indel polymorphisms within goat GHR without the use of any special equipment, thus improving the experimental efficiency and fast obtaining the experimental result for indel genotyping.

Development of a touch-down multiplex PCR method for simultaneously rapidly detecting three novel insertion/deletions (indels) within one gene: an example for goat GHR gene

The aim of the present study was to determine the direct cause of the mutation-induced, increased ovulation rate in Booroola Merino (BB) sheep. Granulosa cells were removed from antral follicles before ovulation and post-ovulation from BB (n=5) and WT (n=12) Merino ewes. Direct immunofluorescence measurement of mature cell surface receptors using flow cytometry demonstrated a significant up-regulation of FSH receptor (FSHR), transforming growth factor beta type 1, bone morphogenetic protein receptor (BMPR1B), and LH receptor (LHR) in BB sheep. The increased density of FSHR and LHR provide novel evidence of a mechanism for increasing the number of follicles that are recruited during dominant follicle selection. The compounding increase in receptors with increasing follicle size maintained the multiple follicles and reduced the apoptosis, which contributed to a high ovulation rate in BB sheep. In addition, we report a mutation-independent mechanism of down-regulation to reduce receptor density of the leading dominant follicle in sheep. The suppression of receptor density coincides with the cessation of mitogenic growth and steroidogenic differentiation as part of the luteinization of the follicle. The BB mutation-induced attenuation of BMPR1B signaling led to an increased density of the FSHR and LHR and a concurrent reduction in apoptosis to increase the ovulation rate. The role of BMPs in receptor modulation is implicated in the development of multiple ovulations.

/pdf/flow-cytometric-analysis-of-fshr-bmrr1b-lhr-and-apoptosis-in-2rda0fwd1w.pdf

Flow cytometric analysis of FSHR, BMRR1B, LHR and apoptosis in granulosa cells and ovulation rate in merino sheep

In this study, the polymorphisms of growth hormone (GH) gene 5′ promoter region and intron 8, exons 4 and 10 of growth hormone receptor (GHR) gene were analyzed in Xinong Saanen goats (SG) and Boer goats (BG). Two alleles (A and B) and three genotypes (AA, AB and BB) were detected at P1 locus of GH gene, and two alleles (G and T) and two genotypes (GG and GT) were detected at P4 locus of GHR gene by PCR-SSCP analysis. In addition, two single nucleotide polymorphisms (SNPs)-A73C (P1 locus) and G114T (P4 locus), were identified by DNA sequencing. The frequencies of alleles A and B in the two goat breeds were 0.61–0.62, and 0.39–0.38, respectively, and the frequencies of alleles G and T in the two goat breeds were 0.82–0.86, and 0.18–0.14, respectively. The SNP loci were in Hardy–Weinberg disequilibrium in both goat breeds (P < 0.05). Polymorphisms of GH and GHR genes were shown to be associated with growth traits in BG breed. AA and GG genotypes were associated with superior growth traits in 1-, 2- and 3-month old individuals. Hence, AA and GG genotypes are suggested to be a molecular marker for superior growth traits in BG breed.

Novel polymorphisms of goat growth hormone and growth hormone receptor genes and their effects on growth traits.

This study reported the analysis of KIT ligand (KITLG) gene polymorphisms in 681 goats of three breeds: Xinong Saanen (SN), Guanzhong (GZ), and Boer (BG). In addition, the study identified three allelic variants: g.769T>C and g.817G>T in SN and GZ breeds, and g.9760G>C in the three goat breeds. The g.769T>C and g.817G>T loci were closely linked (r(2)  > 0.33). All the single nucleotide polymorphism loci were in Hardy-Weinberg disequilibrium (P < 0.05). Significant associations were found for litter size with all three loci. Therefore, these results suggest that the KITLG gene is a strong candidate gene affecting litter size in goats.

Polymorphism identification in the goat KITLG gene and association analysis with litter size.

New genetic polymorphisms of KiSS-1 gene and their association with litter size in goats

Bone morphogenetic proteins (BMPs), belonging to the transforming growth factor-beta family, are crucial factors in follicular growth and development in the mammalian ovary. In this study, we examined the effects of BMP2, 4, 6 and 7 on goat granulosa cells by in vitro culture. The results showed that BMP2, 4 and 7 increased follicle-stimulating hormone receptor (FSHR) level in goat granulosa cells whereas they decreased luteinizing hormone receptor (LHR) level in both transcript and protein levels; however, BMP6 upregulated LHR transcript and protein level in goat granulosa cells, whereas it had no effect on FSHR level. FSHR knockdown increased granulosa cell apoptosis. Our data provided the i¬rst evidence that BMP2, 4, 6 and 7 may inhibit granulosa cell apoptosis through the regulation of FSHR and/or LHR level. These i¬ndings provided new insight into the biological functions of BMP2, 4, 6 and 7 for follicular development in goat ovary.

	 

	Key words: Granulosa cell, bone morphogenetic proteins (BMPs), follicle-stimulating hormone receptor (FSHR), luteinizing hormone receptor (LHR)

/pdf/bone-morphogenetic-proteins-bmp-2-4-6-and-7-affect-ovarian-jcujm841sq.pdf

Bone morphogenetic proteins (BMP) 2, 4, 6 and 7 affect ovarian follicular development through regulation of follicle-stimulating hormone receptor (FSHR) and luteinizing hormone receptor (LHR) expression in goat granulosa cells

The invention discloses a method for detecting the polymorphism of three base mutations of the gene exon 4 of goat gonadotropin releasing hormone and the gene exon 2 of the growth differentiation factor 9. The method comprises the following steps: using the DNA sequence of goat genome as template, using 2 to amplify the primer P1 and P2 under PCR condition in the presence of Taq DNA polymerase, buffer environment, Mg2+ and dNTPs, using agarose gel electrophoresis to judge the size of the target fragments; using polyacrylamide gel electrophoresis to detect the PCR amplified product of the primer P1 and P2 to find that the two amplification sites have three base mutations, then performing genotyping and gene frequency analysis to the SNPs of the two genes, and performing association analysis with the fecundity traits of Boer goat and Xinong Saanen dairy goat. The result shows that the SNPs sites detected by the primer P1 and P2, of the GnRH gene exon 4 (containing partial intron 3 and 3' non-translational region) and the GDF9 gene exon 2, thus the method can be used for molecular marking of fecundity trait selection of goat.

Yihong Cui

Papers

Novel polymorphisms of goat growth hormone and growth hormone receptor genes and their effects on growth traits.

Polymorphism identification in the goat KITLG gene and association analysis with litter size.

New genetic polymorphisms of KiSS-1 gene and their association with litter size in goats

Bone morphogenetic proteins (BMP) 2, 4, 6 and 7 affect ovarian follicular development through regulation of follicle-stimulating hormone receptor (FSHR) and luteinizing hormone receptor (LHR) expression in goat granulosa cells

Method for detecting base mutation polymorphism of goat gonadotropin releasing hormone and growth differentiation factor 9