Synaptonemal complex protein 3 (SYCP3) is a DNA-binding protein located on the lateral elements of the synaptonemal complex. This protein plays an important role in homologous chromosome pairing and is necessary for male meiosis of spermatogenesis. To further understand the SYCP3 gene function and its relationship to cattle-yak male sterility, we investigated the characteristics of the bovine SYCP3 (bSYCP3) gene as well as its transcription level and epigenetic modification status. The bSYCP3 gene encodes a 225-amino acid protein with the Cor1 motif and two coiled-coil-forming regions, homologous with other mammals (59-77% identity overall). Real-time PCR analysis indicated that the expression level of bSYCP3 mRNA in yak testes was significantly higher than that in cattle-yak (p < 0.05). The methylation level of the bSYCP3 promoter (mainly 7th, 13th and 17th CpG sites) in cattle-yaks (40%) was significantly higher than that in yaks (0%) (p < 0.05). This suggests that bSYCP3 plays an important role in meiosis of bovine spermatogenesis; further, interspecific hybridization between yak and cattle might influence bSYCP3 gene expression in cattle-yak testes, which might be influenced by bSYCP3 gene promoter methylation.
Embryonic survival rate, an important factor in the fecundity of sows, is affected by endometrium-secreting histotroph. A higher concentration of calcium ion has been observed in the uterus of highly prolific Erhualian sows (EH) compared with those of less prolific (EL) sows. This suggests that EH sows have better establishment and maintenance of pregnancies, thus increasing embryonic survival rate during the peri-implantation period. To understand the mechanisms of how the endometrium-secreting histotroph affects embryonic survival rate during the Erhualian peri-implantation period, the expression patterns of endometrial mRNA in the EH and EL sows on day 12 of gestation were analyzed using RNA sequencing technology. A total of 164 differentially expressed genes (DEGs) were identified (P adj < 0.05, |log 2 (FC)| 1), including 46 upregulated and 118 downregulated genes in EH compared to EL. Gene Ontology enrichment indicated that a subset of DEGs was involved in calcium ion binding and cell adhesion. Solute carrier family 8 member A3 and solute carrier family 24 member 4, identified as upregulated genes (P adj < 0.05) in EH, were considered key candidate genes expressed in the endometrium affecting embryonic survival rate during the peri-implantation period. The results improve understanding of the genetic mechanism underlying the variation in litter size of Erhualian pigs during the peri-implantation period.
ABSTRACT. The aim of this study was to construct a mammary glandspecific expression vector, pGN, and to validate its function in expressing growth hormone (GH) both in vitro and in vivo. First, the GH gene was amplified and inserted downstream of the b-casein 5'-arm. Next, the neo gene was cloned downstream of the b-casein 3'-arm as a selection marker. To analyze the bioactivity of the pGN plasmid, we expressed pGN in a Bcap-37 cell line and in the goat mammary gland. Quantitative PCR analysis revealed that the expression of GH mRNA in the pGN-transfected group was higher than that of the control group in Bcap-37 cells. Results of a radioimmunoassay and an enzyme-linked immunosorbent assay demonstrated that the pGN-transfected group expressed much more GH protein than the non-transfected group (P < 0.05). Upon injection of the pGN plasmid into the goat mammary gland, GH mRNA and growth hormone receptor mRNA expressions increased 2-fold. In vivo analyses revealed that GH protein expression was higher in the injected group than in the control group. Together, these results strongly demonstrated that the pGN plasmid was constructed correctly and exhibited favorable bioactivity in efficiently expressing GH both in vitro and in vivo.
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