SerpinB6b在大鼠着床期子宫及蜕膜的表达与调控

SerpinB6b是丝氨酸蛋白酶抑制剂亚家族B成员之一。本实验旨在利用荧光定量PCR、原位杂交、免疫组织化学与蛋白质印迹分析等实验技术,研究SerpinB6b mRNA和蛋白在大鼠早期妊娠1~9 d、人工诱导蜕膜化及体外诱导蜕膜化模型子宫中的表达规律。结果显示:SerpinB6b mRNA和蛋白在大鼠早期妊娠1~5 d腔上皮、腺上皮有微弱表达;在6~9 d,表达主要集中在胚胎和蜕膜区,且随蜕膜化程度增加,表达量逐渐增强,且在第8天达到最高;在大鼠人工诱导蜕膜化模型中,SerpinB6b在蜕膜区的表达均显著高于非蜕膜区;在大鼠子宫内膜基质细胞体外诱导蜕膜化模型中,SerpinB6b随着诱导蜕膜化天数的增加呈上升趋势。以上体内和体外实验结果表明,SerpinB6b在大鼠子宫中的表达具有着床相关特异性,SerpinB6b参与大鼠子宫蜕膜化过程。

Large‑scale transcriptomic and genomic analyses reveal a novel functional gene SERPINB6 for chicken carcass traits

Background Carcass traits are crucial indicators of meat production efficiency.However,the molecular regulatory mechanisms associated with these traits remain unclear.Results In this study,we conducted comprehensive transcriptomic and genomic analyses on 399 Tiannong partridge chickens to identify key genes and variants associated with carcass traits and to elucidate the underlying regulatory mechanisms.Based on association analyses with the elastic net(EN)model,we identified 12 candidate genes(AMY1A,AP3B2,CEBPG,EEF2,EIF4EBP1,FGFR1,FOXD3,GOLM1,LOC107052698,PABPC1,SERPINB6 and TBC1D16)for 4 carcass-related traits,namely live weight,dressed weight,eviscerated weight,and breast muscle weight.SERPINB6 was identified as the only overlapping gene by 3 analyses,EN model analysis,weighted gene co-expression network analysis and differential expression analysis.Cell-level experiments confirmed that SERPINB6 promotes the proliferation of chicken DF1 cells and primary myoblasts.Further expression genome-wide association study and association analysis indicated that rs317934171 is the critical site that enhances SERPINB6 expression.Furthermore,a dual-luciferase reporter assay proved that gga-miR-1615 targets the 3′UTR of SERPINB6.Conclusions Collectively,our findings reveal that SERPINB6 serves as a novel gene for chicken carcass traits by promoting fibroblast and myoblast proliferation.Additionally,the downstream variant rs317934171 regulates SERPINB6 expression.These results identify a new target gene and molecular marker for the molecular mechanisms of chicken carcass traits.

SerpinB6b过表达载体的构建及对小鼠子宫基质细胞蜕膜化的影响

试验构建丝氨酸蛋白酶抑制剂亚家族成员6b(Serine peptidase inhibitor,clade B,member 6b,SerpinB6b)基因过表达载体,研究其对小鼠子宫基质细胞蜕膜化的影响。根据Gen Bank中已发表的SerpinB6b基因序列,设计含酶切位点的特异性引物,PCR扩增并与p GEM-T载体连接,经限制性内切酶EcoRⅠ和HindⅢ双酶切后,与pc DNA3. 1(+)载体融合得到SerpinB6b过表达载体。将其转染到体外分离培养的小鼠子宫基质细胞,利用荧光定量PCR法检测子宫基质细胞中SerpinB6b mRNA表达的变化,以及SerpinB6b过表达后蜕膜化标志分子Prl8a2和Prl3c1表达的变化,对照组转染pc DNA3. 1(+)空载体。结果表明:SerpinB6b过表达载体经双酶切鉴定后,结果与目的基因条带吻合,测序结果与Gen Bank中小鼠SerpinB6b基因序列同源性为100%。与对照组相比,转染SerpinB6b过表达载体后,SerpinB6b在小鼠子宫基质细胞中的表达显著升高。同时,过表达SerpinB6b可明显促进蜕膜化标志分子Prl8a2和Prl3c1 mRNA的表达。提示SerpinB6b可能在小鼠子宫基质细胞蜕膜化过程中起重要的调节作用。