生长素对猪孤雌激活胚胎体外发育的影响及其受体基因的表达模式

In vitro development of porcine parthenogenetic embryos in response to ghrelin application and the expression of receptor gene GHSR-1a

为探明生长素及其受体GHSR-1a在胚胎早期发育中的作用,以猪孤雌激活胚胎为研究对象,在其体外发育过程中添加不同浓度生长素,观察胚胎发育速度和发育效率,并用荧光定量PCR检测猪孤雌激活胚胎早期发育过程中生长素特异性受体基因GHSR-1a的表达。结果显示,相对于对照组,添加10.0 mg/L生长素可以显著提高4-细胞胚胎、8-细胞胚胎、桑葚胚的发育率(P<0.05),并能加快4-细胞胚胎、8-细胞胚胎、桑葚胚的发育速度(P<0.05)。GHSR-1a mRNA在2-细胞期到桑葚胚期保持较高表达水平,囊胚期后显著下降(P<0.05);免疫荧光染色表明GHSR-1a在猪孤雌激活胚胎各发育时期均有表达,4-细胞期、8-细胞期和桑葚胚期的表达量显著高于囊胚期(P<0.05)。推测生长素通过提高GHSR-1a的表达来提高猪孤雌激活胚胎的发育速度和效率。

The present study was designed to investigate the roles of exogenous ghrelin and its receptor GHSR-1 a during early in vitro development of porcine parthenogenetic embryos. The developmental rates of 4-cell embryos, 8-cell embryos, and morula of porcine parthenogenetic embryos cultured in PZM3 supplemented with 10. 0 mg/L ghrelin were significantly higher than control group ( ghrelin free) ( P < 0. 05 ) , and the development of 4-cell embryos, 8-cell embryos, and morula were speeded up as well. The mRNA expression of GHSR-1 a was identi-fied at each stage of porcine parthenogenetic embryos (2-cell, 4-cell, 8-cell, morula and blastocyst). Indi-rect immuno-fluorescence revealed that the protein GH-SR-1 a levels were higher at 4-cell stage, 8-cell stage <br> and morula stage. It was suggested ghrelin improved the developmental rate and speed by increasing the receptor gene GHSR-1 a expression.