Department of Animal Science, National Chung Hsing University, Taichung, Taiwan, ROC.
Reproduction. 2011 Jul;142(1):87-97. doi: 10.1530/REP-11-0049. Epub 2011 Apr 13.
We investigated the expression of sonic hedgehog (SHH) receptor PTCH1 and its co-receptor smoothened (SMO) in fertilized porcine embryos. Effects of exogenous SHH on embryonic development and expressions of survival- and pluripotency-related genes were also determined. We found that PTCH1 and SMO are expressed from two-cell to blastocyst embryos. When oocytes or fertilized embryos were respectively cultured in the maturation or embryo culture medium supplemented with SHH (0.5 μg/ml), their blastocyst rates and total cell numbers increased (P<0.05) compared with the untreated control. When cultured simultaneously in the in vitro maturation (IVM) and in vitro culture (IVC) media supplemented with SHH, the oocytes gained increased blastocyst rates and total cell numbers in an additive manner, with reduced apoptotic indices (P<0.05). Interestingly, SHH treatment did not affect the expression of the BCL2L1 (BCL-XL) gene, yet reduced BAX expression. Blastocysts cultured with various SHH regimes had similar pluripotency-related gene (POU5F1 (OCT-4) and CDX2) expression levels, but blastocysts derived from SHH treatment during IVM had higher ZPF42 (REX01) expression (P<0.05). The highest ZPF42 expression was observed in the blastocysts derived from SHH-supplemented IVC and from dual IVM and IVC treatments. The levels of acetylated histone 3 (AcH3K9/K14) increased in the two-cell and the four-cell embryos when IVM and/or IVC media were supplemented with SHH (P<0.05). Our findings indicate that SHH conferred a beneficial effect on preimplantation development of porcine embryos, particularly when both IVM and IVC media were supplemented with SHH, and the effects may be further carried over from IVM to the subsequent embryonic development.
我们研究了 sonic hedgehog (SHH) 受体 patched1 (PTCH1)及其共受体 smoothened (SMO) 在受精猪胚胎中的表达。还确定了外源性 SHH 对胚胎发育和生存及多能性相关基因表达的影响。我们发现 PTCH1 和 SMO 从二细胞期到囊胚期胚胎均有表达。当卵母细胞或受精卵分别在添加 SHH（0.5μg/ml）的成熟或胚胎培养液中培养时，其囊胚率和总细胞数均高于未处理对照组（P<0.05）。当在添加 SHH 的体外成熟（IVM）和体外培养（IVC）培养基中同时培养时，卵母细胞以累加的方式获得更高的囊胚率和总细胞数，同时凋亡指数降低（P<0.05）。有趣的是，SHH 处理并不影响 BCL2L1（BCL-XL）基因的表达，但降低了 BAX 的表达。用不同 SHH 方案培养的囊胚具有相似的多能性相关基因（POU5F1（OCT-4）和 CDX2）表达水平，但在 IVM 期间用 SHH 处理的囊胚 ZPF42（REX01）表达更高（P<0.05）。在添加 SHH 的 IVC 和双重 IVM 和 IVC 处理中获得的囊胚中观察到最高的 ZPF42 表达。当 IVM 和/或 IVC 培养基中添加 SHH 时，二细胞期和四细胞期胚胎中的乙酰化组蛋白 3（AcH3K9/K14）水平增加（P<0.05）。我们的研究结果表明，SHH 对猪胚胎的植入前发育有有益的影响，特别是当 IVM 和 IVC 培养基均添加 SHH 时，这种影响可能从 IVM 延续到随后的胚胎发育。
