CAJ | 학술논문

生长素是促进动物生长发育所必需的激素之一.本研究探索牛生长激素(bovine growth hormone, bGH)对小鼠胚胎发育的影响.将牛生长激素基因连接到慢病毒载体(Lenti-CMV-EF1-eGFP)形成Lenti-CMV-bGH/E F1-eGFP,重组病毒经293T细胞包装后直接感染小鼠胚胎和G1胚胎干细胞(embryonic stem cells, ESCs).阳性表达的胚胎经体外培养,发育到囊胚期的胚胎进行胚胎移植,为直接感染组.感染的小鼠G1胚胎干细胞注射到正常昆明白小鼠囊胚制作嵌合胚胎后进行胚胎移植,为嵌合体组.分别解剖3只妊娠15 d的直接感染组和嵌合组的小鼠,观察受体小鼠妊娠情况.结果发现,在1-细胞期感染胚胎,感染效率可达(74.7依6.7)%,在2-细胞期连续感染,感染效率为(79.4依5.7)%,两组感染方法的效率没有显著差异(P>0.05).1次和2次感染胚胎发育和正常胚胎之间没有显著差异((87.6依3.5)%,(85.4依6.3)%VS (83.0依5.5)%, P>0.05).所有移植的胚胎没有发育到足月,解剖妊娠15 d的小鼠,发现直接感染组没有妊娠,嵌合组胎盘发育正常,但胚胎部分已经死亡和被吸收.因此,本研究证明bGH对小鼠附着前胚胎发育没有影响,但影响小鼠胚胎附着后的发育.这为后期bGH的研究提供参考.
생장소시촉진동물생장발육소필수적격소지일.본연구탐색우생장격소(bovine growth hormone, bGH)대소서배태발육적영향.장우생장격소기인련접도만병독재체(Lenti-CMV-EF1-eGFP)형성Lenti-CMV-bGH/E F1-eGFP,중조병독경293T세포포장후직접감염소서배태화G1배태간세포(embryonic stem cells, ESCs).양성표체적배태경체외배양,발육도낭배기적배태진행배태이식,위직접감염조.감염적소서G1배태간세포주사도정상곤명백소서낭배제작감합배태후진행배태이식,위감합체조.분별해부3지임신15 d적직접감염조화감합조적소서,관찰수체소서임신정황.결과발현,재1-세포기감염배태,감염효솔가체(74.7의6.7)%,재2-세포기련속감염,감염효솔위(79.4의5.7)%,량조감염방법적효솔몰유현저차이(P>0.05).1차화2차감염배태발육화정상배태지간몰유현저차이((87.6의3.5)%,(85.4의6.3)%VS (83.0의5.5)%, P>0.05).소유이식적배태몰유발육도족월,해부임신15 d적소서,발현직접감염조몰유임신,감합조태반발육정상,단배태부분이경사망화피흡수.인차,본연구증명bGH대소서부착전배태발육몰유영향,단영향소서배태부착후적발육.저위후기bGH적연구제공삼고.
Growth hormone (GH) is one of necessary hormones to promote growth and development of animals. In this study, the effects of bovine growth hormone (bGH) on mice embryonic development was investigated. bGH gene was linked to lentiviral vector Lenti-CMV-EF1-eGFP and get a recombinant vector Lenti-CMV-bGH/EF1-eGFP, then the reconstructed lentivirus was packaged by 293T cells. Mice embryos and G1 embryonic stem cells (ESCs) was infected with the reconstructed lentivirus. The positively infected embryos were cultured to blastocyst stage in vitro, then embryonic transfer was carried out, which was used as direct infection group;the positively in-fected ESCs were injected into normal Kunming white (KM) mice blastocysts to make chimerical embryos, then embryonic transfer was carried out, which was used as chimerical group. Three mice in direct infection group and three mice in chimerical group were anatomized after 15 days of embryonic transfer, and pregnant events were ob-served. The results showed that the infection efficiency was (74.7±6.7)%when the embryos were infected at 1-cell stage, and the infection efficiency was (79.4±5.7)%when the embryos were infected from 1-cell stage to 2-cell stage, there was no significant difference between these two groups (P>0.05), and the rate of blastocyst development of these two groups were not significantly different from that of normal embryos ((87.6±3.5)%and (85.4±6.3)%VS (83.0±5.5)%, P>0.05). There were no transferred embryos developing to term in both direct infection group and chimerical group. On 15 day of gestation, anatomy was performed in mice, no pregnant was found in direct infec-tion group. Although the placenta was normal in chimerical group, fetus was dead and absorbed. Therefore, the re-sults revealed that bGH does not impact the pre-implantation embryo development, but it could be harmful for post-implantation embryo development. The results can provide theoretical support and experimental basis for fur-ther research on bovine growth hormone.