牦牛妊娠期卵巢、卵泡及黄体的形态学观测

对87头妊娠母牦牛卵巢及其表面的卵泡、黄体的形态学观测发现,牦牛妊娠后孕侧卵巢大小、质量显著增加(P<0.05),而对侧卵巢则无明显变化(P>0.05).妊娠4个月以后,卵巢逐渐变扁长,以孕侧更为明显.牦牛在妊娠期,卵巢上仍然有规律地出现非排卵卵泡波,但卵泡数比未孕状态下少.在卵巢表面,主要以Ⅰ类卵泡(直径≤5 mm)为主,为3~6个.在孕侧随妊娠的进行Ⅰ类卵泡数呈递增趋势,妊娠1个月为(2.3±1.8)个,妊娠6个月达到(6.6±5.2)个,而空侧数量变化不明显(P>0.05),整个妊娠期其卵泡数保持在5~7个之间.Ⅱ类(5 mm<直径≤10 mm)和Ⅲ类(直径>10 mm)卵泡数很少,妊娠2个月平均数分别为(0.7±0.5)个和(0.3±0.1)个.牦牛妊娠黄体一半以上不突出于卵巢表面,且黄体直径在整个妊娠期基本不变.     

干扰素对附植前后哺乳动物妊娠识别与维持的作用

哺乳动物胚泡附植过程中能产生一种特异的妊娠相关蛋白——干扰素,它对妊娠的识别与维持十分重要。综述了绵羊、牛、猪、红鹿早期胚胎发育期间所产生的干扰素及其作用。     

论犬子宫破裂

犬的子宫体部或子宫角在妊娠期、分娩期或子宫发生异常病变时,发生破裂称为子宫破裂。子宫破裂为生殖系统最严重并发症之一,常引起母体和胎儿的死亡。     

TETs与细胞程序性死亡相关基因在山羊妊娠早期输卵管及子宫角的表达

【目的】TET（ten-eleven translocation）家族蛋白在调控胚胎和胎盘发育等过程中具有重要作用,输卵管与子宫内膜细胞的死亡平衡会影响受精卵及早期胚胎的发育。因此探讨TETs、细胞程序性死亡相关基因和附植相关基因在妊娠早期山羊输卵管及子宫角中的表达规律及其潜在调控作用,以期为研究胚胎发育及附植的分子机制奠定基础。【方法】以合川白山羊为研究对象,采集妊娠19d（P19）山羊子宫角及输卵管组织,以空配19d（C19）为对照,利用H.E染色观察C19组和P19组山羊子宫角组织形态,利用qRT-PCR技术检测了胚胎附植相关基因（WNT5a、OPN、VEGFA）、TETs（TET1、TET2、TET3）、细胞程序性死亡相关基因（凋亡相关基因（BAX、BCL2、Caspase9）、焦亡相关基因（GSDMD、NLRP3、Caspase1）和炎症相关基因（NF-kB、TNF-α））的相对表达,并分析了TETs及细胞程序性死亡相关基因在输卵管和子宫角组织的表达相关性。【结果】H.E染色结果显示,在P19组中山羊子宫角的腺体增多,形态多变。qRT-PCR结果显示,OPN在P19组山羊子宫角中的表达量极显著高于C19组（P＜0.01）,BAX/BCL2的表达量显著增加（P＜0.05）,而WNT5a和 VEGFA表达差异不显著;TET1、TET2、TET3在山羊子宫角及输卵管中均有表达,在C19组和P19组山羊子宫角中TET1及TET2的表达量极显著高于C19组和P19组中TET3的表达（P＜0.01）;在P19组山羊输卵管中TET2（P＜0.01）、NF-kB（P＜0.01）、Caspase1（P＜0.01）、GSDMD（P＜0.05）显著增高。相关性分析表明,输卵管中TET2与TET3（P＜0.01）、NF-kB（P＜0.01）、Caspase9（P＜0.05）显著正相关;子宫角中,TET3与VEGFA（P＜0.05）、WNT5a（P＜0.01）显著负相关,胚胎附植关键基因OPN与Caspase1（P＜0.01）、NLRP3（P＜0.05）、GSDMD（P＜0.05）显著正相关,而细胞焦亡关键基因GSDMD在输卵管与子宫角中均与Bax（P＜0.05）、Caspase1（P＜0.01）、NLRP3（P＜0.05）呈显著正相关。【结论】在妊娠早期山羊子宫角和输卵管中TETs表达与细胞程序性死亡部分基因表达具有一定的相关性,推测TETs与细胞凋亡或焦亡的发生对输卵管、受精卵、早期胚胎及子宫的发育具有一定的调控作用。胚胎附植关键基因OPN与焦亡相关基因显著相关,暗示妊娠早期子宫角中细胞焦亡的发生可能参与胚胎附植。为研究TETs通过细胞程序性死亡相关基因在妊娠早期输卵管及子宫角中的表达规律提供了理论参考。     

采胚方法对波尔山羊胚胎移植效果的影响

采用2种采胚方法研究了64只波尔山羊的胚胎移植效果。结果表明:输卵管采胚法的胚胎回收率极显著高于(P<0.01)子宫角采胚法,且前者的获胚数与可用胚数均显著高于(P<0.05)后者,而两者的黄体数则差异不显著(P>0.05),说明采用输卵管采胚法所采集的胚胎数量与质量均优于子宫角采胚法;而子宫角移胚法的受体妊娠率及产羔率均极显著高于(P<0.01)输卵管移胚法。试验还表明,输卵管采胚法对处理供体术后的发情率有一定的不良影响,故在山羊胚胎移植中应优先采用子宫角采胚法回收胚胎。     

体细胞核移植法获得转人乳铁蛋白基因克隆山羊妊娠的研究

[目的]探讨供体细胞类型、移植胚胎发育阶段、数量及部位对山羊转基因克隆效率的影响。[方法]利用体细胞核移植技术将转染人乳铁蛋白基因hLF的山羊胎儿成纤维细胞(GFF)和乳腺上皮细胞(GMGE)移植到MII期去核卵母细胞内,经电融合、激活、体外培养后,2~8细胞期克隆胚被移植到同期发情山羊的输卵管内,囊胚被移植到子宫角内。[结果]GFF与GMGE的妊娠率相近(输卵管移植妊娠率分别为26.47%及20.00%);在GFF,输卵管移植的妊娠率与子宫角内移植妊娠率接近(分别为26.47%及25.00%),输卵管移植胚胎平均数为21.2组的妊娠率显著高于5.93组和9.64组(40.00%及26.67%,21.43%)。[结论]供体细胞类型、移植胚胎的发育阶段及移植部位对山羊转基因克隆效率的影响不大,但对于输卵管移植,受体羊移植胚胎数量对妊娠率有明显的影响。此外,该研究还提示了利用成年羊乳腺上皮细胞制作转基因动物的可行性。     

体细胞核移植法获得转人乳铁蛋白基因克隆山羊妊娠的研究(英文)

[Objective] The aim of this study is to understand the effects of donor cell type,embryo stage,number and transfer position on the efficiency of goat transgenic clone.[Method] Using somatic cell nuclear transfer technology,the single goat fetal fibroblasts(GFF)and mammary gland epithelial cells(GMGE)harboring human lactoferrin(hLF)gene were transferred to the enucleated oocyte.Reconstructed karyoplast-cytoplast couplets were fused,activated,and cultured in vitro.Embryos at 2-8 cell stage were transferred into oviduct of synchronized recipients,and blastocysts were transferred into uterine horn.[Result] The pregnancy rate was similar between GFF and GMGE(oviduct transfer:26.47% vs.20.00%),and between oviduct transfer and uterine horn transfer(26.47% vs.25.00%)for GFF group;pregnancy rate in the group with the mean number of embryo transferred per recipient of 21.2 was significantly higher than in those the 5.93 group and 9.64 group(40.00% vs.26.67% and 21.43%).[Conclusion] These results indicate that pregnancy rate of goat transgenic clone couldn't be affected by donor cell type,embryo stage and transfer position but be done by the number of embryo transferred per recipient.In addition,the study also suggests the feasibility of making transgenic goat using GMGE as donor cells.     

Pregnancies Resulting from Transgenic Embryos with Human Lactoferrin Gene Produced by Somatic Cell Nuclear Transfer

[Objective] The aim of this study is to understand the effects of donor cell type,embryo stage,number and transfer position on the efficiency of goat transgenic clone.[Method] Using somatic cell nuclear transfer technology,the single goat fetal fibroblasts(GFF)and mammary gland epithelial cells(GMGE)harboring human lactoferrin(hLF)gene were transferred to the enucleated oocyte.Reconstructed karyoplast-cytoplast couplets were fused,activated,and cultured in vitro.Embryos at 2-8 cell stage were transferred into oviduct of synchronized recipients,and blastocysts were transferred into uterine horn.[Result] The pregnancy rate was similar between GFF and GMGE(oviduct transfer:26.47% vs.20.00%),and between oviduct transfer and uterine horn transfer(26.47% vs.25.00%)for GFF group;pregnancy rate in the group with the mean number of embryo transferred per recipient of 21.2 was significantly higher than in those the 5.93 group and 9.64 group(40.00% vs.26.67% and 21.43%).[Conclusion] These results indicate that pregnancy rate of goat transgenic clone couldn't be affected by donor cell type,embryo stage and transfer position but be done by the number of embryo transferred per recipient.In addition,the study also suggests the feasibility of making transgenic goat using GMGE as donor cells.     

体细胞核移植法获得转人乳铁蛋白基因克隆山羊妊娠的研究(英文)

[Objective] The aim of this study is to understand the effects of donor cell type,embryo stage,number and transfer position on the efficiency of goat transgenic clone.[Method] Using somatic cell nuclear transfer technology,the single goat fetal fibroblasts(GFF)and mammary gland epithelial cells(GMGE)harboring human lactoferrin(hLF)gene were transferred to the enucleated oocyte.Reconstructed karyoplast-cytoplast couplets were fused,activated,and cultured in vitro.Embryos at 2-8 cell stage were transferred into oviduct of synchronized recipients,and blastocysts were transferred into uterine horn.[Result] The pregnancy rate was similar between GFF and GMGE(oviduct transfer:26.47% vs.20.00%),and between oviduct transfer and uterine horn transfer(26.47% vs.25.00%)for GFF group;pregnancy rate in the group with the mean number of embryo transferred per recipient of 21.2 was significantly higher than in those the 5.93 group and 9.64 group(40.00% vs.26.67% and 21.43%).[Conclusion] These results indicate that pregnancy rate of goat transgenic clone couldn’t be affected by donor cell type,embryo stage and transfer position but be done by the number of embryo transferred per recipient.In addition,the study also suggests the feasibility of making transgenic goat using GMGE as donor cells.     

胚胎着床期间母-胎对话研究进展

哺乳动物胚胎着床时,母体和胚胎之间进行精密的分子对话和信号交换,以使两者达到协调。母体和胚胎来源的类固醇激素、生长因子、细胞因子、整合素等在胚胎着床及母体妊娠识别过程中发挥着重要的作用,使母体-胎儿间形成一个紧密联系的对话体系。综述了胚胎着床过程中母-胎对话的研究进展。     

ICR小鼠着床障碍模型的建立

为建立ICR小鼠胚泡着床障碍模型,本研究用米非司酮和吲哚美辛对妊娠ICR雌鼠进行抗着床处理,观察其对胚泡着床及子宫内膜的影响。实验分为米非司酮组和吲哚美辛组,每组设对照组。于妊娠第4天分别注射0.3、0.6、0.9、1.2 g.L-1的米非司酮0.1 mL.mouse-1,及0.12、0.13、0.14、0.15 mL吲哚美辛溶液,对照组注射二者相应的溶剂丙二醇和麻油。妊娠第5天检查子宫内膜对台盼蓝的反应点数;妊娠第8天观察胚泡着床情况,统计子宫重量,制作切片,检测子宫内膜形态结构的变化。结果表明,0.09 mg米非司酮组和0.13 mL吲哚美辛组的子宫重量、内膜容受性、发育及蜕膜化均受到明显的抑制。结果表明,米非司酮和吲哚美辛可成功建立ICR着床障碍模型,用于相关研究。     

热应激小鼠胚胎着床模型的建立

模拟夏季高温环境对孕鼠的应激作用,建立热应激影响昆明白小鼠胚胎着床的模型。试验设为对照组(23℃±2℃)、40℃和42℃应激组(小鼠妊娠1～4d,分别应激2h),比较观察妊娠5、6、7d小鼠胚胎着床数及子宫状况和产仔数。结果表明,与对照组相比40℃和42℃应激组妊娠5d(11.00±2.65、10.00±2.12vs15.00±1.58,P0.05)、6d(11.40±3.21、10.60±2.30vs 15.00±0.71,P0.05)、7d(11.00±2.55、11.00±1.58vs 15.60±1.67,P0.05)胚胎着床率显著下降,产仔数(8.60±1.14、8.40±1.82vs 11.40±1.82,P0.05)也显著下降;且相比对照组其着床位点均较少且分布失衡。40℃和42℃应激组之间无显著差异,以42℃应激组影响相对较大。可见,胚胎着床前孕鼠连续4d40℃、42℃应激2h,可显著影响小鼠的繁殖能力。42℃可作为影响小鼠胚胎着床的热应激模型。     

牛胚胎分割及半胚移植研究

对来自黑白花及西门塔尔超排母牛的７～８日龄胚胎，采用手持微玻璃针，在普通解剖镜下进行分割。通过非手术法将裸半胚移入发情后７天的受体牛黄体侧子宫角。在太原市郊奶牛场及太行山区农民户养牛中，共移植受体牛４５头，结果妊娠产犊２５头，移植成功率为５５．６％。     

The effect of the floor plate on pattern and polarity in the developing central nervous system

The effect of floor plate on cellular differentiation in the neural tube of quail embryos was examined. In the developing neural tube the floor plate, which consists of specialized neuroepithelial cells, is located in the ventral midline of the neural tube. When Hensen's node was extirpated the floor plate and notochord did not develop, and the normal differentiation of the ventral horn motor neurons and dorsal and ventral roots did not occur. When one side of the neural tube was deprived of notochord, the ventro-dorsal differentiation took place on both sides. However, when one side of the neural tube was deprived of the floor plate, the ventral horn motor neurons and dorsal and ventral roots did not develop on that side. These observations suggest that the floor plate influences motor neuron differentiation and acts as an intrinsic organizer to establish pattern and polarity in the developing nervous system.     

Expression of Eph-Ephrin A Molecules in Endometrium During Swine Embryo Implantation Examined Using Real-Time RT-PCR

Erythropoietin-producing hepatocellular receptor and its membrane-bound ligands (Eph-Ephrin) system could regulate some mammalian blastocyst attachment and spreading. In order to investigate the involvement of the Eph-Ephrin system in swine embryo attachment, mRNA expression of Eph-Ephrin molecules in endometrium was examined by real-time RT-PCR during embryo implantation in pigs. The results indicated that mRNA expressions of Eph A5, A7 and Ephrin A5 all continually increased from pregnancy day 13 to 24. Ephrin A3 mRNA expression significantly increased from day 13 to 18 and decreased from day 18 to 24, and the expression was the lowest on pregnancy day 13 and the highest on day 18. However, Ephrin A4 mRNA expression was the lowest on pregnancy day 18 and the highest on day 24, and the expression decreased from day 13 to 18 and increased from day 18 to 24. Furthermore, mRNA expressions of Eph A5 and A7 were both found in other tissues, such as brain, muscle, intestine, stomach, etc. These findings suggest that the Eph-Ephrin system may play an important role in regulating the contact between blastocysts and endometrium during swine embryo implantation.     

反刍动物的胚胎着床与妊娠识别过程的研究进展

反刍动物的胚胎着床与妊娠识别是个复杂的生理过程，这一过程受多种细胞因子和生殖激素调控。文中叙述了各种细胞因子和生殖激素对妊娠建立和维持的调控作用。     

Copulatory behavior can inhibit pregnancy in female rats

If female rats received genital stimulation soon enough after their male partners had ejaculated, sperm transport and subsequent pregnancy were inhibited. Manual stimulation by the experimenter or five intromissions by a male rat were sufficient stimuli to reduce the number of sperm found in the uterus and to reduce the number of uterine implantation sites.     

附植前后兔子宫和胚胎总蛋白含量的变化

附植前后哺乳动物子宫和胚胎能分泌多种妊娠相关蛋白，这些蛋白对妊娠的建立了维持十分重要，为了从生化角度进一步研究胚泡附植的机理，从分析附植前后兔子宫和胚胎总蛋白含量着手，探讨了非妊娠和受孕不同时期的子宫和胚胎总蛋白含量的变化，结果表明，非妊娠兔比妊娠兔子宫冲洗液总蛋白含量低，妊娠组中，胚泡附植前后随着妊娠时间的延长，子宫冲洗兴中蛋白质量逐渐增加，妊娠前4d，增加幅度不大，从第4d到第6d增加幅度增大，从妊娠第6d到第9d，蛋白质增加特别明显，对于妊娠第9d胚泡液，大胚泡（直径1cm左右）中胚泡液总蛋白含量高于小胚泡（直径0．5cm左右），说明囊胚直径可以作判断胚胎发育的一个比较胡切的指标。     

水牛胚胎附植过程中瘦素及其受体在子宫内膜中的表达

[目的]探讨瘦素及其受体ob-Rb在水牛子宫内膜中的表达规律及其对水牛胚胎附植的影响。[方法]母水牛经孕马血清促性腺激素(PMSG)超排后与公牛交配,分别在交配后第0、5、7、9和15天各屠宰10头水牛,分别取胚胎附植部位的子宫组织,提取瘦素及其受体总RNA。利用RT-PCR方法检测不同时期瘦素mRNA及其受体表达情况。[结果]瘦素在胚胎附植过程中在水牛子宫内膜都有表达,各时期的表达水平差异不明显(P>0.05),Ob-Rb在胚胎附植过程中亦有表达,但是交配后第7、9和15天的表达水平与0和5天差异显著(P<0.05),并且在妊娠期0～7天内表达水平上升,7天达到峰值,然后下降。[结论]瘦素及其受体Ob-Rb可能不仅参与胚胎的附植,而且也可能参与附植后胚胎的生长发育。