猪胚胎附植部分因子研究进展

产仔数性状是养猪业中一个重要的经济性状,而胚胎附植对猪产仔数的高低有重要的影响。胚胎附植的调控涉及到多个生物学过程,在其中发挥作用的物质（附植因子）很多。作者针对孕酮、孕酮受体、雌二醇、雌激素受体α（ESR1）和促红细胞生成素产生肝细胞受体配体（Eph-ephrin）系统这几种附植因子的相关研究进行了归纳总结,重点阐述了这些因子在母胎对话过程中的时空表达、功能、突变、调控等方面的研究进展。其中,孕酮及其受体在猪胚胎附植活动中全程发挥着重要作用,黄体分泌的孕酮与母猪子宫内膜腔上皮、腺上皮、基质和肌细胞中的孕酮受体结合发挥作用,使这些组织细胞分泌多种附植因子,这些附植因子进一步参与胚胎附植过程。雌二醇是雌激素中含量最多、活性最强的一种激素,主要由卵巢颗粒细胞分泌,雌激素受体α是子宫内雌二醇的主要受体,二者结合可使母猪发情;此外,附植中处于游离状态的胚泡也分泌雌二醇,其是一个让母猪子宫内膜得以识别的信号,允许胚泡附植。Eph-ephrin系统作用广泛,其在人、小鼠和猪的胚胎附植过程中发挥着重要作用。系统中的EphA1、ephrin A1、EphA4等基因在猪的胚胎附植期表达量显著高于空怀猪,它们在子宫内膜附植点的表达量显著高于附植点间的部位,且ephrin A1的抑制表达会降低子宫内膜上皮细胞的迁移和黏附能力。这些附植因子都在猪胚胎附植活动中发挥着重要作用,对它们的深入研究将有利于明确猪胚胎附植调控机理,进一步揭示猪高繁机理。     

哺乳动物胚胎附植与中药保胎作用机制研究进展

附植是指哺乳动物囊胚滋养层细胞与母体子宫上皮细胞之间逐步建立组织生理上联系的过程。成功的附植有赖于胚泡与母体相互识别,胚泡发育与母体子宫变化的同步以及抑制母体的免疫排斥反应和母体接受性等一系列事件。这些事件又受到母体和胚泡的激素以及一些因子的调控。传统中药可通过提高孕酮的含量、类雌激素样作用、治疗子宫内膜炎的作用以及影响白血病抑制因子和白细胞介素1的表达量来促进山羊胚胎附植。文章主要论述了哺乳动物胚胎附植的机制以及中药对胚胎附植的影响,这有助于进一步阐明中药保胎的作用机理。     

白血病抑制因子及其受体与子宫内膜的关系

白血病抑制因子(LIF)是一种多生物活性的细胞因子,研究表明,LIF在人及动物的子宫内膜的腺上皮细胞、妊娠期蜕膜及胎盘上均有表达;胚泡是动物生殖过程中的关键,一些细胞因子起重要作用,它们能够充当子宫内膜微环境的"局部调节者",LIF对哺乳动物胚胎发育到胚泡阶段具有一定的作用,LIF在子宫内膜中特异性表达,促进胚胎生长发育和启动胚泡植入;LIF在生殖周期中影响子宫的功能及调节子宫内膜生长,这可能与LIF作用于子宫使其能发生蜕膜反应有关;LIF通过与LIF受体及gp130蛋白作用而发挥重要的生物学功能。LIF在哺乳动物早期妊娠中的作用机制具有重要意义。文章对LIF及其受体与子宫内膜的关系做了综述。     

Two concepts on the immunological aspect of blastocyst implantation

The process of blastocyst implantation is a series of interactions between the blastocyst and maternal tissues. The purpose of this process is (1) to provide nourishment to the embryo for developmental growth in appropriate physiological and endocrinological environment until a placenta is established, and (2) to protect the (semi-)allogeneic embryo from any attacks from the maternal immune system. To facilitate successful implantation, therefore, these two aspects of the embryonic demand must be satisfied in the embryo-maternal interface throughout the entire process of implantation. The first concept I present in this paper is that blastocyst implantation essential factors (BIEFs) have dual functions: one, for structural and functional modification of the endometrium to accommodate the developing embryo and provide nourishment and suitable environment for its development, and the other, for modulation, directly or indirectly, of the maternal immune system to prevent attacks by the maternal immune system. The second concept is that BIEFs convert the endometrium (or uterus) from an immunologically non-privileged site to a privileged site. This endometrial (uterine) conversion is the immunological aspect of the blastocyst implantation process. When the endometrium has become receptive for blastocyst implantation, it signifies that the immunological conversion of the endometrium by BIEFs has been sufficiently attained to let the embryo start contacting maternal tissues. During the early stages of placentation, as the trophoblast cells differentiate and make their way to the maternal blood vessels to establish the placenta, BIEFs continuously provide nourishment and immunological protection to the developing embryo. The immunological protection of the embryo/fetus from potential attacks by the maternal immune system appears to reach a peak at the time of establishment of the placenta. Thus, clarification of the roles of BIEFs in both the physiological/endocrinological aspect as well as the immunological aspect is essential for understanding the biological process of implantation.     

Kontaktaufnahme zwischen Trophoblast und Uterusepithel während der frühen Implantation beim Rind

Development of contact between trophoblast and uterine epithelium during the early stages of implantation in the cow Implantation in the cow may be divided into precontact, apposition, and adhesion phases. These can be recognized grossly at the site of nidation (18 or 19 days post inseminationem) by changes in the endometrium and in the embryonic chorion, which differs from the extraembryonic chorion. Also light and electronmicroscopical examinations of the trophoblast and uterine epithelium during the course of nidation show typical phase-correlated differences. The surfaces of trophoblastic and uterine epithelia increase and unite by interdigitiation of their microvilli. The functional relationship between blastocyst and uterus at the time of contact, as determined from their structure, is discussed.     

Analysis of integrins and vascular endothelial growth factor isoforms mRNA expression in the canine uterus during perimplantation period

Integrins are the major receptors within the extracellular matrix (ECM) that mediate several functions connected with cell life and metabolism, such as cell adhesion, migration, cytoskeletal organization, proliferation, survival, and differentiation. A vascular endothelial growth factor (VEGF) is one of the most important angiogenic factors. It has been suggested that the expression of this gene may play crucial physiological roles in reproductive organs. All investigated endometrial tissues were isolated on day 10-12 after mating. Control bitches, used in this study, were in metestrus, which was determined according to the vaginal cytology and progesterone level in blood. Early pregnancy was verified by flushing the uterine horns with PBS. Total RNA was isolated from the bitches endometrium by means of the Chomczyński and Sacchi method, treated by DNase I, and reverse-transcribed into cDNA. A quantitative analysis of integrins alpha2b, beta2 and beta3, VEGF 164, 182 and 188 cDNA was performed by RT-PCR. In results we have shown an increased expression of all investigated genes (integrins alpha2b, beta2 and beta3, VEGF 164, 182, and 188) in pregnant bitches uterus as compared to non-pregnant females (P < 0.001). Our results indicated that the expression of genes encoding integrins and vascular endothelial growth factors is different in relation to the time of the embryo implantation and it is increased in the first period of this process. This may be associated with the induction of specific mechanisms responsible for receptivity of uterus following the embryo attachment. In addition, all of investigated genes are up-regulated in a pregnancy-specific manner and the increased expression of these genes may regulate the uterus function during the implantation of canine embryos.     

MicroRNA调控胚胎植入研究进展

植入是胚胎与子宫内膜识别、接触并依附于子宫内膜上或进一步侵入、包埋于子宫内膜中的过程,是哺乳动物生殖生理的重要环节,这一过程对于胚胎能够在子宫持续的发育并最终获得成功的妊娠是必需的。成功的植入需要同步发育的正常胚胎、容受性的子宫内膜及母胎的和谐对话来实现。胚胎植入过程复杂而短暂,参与调控的因素有很多,包括microRNA。论文介绍了与植入相关microRNA的最新研究进展,包括microRNA对胚胎发育、子宫内膜的容受性等方面的作用,将为我们理解植入机制提供新的视角。     

Analysis of uteroplacental-specific molecules and their functions during implantation and placentation in the bovine

In cattle, the mechanisms underlying implantation and placental development are still unclear. Synepitheliochorial placentation in cattle is noninvasive, and thus generates limited interest in terms of degradation and remodeling of endometrial tissues. The overall purpose of this study was three-fold: (1) to examine the gene circuitry around the implantation window, (2) to understand development of the placenta during the peri-implantation period by using a uteroplacental cDNA microarray, and (3) to study the roles of molecules involved in endometrial remodeling. Bovine trophoblastic binucleate cell-specific molecules, such as pregnancy-associated glycoproteins (PAGs), placental lactogen (PL), and prolactin-related proteins (PRPs), were markedly expressed in binucleate cells (BNCs) around implantation. The expression of PRP-1 was specific to the caruncular (CAR) area of the gravid uterine horn. Gelatinases (MMP-2 and -9) in association with heparanase may be central to endometrial remodeling. In situ hybridization analyses of PAGs, PRPs, PL, and heparanase suggested that BNCs expressed these molecules simultaneously. Future studies will further investigate the specific roles of these molecules in placentogenesis. The uteroplacental cDNA microarray presented cascades of molecular signatures not only for the endometrium but also for the intricate dialogue at the level of the feto-maternal interface in cattle. Placentome morphogenesis potentially parallels the dynamic multigenic circuitry and regulates the cell cycle in the endometrium. The roles of BNCs and their secreted molecules remain an enigma, particularly with regard to the adhesion process and endometrial remodeling, which is the focus of this study.     

Uterine secretions from different endometrial classifications affect the viability of early murine embryos cultured in vitro

An interspecies (murine-equine) bioassay system was employed to investigate the effect of equine endometrial secretions on embryonic survival. Uterine fluid was obtained from 15 diestrous mares by lavage with 150 ml of sterile Whitten's medium. Samples were grouped according to their Kenney score for endometrial type. Samples were sterilized and protein concentration was standardized to 4 mg/ml protein with BSA. Controls consisted of either 4 mg/ml BSA in Whitten's medium or 50% BSA-50% heat-treated equine serum. One hundred 2-cell mouse embryos were cultured in each sample and controls. Embryonic development was evaluated for 5 days and was expressed as the percent reaching the 4-cell, 8-cell, morula, blastocyst, expanded blastocyst, and hatched blastocyst stages. Embryonic development in Type I mare uterine fluid did not differ from controls. Development to the 4 cell, 8 cell, morula, blastocyst and expanded blastocyst stages in the Type II mare media was less (p<.01) than the controls and other endometrial types. Development to all these stages was also lower than controls and Type I incubates in Type III mare media (p<.05), although development was higher in Type III compared with Type II. Leukocyte infiltration in the endometrium was assessed to determine the level of inflammation present in these mares. Basophil and neutrophil numbers were not different (p>.05) between endometrial types. Lymphocyte numbers were significantly different (p<.05) between each endometrial type. The quantity of lymphocyte infiltration was inversely proportional to the rate of in vitro embryo development in Type I, Type II and Type III endometria.     

Comparative analysis between endometrial proteomes of pregnant and non-pregnant ewes during the peri-implantation period

Background:Early pregnancy failure has a profound impact on both human reproductive health and animal production.2/3 pregnancy failures occur during the peri-implantation period;however,the underlying mechanism(s)remains unclear.Well-organized modification of the endometrium to a receptive state is critical to establish pregnancy.Aberrant endometrial modification during implantation is thought to be largely responsible for early pregnancy loss.Result:In this study,using well-managed recipient ewes that received embryo transfer as model,we compared the endometrial proteome between pregnant and non-pregnant ewes during implantation period.After embryo transfer,recipients were assigned as pregnant or non-pregnant ewes according to the presence or absence of an elongated conceptus at Day 17 of pregnancy.By comparing the endometrial proteomic profiles between pregnant and non-pregnant ewes,we identified 94 and 257 differentially expressed proteins(DEPs) in the endometrial caruncular and intercaruncular areas,respectively.Functional analysis showed that the DEPs were mainly associated with immune response,nutrient transport and utilization,as well as proteasome-mediated proteolysis.Conclusion:These analysis imply that dysfunction of these biological processes or pathways of DEP in the endometrium is highly associated with early pregnancy loss.In addition,many proteins that are essential for the establishment of pregnancy showed dysregulation in the endometrium of non-pregnant ewes.These proteins,as potential candidates,may contribute to early pregnancy loss.     

外泌体在哺乳动物胚胎附植中的作用研究进展

外泌体的分泌是不同类型的细胞间信息交流的一种重要方法,近几年发现外泌体在胚胎附植中介导胚胎与母体的信息交流起重要作用。本文综述了外泌体的主要释放机制和分离鉴定方法、子宫内膜细胞和胚胎来源的外泌体在哺乳动物中调节胚胎附植的重要作用,为提高动物胚胎附植的成功率提供新依据,也为人类体外受精、辅助生殖技术的改进提供参考资料。     

水貂胚胎滞育的影响因素及调节机制

水貂胚胎滞育是影响水貂养殖经济效益的一个重要因素,国内外学者对此进行了大量研究。作者对水貂胚胎着床的影响因素、调控机制及克服胚胎滞育、缩短妊娠期的方法进行了综述,并提出了水貂胚胎滞育研究存在的问题及进一步研究方向,为今后研究水貂胚胎着床机理及克服胚胎滞育方法提供科研思路。     

Expression and Hormonal Regulation of E‐Cadherin in Canine Uterus During Early Pregnancy

E‐cadherin, a Ca^{2 +} ‐dependent cell adhesion molecule, is necessary for endometrial receptivity to blastocyst implantation. The aim of this study was to investigate the differential expression of E‐cadherin in canine uterus during early pregnancy and its regulation under different conditions by in situ hybridization. E‐cadherin mRNA expression was at a low level in the glandular epithelium on days 6, 12 and 17 of pregnancy. On days 20 and 23 of pregnancy, E‐cadherin mRNA was highly expressed in the glandular epithelium surrounding the embryo, but not in the luminal epithelium and declined in villi and placenta on day 28 of pregnancy. During oestrous cycle, a moderate level of E‐cadherin mRNA expression was found in the luminal and glandular epithelium of canine uteri at oestrus stage. The same expression was also found at anoestrus stage. Progesterone slightly induced the expression of E‐cadherin mRNA in the luminal and glandular epithelium of ovariectomized canine uterus. These results suggest that E‐cadherin expression is closely related to canine implantation and can be up‐regulated by progesterone.     

Immunolocalization of angiogenic growth factors in the ovine uterus during the oestrus cycle and in response to Steroids

The vascular changes associated with endometrial maturation in preparation for embryo implantation depend on numerous growth factors, known to regulate key angiogenic events. Primarily, the vascular endothelial growth factor (VEGF) family promotes vascular growth, whilst the angiopoietins maintain blood vessel integrity. The aim was to analyse protein levels of VEGFA ligand and receptors, Angiopoietin‐1 and 2 (ANG1/2) and endothelial cell receptor tyrosine kinase (TIE‐2) in the ovine endometrium in the follicular and luteal phases of the oestrus cycle and in response to ovarian steroids. VEGFA and its receptors were localized in both vascular cells and non‐vascular epithelium (glandular and luminal epithelium) and stroma cells. VEGFA and VEGFR2 proteins were elevated in vascular cells in follicular phase endometrium, compared to luteal phase, most significantly in response to oestradiol. VEGFR1 was expressed by epithelial cells and endothelial cells and was stimulated in response to oestradiol. In contrast, Ang‐1 and Ang‐2 proteins were elevated in luteal phase endometrium compared to follicular phase, and in response to progesterone, evident in vascular smooth muscle cells and glands which surround TIE‐2‐expressing blood vessels. Our findings indicate that VEGFA is stimulated by oestradiol, most predominantly in follicular phase endometrium, and Ang‐1 and 2 are stimulated by progesterone and were increased during the luteal phase of the oestrus cycle, during the time of vascular maturation.