猪妊娠过程中胎盘发育及其调控基因研究进展

胎盘是母体与胎儿进行营养物质、气体及废弃物交换的重要器官。妊娠过程中,猪胎盘功能是影响母猪产仔数、死胎数及断奶前死亡率的最重要因素之一。作者介绍了猪妊娠早期胎盘的建立过程及形态变化、妊娠中期胎盘褶皱的形成、妊娠后期胎盘的进一步发育,阐述了这3个妊娠阶段猪胎盘的形态变化与其对应的胎盘功能之间的关系,并介绍了目前所发现的调控猪胎盘建立和发育的相关基因,包括透明质酸酶(HYAL)、组织蛋白酶(CTSB和CTSL1)及乙酰肝素酶(HPSE)基因,为提高母猪胎盘效率、增加母猪繁殖力提供科学依据。     

氨基葡萄糖调控仔猪初生重和均匀度的应用及机理

养猪生产中产（活）仔数和胎儿初生重是影响母猪繁殖生产效率的主要因素,但产仔数和初生重呈负相关关系,近年来母猪繁殖生产的研究重点之一便是如何提高仔猪初生重并降低初生重变异。氨基葡萄糖是一种饲料原料,其制作原料广泛存在于自然界中,近期研究发现氨基葡萄糖应用于母猪妊娠后期可调控仔猪的胎盘发育和初生重。文章介绍了近年来有关氨基酸和蛋白质、能量、功能性添加剂在调控仔猪初生重和均匀度中的应用及其效果,简述了氨基葡萄糖的性质、来源及合成方法,重点阐述了母猪妊娠后期使用氨基葡萄糖在维持胎儿果糖浓度稳定、促进母猪胎盘发育和改善胎盘功能等方面的效果,分析了氨基葡萄糖通过促进胎盘基质和胎褶双分子层发育、刺激胎盘滋养层细胞增殖、增强胎盘功能等途径调控胎盘发育、仔猪初生重和均匀度可能的机理;提出了氨基葡萄糖在调控仔猪初生重和均匀度方面未来的研究方向,以期为在生产实践中调控仔猪初生重和均匀度提供理论参考和实践依据。     

猪胎盘特异基因1(PLAC1)的克隆、组织表达及遗传方式研究

旨在研究猪胎盘特异性基因PLAC1的表达规律,并对其基因结构、生物功能和遗传方式进行初步鉴定。本研究以妊娠26、50和95天的猪胎盘组织为材料,利用RACE-PCR及RT-PCR技术克隆猪PLAC1基因及其不同转录本的全长序列,同时通过原位杂交验证其在胎盘中的表达模式。并检测猪PLAC1基因的序列变异。结果表明,猪PLAC1基因存在3种不同的转录本,3种转录本的CDS区完全相同,长525bp,编码174个氨基酸;组织表达谱和原位杂交结果显示,PLAC1基因特异性表达于猪胎盘绒毛膜上皮细胞中,并在不同妊娠时期差异表达,妊娠50和95天PLAC1mRNA的表达显著高于妊娠26天(P<0.01)。猪群中PLAC1基因的SNP基因分型检测结果表明该基因遵循半合基因的遗传方式。本试验结果为研究猪PLAC1基因在胎盘发育过程中的生物功能奠定了基础。     

微小RNA调控母猪胎盘发育的研究进展

微小RNA(miRNA)是内源性非编码的小分子RNA,它们存在于动物、植物和病毒中，主要通过调控靶基因的翻译和mRNA稳定性而参与多种生理过程。胎盘是哺乳动物妊娠过程中存在的一个临时性的内分泌器官，对胎儿的发育至关重要。而母猪胎盘的不良发育不仅会影响到妊娠期仔猪的发育，还会对生产效益造成一定的损失。研究表明，miRNA对哺乳动物的胎盘滋养层细胞和血管生成等方面具有重要的调控作用。本文综述了miRNA生成过程及其功能，以及目前关于miRNA对母猪胎盘发育的调控作用，旨在为提高母猪繁殖效率、提高生产效益提供理论参考。     

母猪妊娠后期日粮能量、蛋白质及氨基酸水平对乳腺发育及胎猪生长的影响

母猪妊娠后期是母猪乳腺及胎猪快速发育的重要时期,为处于这一时期的母猪配制营养水平适宜的日粮对提高母猪繁殖效率尤为重要。妊娠后期母猪乳腺发育及胎猪的生长对能量、蛋白质及氨基酸的需求不同,如何给妊娠母猪提供合理的营养,在满足胎猪生长发育的同时满足乳腺发育的需要研究还比较少。作者主要介绍了母猪妊娠后期日粮中的能量、蛋白质及氨基酸水平对乳腺发育及胎猪生长的影响,同时提出母猪妊娠后期能量、蛋白质及氨基酸的需要量及合理比例,以期为养猪生产者提供参考。     

猪胎盘屏障的研究进展

妊娠是一个微妙而复杂的生理过程,胎盘的建立为母胎之间的物质交换奠定了基础。对于猪来说,胎盘功能障碍会降低胎盘效率,引起猪宫内发育迟缓、窝内体重变异增大,给养猪业的经济效益带来巨大损失。胎盘是母胎之间的一道天然屏障,其功能较为复杂,有物质交换、排泄、免疫保护、合成和分泌等多种功能。鉴于胎盘屏障的重要作用,本文综述了猪胎盘屏障的结构、功能及影响因素,以期为相关研究和养猪生产提供参考。     

大足黑山羊妊娠不同阶段胎盘组织线粒体酶活性和超微结构变化

山羊的胎盘属于子叶型胎盘,胎盘子叶性状和结构与山羊繁殖性能具有较强的相关性。胎盘子叶组织线粒体发挥正常功能是保证胎儿生长发育,进而维持正常妊娠的关键。本试验采用大足黑山羊妊娠60,90,120d的胎盘和胎儿组织为研究对象,检测胎盘子叶及胎儿心、肝、脾、肺、肾组织的线粒体细胞色素C氧化酶(CCO)、琥珀酸脱氢酶(SDH)、Ca++-Mg++-ATP酶和Na+-K+-ATP酶的活性变化,并通过透射电镜技术观测胎盘子叶组织线粒体的形态结构。结果发现,胎盘子叶及胎儿心、肝、脾、肺、肾的线粒体CCO、SDH及ATP酶的活性都有随着妊娠时间的增加而显著提高的趋势,且在妊娠90~120d的上升趋势高于妊娠60~90d,表明妊娠中后期的胎儿生长发育所需要的能量较多;在同一妊娠阶段,胎盘子叶线粒体酶活性均高于胎儿各组织,且差异显著(P<0.05),说明胎盘子叶是妊娠过程中的重要部位,胎儿组织器官在发育过程中存在先后顺序,并且与该器官的功能作用有关;透射电镜观察发现,随着妊娠时间的增加胎盘线粒体数量增多,密度增大且主要聚集在子叶绒毛附近,这体现了胎盘子叶的营养交换作用。     

氨基葡萄糖调控仔猪初生重和均匀度的应用及机理

养猪生产中产(活)仔数和胎儿初生重是影响母猪繁殖生产效率的主要因素,但产仔数和初生重呈负相关关系,近年来母猪繁殖生产的研究重点之一便是如何提高仔猪初生重并降低初生重变异。氨基葡萄糖是一种饲料原料,其制作原料广泛存在于自然界中,近期研究发现氨基葡萄糖应用于母猪妊娠后期可调控仔猪的胎盘发育和初生重。文章介绍了近年来有关氨基酸和蛋白质、能量、功能性添加剂在调控仔猪初生重和均匀度中的应用及其效果,简述了氨基葡萄糖的性质、来源及合成方法,重点阐述了母猪妊娠后期使用氨基葡萄糖在维持胎儿果糖浓度稳定、促进母猪胎盘发育和改善胎盘功能等方面的效果,分析了氨基葡萄糖通过促进胎盘基质和胎褶双分子层发育、刺激胎盘滋养层细胞增殖、增强胎盘功能等途径调控胎盘发育、仔猪初生重和均匀度可能的机理;提出了氨基葡萄糖在调控仔猪初生重和均匀度方面未来的研究方向,以期为在生产实践中调控仔猪初生重和均匀度提供理论参考和实践依据。     

母猪胎盘发育的营养调控北大核心CSCD

胎盘发育是一个复杂且精细的过程,胎盘发育及其功能的完整性是胎儿正常生长发育的首要保障。母猪胎盘功能异常将会导致母胎间养分运输的滞后,从而引发宫内发育迟缓现象。本文综述了几种主要营养物质(氨基酸、脂肪酸、维生素和矿物质)对母猪胎盘发育和功能的影响及机制,为充分发挥母猪繁殖潜能提供营养策略。     

应用荧光定量PCR方法研究RPL29基因在通城猪和长白猪不同时期胚胎骨骼肌中的表达

为了研究蛋白合成相关基因RPL29在骨骼肌发育中的作用,采用SYBRGreen染料建立该基因的实时荧光定量PCR（QPCR）分析方法,并分析该基因在猪胚胎骨骼肌中的表达变化。以不同浓度稀释样本为标准品,建立标准曲线方程：y=-3.303X＋43．077（决定系数R^2=0．999）。分析表明：该方法具有较大的检验范围（PCR扩增Q值13～35）,很好的扩增效率为100％,熔解曲线显示产物特异的单峰,其Tm值为86．5℃。本研究以H3F3A基因为内参,用所建立的实时定量PCR方法,分析RPL29在妊娠33、65和90d的通城猪和长白猪胚胎骨骼肌中的表达变化。研究发现：RPL29基因在通城猪和长白猪胚胎骨骼肌发育过程中分别呈现下调和上调表达模式;妊娠33d时,通城猪中的表达水平高于长白,而妊娠90d时在长白猪中高表达;妊娠65d时两品种间无显著差异。结果显示RPL29基因可能与猪骨骼肌的生长发育有关,并可能影响产肉性状。     

Morphometric analysis of Hofbauer cells in normal placenta and chorioamnionitis in humans

Hofbauer cells are macrophages residing in the stroma of placental villi and play a number of roles during normal pregnancy, as well as pathological conditions. A morphometric analysis of Hofbauer cells, in particular to investigate the number of cells, their size and shape in samples of normal human placenta from 1st trimester, term and with chorioamnionitis was performed. Tissue samples were immunostained for CD206 antigen and evaluated using ImageJ software. We detected significant changes in number and morphology of HBCs between normal placenta and placenta with chorioamnionitis samples. In chorioamnionitis, the cells were unevenly distributed within the villi, generally present in higher numbers, larger and more elongated than those in normal 1st trimester and term placenta.     

Differences in placental structure during gestation associated with large and small pig fetuses

The efficiency of nutrient transport from the pregnant female pig to the developing fetus depends on the size and function of the placenta. It has been reported that maternal and fetal blood vessels are arranged in a cross-countercurrent arrangement within placental microscopic folds. Thus, the blood supplies are in close apposition to each other within these microscopic folds, and maternal and fetal blood flows in approximately opposite directions perpendicular to the plane of the placenta. This arrangement indicates that the width of the microscopic folds influences placental efficiency. The objective of this study was to determine whether differences in pig placental microscopic fold development are associated with differences in fetal size or are influenced by selection for ovulation rate or uterine capacity. Gilts from a randomly selected control line, a line selected for ovulation rate, and a line selected for uterine capacity were slaughtered, and uterine wall samples were collected within the placentas associated with the largest and smallest fetuses in each litter on d 45, 65, 85, and 105 of gestation. The uterine wall samples were processed for histology and analyzed using computer-assisted morphometry. Average width of the placental folds and average width of the placental stroma above the folds were measured. To measure fold complexity, the length of the epithelial bilayer for a given length of placenta was also measured. The width of the folded bilayer increased significantly from d 65 to 105 and was greater in placentas associated with small fetuses compared with large fetuses on d 105 of gestation. In contrast, the width of the placental stroma above the folded bilayer decreased with gestation and decreased more rapidly in placenta associated with the smallest compared with the largest fetus. These results indicate that the width of the microscopic folds of the placental trophoblast/endometrial epithelial bilayer is increased in placenta associated with small fetuses, which we hypothesize will increase the surface area for interaction between maternal and fetal blood supplies, thus improving placental efficiency in response to reduced placental size.     

The Ultrastructure of the Trophoblast Fossal Regions in the Pig Placenta During Pregnancy: Its Steroidogenic Features*

The fine structural changes of the trophoblast covering the fossae of the porcine placenta between the 21st day of pregnancy and term are described. Considering the characteristic features of cells, six morphological stages in the development of the fetal placental fossae were distinguished. Three types of trophoblast cells were recognized: light, dark and intermediates. The light cells were characterized by whorls of strongly convoluted SER channels, dense granules were sometimes associated with these whorls and by lipid droplets. They were particularly numerous in the second month and the last week of pregnancy. With the ultrastructural investigation, ultracytochemical and histochemical methods were used for the detection and localization of 3 β-hydroxysterols and 3 β-hydroxysteroid dehydrogenase activity. The localization of 3 β-hydroxysterols was studied by the digitonin reaction adapted to electron microscopy by Ökrös (1968). The digitonin 3 β-hydroxysteroid complexes were most numerous during the second month and last week of pregnancy. They were located mainly in contact with granules in the region of the SER whorls and in the matrix of mitochondria. The 3 β-hydroxysteroid dehydrogenase activity was investigated with the use of dehydroepiandrosterone and dehydroisoandrosterone as substrates. The enzyme activity in the trophoblast determined with dehydroepiandrosterone increased in the 52nd and 106th day of pregnancy. The cells of the fossae showed higher activity than that observed on the sides and tops of the folds. Lower activity was exhibited by dehydroepiandrosterone. The finding suggest that the trophoblast of the fossae is a region where synthesis of steroid hormones takes place and that the light cells are involved in this process. The results correspond to biochemical findings regarding progesterone and estradiol concentrations in the maternal and fetal circulation and estrogen distribution in fetal organs and placenta.     

山羊妊娠早期胎儿胎盘血管形成及相关基因的表达

旨在探究大足黑山羊妊娠早期胎儿胎盘血管发育、分布和血管生成相关因子表达状况,并对血管生成相关因子表达之间和血管发育进行相关性分析。本研究随机选取15只8月龄、体格相近、身体健康的国家级畜禽遗传资源—大足黑山羊青年母羊,经同一种公羊自然交配后(以末次配种当天记为0 d),经剖腹产手术采集妊娠第20、25和30天的胎儿和胎儿胎盘以及经屠宰采集妊娠第45和60天的胎儿和胎儿胎盘作为研究对象,通过免疫组化法评估胎盘血管发育和分布情况,采用qRT-PCR技术检测主要血管生成相关基因的mRNA表达水平,分析血管生成相关基因之间以及与胎盘血管发育分布的相关性。结果显示,胎儿体重和体长随着妊娠的进行逐渐增加,且血管内皮细胞阳性率处于较高水平(>26%)。山羊胎儿胎盘毛细血管总面积/组织区域面积(CAD)逐渐升高;毛细血管总数/单位组织区域面积(CND)在妊娠25~30 d增加,30~60 d降低,毛细血管总面积/毛细血管根数(APC)呈相反趋势;毛细血管总周长/单位组织区域面积(CSD)无显著变化。妊娠早期胎儿胎盘VEGFA的表达量随着妊娠的进行逐渐升高,且与血管分布存在一定的相关性,其受体FLT1和KDR表达量呈先增高后降低的趋势,分别在30和45 d达到峰值。血管生成相关基因ANGPT1/2及其受体TEK和FGF2及其受体FGFR2的表达均在妊娠30 d时较高。综上表明,在妊娠30 d前,胎儿胎盘主要通过毛细血管形成分支促进血管面积的增加;而在30~60 d,主要通过单位血管的增粗促进血管面积的增加,并且血管生成相关基因的表达在妊娠30 d时较高,这可能和子叶开始形成有关。山羊妊娠早期胎儿胎盘中VEGFA可能通过其受体结合与其他血管生成因子共同作用调控胎儿胎盘血管形成。     

Toxicological pathology in the rat placenta

The placenta grows rapidly for a short period with high blood flow during pregnancy and has multifaceted functions, such as its barrier function, nutritional transport, drug metabolizing activity and endocrine action. Consequently, the placenta is a highly susceptible target organ for drug- or chemical-induced adverse effects, and many placenta-toxic agents have been reported. However, histopathological examination of the placenta is not generally performed, and the placental toxicity index is only the placental weight change in rat reproductive toxicity studies. The placental cells originate from the trophectoderm of the embryo and the endometrium of the dam, proliferate and differentiate into a variety of tissues with interaction each other according to the development sequence, resulting in formation of a placenta. Therefore, drug- or chemical-induced placental lesions show various histopathological features depending on the toxicants and the exposure period, and the pathogenesis of placental toxicity is complicated. Placental weight assessment appears not to be enough to evaluate placental toxicity, and reproductive toxicity studies should pay more attention to histopathological evaluation of placental tissue. The detailed histopathological approaches to investigation of the pathogenesis of placental toxicity are considered to provide an important tool for understanding the mechanism of teratogenicity and developmental toxicity with embryo lethality, and could benefit reproductive toxicity studies.     

Streptozotocin-induced diabetes decreases placenta growth factor (PlGF) levels in rat placenta

The placenta produces several growth factors, including placenta growth factor (PlGF), which are essential for placenta growth and fetal growth. Diabetic pregnancy induces the abnormal placental growth and fetal development. This study investigated whether diabetes in pregnant rats induces changes in PlGF expression in the placenta. Diabetes was induced by a single intravenous injection of streptozotocin (35 mg/kg body weight) on day 0 of pregnancy, blood and tissue samples were collected on day 20 of pregnancy. In the diabetic group, maternal body weight and fetal weight significantly decreased compared to controls. RT-PCR and Western blot analyses showed that expression of PlGF was significantly decreased in placenta by streptozotocin treatment. Immunohistochemical study showed that the positive signal of PlGF in trophoblast cells was decreased in the diabetic group compared to controls. These findings demonstrate the decline of PlGF in the placenta in diabetic pregnancy.     

The effects of maternal age and parity on placental and fetal development in the mare

REASONS FOR PERFORMING STUDY: The normality of equine placentation is essential for fetal health and development. Substantial information exists on the gross morphological status of the placenta but few studies have addressed the problem of degenerative lesions that interfere with placental morphology and placental efficiency. HYPOTHESIS: Degenerative changes in the endometrium with increasing age and parity are reflected in the morphology and density of the placental microcotyledons. OBJECTIVE: To assess placental efficiency on the basis of foal birthweight as a function of total microscopic area of fetomaternal contact. METHODS: Stereology was used to examine the placentae of 84 Thoroughbred mares grouped on the basis of age and parity. Placental efficiency, assessed by expressing foal birthweight as a function of total microscopic area of fetomaternal contact, was also determined. RESULTS: Mare age and parity influenced the development of the microcotyledons and microcotyledon surface density (Sv) was lowest in aged multiparous mares, presumably due to degenerative changes in their endometrium. However, primiparous mares also showed significantly lower Sv values than young multiparous mares, despite the virginal endometrium of the former group. This apparent 'priming' effect of a first pregnancy on microcotyledonary Sv was illustrated further by 11 maiden mares followed in 2 successive pregnancies. They all showed significant increases in Sv values in their second parity, with equivalent improvements in foal birthweight. CONCLUSIONS: Foal birthweight is a reflection of the balance between fetomaternal contact and placental efficiency. Increases in fetomaternal contact are correlated to reductions in placental efficiency, which may reflect the ability of the placenta to modify its exchange capabilities. POTENTIAL RELEVANCE: Further work is needed to elucidate how such changes in microcotyledon morphology and efficiency are brought about.     

The secretory pattern and source of immunoreactive prolactin in pregnant African (Loxodonta africana) and Asian (Elephas maximus) elephants

The objective of the present study was to define the secretion of prolactin (PRL) in pregnant African and Asian elephants. Levels of immunoreactive (ir-) PRL in serum and placental homogenates were measured by a heterologous radioimmunoassay (RIA) based on an ovine and human RIA system, and the localization of ir-PRL in the placenta was detected by immunohistochemistry using anti-human PRL. Circulating ir-PRL clearly showed a biphasic pattern during pregnancy in African and Asian elephants. Serum levels of ir-PRL started to increase from the 4 - 6th month of gestation and reached the first peak level around the 11-14th month. A second peak of circulating ir-PRL levels was observed around the 18-20th month of gestation followed by an abrupt decline after parturition. In contrast, in a case of abortion of an African elephant, the second peak of ir-PRL was not observed, and the levels remained low for about four months until parturition. The weight of the fetus delivered at the 17th month of gestation was 23.5 kg, which was quite small compared with normal fetuses in previous reports. Ir-PRL was detected in placental homogenates, and immunolocalization was observed in trophoblasts in both the African and Asian elephants, indicating that the placenta is the source of ir-PRL during pregnancy in elephants. The present results clearly demonstrated that circulating ir-PRL shows a biphasic pattern during normal pregnancy and that the placenta appears to be an important source of circulating ir-PRL during pregnancy in both African and Asian elephants.