生长分化因子-9在大鼠卵巢中的表达

目的：探讨生长分化因子-9(GDF-9)在大鼠卵巢中的表达部位。方法：选择不同发育时期的大鼠卵巢,采用免疫组化和Western blot技术检测大鼠卵泡各发育阶段GDF-9蛋白表达情况及分布规律。结果：免疫组化方法显示,出生0～2d卵巢中未见GDF-9表达,出生3d卵巢中见部分原始卵泡卵母细胞GDF-9表达阳性,以后从初级卵泡到发育成熟的卵泡卵母细胞均明显可见GDF-9蛋白表达。Western blot技术进一步提示,出生0-2d卵巢中无GDF-9蛋白形成,第3d开始有GDF-9蛋白产生,以后在卵巢发育的各时期均有GDF-9蛋白分泌。结论：GDF-9在卵巢发育的早期即开始产生,以后在各阶段卵巢中持续表达,提示卵母细胞通过产生GDF-9等因子对卵泡的发育进行调节。     

国人胎儿卵巢组织发生的形态学观察

在16～38周41例胎儿中,取右侧卵巢固定于Bouin液中,制成切片,HE染色,在显微镜下观察了卵巢的发育过程。见到卵巢表面上皮随胎龄变化而发生变化;20周胎儿卵巢出现白膜,到36周时发育完整;16周时在卵巢皮质和髓质交界区出现原始卵泡,20周后大量原始卵泡存在于皮质中且可观察到少数生长卵泡;36周时生长卵泡出现卵泡腔。     

氯化汞对小白鼠卵巢损伤的显微镜及超微结构研究

对氯化汞染毒小白鼠卵巢进行了显微和超微结构研究。结果表明,氯化汞对小白鼠卵巢发育有明显的毒性作用,主要表现出卵巢内发育的卵泡（原始卵泡、生长卵泡、成熟卵泡）数量随剂量的增高依次性减少,而闭锁卵泡的数量则相反,为依次性增高。超微结构显示,从中剂量组开始,卵照相结构表现出病理学变化,主要表现为卵细胞核膜皱缩,染色质凝聚,胞质内线粒体肿胀、变形,外膜和内嵴受损,次级溶酶体及残余小体增多等现象。     

骨形态发生蛋白7在兔卵巢中的表达分析

为探讨BMP7蛋白在卵泡发育、黄体化和排卵过程中的作用,本研究采用免疫组织化学方法检测了BMP7在兔卵巢中的表达定位情况.结果显示, BMP7蛋白在兔卵巢原始卵泡和初级卵泡中未检测到表达;在次级卵泡卵母细胞及有腔卵泡的各种细胞中都有表达,但只在部分内膜细胞中表达;在间质细胞、黄体及表面上皮中也有表达.由此表明,BMP7没有参与兔原始卵泡的激活,而在初级/次级卵泡转化以及其后的卵泡成熟过程中发挥了作用;BMP7不仅是卵泡内膜细胞分泌因子,而且也是卵母细胞和颗粒细胞分泌因子.对BMP7的表达来说卵泡内膜细胞存在功能性差异.BMP7可能参与了兔黄体、排卵等活动的调节.     

中药复方液对麻黄素损伤小鼠卵巢的保护作用

为了研究中药对麻黄素损伤小鼠卵巢的影响,将60只受孕小鼠随机分为对照组、麻黄素组与中药组.对照组腹腔连续注射生理盐水,麻黄素组与中药组均腹腔注射0.2mL 6g·L-1麻黄素溶液,中药组在注射麻黄素溶液1h后灌胃0.3mL的中药复方液.在处理15、20、25d后,分别用ELISA法测定小鼠血浆中肾上腺素(EPI)、雌二醇(E2)含量的变化,用比色法检测卵巢组织SOD活性及MDA含量的变化,称量卵巢重量,用生物显微技术观察卵巢组织的显微结构,计数次级卵泡与闭锁卵泡数量,测量次级卵泡直径,用免疫组织化学法检测Bax蛋白在卵巢组织中表达的变化.结果显示,麻黄素组小鼠血浆EPI浓度升高,E2浓度降低,卵巢组织SOD活性降低,MDA含量升高,次级卵泡与闭锁卵泡数量增多,次级卵泡直径减小,卵巢组织受损,结构紊乱,颗粒细胞排列松散、降解,黄体细胞核固缩,Bax蛋白的表达增强.与麻黄素组相比,中药组小鼠血浆中EPI浓度降低,E2浓度升高,卵巢组织SOD活性明显升高,MDA含量降低,卵巢结构清晰,次级卵泡数量减少,颗粒细胞排列紧密整齐,崩解减少,Bax蛋白的阳性表达明显减少.研究结果表明,中药复方液对麻黄素损伤小鼠卵巢有一定的保护作用,这可能与SOD活性和Bax蛋白的表达有关.     

中华绒螯蟹(Eriocheir sinensis)雌性生殖系统的组织学研究

以组织切片方法研究中华绒螯蟹雌性生殖系统结构以及卵子发生和卵巢发育的周年变化表明:卵巢壁由外膜及内生殖上皮组成;输卵管壁由外膜及管壁上皮组成;受精囊分为伸展腔和皱缩腔,二者囊壁上皮结构不同,并有周期性变化。卵子发生根据卵细胞、细胞核及核仁的大小形态、核质比(NP)和卵黄积累情况可分为:1.卵原细胞期;2.小生长期;3.大生长期;4.成熟前期和5.成熟期。卵巢发育则根据第一批卵细胞的发育情况、卵巢长度及颜色分为五期,大致可归纳为三个阶段:5～7月为延缓期;8～11月为对数增长期;12～4月为稳定期。     

中华绒螯蟹(Eriocheir sinensis)雌性生殖系统的组织学研究

以组织切片方法研究中华绒螯蟹雌性生殖系统结构以及卵子发生和卵巢发育的周年变化表明:卵巢壁由外膜及内生殖上皮组成;输卵管壁由外膜及管壁上皮组成;受精囊分为伸展腔和皱缩腔,二者囊壁上皮结构不同,并有周期性变化。卵子发生根据卵细胞、细胞核及核仁的大小形态、核质比(NP)和卵黄积累情况可分为:1.卵原细胞期:2.小生长期;3.大生长期;4.成熟前期和5.成熟期。卵巢发育则根据第一批卵细胞的发育情况、卵巢长度及颜色分为五期,大致可归纳为三个阶段:5～7月为延缓期;8～11月为对数增长期:12～4月为稳定期。     

蛤蚧卵巢结构和卵泡发育的研究

利用光镜和电镜对蛤蚧卵巢结构和卵泡发育进行了观察研究，成年蛤蚧卵巢由生殖基，卵巢壁和不同发育阶段的卵泡所组成，卵原细胞位于生殖基内，每一卵巢仅有一个生殖然，卵泡发育整个过程经历卵原细胞期，初级卵泡期，生长卵泡期的成熟卵泡期４个时期。在生长儿泡期，同形滤泡细胞转变成为形态和各功能各异的３种细胞－小细胞，喑细胞和梨形细胞；随后再由异形细胞变为同形细胞，这种现象是鳞类爬行动物独有的特征。     

分离方法和卵巢发育状况对猪腔前卵泡分离及体外培养的影响

以直径在200～300μm的腔前卵泡为研究对象，研究了不同分离方法及卵巢不同发育状况对猪腔前卵泡分离及体外培养的影响。结果表明，显微分离法每个卵巢可采集的卵泡数极显著高于剪碎过滤法（24．75±2．99vs2．60±0．35，P〈0．01），显微分离法分离每个腔前卵泡所用的时间极显著低于剪碎过滤法（5．40±0．44vs15．52±1．18，P〈0．01）；有黄体和无黄体的卵巢对卵泡分离的影响差异不显著（26．10±2．02vs24．35±2．32，P〈0．05），但有黄体的卵巢分离的卵泡数稍多于无黄体的卵巢。此外，对两种分离方法分离出的腔前卵泡进行培养研究表明，两种分离方法对卵泡体外生长和存活，以及正常卵母细胞比率均没有显著影响（P〉0．05）。     

大鳍鳠卵巢滤泡细胞发育的组织学研究

用组织学及组织化学的方法对大鳍鳠卵巢滤泡细胞的发育及精孔细胞的形成进行研究,结果为:①大鳍鳠卵巢滤泡细胞来源于卵巢基质细胞,由围在卵细胞外围的卵泡膜细胞层和其内侧的颗粒细胞层组成,滤泡细胞从发生到退化的全过程可分为7个时期;②大鳍鳠仅有1个受精孔一个精孔细胞,精孔细胞由1个滤泡细胞发育而成.     

Follicular growth,differentiation and atresia

Only limited numbers of primordial follicles in mammalian ovary grow and differentiate to reach the stage of dominate follicles and ovulate. 99% of the follicles in the ovary undergo atresia at various stages of development.Regulation of follicular growth, development and atresia is a complex process and involves interactions between endocrine factors and intraovarian regulators. This review summarized: i ) FSH may not be a survival factor in regulating slow-growing preantral follicles. Some locally produced growth factors, activin and orphan receptors might play a more important role at this stage, ii ) Estrogen, activin/inhibin and follistatin coordinate with FSH to regulate and control follicle differentiation, iii) There are two types of follicular atresia induced by apoptosis which originates from GC or oocyte, respectively. Early translation of tPA mRNA into tPA protein in oocyte may be associated with oocytea apoptosis.     

Germline stem cells and follicular renewal in the postnatal mammalian ovary

A basic doctrine of reproductive biology is that most mammalian females lose the capacity for germ-cell renewal during fetal life, such that a fixed reserve of germ cells (oocytes) enclosed within follicles is endowed at birth. Here we show that juvenile and adult mouse ovaries possess mitotically active germ cells that, based on rates of oocyte degeneration (atresia) and clearance, are needed to continuously replenish the follicle pool. Consistent with this, treatment of prepubertal female mice with the mitotic germ-cell toxicant busulphan eliminates the primordial follicle reserve by early adulthood without inducing atresia. Furthermore, we demonstrate cells expressing the meiotic entry marker synaptonemal complex protein 3 in juvenile and adult mouse ovaries. Wild-type ovaries grafted into transgenic female mice with ubiquitous expression of green fluorescent protein (GFP) become infiltrated with GFP-positive germ cells that form follicles. Collectively, these data establish the existence of proliferative germ cells that sustain oocyte and follicle production in the postnatal mammalian ovary.     

小鼠阑尾淋巴组织的发生

对不同日龄幼鼠阑尾淋巴组织的发生、T和B细胞在阑尾中出现的时间及分布的系统研究表明:小鼠阑尾淋巴组织是在出生后逐渐发育形成的.幼鼠出生当日,在其阑尾内固有膜增厚,在淋巴滤泡原基中已具有成熟的T淋巴细胞和较成熟的B淋巴细胞.出生后随着淋巴滤泡的发育及肠道内抗原的刺激,T和B淋巴细胞数目逐渐增多,最终分别定位分布于淋巴滤泡的滤泡间区与滤泡区内.     

大熊猫（Ailuropoda Melanoleuca）卵巢的年龄变化

对比观察了5只不同年龄大熊猫(包括青年、中年和老年)的卵巢皮质石蜡切片。可见皮质结构有较大的年龄变化。主要表现在:性成熟后,卵巢皮质的厚度随年龄的增长有减薄的趋势。成年的各级正常卵泡数量较少,初级和次级的更少;闭锁卵饱和间质腺较多,并呈现出随年龄增长而增多的动向。老年卵巢结构变化明显,皮质纤维化,卵泡全部消失,仅有较多囊泡,小动脉管壁增厚,管腔变小。老年卵巢的外形、体积和重量等方面与成年之间未见明显年龄变化。     

Abnormal gonad development in Kit W-2Bao mice caused by a Kit gene missense mutation

Kit ^W-2Bao mice are single-gene autosomal dominant mutation mice with a B6 background that were bred in our laboratory. Heterozygotes had morphological characteristics including albinism of the abdomen, extremities, and tail, whereas the homozygotes had albinism of the body, black eyes, and infertility. The homozygous mutants showed small, structurally abnormal gonads, and lacked germ cells. Heterozygous male mice lacked germ cells in some contorted seminiferous tubules. This mutation has been mapped at 43.8 cM from the centromere in chromosome 5 by linkage analysis and Kit has been identified as the candidate gene. After Kit full-length mRNA amplification, it was found that a G to T conversion at position 1228 in the ORF changed the 410th amino acid from V to F. This amino acid change could affect the protein’s secondary structure. Heterozygous mutant mice were intercrossed and homozygous mutant mice were bred and genotyped. We found that no primordial germ cells (PGCs) appeared in the urogenital ridge area at fetus day 11.5 in the homozygotes. The number of PGCs also significantly decreased in heterozygotes. At fetus day 15.5, the differentiation of the testis tubule structure was unclear; as well, they contained no spermatogonia. Female homozygotes contained no primordial follicles in the ovary. The numbers of PGCs and primordial follicles were significantly decreased in heterozygous mice. W ^?2Bao is the only mutated site in the extracellular 4th Ig-like domain and this mutant mouse model provides new material for the study of the mechanism of reproductive system development.     

miR-26b和miR-224在梅山与杜洛克猪卵泡中的差异表达分析

为了探讨miRNAs在猪卵泡和繁殖相关组织中的表达,采用荧光定量RT-PCR法和组织表达谱分析法,对miR-26b和miR-224在梅山和杜洛克猪卵泡及其他繁殖相关组织中的表达进行了研究。组织表达谱分析结果表明:miR-26b和miR-224在下丘脑、垂体、子宫、输卵管、卵巢、子宫角、黄体组织中均有表达。RT-PCR结果表明:miR-26b在L卵泡中的表达量梅山猪是杜洛克猪的2倍(P0.01);而在S卵泡中的表达量杜洛克是梅山猪的10倍(P0.01);两品种在M1和M2卵泡中的表达量差异不显著。miR-224在S卵泡中的表达量杜洛克是梅山的7.79倍(P0.01);而在M2卵泡中表达量梅山是杜洛克的1.84倍(P0.05);两品种在M1和L卵泡中的表达量差异不显著。在猪种内大卵泡L、中等卵泡M2、中等卵泡M1、小卵泡S的相对表达量也有不同程度的差异。此研究表明,miR-26b和miR-224在梅山和杜洛克猪中表达趋势相似,它们的表达差异可能影响卵泡发育和成熟,为深入了解miR-26b和miR-224在卵泡发育中的作用奠定了基础。     

南方鲇卵巢滤泡细胞和卵膜生成的超微结构研究

滤泡细胞来源于卵巢基质细胞,其发育过程分为:零散卵泡膜细胞期、单层扁平卵泡膜细胞期、多层扁平 卵泡膜细胞期、立方形颗粒细胞期、柱状颗粒细胞期、颗粒细胞分泌期6个时期.多层细胞的滤泡可以分为外膜 层、鞘膜细胞层、粗纤维层、细纤维层、滤泡细胞层5个亚层.颗粒细胞具丰富的微丝、线粒体、内质网、核糖 体,高尔基体也丰富而发达.滤泡细胞具有生成次级卵膜、合成卵黄蛋白并加工成卵黄前体颗粒和中间颗粒、产 生类固醇激素和协助排卵等作用. 卵膜包括初级卵膜和次级卵膜.初级卵膜分为3个亚层,由卵母细胞依次产生的3个亚层最终融合并致密化 形成放射带.而次级卵膜在卵黄合成后期由滤泡细胞产生.     

卵泡分化排卵和黄体发生的分子基础

本文综述了近二十年来在雌性生殖研究领域中五个方面的研究进展。（1）原始卵泡的启动生长和分化：原始卵泡生长启动可能受卵巢内在因子,像生长因子的调控,而不是人所想象的FSH;启动生长的卵泡受生长因子,FSH,激活素,孤儿受体和其他卵巢旁分泌因子调控,并发展成“优势卵泡”;（2）优势卵泡和闭锁卵泡：FSH,激活素／抑制素,卵泡抑素和雌激素调节卵泡分化的趋向,它们的相互作用和平衡决定“优热卵泡”和“闭锁卵泡”的形成,其中雌激素在此起关键性作用;它一方面促进FSH对LH受体的诱导作用,增加GC上的LH受体数目;另一方面可促进GC芳香水酶的产生,使此卵泡进入良性循环,发展为优势卵泡;（3）两种细胞两种促性尕激素学说：在FSH和LH作用下,GC产生的孕酮可被膜细胞利用转化为雄激素;因此GC和TC相互作用是卵巢雌激素产生的前提;（4）排卵:GC主要产生tPA,而TC主要表达tPA的抑制因子PAI－1,在激素作用下,GC的tPA和TC的PAI－1相互作用,形成一个局部蛋白水解窗口,它对卵泡破裂起决定性作用;（5）黄体形成和萎缩：uPA和MMP／TIMP系统对黄体发生起重要作用;而tPA对黄体萎缩起关键作用。     

黄边糙鸟蛤生殖腺的组织学研究

采集黄边糙鸟蛤成年个体,并进行活体解剖和显微镜观察.结果表明:性成熟个体为雌雄两性功能同体存在的同步性雌雄同体.性腺由树枝状分枝的生殖管、输精管和输卵管组成.生殖管反复分枝成生精小管、生卵小管及其末端膨大的生殖泡囊.性腺由外膜及其内生精小管和生卵小管盘绕交错排列构成.外膜含单层上皮及薄层肌肉,生殖小管和泡囊皆由位于基膜上的生殖上皮构成,生殖上皮不断向腔内增殖各阶段生殖细胞.本文详细描述了精原细胞、卵原细胞、精母细胞、卵母细胞以及精子和卵子的形态结构.此外,还报道了极少数黄边糙鸟蛤性腺存在一些雄性滤泡转化为雌性滤泡现象.     

Wnt-dependent de novo hair follicle regeneration in adult mouse skin after wounding

The mammalian hair follicle is a complex 'mini-organ' thought to form only during development; loss of an adult follicle is considered permanent. However, the possibility that hair follicles develop de novo following wounding was raised in studies on rabbits, mice and even humans fifty years ago. Subsequently, these observations were generally discounted because definitive evidence for follicular neogenesis was not presented. Here we show that, after wounding, hair follicles form de novo in genetically normal adult mice. The regenerated hair follicles establish a stem cell population, express known molecular markers of follicle differentiation, produce a hair shaft and progress through all stages of the hair follicle cycle. Lineage analysis demonstrated that the nascent follicles arise from epithelial cells outside of the hair follicle stem cell niche, suggesting that epidermal cells in the wound assume a hair follicle stem cell phenotype. Inhibition of Wnt signalling after re-epithelialization completely abrogates this wounding-induced folliculogenesis, whereas overexpression of Wnt ligand in the epidermis increases the number of regenerated hair follicles. These remarkable regenerative capabilities of the adult support the notion that wounding induces an embryonic phenotype in skin, and that this provides a window for manipulation of hair follicle neogenesis by Wnt proteins. These findings suggest treatments for wounds, hair loss and other degenerative skin disorders.