水牛FGF10基因的克隆分析及其在不同组织中表达情况的研究

本实验旨在对水牛成纤维细胞生长因子10(FGF10)基因进行克隆、生物信息学分析,同时探讨其在不同组织中的表达模式。以水牛卵巢的总RNA为模板,通过RT-PCR方法克隆得到FGF10基因CDS区全长642 bp,编码213个氨基酸。多重序列分析显示,水牛FGF10基因核苷酸序列与黄牛、羊、猪、人、小鼠同源性分别达到了99%、99%、93%、91%和89%。系统进化树分析表明,水牛和黄牛亲缘性最近,并且在不同物种进化过程中具有高度保守性。QRT-PCR结果表明,FGF10在睾丸中的表达量最高,卵巢和脾脏次之,肝脏和心脏中最低。本研究成功克隆水牛FGF10基因,并检测其在水牛不同组织中的差异表达,为研究FGF10基因在水牛卵泡发育过程中的作用奠定基础。     

FGF21基因重要生理学功能及潜在育种价值研究进展

成纤维细胞生长因子21(Fibroblast Growth Factor 21,FGF21)是成纤维细胞生长因子家族成员之一,是一种非典型的成纤维细胞生长因子,主要作为内分泌激素发挥作用。FGF21作为一种能量稳态和代谢调控因子,在动物机体内发挥着调节糖脂代谢、促进骨骼肌发育、改善动物繁殖性状等重要生理学功能。近年来大量研究证实,FGF21广泛参与骨骼肌生长发育过程,影响畜禽肌肉的产量和质量,因此有必要深入研究FGF21基因作用的分子机制。本文主要从FGF家族基因组成、FGF21分子生物学特征、FGF21重要生理学功能等方面进行综述,为后续研究FGF21功能以及应用于畜牧生产、加强畜禽遗传资源保护等各方面提供理论基础。     

FGF10基因的生理功能及其在畜牧生产中的应用研究进展

成纤维细胞生长因子10(FGF10)是成纤维细胞生长因子家族成员之一,在机体的肝、肠、肺等器官都有表达,FGF10能够与其受体结合发挥重要的生理功能,调节胚胎发生、血管生成、伤口修复、组织稳态等多种生物学过程,还能促进细胞增殖,并且有助于细胞分化。FGF10特异性表达于白色脂肪,可以调节脂肪细胞的发育和代谢。近年来的一些研究表明,FGF10在脂肪生成及脂肪沉积等方面发挥作用,有利于改善畜禽的肉品质性状,提高畜产品的质量。本文主要综述了FGF10基因的结构特征、生理功能以及在畜牧等方面的研究进展,为进一步研究该基因的功能及其应用于畜牧生产提供依据。     

DMRT1和FOXL2基因在动物性别决定中的功能研究进展

性别决定(Sex determination)是指有性生物个体在性别决定基因的作用下,经过一系列生物发育过程后,原始性腺发育成卵巢或睾丸并使生物体发育为雌性或雄性的过程。在胚胎发育过程中,性别分化由少数的关键基因决定,当一个或多个关键基因出现功能失调时,可能会导致性腺发育异常甚至发生性别逆转的现象。近年来,大量数据证明,DMRT1和FOXL2在多个物种中被证实为与性别决定相关的关键基因,然而两者在性别调控过程中的具体作用尚不十分清楚。因此,文章对DMRT1和FOXL2基因在多个物种性别决定中的研究进展进行综述与展望,为进一步探索与研究家禽性别的分化提供新思路。     

绵羊繁殖学研究进展

通过国外对绵羊在胚胎发育过程中的基因调控、激素分泌、排卵机理与妊娠的分子生物学研究,部分解答了绵羊生长发育的调控过程与机理.叙述了绵羊的性腺决定于胎儿发育的第30～35天之前,因此,区别胎儿睾丸和卵巢之间的早期形态,垂体促性腺激素细胞在妊娠的49～50d就可以鉴别,并表明是睾丸而不是卵巢.在妊娠中期对下丘腺垂体产生负反馈.与胎儿睾丸相比,胎儿卵巢发育在妊娠后半期,并不完全依赖于促性腺激素的刺激.发情期、发情活动,季节性及产后间隔的合并效应,决定了绵羊群体的繁殖力水平,好的管理和营养条件会使繁殖力达到最大值,这是影响肉羊生产性能的重要指标之一.     

DMRT1基因通过剂量机制调控鸡胚性腺分化

正DMRT1基因编码的保守转录因子对性腺功能发挥具有重要的作用。在家禽中,DMRT1基因定位于Z染色体上,最适合用于研究家禽性腺分化的调控因子。先前的研究显示在性别分化期间敲除该基因会诱导雄性鸡胚性腺出现雌性化,因此这个基因为鸡的睾丸正常发育所必需,但它是否足以诱导睾丸分化则不甚清楚。澳大利亚、美国、日本3个国家的科研人员联合进行的一项研究发现,过量表达的DMRT1可诱导     

miRNA在哺乳动物性腺发育中的功能

MicroRNA(miRNA)作为一种内源性的非编码RNA,以影响靶基因表达的方式来调节机体功能。miRNA能调控哺乳动物性腺(睾丸和卵巢)的发育,促进精子与卵母细胞的分化成熟,影响受精卵的发育过程,并可能作为诊断生殖疾病卵巢癌的重要指标。本文对miRNA的生物合成过程,miRNA在哺乳动物性腺发育以及卵巢癌变过程中的调控功能进行了综述,提出该领域主要发展方向是miRNA在哺乳动物性腺发育中的体内功能研究,希望以分子生物学和生物信息学手段,从分子、系统等不同角度阐明单一某种miRNA在哺乳动物性腺发育及卵巢癌变过程中的具体调节机制和信号通路情况。这为研究哺乳动物生殖疾病,选育畜牧生产中优质品种提供了新的思路和途径。     

FGFs/FGFRs及其介导信号通路基因的异常表达影响犏牛未分化精原细胞增殖活性

旨在研究FGFs(fibroblast growth factors)/FGFRs(fibroblast growth factor receptors)及其介导的Ras和MAPK信号通路基因在牦牛和犏牛未分化精原细胞的差异表达，以探讨犏牛生精停滞的原因。本研究采集健康的24月龄3头公麦洼牦牛和3头F1代公犏牛的睾丸组织，分为牦牛和犏牛两个样品组，每组3个生物学重复。采用HE染色检测牦牛和犏牛精子发生的差异；采用细胞群体倍增时间、CCK-8和EDU染色检测牦牛和犏牛未分化精原细胞增殖能力的差异；采用荧光定量PCR检测6种FGF家族成员、3种FGFR、FGFs/FGFRs介导的Ras和MAPK信号通路基因在牦牛和犏牛未分化精原细胞中的差异表达。与牦牛相比，犏牛生精小管发育异常，生殖细胞类型单一，未分化精原细胞增殖活性显著低于牦牛(P<0.05)。FGF2、FGF4、FGF5、FGF8、FGF9和FGF21在犏牛睾丸组织及FGFs受体FGFR1、FGFR2和FGFR3在犏牛未分化精原细胞的表达都显著低于牦牛(P<0.01)。FGFs/FGFRs介导Ras信号通路基因中，除HRa...     

在缺少SRY基因的男性个体中SOX9的表达上调

在哺乳动物中,正常的性别是由是否存在Y-染色体基因SRY所决定的。SRY基因在未分化的性腺中表达以后,大量与睾丸分化相关的转录因子和生长因子基因开始特异性表达。然而,在XX雄性个体中这些基因一定在缺少SRY基因的情况下得到上调,但是XX雄性个体中缺少SRY基因的原因还不清楚。作者检测了标记的典型性腺基因在缺少SRY基因的XX雄性睾丸中的表达。结果发现,RT-PCR和免疫组化研究结果显示在缺少SRY基因的XX雄性的睾丸组织中SOX9,DAX-1,Ad4BP/SR-1,WT-1,GATA-4和MIS得到了表达。这些病人睾丸组织中SOX9的表达水平是正常XY个体睾丸组织中的1.9倍,而Ad4BP/SF-1,DAX-1和MIS的表达水平SRY缺少的XX个体睾丸低于XY个体的睾丸。所有的XX病人被发现在每个二倍体基因组中带有两个拷贝的SOX9基因,如同正常的XX雌性和XY雄性。XX雄性病人带有两个拷贝的DAX-1基因和正常的XX雌性一样,而正常XY雄性带有1个DAX-1基因。资料表明Lesions影响SOX9的表达是缺少SRY基因的XX雄性性别决定的关键因素,并且Ad4BP/SF-1,DAX-1和MIS表达水平的降低有...     

FGF22及其受体在庆阳黑山羊正常睾丸与隐睾中的表达比较

试验旨在研究成纤维生长因子22（fibroblast growth factor 22,FGF22）及其受体2（fibroblast growth factor receptor 2,FGFR2）、硫酸乙酰肝素糖蛋白（heparan sulfate proteoglycans,HSPG）在庆阳黑山羊正常睾丸与隐睾中的分布与表达,探究其在山羊睾丸发育和隐睾形成中的作用。采用HE和特殊染色观察其组织学结构特征,进而以免疫组织化学及免疫荧光法结合形态计量学统计研究FGF22、FGFR2和HSPG在山羊正常睾丸及隐睾中的定位。结果表明,山羊隐睾较正常睾丸生精小管缩窄,腔内各级生精细胞排列紊乱,间质的胶原纤维和网状纤维增多,糖原类物质阳性反应较弱,FGF22在隐睾组织的Leydig、Sertoli细胞、管周肌样细胞及血管内皮细胞整体表达密度相较于正常睾丸显著减弱（P<0.05）。HSPG在正常睾丸表达显著强于隐睾（P<0.05）,间质组织变化尤其明显。FGFR2在隐睾组表达显著增高（P<0.05）,且以Sertoli细胞强阳性表达为主。庆阳黑山羊隐睾较正常睾丸发育异常,间质组织有纤维化趋势,糖原类物质含量减少;FGF22及HSPG表达降低应与隐睾局部环境温度变化密切相关;FGFR2在隐睾组表达增高提示其在发生隐睾时可能通过Sertoli细胞进行适应性调节。     

The Case of an Sry‐Negative XX Male Pug With an Inguinal Gonad

A case of intersexuality in a Pug that was bought as a male in a pet shop is described. The dog was presented at the Veterinary Teaching Hospital, University of Turin, for a reddish mass protruding from the prepuce. The mass had the aspect of an enlarged clitoris, with a caudoventral direction and a dorsal urethral ostium. A gonad was palpable in the left inguinal region. Laparotomy confirmed ultrasound detection of an abdominal uterine structure together with the right gonad. The histology of both gonads was similar, showing an exclusively masculine character, with seminiferous tubules lined only by Sertoli cells; the uterus showed a normal histological structure. Karyological analysis revealed a female karyotype (78,XX), and polymerase chain reaction showed the absence of Sry. The diagnosis was an XX male. The pathogenesis of the XX sex reversal syndrome in dogs is not completely understood, as Sry, the master gene regulating testis differentiation, is not present; to date, no genetic cause has been identified for this phenotypic condition in dogs. This case is unusual because the dog showed an inguinal testis, implying a partial activity of the mechanisms leading to abdominal testis translocation along a gubernaculum and transinguinal migration.     

豪猪雄性生殖腺(睾丸)的解剖及组织结构研究

以豪猪睾丸为研究对象,观察豪猪睾丸的解剖及组织结构,研究豪猪的雄性生殖腺,为豪猪的人工繁殖研究提供理论依据。结果表明:雄性豪猪没有阴囊,其睾丸位于腹股沟内;睾丸的重量为(2.76±0.08)g,其纵径为(3.93±0.09)cm,横经为(1.38±0.06)cm;睾丸小隔不发达,每个睾丸小叶内有1~4条曲精小管,曲精小管的管壁中可见许多不同发育阶段的生精细胞,其形态多样,但精原细胞和精子细胞数量较少,未见次级精母细胞。     

A Proposed Role for VEGF Isoforms in Sex-Specific Vasculature Development in the Gonad

Many scientists have expended efforts to determine what regulates development of an indifferent gonad into either a testis or ovary. Expression of Sry and upregulation of Sox9 are factors that initiate formation of the testis-specific pathway to allow for both sex-specific vasculature and seminiferous cord formation. Migration of mesonephric precursors of peritubular myoid cells and endothelial cells into the differentiating testis is a critical step in formation of both of these structures. Furthermore, these events appear to be initiated downstream from Sry expression. Sertoli cell secretion of growth factors acts to attract these mesonephric cells. One hypothesis is that a growth factor specific for these cell linages act in concert to coordinate migration of both peritubular and endothelial cells. A second hypothesis is that several growth factors stimulate migration and differentiation of mesonephric 'stem-like' cells to result in migration and differentiation into several different cell lineages. While the specific mechanism is unclear, several growth factors have been implicated in the initiation of mesonephric cell migration. This review will focus on the proposed mechanisms of a growth factor, Vascular Endothelial Growth Factor, and how different angiogenic and inhibitory isoforms from this single gene may aid in development of testis-specific vascular development.     

哺乳动物SRY基因研究进展

尽管SRY基因的发现已有10多年，但到目前为止，人们还没有弄清楚SRY基因是如何调控胚胎双性腺发育成睾丸的。经过10多年的研究，人们对SRY基因有了新的认识。作者综述了SRY基因的功能，SRY蛋白以及SRY基因在性腺中表达的最新研究进展。     

Hypoxia induces FGF2 production by vascular endothelial cells and alters MMP9 and TIMP1 expression in extravillous trophoblasts and their invasiveness in a cocultured model

The role of fibroblast growth factor 2 (FGF2) secretion by vascular endothelial cells during trophoblast invasion was assessed. The human extravillous trophoblast cell line, TEV-1, and umbilical vein endothelial cell line, HUVE-12, were cocultured under normal and hypoxic conditions. FGF2 expression in HUVE-12 cells and matrix metalloproteinase 9 (MMP9) and tissue inhibitor of metalloproteinase 1 (TIMP1) expression in TEV-1 cells were analyzed using quantitative RT-PCR and Western blot analyses. TEV-1 cell invasion was also examined. FGF2 expression in the HUVE-12 cells cocultured with TEV-1 cells was significantly increased under hypoxic conditions. In the TEV-1 cells cocultured with HUVE-12, hypoxia reduced MMP9 expression and increased TIMP1 expression; it also reduced cell invasion by 43%. However, the expression of MMP9 and TIMP1 and ratio of MMP9/TIMP1 were increased when the TEV-1 cells were cultured alone under hypoxic conditions. These findings suggest that FGF2 release by stressed endothelial cells of uterine spiral arteries play roles in decreasing MMP9 and increasing TIMP1 production in extravillous trophoblasts (EVT) in response to stress, resulting in reduced EVT invasion and possibly shallow implantation of the placenta.     

家蚕幼虫性腺表达序列标签分析(英文)

分别以5龄幼虫的精巢和卵巢构建了2个家蚕cDNA文库.利用表达序列标签方法分析参与家蚕性腺发育的基因,通过测序获得 16 737条ESTs(精巢 8 407条,卵巢 8 330条),拼接这些ESTs共得到 2 107个重叠群(contig)和 3 532个单拷贝(singlet).将这些转录物与GenBank非冗余蛋白质库比较,结果显示约34.3%的基因与数据库中蛋白质具有相似性.功能注释表明蛋白质合成相关基因、精巢特异基因等在文库中大量存在.基因本体(gene ontology)功能分类显示精巢与卵巢基因表达谱差异较大.研究结果为理解家蚕性腺发育提供了信息.     

Preventive effect of zinc against cadmium-induced oxidative stress in the rat testis

The aim of this study was to investigate the antioxidant role of zinc (Zn) in the Cd-exposed testes of Wistar rats. Subchronic exposure to Cd (CdCl(2), 40 mg/l, per os) for 30 days resulted in a significant reduction in growth rate (-11%) and relative weights of testes (-36%) and seminal vesicles (-80%). Treated rats displayed a decrease in testicular and plasma testosterone levels, respectively (-70%, P<0.05; -48%, P<0.05), epididymal sperm count (-22%, P<0.05), and spermatozoa motility (-35%, P<0.05). In contrast, Cd increased the malondialdehyde (+46%, P<0.05), metallothionein (+200%, P<0.05), and 8-oxodGuo concentrations (+71%, P<0.05) in the testis. In the gonad, Cd decreased the GPx (-30%, P<0.05), CAT (-32%, P<0.05), mitochondrial Mn-SOD (-34%, P<0.05), and cytosolic CuZn-SOD (-32%, P<0.05) activities. Zinc supplementation (ZnCl(2), 40 mg/l, per os) in the Cd-exposed rats restored the activities of GPx, CuZn-SOD, and Mn-SOD in the testes to the levels of the control group. Moreover, zinc administration was capable of reducing the elevated levels of malondialdehyde in the testis. Interestingly, zinc supplementation attenuated DNA oxidation induced by Cd in the gonad and restored the testosterone level and sperm count to the levels of the control group. Zinc administration minimized oxidative damage and reversed the impairment of spermatogenesis and testosterone production induced by Cd in the rat testis.     

鸡早期胚胎精原细胞和睾丸发生发育关系的研究

采用连续切片技术，研究了15～45期(孵化50h～19d)鸡早期胚胎发育过程中精原细胞与睾丸发生发育的关系。结果显示：孵化第22～28期(第3．5～5天)，原始生殖腺内的原始生殖细胞(PGCs)胞质内的糖原颗粒开始分解；第29期(孵化第6天)，鸡胚性腺开始分化，PGCs的糖原颗粒进一步分解；第31期(孵化第7天)，PGCs内的糖原颗粒完全消失。在雄性，性腺开始显示睾丸特征，PGCs分化为精原细胞；第34期(孵化第8天)，睾丸内曲精细管索开始形成，呈实心状，精原细胞位于其中；第35～37期(孵化第9～11天)，曲精细管索数量逐渐增加，索内精原细胞体积较大，呈圆形单层排列，且已分化出支持细胞，但数量不多，形态不易分辨，间质内有少量间质细胞；第38～40期(孵化第12～14天)，可见到典型的曲精细管，精原细胞数量增加，支持细胞亦增多；曲精细管间的间质细胞成群分布。第40～45期(孵化第16～19天)，两侧睾丸大小略有差异，右侧稍大于左侧，精原细胞数量明显增多，呈葡萄串状分布在曲精细管的中央；曲精细管的管腔已经形成，精原细胞已分层排列。     

Malignant seminoma with gross metastases in a dog

A 10-year-old male crossbred dog developed a non-painful unilateral testicular swelling; the other gonad was reduced in size. Both gonads were removed surgically; the larger contained a diffuse seminoma, the smaller atrophic testis contained an interstitial cell adenoma and several intratubular seminomas. Six months after being castrated the dog developed palpable intraabdominal masses. Necropsy revealed gross metastatic malignant seminoma in para-aortic lymph nodes. No histological features were identified to correlate with the malignant behaviour of this usually benign tumour.