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1.
《畜牧兽医学报》2017,(8)
旨在用管道铸型技术制作绵羊卵巢动脉立体构筑标本,观察其形态特征及分布规律,分析其功能相关性,为绵羊生殖生理及卵巢解剖学研究奠定基础。采集20只藏绵羊子宫及卵巢样本,用8%丙烯腈-丁二烯-苯乙烯共聚物(ABS)铸型剂通过卵巢动脉进行灌注,获得卵巢动脉风干及腐蚀标本,通过观察或图片采集分析其解剖学特性。研究发现,绵羊的卵巢动脉包括卵巢支、子宫支及输卵管支,并有更小分支向系膜供血;卵巢支螺旋动脉可分为重螺旋、螺旋及轻螺旋3种不同形态,部分个体有分支螺旋动脉伴行;进入卵巢后,卵巢微动脉在卵巢门髓质部呈拳头状重螺旋折叠,末端发出卵泡或黄体微动脉。结果表明,绵羊卵巢动脉及其分支与普通牛类似,其卵巢支呈复杂的螺旋形盘绕特征,推测能降低血压、维持卵巢供血稳定性及促进优势卵泡成熟。 相似文献
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试验旨在克隆获得水牛脂素1(LPIN1)基因,并对其进行生物信息学分析,为揭示该基因在水牛脂肪沉积、生殖发育和泌乳调控中的作用奠定基础。本研究以水牛卵巢组织cDNA为模板,PCR扩增获得了LPIN1基因CDS区全长后测序,并结合生物信息学分析方法预测及分析蛋白质理化性质、二级结构及三级结构等。结果表明,水牛LPIN1基因编码区长2 793bp,编码930个氨基酸。MegAlign软件分析显示,水牛LPIN1基因核苷酸序列与水牛(预测)、牦牛、黄牛、山羊、藏羚羊、绵羊、猪、骆驼、人和小鼠LPIN1基因的同源性分别为99.6%、97.9%、97.7%、97.5%、97.4%、97.1%、89.9%、89.8%、86.2%和83.5%;水牛lipin1蛋白氨基酸序列与黄牛、牦牛、山羊、藏羚羊、骆驼、猪及人的同源性分别为99%、99%、99%、99%、94%、94%及90%。应用Mega 5.0软件构建系统进化树发现,水牛与黄牛的亲缘关系最近,其次为绵羊和山羊,LPIN1基因在不同物种及进化的过程中具有高度保守性。对lipin1蛋白分析发现,其二级结构由α-螺旋、β-折叠、T-转角和无规则卷曲组成;蛋白呈弱酸性,无信号肽,亚细胞主要定位于细胞核中,存在Lipin_N、LNS2和AF1Q等结构域,其中Lipin_N、LNS2为保守结构域。 相似文献
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试验旨在克隆获得水牛脂素1（LPIN1）基因，并对其进行生物信息学分析，为揭示该基因在水牛脂肪沉积、生殖发育和泌乳调控中的作用奠定基础。本研究以水牛卵巢组织cDNA为模板，PCR扩增获得了LPIN1基因CDS区全长后测序，并结合生物信息学分析方法预测及分析蛋白质理化性质、二级结构及三级结构等。结果表明，水牛LPIN1基因编码区长2 793 bp，编码930个氨基酸。MegAlign软件分析显示，水牛LPIN1基因核苷酸序列与水牛（预测）、牦牛、黄牛、山羊、藏羚羊、绵羊、猪、骆驼、人和小鼠LPIN1基因的同源性分别为99.6%、97.9%、97.7%、97.5%、97.4%、97.1%、89.9%、89.8%、86.2%和83.5%；水牛lipin1蛋白氨基酸序列与黄牛、牦牛、山羊、藏羚羊、骆驼、猪及人的同源性分别为99%、99%、99%、99%、94%、94%及90%。应用Mega 5.0软件构建系统进化树发现，水牛与黄牛的亲缘关系最近，其次为绵羊和山羊，LPIN1基因在不同物种及进化的过程中具有高度保守性。对lipin1蛋白分析发现，其二级结构由α-螺旋、β-折叠、T-转角和无规则卷曲组成；蛋白呈弱酸性，无信号肽，亚细胞主要定位于细胞核中，存在Lipin_N、LNS2和AF1Q等结构域，其中Lipin_N、LNS2为保守结构域。 相似文献
4.
卵母细胞成熟质量不仅是哺乳动物繁殖能力的基础，更能直接决定后代的优劣。卵巢早衰通常引起卵子数量和质量下降，如何提高其成熟质量成为生殖衰老领域的研究热点。近年来，烟酰胺腺嘌呤二核苷酸(NAD+)依赖性去乙酰化酶家族Sirtuins(SIRT1-7)在生殖衰老中的功能愈发受到关注，尤其抗衰老因子SIRT2的乙酰化底物直接与卵母细胞成熟事件相关。本文从乙酰化修饰调控卵母细胞衰老与成熟的全新视角，重点综述了NAD+/SIRT2通过减数分裂、能量代谢、线粒体氧化应激、线粒体质量控制等重要生理环节改善衰老卵母细胞成熟质量的最新研究进展，以期为提高老龄母畜的卵母细胞质量及延长繁殖利用年限提供新思路。 相似文献
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放牧是草地利用与管理的主要方式,不同的放牧方式对草地优势物种存在不同的影响。探究不同放牧方式下草地植物营养枝与生殖枝的权衡关系,对研究草地植被恢复具有一定的科学意义。本研究依托于青藏高原高寒草地-家畜系统适应性管理技术平台,在中度放牧压力下,设置了牦牛单牧、藏羊单牧、牦牛藏羊1:2混牧、牦牛藏羊1:4混牧、牦牛藏羊1:6混牧以及不放牧处理,分析了高寒草地退化指示物种星毛委陵菜(Potentilla acaulis)营养枝与生殖枝的高度、数量、重量及权衡关系。研究结果显示:1)在星毛委陵菜营养枝与生殖枝高度、数量和重量的相关指标变化中,牦牛与藏羊1:6混牧的植株总体显著高于其他处理,牦牛与藏羊1:4混牧的植株数据总体低于其他处理;2)在不同放牧方式下星毛委陵菜营养枝与生殖枝的权衡指数,牦牛与藏羊1:4混牧是除不放牧外时最低的,牦牛与藏羊1:6混牧的权衡指数倾向于营养枝,牦牛单牧、藏羊单牧和牦牛藏羊1:2混牧处理的权衡指数倾向于生殖枝,不放牧和牦牛与藏羊1:4混牧时营养枝与生殖枝之间协同生长。结果表明,高寒草地牦牛与藏羊1:4的家畜组合是较为不利于草地健康的放牧方式。本研究提出有利于草原可... 相似文献
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7.
《动物医学进展》2021,42(9)
为探究正常生理下条件下溶血磷脂酸受体(LPAR)1-3在牦牛不同时期卵巢(卵泡期、黄体期、妊娠期)表达及生物学作用,利用RT-qPCR检测LPAR1-3基因在不同时期卵巢上的表达,同时利用蛋白质免疫印迹、免疫组织化学(IHC)等方法对LPAR1-3基因和蛋白的表达进行分析和定位。结果显示,LPAR1-3在牦牛不同时期卵巢上均有表达,其中LPAR1-3的表达量在妊娠期显著高于卵泡期和黄体期(P0.05),卵泡期表达量最低;在卵泡期、黄体期、妊娠期LPAR3的表达量均显著高于LPAR1和LPAR2(P0.05),LPAR2的表达量最低;IHC结果显示,LPAR1-3在卵巢生殖上皮、颗粒细胞、卵泡液、卵泡膜和黄体细胞均有表达。研究结果提示LPAR1-3可能在卵泡生长、发育、排卵以及维持妊娠等一系列生殖过程中发挥重要的作用,其中LPAR3可能在这一过程中起主要作用,该结果有助于对牦牛繁殖机能的进一步研究,可为高原哺乳动物生殖生理的研究提供理论依据。 相似文献
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近年来,菟丝子在临床上经常用于治疗生殖内分泌类疾病,且作用效果显著。菟丝子是旋花科植物,具有毒副作用小、无耐药性、富含多种营养物质、成本低廉等优点,是纯天然的药用植物。菟丝子总黄酮是菟丝子中最有效的成分,具有止泻、滋补肝肾、益精壮阳和安胎等功效,不仅对雌雄动物生殖内分泌活动有调节作用,还对免疫、心脑血管等多个系统具有药理作用。动物的生殖内分泌活动主要由下丘脑-垂体-性腺轴调控,包括生殖激素的分泌以及精子的发生、卵泡的发育,对动物生殖繁育起着重要作用。绝大多数动物的生殖内分泌疾病都与下丘脑-垂体-性腺轴所调节的生殖激素以及生殖器官的发育状态有关,生殖激素可以通过直接或间接作用引起下丘脑-垂体-性腺轴功能性障碍疾病。鉴于目前经济动物禁用激素类药物,因此迫切需要研究安全有效低毒的中草药的作用及其机理,为其广泛应用奠定基础。文章介绍了菟丝子总黄酮主要的药理作用,并详细阐述了菟丝子总黄酮对下丘脑-垂体-性腺轴不同级内分泌活动的调控作用,为今后进一步研究菟丝子总黄酮对下丘脑-垂体-性腺轴调控的作用靶点和作用机理提供参考。 相似文献
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NPR1是SA介导的植物抗病通路中的转录共激活因子。本实验采用RT-PCR技术从杧果（Mangifera indica）中分离得到了MiNPR1-1和MiNPR1-2。序列分析显示：MiNPR1-1基因编码471个氨基酸,MiNPR1-2基因编码445个氨基酸。2种MiNPR1蛋白皆含BTB/POZ结构域和锚蛋白重复序列,非跨膜蛋白且无信号肽,为不稳定酸性亲水蛋白。蛋白结构预测表明：MiNPR1蛋白由α-螺旋、无规则卷曲、延伸链和β-折叠构成,三级结构与二级结构相符。进化关系得出：MiNPR1-1为单独分支起源,MiNPR1-2蛋白与阿月浑子（Pistacia vera）的XP_031280798.1聚类为一簇。本研究克隆了杧果NPR1基因,预测其编码蛋白特征,为后续研究该基因在杧果上的功能及培育杧果优良抗病品种提供参考。 相似文献
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本文以保幼激素类似物ZR-515和20-羟蜕皮酮添食四、五龄期的家蚕,探讨其生殖效应(造卵)。结果表明:(1)JHA和MH不仅能显著影响茧质性状,对生殖生物量也有显著效果。四龄第1日和五龄前4日施用JHA,对总造卵生物量具有明显的增值效应;五龄后期(第6—7日)使用MH,具有显著的减值效应。两种激素的这种生殖效应与雌蛹体重具有较好的一致性。(2)根据对生殖指标的分析,激素类在幼虫期使用,其生殖效应以改变成熟卵粒数为主,影响卵重为辅。(3)经对成熟卵蛋白和糖元含量测定发现;卵蛋白含量和卵糖元含量与激素的生殖效应相平行。 相似文献
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1. The present study was carried out to determine whether primordial germ cells isolated from embryonic blood can enter the bloodstream and successfully migrate to the germinal ridges of recipient embryos after transfer to stage X blastoderms, and also whether they can differentiate into blood cells, as is suggested in mice. 2. Primordial germ cells were transfected in vitro by lipofection and then transferred to stage X blastoderms. The introduced GFP gene was efficiently expressed in the gonads of 6-d incubated embryos. 3. Freshly collected primordial germ cells were transferred to stage X blastoderms. The fate of the transferred primordial germ cells was traced by detecting the single nucleotide polymorphism in the D-loop region of the mitochondrial DNA in White Leghorn and Barred Plymouth Rock chickens used in this study. The transferred donor primordial germ cell-derived cells were detected in the gonads, but not in the blood cells, of 17-d incubated embryos by PCR. 4. This procedure for primordial germ cell manipulation could provide a novel method of producing germline chimaeric chickens. 5. In conclusion, our findings indicate that primordial germ cells isolated from embryonic blood can migrate to the germinal ridges of recipient embryos after being transferred to stage X blastoderms. Although these transferred primordial germ cells differentiated into germ cells, no differentiation into blood cells was observed. 相似文献
12.
体外培养山羊50~68日龄雄性胎儿生殖细胞,并检测它们的碱性磷酸酶(AP)活性和Oct-4蛋白,探讨性别分化后的生殖细胞用于建立干细胞系的可行性及检测指标。当山羊胎儿睾丸细胞体外培养时,生殖细胞及其来源的细胞克隆均呈AP阴性和Oct-4蛋白阴性,其中有部分细胞克隆表现为AP假阳性。山羊胎儿生殖细胞克隆呈隆突状生长,多为圆形,与周围细胞界限分明,但克隆内细胞间界限不清。细胞克隆至少可以培养3代以上。研究结果显示,山羊雄性胎儿生殖细胞可以用于建立生殖系来源的干细胞系;AP和Oct-4蛋白不适宜用来检测体外培养的山羊胎儿生殖细胞及其来源的细胞系。 相似文献
13.
Isolation and culture of rabbit primordial germ cells 总被引：2，自引：0，他引：2
Kakegawa R Teramura T Takehara T Anzai M Mitani T Matsumoto K Saeki K Sagawa N Fukuda K Hosoi Y 《The Journal of reproduction and development》2008,54(5):352-357
Primordial germ cells (PGCs) are embryonic precursors of the gametes of adult animals and are considered stem cells of the germline. Since their proliferation in vitro correlates well with the schedule of developmental changes in vivo, they might be interesting research tools for genomic imprinting, germ-cell tumors and fertility. Furthermore, once primordial germ cells are separated and placed on a feeder layer with cytokines, they become cultured pluripotent cell lines called embryonic germ (EG) cells. EG cells share several important characteristics with embryonic stem (ES) cells as they can also contribute to the germ line of chimeras. To investigate the characteristics of PGCs and establish rabbit EG (rEG) cells, we cultured rabbit PGCs (rPGCs) in vitro with various combinations of leukemia inhibitory factor (LIF), basic fibroblast growth factor (bFGF) and forskolin on inactivated mouse embryonic fibroblast (MEF) feeder layers. The present study found PGC proliferation in early cultures and induction of rEG-like colonies. These cells expressed pluripotent markers, such as alkaline phosphatase activity, OCT-4, Sox-2 and SSEA-1, in the undifferentiated state; however, the cells did not develop into a teratoma when injected into the kidney capsules of SCID mice, although the restricted differentiation potentials to neural cells were determined via embryoid body formation. From these characteristics and further characterization of the germ stem cell markers Vasa, SCP-1 and SCP-3, we suggested that these were hybrid cells with characteristics somewhere between PGC and EG cells. 相似文献
14.
Transplantation of bovine germinal cells into mouse testes 总被引：5，自引：0，他引：5
Oatley JM de Avila DM McLean DJ Griswold MD Reeves JJ 《Journal of animal science》2002,80(7):1925-1931
To develop techniques for spermatogonial transplantation in bulls, it is essential to have an effective bioassay procedure to evaluate the transplantation efficiency of spermatogonial stem cell collection, purification, and culture techniques. The objective of the present study was to develop a mouse bioassay model to evaluate transplantation efficiency of fresh and cultured bovine germ cells. Bull calves of four ages (1, 2, 3, and 4 mo) were used as a source of donor testes cells. Two calves were used for each age point, one calf was experimentally made cryptorchidistic at 1 wk of age and the other left normal. A STO (mouse fibroblast) feeder cell line was used to culture bovine testes cells for 2 wk preceding transfer into recipient testes. Immunodeficient nude mice (nu/nu) in which endogenous spermatogenesis had been abolished by busulfan treatment served as recipient animals for transplantation. Donor bovine germ cells were microinjected into mouse seminiferous tubules. Mouse testes were analyzed 2 wk after transplant with the use of a bovine-specific antibody and whole-mount immunohistochemistry for the presence of bovine donor germ cells. Bovine testis cells were present in all recipient mouse testes analyzed. Fresh bovine testes cells were observed as colonies of round cells within mouse seminiferous tubules, indicating spermatogonial expansion and colonization; however, cultured bovine testes cells appeared as fibrous tissue and not as spermatogenic colonies. The average number of colonies resulting from donor cryptorchid testes was not different (P > 0.05) from noncryptorchid, 56+/-4 and 78+/-7, respectively. Fresh donor cells from calves older than 1 mo gave rise to a greater average number of colonies within recipient testes (P <0.05) (1 mo, 33+/-4; 2 mo, 70+/-8; 3 mo, 63+/-6; 4 mo, 87+/-9). Fresh bovine germ cells are capable of colonization in the busulfan-treated nude mouse testis, making it a suitable model for evaluation and development of spermatogonial transplant techniques in bulls. 相似文献
15.
The removal of endogenous germ cells of recipient stallions is a key step to produce donor germ cell-derived sperm using the germ cell transplantation technique. Six Thoroughbred stallions were divided into a treatment (n = 3) and a control group (n = 3), and 70% glycerin (1, 2, 3-trihydroxypropane, 40 mL per testis) or phosphate-buffered saline, respectively, was locally injected into testes. General semen evaluation, libido, and testicular volume were performed weekly from 3 weeks before to 10 weeks after treatment. The number of round germ cells in the ejaculate was counted using a hemocytometer. The hematoxylin and eosin staining was performed on the cross sections of testicular tissue obtained 11th week of treatment. Plasma testosterone levels in blood collected weekly were measured using a colorimetric competitive enzyme immunoassay kit. The sperm number was significantly lower than that of the control group at 5 and 10 weeks after glycerin injection. No differences in the status of spermatogenesis in the cross sections of seminiferous tubules and testicular volume were found between the two groups. The 70% glycerin-treated stallions had reduced total and progressively motile sperm and exhibited a significantly higher population of round germ cells in the ejaculate. Testosterone levels, testicular volumes, and libido of stallions were not significantly different between the groups. In conclusion, although intratesticular injection of 70% glycerin may have caused disassociation of some germ cells in the seminiferous tubules for several weeks, it did not significantly ablate germ cells in the tubules at 11 week in stallions. 相似文献
16.
V. J. Götzens L. Dominguexz M. Gimeno S. Climent 《Anatomia, histologia, embryologia》1980,9(3):193-197
The presence of annulate lamellae in the germ cells of the chick embryo, in stage 9 of H amburger /H amilton (1951), as well as in gonadal germ cells of male chick embryos which had been incubated for 81/2 days, both in normal and experimental conditions, suggests that these formations may be associated with some specific function in the development of this cell line. 相似文献
17.
体外胚胎干细胞（embryonic stem cells,ESCs）向生殖细胞分化可用于治疗不育症,同时也为揭示种系世代的分子机制提供最佳模型。试验旨在探讨视黄酸（retinoic acid,RA）诱导鸡胚胎干细胞（chicken embryonic stem cells,cESCs）向雄性生殖细胞（male germ cells,MGCs）分化的作用效果。利用胰蛋白酶消化法从新鲜种蛋X期鸡胚中分离胚胎干细胞,以鸡胚成纤维（chicken embryo fibroblast,CEF）细胞为滋养层,进行体外培养,利用形态法、碱性磷酸酶（alkaline phosphatase,AKP）染色和胚胎阶段特异性表面抗原（embryo specific surface antigen 1,SSEA-1）检测对获得的胚胎干细胞进行鉴定。结果表明,获得典型的呈巢状或岛状的cESCs克隆,细胞AKP染色呈蓝紫色,表明其具有较高的内源性AKP活性;SSEA-1鉴定结果呈阳性,显示cESCs克隆具有多能性。采用10-5 mol/L RA诱导鸡胚胎干细胞向雄性生殖细胞分化,镜下观察细胞形态变化,分别于诱导第0、2、4、6、8、10天提取细胞总RNA,反转录成cDNA,用于实时荧光定量PCR检测生殖细胞标志基因的表达。结果表明,在此诱导过程中,作为胚胎干细胞标志基因Nanog、Sox2表达量持续显著下降,而生殖细胞特异性基因Dazl、Stra8、c-kit、integrin α6表达量呈持续上升趋势;免疫细胞化学检测可观察到特异基因相关蛋白的阳性克隆。本研究成功分离出cESCs,可体外培养并保持未分化状态及多能性。10-5 mol/L RA能够促进cESCs向雄性生殖细胞方向分化,可以引起生殖细胞相应基因的表达,为进一步研究雄性生殖细胞的形成和调控机制提供参考。 相似文献
18.
1. Germline chimaeric chickens were produced by the transfer of primordial germ cells, and the generation of donor-derived offspring was examined for a maximum of 146 weeks. 2. The frequencies of donor-derived offspring from the chimaeras were 47% to 97%, and no apparent changes in frequency were observed with increasing age during the test period. 3. Differentiation of donor primordial germ cells into functional gametes appeared to be restricted to a degree at some developmental stage in the gonads of chimaeric chickens of the opposite sex. 相似文献
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Li GX Kang KS Lee YS 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2001,63(4):373-382
2-Bromopropane (2-BP) causes testicular toxicity in humans and rats. However, the germ cell degeneration of testicular toxicity by 2-BP has not been understood. 2-BP at doses of 135, 405, and 1,355 mg/kg/day was daily injected subcutaneously into Sprague-Dawley rats for 28 days. At the dose of 1,355 mg/kg/day, 2-BP significantly decreased the weights of body and testes, eipididymis, seminal vesicle, and prostate, as well as daily sperm production. Atrophy of seminiferous tubules accompanied with degeneration of germ cells such as spermatogonia, spermatocytes, and elongated spermatids was observed in the testes of rats exposed to the 405 mg/kg/day and 1,355 mg/kg/day of 2-BP. TUNEL-positive germ cells were appeared in the 405 and 1,355 mg/kg/day of 2-BP-treated groups. In addition, ultrastructure alterations of apoptotic germ cells were observed by the electron microscopy study. Dead elongated spermatids were observed at 1,355 mg/kg/day after 28 days exposure. These results suggest that 2-BP impair spermatogenesis may result from apoptotic germ cell death. 相似文献