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采用玻璃化冷冻方法对野外采集的牦牛睾丸组织进行冷冻,通过HE(hematoxylin-eosin)染色分析发现玻璃化冷冻后的牦牛睾丸组织曲细精管结构保存较完好,曲细精管可见大量形态完好的各类生精细胞。采用台盼蓝染色检测细胞活率,发现冷冻复苏后细胞活率可达80.20%。分别通过生精细胞和精原干细胞标志蛋白DDX4和GFRA1免疫荧光染色发现复苏后培养14天后的生精细胞和精原干细胞的数量明显减少。通过RT-qPCR对复苏后不同实验处理组牦牛睾丸细胞标志基因的表达分析,发现培养30天的生精细胞中的精原干细胞标志基因Thy1和UCHL1的表达量显著增高。因此,玻璃化冷冻保存的牦牛睾丸组织中曲细精管及其生精细胞得到了较好的保护,该方法对于其他哺乳动物生精细胞的长久有效保存具有重要的参考价值。 相似文献
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本实验旨在探究鸟苷酸交换因子15(ARHGEF15)基因在小鼠睾丸中的表达模式及其对精子发生的影响，本研究通过免疫组化和免疫荧光双染色实验检测ARHGEF15蛋白在5日龄和成年（8～9周龄）小鼠睾丸中的表达模式，并利用Cre-LoxP系统特异性敲除小鼠生精细胞内的ARHGEF15基因。结果显示：在5日龄小鼠睾丸中，ARHGEF15蛋白主要表达于精原细胞的细胞质和/或细胞膜，在成年小鼠睾丸中ARHGEF15蛋白主要定位于支持细胞的细胞核；条件性敲除（cKO）组小鼠睾丸的形态大小、组织学结构、精子的运动性能及形态与对照组相比无显著差异，且cKO组小鼠的繁育力也不受影响。因此，ARHGEF15基因主要在发育早期小鼠睾丸的生精细胞中表达，敲除生精细胞内的ARHGEF15基因不影响小鼠的繁殖能力和精子发生。 相似文献
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小公牛提睾后睾丸体积变小、质地变硬、平均每只重约 82 .5g(n=10 )。组织学观察发现 ,其细精管细小 ,管腔狭窄 ,管壁以支持细胞为主 ,生精细胞的数量很少 ,管腔内没有或仅有少量精子样物。整个睾丸呈萎缩的退行性变化。 相似文献
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本文利用生物体视学的技术和方法对性成熟的鹌鹑和鸡睾丸组织进行了１５项定量组织学指标测定及比较。结果表明，两者睾丸的曲细精管直径、生精细胞和内分泌细胞的数量密度和直径均存在显著的种间差异（Ｐ＜０．０１；０．００１），生精细胞的大小与其数量密度成反比，与曲细精管直径和生精上皮高度成正比。 相似文献
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《中国兽医杂志》2016,(6)
为了观察小鼠氟中毒睾丸形态变化特点,研究氟对睾丸的毒性影响,选用45只健康性成熟雄性昆明系小鼠,随机分为对照组、低氟组(25 mg/L Na F)、高氟组(100 mg/L Na F),连续饲喂60 d,制作睾丸石蜡组织切片,H.E.染色后,显微镜下观察曲细精管形态变化,通过定量分析方法探讨氟对睾丸的毒性影响。结果表明,与对照组相比,低氟组和高氟组曲细精管半径、面积以及曲细精管管腔半径、面积均无显著性差异(P0.05);高氟组小鼠曲细精管细胞层数和厚度极明显低于对照组(P0.01);形态学观察显示,25 mg/L氟中毒小鼠睾丸生精细胞减少,100 mg/L Na F可导致生精细胞排列紊乱、各级生精细胞减少等。本试验可为氟的雄性生殖毒性研究提供直观、科学的参考。 相似文献
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《畜牧兽医学报》2016,(5)
比较观察同年龄(2岁)双峰驼(Camelus bactrianous)正常睾丸与隐睾的显微及超微结构特点。采用光镜和电镜制片及组织化学染色技术,比较双峰驼隐睾与正常睾丸的组织结构特点,进而用IPP图像分析软件进行定量分析。结果:光镜下双峰驼隐睾生精上皮由1~3层细胞组成,生精小管平均直径显著减小(P0.05),间质组织面积极显著增加(P0.01),间质/管腔面积比较正常组极显著增大(P0.01)。电镜观察,双峰驼正常睾丸生精小管固有膜层发育良好,相邻Sertoli细胞之间形成典型连接复合体;生精细胞之间多处形成细胞间小管和桥粒结构。隐睾生精上皮基膜增生明显,其上半桥粒结构不清晰,发育不成熟的Sertoli细胞相邻细胞膜间存在不完全的桥粒结构,可见相邻精母细胞间的胞质桥及Sertoli细胞与初级精母细胞之间桥粒连接;双峰驼正常睾丸Leydig细胞内滑面型内质网和粗面型内质网相延续;隐睾Leydig细胞形态不规则,胞内肿胀线粒体数量较多,内质网不发达;隐睾间质毛细血管基膜不清晰;血管内皮细胞之间可见缝隙连接。双峰驼隐睾生精小管组织结构主要变化为Sertoli细胞异常分化及精子发育阻滞;隐睾Sertoli细胞多为幼稚型,其与基膜连接的改变以及Sertoli细胞外质特化区形态结构及桥粒结构不完整影响了血-睾屏障构成;Leydig细胞内质网发育不均衡为其分泌功能与机体发育阶段关系研究提供思路。 相似文献
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采用单侧睾丸动脉结扎／再通以研究其对小鼠睾丸生精功能以及血清中抗精子抗体IgG、IgM含量的影响。实验选择健康成年BALB／c雄性小鼠于显微镜下行单侧睾丸动脉结扎。再于2h、4h、6h后开通血管。ELISA法检测血清抗精子抗体IgG、IgM水平,光镜观察睾丸生精小管内变化。结果表明：与非术侧相比,术侧睾丸生精小管内生精细胞不同程度地凋亡坏死,生精上皮脱落。各手术组血清中抗精子抗体IgG、IgM含量均有明显增长（P〈0．05）。实验结论：单侧睾丸动脉结扎／再通可引起睾丸生精细胞的凋亡坏死,同时血清抗精子抗体IgG、IgM含量增加。 相似文献
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以新生犊牛睾丸为实验对象,应用组合酶法进行支持细胞分离培养,并研究了冷冻保存后支持细胞的生长特性。结果表明:在细胞分离时,消化睾丸组织,分离曲细精管法所获得的细胞悬液中的有效细胞数高于组织剪碎法。支持细胞体外培养,4 h后开始贴壁,3~4 d铺满培养皿底壁,传代后细胞生长较快,2 d即可增殖一代。HE染色,胞质染色较淡,而细胞核染色较深,呈圆形或椭圆形位于细胞质中央或偏位,核仁明显。采用10%FBS+10%DMSO的DMEM液做冷冻液,对细胞进行冷冻保存时,支持细胞的复苏率在65%以上。解冻后的支持细胞体外培养,4h开始有细胞贴壁,24h后大部分细胞贴壁,3~4d铺满培养皿底壁。 相似文献
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为评价增精宝对雄性动物生精作用的影响,采用环磷酰胺致小鼠生精障碍为动物模型,考察不同剂量增精宝对环磷酰胺致小鼠生精障碍的保护作用。结果显示,高剂量组对小鼠的附睾、睾丸、储精囊和前列腺均有较显著的促进作用;低剂量组精子总密度和精子活率均差异显著(P<0.05);高剂量组精子活率差异显著(P<0.05);对小鼠睾丸各倍体生精细胞比例的影响,高剂量组小鼠睾丸生精小管较为饱满,管腔内存在大量精子,生精上皮较厚,可见各级生精细胞排列整齐,界膜明显,睾丸间质细胞排列整齐。结论表明,增精宝两种剂量均能够显著促进环磷酰胺导致小鼠生精障碍的保护作用,高剂量组对雄性小鼠的作用尤为显著。 相似文献
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This study aimed to assess the biometrics of the testes and the morphology of the seminiferous tubules of Crioulo horses. We studied 10 sexually mature stallions (3–6 years of age). After orchiectomy, testes were perfused with Karnovsky's solution and then embedded in glycol methacrylate. Testis sections (4 μm) were cut and stained with toluidine blue and a solution of 1% sodium borate. The histological images were digitized, and the morphometric analysis was performed using ImageJ software. The average weight of the stallions was 377.5 kg, and the average weight of both testicles was 162.9 g. The percentage of testicular parenchyma occupied by the seminiferous tubules and the intertubular tissue was 77.97% ± 6.34% and 22.03% ± 6.34%, respectively. The average tubular diameter was 205.00 ± 36.91 μm, whereas the average height of the seminiferous epithelium was 70.56 ± 2.82 μm. Average tubular length per testicle and average tubular length per gram of testicle were 4,085.10 ± 1,170.68 m and 26.09 ± 10.63 m/g, respectively. The characteristics of the eight stages of the seminiferous epithelium cycle were similar to those reported in other horse breeds. We conclude that the morphometry of the seminiferous tubules of Crioulo horse resembles what has been reported in other horse breeds. The volumetric proportion of the seminiferous tubules and the Leydig cells of the Crioulo horse is one of the highest ever reported for stallions. 相似文献
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试验旨在探究初情期前后生精上皮周期差异及睾丸发育过程中的形态学变化。通过测定15、30、60和90 d睾丸相关指数，结合睾丸组织形态学特征，判断香猪初情期，划分从江香猪生精上皮周期。结果显示，30 d的睾丸指数较15 d极显著升高（P<0.01），睾丸重、长轴及短轴的增长率分别为298.05%、66.42%和65.45%，60和90 d两个阶段睾丸重增长率相对稳定。形态学观察表明，从江香猪30 d时生精小管出现游离精子，完成第一次生精并进入初情期；与15 d相比，30 d生精小管面积和生精上皮厚度极显著增加（P<0.01），增长率分别为136.12%和40.19%，在60和90 d均处于稳定增长状态。睾丸细胞数统计显示，日龄增加不影响支持细胞（setoli cells，SC）数量（P>0.05），而30 d生殖细胞数（germ cells，GC）较15 d极显著增加（P<0.01）。相关分析结果发现，生殖细胞数量增加与生精小管面积增大、生精上皮厚度变化之间呈明显正相关（r=0.994；0.96）。根据生殖细胞组合形式差异，将初情期前后生精上皮分为3和8个阶段。初情期前生殖细胞以第一次减数分裂前期为主，A、B型精原细胞、SC、初级精母细胞（primary spermatocyte，Ps）、前细线期（preleptotene，PI）、细线期（leptotene，L）等生殖细胞在初情期前后生精上皮中均存在，而圆形精子（round spermatids，R）、延伸精子（elongating spermatid，E）、精子细胞（spermatozoa，S）仅存在于初情期后的生精上皮。本研究结果表明，从江香猪30 d初情，睾丸发育以生殖细胞和生精小管面积的迅速增加为主，该结果对从江香猪早熟性状挖掘、种猪选育及开发利用等有重要的指导意义。 相似文献
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Morphological features of the seminiferous epithelium in cat (Felis catus, L. 1758) testes 总被引：1，自引：0，他引：1
The aim of the present study was to assess the changes in the germinal epithelium in cats of different ages. Routine histological staining was applied to perform morphological and stereological examinations. The animals were divided into five groups according to age: under 8 months (n=28), 8-12 months (n=30), 12-36 months (n=33), 3-6 years (n=14) and older than 6 years (n=13). The appearance of the gonads of the males in the first group varied the most. The seminiferous tubules of the youngest cats consisted of a monolayer of supporting cells and a few spermatogonia. No tubular lumina were present, and the diameters of the seminiferous tubules reached 132.5 microm. We noted the typical arrangement of gametogenic cells with a tubule diameter of 191.83 microm in the second group. We observed multilayer germinal epithelia with the most significant production of gametes and a seminiferous tubule diameter of 202.61 microm in the third group. The diameters of the seminiferous tubules of the forth and fifth groups were 193.38 microm and 191.84 microm, respectively. The obtained data revealed that the most intensive morphological diversification of the seminiferous epithelium in cats occurs at about 7-8 months of age. The diameters of seminiferous tubules were highest in the third group of cats, and the activity of spermatogenesis of this group, expressed as the number of sperm per 10 mm(2), was also the most distinctive. The spermatogenesis process was most evident in cats between 12 and 36 months of age, which was also when the sperm concentration was highest per estimated surface. 相似文献
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60只雄性小鼠随机分为4组,每天分别腹腔注射0.2mL的0.9%生理盐水或7.5、15、30g/L CA,于14、21d测定曲细精管精子的生成及附睾内精子特性的相关数据。结果显示,随柠檬酸处理剂量的增加精子的生成数减少。14d时,对照组与15、30g/L CA组间、7.5、30g/L CA组间曲细精管内精子的生成差异显著（P〈0.05）;21d时,对照组与柠檬酸处理组间及7.5、15g/L CA与30g/L CA处理组间曲细精管内精子的生成有显著差异（P〈0.05）。柠檬酸对附睾精子品质的影响表现为：对照组与柠檬酸处理组间及柠檬酸处理组间精子密度和活动率差异均显著（P〈0.05）;对照组和15、30g/L CA处理组及15g/L CA与30g/L CA处理组间精子活力差异显著（P〈0.05）;精子的畸形率随柠檬酸浓度的增加而增加,15g/L CA处理组精子畸形率显著高于对照组（P〈0.05）,30g/LCA处理组显著高于对照组和7.5g/L CA（P〈0.05）处理组。组织形态学观察结果表明柠檬酸能破坏睾丸正常组织形态结构。柠檬酸高剂量组的生精小管内生精细胞排列疏松、紊乱,生精细胞间出现空隙;生精小管中的生精细胞脱落或退化,精原细胞、精母细胞和精子数量明显减少。研究表明,外源柠檬酸雄性小鼠具有显著的生殖遗传毒性。 相似文献
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牦牛和普通牛种间杂种公牛睾丸的组织学观测与研究 总被引：10，自引：1，他引：9
通过对牦牛和黄牛种间杂种公牛及未成年公牦掉睾丸的比较组织学观测，结果表明：杂种牛雄性不育的生殖隔离机制与生精小管及生精上皮细胞的发育状况、生精小管基膜的厚度、生精小管的间距、间质组织细胞的发育状况、间质组织与生精小管间形成大量组织液等因素有关。随种间杂交代数增加，睾丸组织的发育状况逐渐改善，精原细胞的死亡率相应降低，杂种3代（F3）已能完成减数分裂产生精子，并显示出精原细胞的死亡率的逐代减少与父本基因含量逐代增多有一定的相关性。 相似文献
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2018年4～6月,在武汉市黄陂区境内对青头潜鸭的繁殖行为进行了初步观察,以了解青头潜鸭在该区域的繁殖生境、繁殖行为及繁殖受胁情况等。采用定点观察法观察繁殖前期行为,对2巢青头潜鸭采用录像监控法记录孵化行为及出雏情况。测量了2巢及卵特征参数。发现青头潜鸭4月中旬开始分群,求偶交配,5月初基本完成配对,6月下旬出雏。巢为圆形碗状水面浮巢,巢材主要为草叶及草茎,巢内径(13. 5±7. 78) cm,巢外径(35. 5±6. 36) cm,巢高(21. 5±4. 95) cm,巢深(9. 5±0. 71) cm,窝卵数8～9枚,平均(8. 5±0. 71)枚(n=2);平均卵重(37. 32±4. 82) g,卵长径(50. 94±1. 75) mm,卵短径(37. 32±1. 41) mm(n=17)。孵卵期23～27 d,孵化率为17. 6%。青头潜鸭繁殖受人类活动干扰威胁较为严重,应当加大保护力度。 相似文献
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Lydka M Kotula-Balak M Kopera-Sobota I Tischner M Bilińska B 《Equine veterinary journal》2011,43(2):184-189
Reasons for performing study: Specific patterns of cytoskeletal filaments reflect a functional state of the cell. In testicular cells intermediate filaments (IFs) are of the vimentin type. Since it is known that Sertoli cells regulate the spermatogenic function in the male gonad, it became important to propose a system that could quantify the state of seminiferous tubular quality. To date, a Johnsen score system has never been used to equine testes. Objectives: To demonstrate the expression pattern of vimentin in testes of mature Arabian stallions and correlate its distribution with grade of seminiferous tubule impairment as indicated by a Johnsen score. Methods: For histological examination by the Johnsen method, routine haematoxylin‐eosin staining was used. Vimentin expression and its presence in testicular sections and testicular homogenates were detected by immunohistochemistry and western blot, respectively. Both analyses were performed qualitatively and quantitatively and further validated by ANOVA tests. Results: Distinct morphology of seminiferous tubules was found in testes harvested from 3 stallions. Vimentin in IFs was immunolocalised to the cytoplasm of Sertoli, Leydig and peritubular‐myoid cells. The intensity and pattern of the IFs staining was different in individual seminiferous tubules suggesting a correlation between vimentin expression and the severity of tubule degeneration. Qualitative results by immunohistochemistry and western blot were confirmed by further quantitative analyses. Conclusions: In equine testes, differential expression of vimentin was found to be correlated with the impairment of seminiferous tubules indicated by a decrease in Johnsen score. Potential relevance: The Johnsen score system may be a useful method to facilitate the identification of tubular alterations in the stallion testes. Combined histological and immunohistochemical approach may provide a detailed phenotypic classification of stallions with decreased fertility. 相似文献