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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(P0.01),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(P0.01),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(P0.05),而30 d生殖细胞数(germ cells,GC)较15 d极显著增加(P0.01)。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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T1R1和T1R3在从江香猪附睾发育中的表达模式 总被引：1，自引：0，他引：1
为研究味觉受体第一家族亚型1（T1R1）和3（T1R3）在从江香猪附睾发育过程中的表达模式,探讨味觉受体在哺乳动物雄性生殖机能中可能发挥的作用及潜在医学价值,本试验以从江香猪附睾组织为研究对象,分析附睾发育4个关键时期：初情前（15 d）、初情时（30 d）、初情后（60 d）和性成熟期（180 d）T1R1与T1R3的差异表达。采用实时荧光定量PCR、免疫组织化学（IHC）和Western blot检测两个味觉受体在不同日龄从江香猪附睾组织中转录、翻译水平的变化及其分布情况。RT-qPCR结果表明：TAS1R1与TAS1R3 mRNA在从江香猪附睾初情前（15 d）至性成熟期（180 d）表达量逐渐增加,且任意两个时期间差异极显著（P<0.01）。Western blot结果显示,T1R1/T1R3蛋白在180 d表达量最高,在15 d表达量最低,两者之间差异显著（P<0.05）,平均表达丰度依次为180 d > 30 d > 60 d > 15 d。IHC结果显示,T1R1和T1R3蛋白在各日龄组从江香猪附睾组织均有分布,其中T1R1蛋白主要在上皮细胞膜上,尤其是基细胞和窄细胞;而T1R3蛋白主要在微绒毛、环状空泡和精子呈强阳性表达。综上,本研究发现不同日龄从江香猪附睾的T1R1/T1R3表达从15 d逐渐增加,至性成熟达到峰值,这一表达变化与附睾上皮基细胞和窄细胞及微绒毛的T1R1/T1R3的差异表达有关,这些特殊的表达模式与附睾生理功能存在时间关联,故推测T1R1和T1R3参与附睾内精子成熟和储存的调节过程。 相似文献
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细胞凋亡可及时清除机体内多余和受损伤的细胞，以维持组织器官的稳定性。为探讨凋亡过程在公猪附睾发育过程中发挥的潜在作用，本实验以从江香猪为研究对象，利用实时荧光定量PCR(qRTPCR)、免疫组织化学（IHC）和WesternBlot技术分析了附睾15d（初情前期）、30d（初情期）、60d（初情后期）和180 d（性成熟期）4个时期凋亡相关因子Caspase-3、Bax和Bcl-2的差异表达模式。qRTPCR结果显示：Casp3和Bax基因mRNA在从江香猪60d表达量最高，Casp3在15d表达量最低，15d和180 d差异不显著；Bax在180 d表达量最低，且15 d、60 d和180 d差异不显著；Bcl-2在60 d表达量最低，在15 d表达量最高，4个日龄段差异显著。免疫组化结果显示：Caspase-3蛋白在30 d主要表达在附睾管腔的微绒毛，而在180 d主要表达在管腔中的精子上。Western Blot结果显示：Caspase-3蛋白表达丰度依次为60 d>30 d>15 d>180 d，且各日龄段差异显著。综上，本实验发现不同日龄从江香猪附睾凋亡相关... 相似文献
4.
睾丸是雄性动物的生殖器官，具有产生雄激素和雄性生殖细胞的功能。因此，睾丸的发育形态对猪养殖生产具有重大影响。本实验选取DLY公猪胚胎期（妊娠105 d）和成年期（日龄200 d）睾丸为研究对象，对睾丸组织进行形态学观察和高通量转录组测序，并对差异表达基因进行功能富集分析。形态学观察结果表明：相较于雄性胎猪，成年公猪生精小管直径更大，生精上皮更厚，已有完整的生精结构，并可见各发育阶段精子细胞。转录组测序结果表明，共鉴定出6 296个差异表达基因。差异基因显著富集在精子发生、多细胞生物发育和细胞分化等生物过程，可能在代谢、细胞黏附、细胞周期和AMPK等通路发挥作用。本研究结果揭示了公猪胚胎期和成年期睾丸组织的形态学和转录组学的差异，为后期猪睾丸发育研究提供参考。 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
7.
【目的】探究不同日龄伊拉兔睾丸和附睾组织在性成熟前的变化规律,为其初情期及性成熟的判定提供组织学依据。【方法】将45只伊拉兔随机分为9组,每组5只,从30日龄开始每隔15日采集1组试验兔的睾丸及其附睾,运用形态学与组织解剖学方法对其生长发育规律进行研究分析。【结果】随着日龄的增加,睾丸指数、睾丸重、睾丸长径、睾丸短径及厚径等指标逐步增加,且150日龄时各指标均显著高于其他组（P＜0.05）;30、45、60日龄睾丸内生精小管排列稀疏,75、90、105日龄时睾丸间质成分逐渐增多,管腔逐步形成,120、135、150日龄时间质细胞进一步增生并成群分布,90日龄起出现圆形和长形未成型精子,120日龄的睾丸生精小管、附睾管腔中出现少量成型精子,150日龄时有大量精子密集分布于睾丸管腔内,且在120、135、150日龄时生精小管面积、上皮细胞厚度、支持细胞数均显著大于其他组（P＜0.05）;30日龄以上各组间质细胞个数在各组间差异不显著（P＞0.05）,但均显著高于30日龄;120、135、150日龄时附睾头、体、尾的直径均极显著高于其他组（P＜0.05）,而附睾头的柱状上皮厚度和纤毛长度则从105日龄起就显著高于30、45、60、75和90日龄组（P＜0.05）。【结论】伊拉兔睾丸和附睾随日龄增加同步发育,在120日龄时达到初情期,在150日龄时睾丸和附睾已发育成熟,基本达到性成熟。 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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《中国畜牧杂志》2021,(7)
为探讨来曲唑对香猪睾丸细胞波形蛋白表达及分布的影响,试验将14日龄、体重3 kg的12头雄性香猪随机分为3组,对照组不做任何处理,试验组分别经口投喂0.1、0.2 mg/kg来曲唑,每3 d投喂1次,连续投喂15次,最后一次投喂7 d后取左侧睾丸,应用免疫组织化学方法检测波形蛋白在香猪睾丸细胞中的表达与分布。结果表明:波形蛋白在睾丸生精小管细胞胞浆中表达,试验组睾丸生精小管细胞总数较对照组减少(P0.01);而波形蛋白表达细胞阳性率上升(P0.01),且随着来曲唑剂量的增加而增加(P0.01)。来曲唑可通过降低雌激素水平来调节睾丸生精小管细胞数及波形蛋白表达,说明来曲唑可能促进了香猪睾丸生精小管细胞中波形蛋白的表达,进而影响睾丸生精小管细胞的生长发育,但其作用机制机理还需进一步研究。 相似文献
10.
贵州香猪睾丸发育中支持细胞和生精细胞数量变化观察 总被引：1，自引：0，他引：1
为了解香猪睾丸发育过程中生殖细胞和支持细胞数变化规律,用手术取出30,40,50,70,90和110日龄(每个年龄组n=3～4)香猪右侧睾丸,经中性多聚甲醛固定,石蜡包埋,组织切片采用免疫组化SP法,用单克隆抗体GATA-4检测睾丸支持细胞的特异生长转录因子-4,经DAB显色、苏木素复染。光镜下核呈棕色者为支持细胞,核呈蓝色者则为生殖细胞;经显微照相并用Scion image软件测量生精小管及管壁面积。结果:30～110日龄睾丸支持细胞数维持在稳定水平(P>0.05),而生殖细胞数随日龄增加而增多,70日龄生殖细胞数快速增多(P<0.05),持续到110日龄。同样,从70日龄开始睾丸生精小管和管壁面积显著性增大(P<0.05)。香猪睾丸支持细胞快速增殖发生在30日龄前,而生殖细胞数随着日龄的增长而增多。 相似文献
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The purpose of this study was to analyze the distribution and expression of peptidergic neurotransmitters protein gene product 9.5 (PGP9.5) and neuropeptide Y (NPY) in cryptorchidism and testicular tumors of dogs,compare them with normal testicular tissues of the same age,and provide reference for clinical diagnosis of malignant transformation in testicular tumors of dogs.HE staing,Masson trichrome staining,Gomori silver staining and toluidine blue staining were used to observe the tissue characteristics of reticular fibers,collagen fibers and mast cells.Immunohistochemical SP method and immunofluorescence combined with IPP were used to analyze the expression and localization of PGP9.5 and NPY in tissues.The results showed that the seminiferous epithelium of normal dog testis was composed of 4-7 layers of spermatogenic cells and Sertoli cells,and the distribution of collagen fibers and reticular fibers in interstitial tissue was sparse.The thickness of collagen fibers in the basement membrane of cryptorchidism seminiferous tubules increased,the nucleus of Sertoli concentrated at the base of seminiferous tubules,and the interstitial reticular fibers increased.The tissue structure of testicular tumor was unclear,collagen fibers and reticular fibers were irregularly distributed,and mast cells increased significantly compared with normal and cryptorchid groups.The immunofluorescence results showed that PGP9.5 was moderately positive in Leydig cells of normal testis,no significant expression in spermatogenic cells,strongly positive in Leydig cells and spermatogenic cells of cryptorchidism,and occasional expression in testicular tumors.NPY was occasionally expressed in normal testicular Leydig cells,but not in spermatogenic cells,strong positive expression in Leydig cells and seminiferous epithelium of cryptorchidism,high density and strong positive expression in interstitial vessels,and no obvious expression in testicular tumors.Immunohistochemical statistics showed that the expression of PGP9.5 and NPY in testicular tumor tissue were extremely significantly lower than that in normal group (P<0.01),while the expression of PGP9.5 and NPY in cryptorchidism group were significantly or extremely significantly increased (P<0.05;P<0.01).Therefore,the expression of PGP9.5 and NPY in cryptorchidism of dogs was increased suggesting that the cryptorchidism of dogs had a tendency to develop into a tumor,and was related to the degree of malignant transformation of tumor. 相似文献
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试验旨在分析肽能神经递质蛋白基因产物9.5（protein gene product 9.5,PGP9.5）和神经肽Y（neuropeptide Y,NPY）在犬隐睾及睾丸肿瘤中的分布和表达,并与同年龄正常睾丸组织进行比较,为认识犬睾丸肿瘤恶变临床诊断提供参考。应用HE染色、Masson三色染色、Gomori银浸染、甲苯胺蓝染色观察各组织中网状纤维、胶原纤维及肥大细胞等组织特征,采用免疫组织化学SP法及免疫荧光法结合IPP统计分析PGP9.5和NPY在组织中的表达及定位。结果显示,正常犬睾丸生精上皮由4~7层生精细胞及Sertoli细胞构成,间质组织胶原纤维和网状纤维分布稀疏。隐睾生精小管基底膜胶原纤维厚度增加,Sertoli细胞核浓缩位于生精小管基底,间质网状纤维增多。睾丸肿瘤组织结构不清晰,胶原纤维和网状纤维无规则分布,肥大细胞较正常组及隐睾组显著增多。免疫荧光定位表明,PGP9.5在正常睾丸Leydig细胞中呈中等阳性表达,生精细胞中无明显表达;隐睾Leydig细胞及生精细胞中呈强阳性表达;睾丸肿瘤中偶有表达。NPY在正常睾丸Leydig细胞中偶见阳性表达,生精细胞中无表达;隐睾Leydig细胞及生精上皮中无表达,间质小血管管壁呈高密度强阳性表达;睾丸肿瘤组织中无明显表达。免疫组化统计表明,睾丸肿瘤组织中PGP9.5和NPY较正常组极显著降低（P<0.01）,隐睾组PGP9.5和NPY表达显著或极显著增加（P<0.05;P<0.01）。因此,犬隐睾时PGP9.5及NPY的表达增高,提示犬隐睾时已有发展为肿瘤的趋势,且与肿瘤恶变程度相关。 相似文献
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MENG Lijie WANG Weiyong YANG Yi XU Yongjian FENG Xianzhou HUANG Yong GAO Yi GONG Ting 《畜牧兽医学报》1956,51(11):2720-2730
The aim of this experiment was to study the expression pattern of taste receptor family 1 subtypes 1 (T1R1) and 3 (T1R3) during epididymal development of Congjiang Xiang pig, and to explore the possible role of these taste receptors in mammalian male reproductive function and its potential medical value. In this study, the differential expressions of T1R1 and T1R3 in epididymis at 4 key developmental periods (neonatal (15 d), peri-puberty (30 d), puberty (60 d) and sexual maturity (180 d)) of Congjiang Xiang pigs were analyzed. RT-qPCR, immunohistochemistry (IHC) and Western blot were used to detect the changes and distribution of the two taste receptors in epididymis of Congjiang Xiang pigs at different ages. The results of RT-qPCR showed that the expression of TAS1R1 and TAS1R3 mRNA increased gradually from neonatal (15 d) to sexual maturity (180 d), and there was a significant difference between each period (P<0.01). The results of Western blot showed that the expression of T1R1/T1R3 protein was the highest on the 180 d and the lowest on the 15 d. The average protein abundance of T1R1/T1R3 was as follows: 180 d > 30 d > 60 d > 15 d. The results of IHC showed that T1R1 and T1R3 proteins were distributed in the epididymis of Congjiang Xiang pigs at 4 periods, in which T1R1 protein was mainly concentrated in epithelial cell membrane, especially in basal and narrow cells, while T1R3 protein was strongly positive in stereocilia, annular vacuoles and spermatozoa. In summary, the expression of T1R1/T1R3 in the epididymis of Congjiang Xiang pigs increased gradually from 15 d to the peak of sexual maturation, which was related to the differential expression of T1R1/T1R3 in epithelial basal cells, narrow cells and stereocilia of epididymis. These special expression patterns were time related to the physiological function of epididymis, so it is speculated that T1R1/T1R3 are involved in the regulation of sperm maturation and storage in epididymis. 相似文献
14.
Cystic testicular degeneration was induced in groups of ducklings by feeding furazolidone at 250, 400, 550, or 750 mg per kilogram of feed (ppm) for 4 weeks. In normal ducklings, tall, columnar Sertoli cells predominated in the seminiferous epithelium. In treated ducklings, the mildest ultrastructural alteration was cytoplasmic vacuolation of Sertoli cells. In birds with more severely affected tubules, cytoplasmic vacuolation was more severe and the cells were cuboidal or rounded. The rounded cells desquamated into the tubular lumens, eventually undergoing cytolysis. In birds with the most severely affected tubules, only extensively flattened epithelial cells lined the extremely dilated seminiferous tubules. The interstitium was edematous and compacted between the expanded tubules. Many of the testicular ultrastructural alterations in furazolidone-intoxicated ducklings were similar to those described in the testicles of sodium-intoxicated cockerels. 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.
Hiroyuki Kaneko Kazuhiro Kikuchi Nguyen Thi Men Junko Noguchi 《Animal Science Journal》2019,90(2):158-166
Testicular xenografting, combined with cryopreservation can assist conservation of the genetic diversity of indigenous pigs by salvaging germ cells from their neonatal testes. Using Meishan male piglets as an example, we examined whether testicular tissue would acquire the ability to produce sperm after cryopreservation and grafting into nude mice (MS group). For comparison, testicular tissue from neonatal Western crossbreed male piglets was used (WC group). Sixty days after xenografting (day 0 = grafting), MS grafts had already developed seminiferous tubules containing sperm, whereas in the WC grafts, sperm first appeared on day 120. The proportion of tubules containing spermatids and sperm was higher in the MS group than in the WC group between days 90 and 120. Moreover, in vitro‐matured porcine oocytes injected with a single sperm obtained from the MS group on day 180 developed to the blastocyst stage. The blastocyst formation rate after injection of the xenogeneic sperm was 14.6%, whereas the ratio in the absence of such injection (attributable to parthenogenesis) was 6.7%. Thus, cryopreserved Meishan testicular tissue acquired spermatogenic activity in host mice 60 days earlier than Western crossbreed tissue. Such xenogeneic sperm are likely capable of generating blastocysts in vitro. 相似文献
17.
为了探索抑制素α基因(inhibin-α,INHA)与从江香猪繁殖性状之间的相关性,试验采用特异性聚合酶链式反应(PCR)技术克隆从江香猪INHA基因,测定其核苷酸序列,通过等位基因特异性PCR(allele-specific PCR,AS-PCR)方法检测从江香猪低产群与高产群之间INHA基因的多态性变化,以实时荧光定量PCR技术检测高产、低产从江香猪卵巢组织中INHA基因的表达量。结果表明,从从江香猪基因组中成功克隆了INHA基因,编码区完整,全长1095 bp,编码364个氨基酸;经比对发现, INHA基因外显子2序列中存在2个候选SNPs位点(G359A和A373G)。经大样本检测,从江香猪高产群与低产群之间2个候选SNPs位点的基因频率没有明显差异;相比之下,高产从江香猪卵巢中INHA基因的表达量较高。研究结果提示,从江香猪INHA基因结构保守,可能主要通过基因的表达量变化调节从江香猪卵巢的生长和卵泡的发育。 相似文献
18.
To reveal the relationship between inhibin-α(INHA) gene and the reproductive traits of Congjiang Xiang pig, INHA gene was cloned and sequenced taking the genomic DNA of Congjiang Xiang pigs as templates by polymerase chain reaction(PCR) method.The polymorphisms of INHA gene were tested in Congjiang Xiang pig populations with high-litter size and low-litter size using allele-specific PCR(AS-PCR) method.The expression profile of INHA gene in ovaries was detected from Congjiang Xiang pigs with high-litter yiled or low-litter yiled by Real-time PCR method.The complete coding region of INHA gene was 1095 bp in length, which coded for 364 amino acid residues.Compared with the known sequence, two candidate sites, G359A and A373G, were found out from exon 2 region of INHA gene in Congjiang Xiang pig.After investigation for the two sites in a large population, the frequency of alleles between two populations was not significant and without obvious relativity with the litter yiled of Congjiang Xiang pig.However, the INHA mRNA level in the ovary of Congjiang Xiang pig with high-litter yiled was higher than that with low-litter yiled.It suggested that INHA gene was much conserved, INHA gene expression level might be concerned for the regulation of ovary growth and follicle development in Congjiang Xiang pig breed. 相似文献