大鼠睾丸支持细胞发育的形态学研究

对７０例不同胎龄及生后不同发育阶段Ｗｉｓｔａｒ大鼠睾丸支持细胞的发育进行了光镜、电镜研究。结果：１．依形态学特点，将支持细胞的发育过程初步分为４期：分化前期（胚胎１１～１５ｄ）、分化期（胚胎１５ｄ～生后１至２周）、成熟前期（生后２周～青春期前）、成熟期（青春期后）。２．胚胎及新生时，在支持细胞间见到两侧胞膜紧密相贴及不典型的桥粒连接等；自生后４周，见到紧密连接。３．自胚胎１７ｄ至成熟以后，在支持细胞与其周围的生殖细胞间见到大量类似桥粒和类似中间连接等不典型的细胞连接。４．胚胎１９ｄ及新生期，睾丸索内除支持细胞及生殖细胞外，尚可见一种特殊的细胞。     

咖啡因对胎鼠及乳鼠睾丸生殖细胞数量和PCNA表达的影响

为了深入了解咖啡因对胎儿、新生儿生殖细胞数量及其ＰＣＮＡ（增殖细胞核抗原）表达的影响，本实验采用低（０．３ｍＭ），中（０．６ｍＭ）、高（１．２ｍＭ）浓度Ｃａｆ．体外培养ＳＤ孕１８天胎鼠、０天及４天乳鼠睾丸组织块，培养时间分别为１周、２周、３周、观察Ｃａｆ．对睾丸内生殖细胞数量及其表达ＰＣＮＡ的影响。结果如下：（１）１８天胎鼠睾丸培养组织内生殖细胞数量受Ｃａｆ．影响最小，０天乳鼠次之，４天习受影响较     

咖啡因对胎鼠及乳鼠睾丸生殖细胞DNA合成影响的体外研究

目的：了解咖啡因对胚胎及新生时期生殖细胞合成ＤＮＡ的影响及其机制。方法：采用低（０３ｍｍｏｌ／Ｌ）、中（０６ｍｍｏｌ／Ｌ）、高（１２ｍｍｏｌ／Ｌ）浓度咖啡因体外培养ＳＤ孕１８ｄ胎鼠、０ｄ及４ｄ乳鼠睾丸组织块,培养时间分别为１、２、３周,用放射自显影、计算机图像分析的方法观察咖啡因对生殖细胞摄取［３Ｈ］－ＴｄＲ及ＤＮＡ含量的影响。结果：（１）１８ｄ胎鼠、０ｄ乳鼠、４ｄ乳鼠睾丸培养组织内生殖细胞受咖啡因影响依次增加。（２）浓度越高、培养时间越长,咖啡因对生殖细胞影响越明显。（３）生殖细胞数量的减少往往伴随ＤＮＡ含量和［３Ｈ］－ＴｄＲ摄取的降低。结论：高浓度咖啡因长时间培养后使生殖细胞数量减少可能与干扰细胞摄取ＤＮＡ前体物质,降低细胞内ＤＮＡ含量有关。     

淫羊藿苷调节雄性大鼠生殖功能

目的:探讨淫羊藿苷在环磷酰胺诱导的大鼠睾丸生精障碍动物模型中的作用,研究淫羊霍苷对睾丸功能的影响.方法:分为空白对照组、阴性对照组、模型组、淫羊藿苷治疗组;H-E染色观察睾丸组织结构变化,TUNEL方法检测睾丸生殖细胞凋亡,放射免疫法检测血清睾酮.结果:睾丸H-E染色切片观察显示模型组睾丸生精小管直径缩小,间距增宽,生精上皮变薄,生殖细胞数量减少,生精小管多未见精子形成,与空白对照组比较其生精小管结构变化显著;淫羊藿苷治疗组与模型组比较生精小管壁增厚,含有精子的生精小管明显增多.睾丸生精小管中生殖细胞凋亡的观察模型组与空白对照组比较其生殖细胞凋亡增多,变化显著;淫羊藿苷治疗组与模型组比较生精小管中生殖细胞凋亡数量明显减少.模型组与空白对照组比较血清睾酮明显降低;淫羊藿苷治疗组与模型组比较其血清睾酮明显增加.结论:淫羊藿苷对环磷酰胺诱导生精障碍的睾丸具有改善睾丸生精小管结构、减少生殖细胞凋亡、促进精子发生和间质细胞分泌睾酮的功能.     

小白鼠睾丸发育分化的组织学及组织化学观察

本文报道用组织学和组织化学方法观察小鼠睾丸胚胎天时睾丸即已有SDH活性,生后以间质细胞和精母细胞反应最强。4.AlP反应在胚胎时以生殖母细胞较强,生后以界膜反应较强。5.支持细胞、间质细胞AcP活性较强。6.生殖细胞和支持细胞5′-Nase呈阳性,间质反应极微。7.胚胎时,睾丸各种细胞ATPase反应均为阴性。生后界膜、支持细胞和生殖细胞渐出现活性且不断增强。8.胚胎14天睾丸内NSE即有微弱的反应,生后主要以间质细胞反应强烈。至生后的发育。结果表明:1.核糖核酸以代谢旺盛的生殖母细胞最为丰富,支持细胞也较多,精子细胞较少。2.胚胎14天时睾丸各种细胞均有丰富的糖原颗粒,16天起仅分布于间质,性索内糖原阴性。3.胚胎14     

淫羊藿苷对环磷酰胺诱导生精障碍大鼠下丘脑垂体睾丸轴的影响

目的:探讨淫羊藿苷在环磷酰胺诱导的大鼠睾丸生精障碍动物模型中的作用,研究淫羊藿苷对下丘脑-垂体-睾丸轴的影响。方法:大鼠分为空白对照组、阴性对照组、模型组、淫羊藿苷治疗组;H-E染色观察睾丸组织结构变化,TUNEL检测睾丸生殖细胞凋亡,放射免疫法检测血清睾酮、卵泡刺激素(FSH)、黄体生成素(LH),SABC免疫组织化学研究下丘脑正中隆起促性腺激素释放激素(GnRH)神经纤维。结果:睾丸H-E染色切片观察显示模型组睾丸生精小管直径缩小,间距增宽,生精上皮变薄,生殖细胞数量减少,生精小管多未见精子形成,与空白对照组比较其生精小管结构变化显著;治疗组与模型组比较生精小管壁增厚,含有精子的生精小管明显增多。模型组与空白对照组比较,睾丸生精小管中生殖细胞凋亡显著增多;治疗组与模型组比较生精小管中生殖细胞凋亡数量明显减少。血清睾酮检测结果显示模型组与空白对照组比较血清睾酮明显降低;治疗组与模型组比较其血清睾酮明显增加;血清FSH、LH水平各组间无差异。与空白对照组比较,模型组正中隆起GnRH阳性纤维增多,染色深,光密度显著增高;治疗组与模型组比较其GnRH阳性纤维明显减少,染色浅。结论:淫羊藿苷具有改善睾丸生精小管结构、减少生殖细胞凋亡、促进精子发生和间质细胞分泌睾酮的功能,降低GnRH在正中隆起的蓄集。     

人胎睾丸的组织发生

用7～38周人胎35例,取睾丸固定于 Carnoy 液,石蜡包埋,以 HE、PAS 反应及甲绿哌喏宁法染色。7周初的性腺尚未分化,可见散在的原始生殖细胞。13周睾丸特征已明显,白膜厚,睾丸素清楚,索间有密集的嗜酸性间质细胞。14.5周部分睾丸索出现小腔,而成管状。睾丸索包含大而着色淡的原始生殖细胞及色深的原始支持细胞。胎早期原始生殖细胞含有丰富的糖原颗粒。睾丸门、白膜及表面上皮内均见散在或成群的生殖细胞。睾丸间质细胞分幼稚型、成熟型及退化型。13～15周以幼稚型为主;16～18周以成熟型为主,成熟型的胞质内含有 RNA 颗粒;20周后退化型增多,成熟型减少。38周时,睾丸间质细胞单个或成行存在,数量大为减少。     

超声波对小白鼠生精上皮的影响

用功率为５Ｗ／ｃｍ２、频率为１．１０ＭＨｚ的超声波照射小白鼠睾丸５分钟（实验组１）、１０分钟（实验组Ⅱ），分别于处理后２４小时，４８小时及７天时间切取睾丸组织，制作石蜡切片，在光镜下观察生精上皮的组织学变化并与对照组进行比较。结果显示：（１）小白鼠睾丸经超声波照射后，曲细精管萎缩，管径变小；生精上皮变薄，精子发生时相消失；生精细胞减少，没有精子形成。（２）超声波照射１０分钟对生精上皮的损伤比照射５分钟更为严重。（３）超声波照射后２４小时，生精上皮即受到破坏，精子细胞减少；照射后４８小时，上皮受损伤的程度增大；照射后７天时间，曲细精管的组织学结构开始恢复，但仍无精子形成。（４）超声波对生精上皮的影响主要限于精母细胞、精子细胞和精子，而精原细胞与支持细胞没有明显变化。上述结果表明，超声波能够抑制小白鼠的精子发生，该抑制作用可能可逆。     

大鼠睾丸间质细胞发育的形态学研究及立体学定量分析

对６３例胚胎期至生后不同发育时期Ｗｉｓｔａｒ大鼠睾Ｌｅｙｄｉｇ细胞进行光镜、电镜观察及立体学分析。结果表明：１．大鼠睾丸Ｌｅｙｄｉｇ细胞从胚胎１５ｄ出现，１７ｄ初具分泌类固醇激素的结构特点。２．Ｌｅｙｄｉｇ细胞存在两个细胞增殖峰，一个在胚胎１７￣１９ｄ，另一个在生后４周。两峰之间有一个低谷（约在生后第２周）。从胚胎至生后，细胞核的体密度逐渐减小，细胞质、线粒体及滑面内质网的体密度逐渐增大，脂滴的体     

乙型肝炎病毒携带者睾丸中HBV DNA的检测研究

目的检测乙型肝炎病毒携带者的睾丸活检组织是否携带乙肝病毒。方法11例乙型肝炎病毒携带者的睾丸活检组织制成石蜡切片,应用特异性乙肝病毒DNA（HBV DNA）的探针以原位杂交法检测这些患者的睾丸组织切片。结果11例患者中有5例检测到HBV DNA,分布于间质细胞、支持细胞及各级生精细胞的细胞核。结论男性乙肝病毒携带者存在经生殖细胞向子代垂直传播乙肝的可能。     

Male germ cell death in mouse testes: possible involvement of Fas and Fas ligand

Loss of germ cells is very common during various stages of mammalian spermatogenesis. Although cell death, particularly apoptosis, has been implicated, our understanding of the mechanisms underlying germ cell death is still limited. In order to elucidate the extent and mechanism of germ cell death, this review first covers what is known of germ cell degeneration in the normal testes of fetal, neonatal, and adult mice from electron microscopy (EM) and from terminal dUTP nick-end labeling (TUNEL) staining. The issue of whether the Fas and Fas ligand (FasL) system is involved in the induction of germ cell apoptosis in normal and damaged testes is then addressed, including consideration of both the ischemia-reperfusion model of testicular torsion and the estrogen-treated testis model of environmental endocrine disruption. Finally, this review proposes that different molecular pathways may be triggered to induce male germ cell apoptosis, depending upon the physiological and pathological states of the germ cells.     

Germ cell apoptosis and its molecular trigger in mouse testes

Germ cell apoptosis is very common during various stages of mammalian testicular development. However, our understanding of the mechanisms underlying male germ cell apoptosis is still limited. This review firstly covers the general features of germ cell death in normal testes of fetal, neonatal, and adult mice from electron microscopy (EM) and terminal dUTP nick-end labeling (TUNEL) staining. The issue of whether the Fas and Fas ligand (FasL) system and/or the Bax and Bcl-2 system is involved in the induction of germ cell apoptosis in normal and damaged testes will then be addressed, including a special consideration of the ischemia-reperfusion model, the endocrine disruptor-treated model, and others. Finally, this review will propose that the process of normal spermatogenesis seems skillfull in taking advantage of apoptotic processes of germ cells and that different molecular pathways may be triggered to induce male germ cell apoptosis, depending upon the physiological and pathological states of germ cells.     

蝌蚪提取液诱导HL60细胞分化和凋亡的实验研究

目的 为了寻找新的肿瘤细胞分化诱导剂,探讨蝌蚪醚提取液（Ｔ８７１２）的抗肿瘤作用。方法 将ＨＬ６０细胞培养于Ｔ８７１２和ＲＰＭＩ１６４０按１：２００（ｖ／ｖ）混合的培养基中。结果 Ｔ８７１２能诱导ＨＬ６０细胞向单核／巨噬细胞方向分化,表现为生长抑制,ＮＢＴ还原能力提高,酸性非特异性酯酶（ＡＮＡＥ）活性增加,形态学观察类似单核／巨噬细胞。当Ｔ８７１２与ＲＰＭＩ１６４０培养基按１：１００（ｖ／ｖ）混合     

小鼠肾脏发育中的细胞凋亡

目的：研究小鼠肾脏发育过程中的细胞凋亡规律及形态学特点,方法：应用光镜、电镜技术和TUNEL法分别对不同胚龄、生后日龄小鼠肾脏细胞凋亡进行了观察。结果：皮质凋亡细胞多出现在生肾区S小体之间和是肾小体内,凋亡高峰期在胚龄14－18d之间,髓质凋亡细胞出现在肾小鼠管上皮内,凋亡高峰期在生后7d 左右。超微结构观察可见皮质和髓质凋亡细胞主要表现为核固缩,染色质凝集,细胞皱缩。皮质和髓质凋亡细胞结局为,被邻近细胞吞噬,或脱落到肾小管腔内,结论：小鼠肾脏发育过程中确有细胞凋亡;皮质中细胞凋亡与生肾区的出现和肾小体发育完善有关,髓质中细胞凋亡与髓质中肾小管和集合小0管的有发育完善有关。     

Induction of apoptosis involving multiple pathways is a primary response to cyclin A1-deficiency in male meiosis.

The meiotic arrest in male mice null for the cyclin A1 gene (Ccna1) was associated with apoptosis of spermatocytes. To determine whether the apoptosis in spermatocytes was triggered in response to the arrest at G2/M phase, as opposed to being a secondary response to overall disruption of spermatogenesis, we examined testes during the first wave of spermatogenesis by terminal deoxynucleotidyl transferase-mediated deoxyuridinetriphosphate nick end-labeling (TUNEL) staining. We observed enhanced apoptosis coinciding with the arrest point in postnatal day 22 tubules, with no overt degeneration. Along with activation of caspase-3, an increase in the levels and change of subcellular localization of Bax protein was observed in cyclin A1-deficient spermatocytes, which coincided with the detection of apoptosis. As p53 is implicated in the activation of Bax-mediated cell death, we generated mice lacking both cyclin A1 and p53. Although the absence of p53 did not rescue the meiotic arrest, there was a decrease in the number of apoptotic cells in the double-mutant testes. This finding suggested that p53 may be involved in the process by which the arrested germ cells are removed from the seminiferous tubules but that other pathways function as well to ensure removal of the arrested spermatocytes.     

遗传性视网膜变性rd小鼠及其感光细胞凋亡研究

目的 研究遗传性视网膜变性rd小鼠感光细胞层的发育变化及细胞凋亡。方法 对出生后5d到40d的rd小鼠及对照小鼠视网膜感光细胞层进行光镜及超微结构观察、TUNEL法检测及形态计量学分析。结果 与同龄对照鼠相比,rd小鼠出生后第10d视网膜开始变性,尔后1周内感光细胞迅速减少,第18d时只残留一层视椎细胞。rd小鼠出生后第10d感光细胞层开始出现TUNEL染色阳性细胞,第14d及16d达到高峰。电镜下变性高峰期rd小鼠视网膜感光细胞层可见大量浓缩核、染色质边聚及凋亡小体。结论 rd小鼠视网膜感光细胞在发育过程中变性,并通过凋亡的方式死亡。     

小鼠出生后肾脏发育过程中的细胞增殖与凋亡

目的：观察小鼠出生后肾脏发育过程中增殖细胞核抗原(PCNA)的表达及细胞凋亡的特征,探讨出生后小鼠肾脏发育过程中细胞增殖与凋亡的规律及其关系。方法：应用免疫组织化学技术和原位末端标记法(TUNEL法)分别检测小鼠出生后1～70d肾脏中PCNA阳性的细胞和凋亡细胞。结果：小鼠出生后1～70d,皮质中的肾小体、肾小管、髓放线以及髓质中的肾小管和集合管的细胞,早期增殖活跃,随着肾脏发育成熟而表达逐渐减弱。同时,也存在着细胞凋亡现象,且凋亡高峰一般出现在增殖高峰之后。结论：细胞增殖与凋亡在小鼠生后肾脏发育的整个过程中普遍存在,生后1～7d细胞增殖旺盛,增殖高峰之后出现凋亡高峰,生后28～70d两者活动均减弱。     

饮酒对小鼠睾丸生精小管一氧化氮合酶、增殖细胞核抗原表达及细胞凋亡影响的研究

目的　探讨酒精对小鼠睾丸的组织结构、内皮型一氧化氮合酶 (eNOS)、增殖细胞核抗原 (PCNA)及细胞凋亡的影响。　方法　用 5 %、10 %及 15 % 3种不同浓度的酒精作用于 2 2d龄小鼠 ,取睾丸做石蜡切片、HE染色 ;用免疫组织化学方法检测睾丸eNOS、细胞增殖的变化 ;TUNEL法检测细胞凋亡的变化 ,并进行统计学分析。　结果　随着酒精浓度的增大 ,睾丸组织结构发生明显改变 ,生精小管的直径逐渐减小 ,eNOS阳性细胞面积密度逐渐增大 ,单位面积内PCNA阳性细胞和凋亡细胞数目增加 ,高浓度酒精组与其他组差异极显著 (P <0 0 1)。　结论　酒精可使生精小管的直径变小 ,eNOS及PCNA表达增强 ,凋亡细胞增加并随酒精浓度的增大而变化加重。这可能是过量饮酒导致生精细胞减少 ,生殖能力降低的重要因素。     

Diverse cell death pathways result from a single missense mutation in weaver mouse.

Neuronal death affects selectively granule cell precursors of the cerebellum and the dopaminergic neurons of midbrain in the weaver mutant mouse. The weaver phenotype is associated with a missense mutation in the gene coding for the GIRK2 potassium channel, which results in chronic depolarization. Using DNA gel electrophoresis, electron microscopy (EM), the in situ end-labeling (ISEL) technique at the light and EM level, and immunohistochemistry for apoptosis-related proteins c-Jun and proliferating cell nuclear antigen (PCNA), we have investigated the mechanisms of cell death in cerebellum and substantia nigra. Between postnatal day P1 and P21, in the external germinal layer of the cerebellum, most degenerating granule cell precursors were found to aggregate to form clusters. Degenerating cells exhibited strong nuclear staining for ISEL, c-Jun, and PCNA and had a typical apoptotic morphology by EM. Increased c-Jun and ISEL staining were also occasionally seen in Purkinje cells. Between P14 and P21, when dopaminergic neurons start to degenerate, staining for ISEL, c-Jun, and PCNA in weaver substantia nigra was the same as in controls. By EM, however, we found only in weaver mice numerous dopaminergic cells that showed extensive vacuolar and autophagic changes of cytoplasm, preservation of membrane and organelle integrity, and absence of chromatin condensation or DNA fragmentation by EM-ISEL. The combination of vacuolar and autophagic changes identifies a novel type of non-necrotic, nonapoptotic cell death. After biochemical analysis of DNA, a clear-cut laddering, suggestive of oligonucleosomal fragmentation, was present in samples from weaver cerebellum. Cell death diversity appears to be influenced by specific features of target cells. These findings may be relevant for understanding the mechanisms of cell death in neurodegenerative diseases.     

Apoptosis of the hyaloid artery in the rat eye

The hyaloid artery is a vestigial vessel situated on the optic nerve extending to the posterior surface of the lens in the vitreous cavity of the eye. We studied the nature and pattern of cell death during regression of the hyaloid artery. The cells comprising the hyaloid artery appear to be alive for 20 days after birth in the rat, and cell death during regression of the hyaloid artery has the characteristics of apoptosis. We observed apoptotic bodies containing condensed chromatin and identified the hyaloid vessels as targets of macrophage-mediated remodelling. Using the “TUNEL method” for labeling fragmented DNA in vascular cells, we assessed the pattern of apoptotic cell death during hyaloid artery regression. Our study demonstrated the appearance of apoptosis in pericytes as well as endothelial cells during regression in the vasculature. In pericytes, apoptosis appeared to begin or to occur more frequently than in endothelial cells. Both morphological and TUNEL analyses indicated that capillary apoptosis occurs mainly from day 10 to day 20 after birth rather than from the 1st day. Macrophages were present near the hyaloid artery and these may influence apoptosis.