羊精子在成熟和获能过程中表面的凝集素标记变化

用辣根过氧化物酶结合的麦芽凝集素和大豆凝集素，对绵羊睾丸和附睾内精子及体外获能精子细胞化学标记，麦芽凝集素在睾丸内精子的顶体区有强标记，在附睾头前段精子顶体区的标记减弱，但在附睾头后段精子顶体区的标记又增强，且尾部也出现弱标记，获能后部分精子的顶体后区出现弱至中等标记。大豆凝集素在睾丸内精子的顶体区有中等标记，在附睾体出现强标记，但获能后标记减至很弱，结果提示，麦芽凝集素记糖复合物可能与肥精有关。     

猕猴精子获能和顶体反应过程中Ca^2＋和Ca^2＋—ATPase定位及咖啡因…

用焦锑酸钾原位沉淀法和枸橼酸铅捕获研究了猕猴精子获能和顶体反应过程中的Ｃａ＾２＋分布和Ｃａ＾２＋－ＡＴＰａｓｅ活性。获能前精子头部Ｃａ＾２＋主要分布于顶体区质膜内外、顶体外膜和顶体内膜内侧，尾部Ｃａ＾２＋主要分布于线粒体基质，在质膜、致密纤维和轴丝处也有一定分布。在获能过程中Ｃａ＾２＋进入精子内部，并在头部结合于顶体区质膜内侧和顶体外膜外侧。顶体反应时Ｃａ＾２＋结合于顶体内膜外侧和由顶体外膜囊泡化     

猕猴精子获能和顶体反应过程中Ca~（2＋）和Ca~（2＋）－ATPase定位及咖啡因和dbcAMP对顶体反应的影响

用焦锑酸钾原位沉淀法和枸橼酸铅捕获法研究了猕猴精子获能和顶体反应过程中的Ｃａ~（２＋）分布和Ｃａ~（２＋）－ＡＴＰａｓｅ活性。获能前精子头部Ｃａ~（２＋）主要分布于顶体区质膜内外、顶体外膜和顶体内膜内侧，尾部Ｃａ~（２＋）主要分布于线粒体基质，在质膜、致密纤维和轴丝处也有一定分布，在获能过程中Ｃａ~（２＋）进入精子内部，并在头部结合于顶体区质膜内侧和顶体外膜外侧。顶体反应时Ｃａ~（２＋）结合于顶体内膜外侧和由顶体外膜囊泡化形成的囊泡内外，另外还有一些Ｃａ~（２＋）分散存在于顶体内容物中。在ｐＨ９．０条件下，头部Ｃａ~（２＋）－ＡＴＰａｓｅ活性存在于顶体外膜外侧和顶体区质膜外侧，顶体后区质膜无此酶活性。尾部Ｃａ~（２＋）－ＡＴＰａｓｅ存在于质膜、线粒体膜及致密纤维和轴丝处。各处的Ｃａ~（２＋）－ＡＴＰａｓｅ活性在获能和顶体反应过程中一直存在。咖啡因和ｄｂｃＡＭＰ可提高精子运动能力，前者还可显著增加顶体反应率。     

考马期亮蓝染色法检测人精子顶体反应的评价

为建立一种检测人检测人精子顶体反应的简便实用的新技术，用３．５％高氧酸水溶液本制０．０５％考马斯亮蓝染色液浸染人精子３０ｍｉｎ，顶体完整者顶体区染成紫蓝色，顶体反应者则不梁。对获能前后和钙离子载体Ａ２３１８７诱导顶体反应的精子进行染色并与经典的顶体反应检测技术ＰＳＡ法对照，两种方法检测顶体反应率无显著差异。     

附睾的胚胎发生、结构及其功能

本文就哺乳动物（包括人）附睾的解剖分区、胚胎发生、结构和功能进行了综述。附睾由输出小管和附睾管构成。前者包含高柱状纤毛细胞和低柱状非纤毛细胞；后者包含主细胞、基细胞、顶细胞、窄细胞、亮细胞和晕细胞等。最近，我们在３３周人胚胎附睾的头段观察到淋巴细胞，体段观察到可能有内分泌功能的细胞，同时对各期胎儿附睾做了ＡＫＰ、ＡＣＰ、ＳＤＨ和ＰＡＳ反应的组化研究。附睾具有吸收、分泌和参与精子成熟的功能。近年来，相继发现了数十种与精子成熟密切相关的蛋白成分，它们为揭开精子成熟的奥秘奠定了基础。     

小鼠精子发生过程中dysbindin-1的表达

目的 探讨精子发生过程中dysbindin-1的表达及dysbindin-1对精子顶体形态的影响。 方法 取生后7d、14d、21d、28d、35d和3月龄的雄性小鼠各3只,用免疫印迹法检测睾丸组织dysbindin-1的表达;以dysbindin-1缺失突变的sdy小鼠为研究对象,收集附睾尾部精子,一部分制备精子涂片,HE染色显示精子形态,用异硫氰酸荧光素-豌豆凝集素（FITC-PSA）和抗精子蛋白56（sp56）单克隆抗体进行荧光染色显示顶体的结构;另一部分采用免疫印迹法检测精子中dysbindin-1的表达。 结果 不同发育阶段小鼠睾丸组织及精子中均只有dysbindin-1A,无dysbindin-1C的表达;sdy小鼠的精子及顶体的形态未见明显异常。 结论 Dysbindin-1A表达于小鼠精子发生的不同时期,dysbindin-1A对精子形态维持不起关键作用。     

微波局部照射小鼠睾丸对睾丸,附睾和精子凝集素受体含量和分…

本实验应用２４５０ＭＨｚ微波，局部照射ＢＡＬＢ／ｃ小鼠睾丸，在肛温达４１±０．５℃后维持１５分钟，分别照射１或３次，以凝集素（ＷＧＡ、ＳＢＡ、Ｃｏｎ－Ａ）作为分子探针，于动物受照后１周、２和５个月观察微波辐射对各级生精细胞、附睾及精子糖脂和糖蛋白含量及分布的影响。结果表明，生精细胞中ＷＧＡ，Ｃｏｎ－Ａ受体的含量均显著下降（Ｐ＜０．０１），形态异常的精子中三种凝集素受体的含量均显著增加，且分布特性有     

微波局部照射小鼠睾丸对睾丸、附睾和精子凝集素受体含量和分布的影响

本实验应用２４５０ＭＨｚ微波，局部照射ＢＡＬＢ／ｃ小鼠睾丸，在肛温达４１±０．５℃后维持１５分钟，分别照射１或３次，以凝集素（ＷＧＡ、ＳＢＡ、Ｃｏｎ－Ａ）作为分子探针，于动物受照后１周、２和５个月观察微波辐射对各级生精细胞、附睾及精子糖脂和糖蛋白含量及分布的影响。结果表明，生精细胞中ＷＧＡ、Ｃｏｎ－Ａ受体的含量均显著下降（Ｐ＜０．０１），形态异常的精子中三种凝集素受体的含量均显著增加，且分布特性有改变。受照睾丸内呈增生的血管内皮ＳＢＡ反应为阳性。本研究结果证明，微波确可引起生殖细胞的损伤。     

考马斯亮蓝染色法检测人精子顶体反应的评价

为建立一种检测人精子顶体反应的简便实用的新技术，用３．５％高氯酸水溶液配制０．０５％考马斯亮蓝（Ｒ２５０）染色液浸染人精子３０ｍｉｎ，顶体完整者顶体区染成紫蓝色，顶体反应者则不染。对获能前后和钙离子载体Ａ２３１８７诱导顶体反应的精子进行染色并与经典的顶体反应检测技术ＰＳＡ法对照，两种方法检测顶体反应率无显著差异。方法学检验证明新技术结果可靠。本法操作简便、经济，普通显微镜即可检测，易于推广，克服了原有技术复杂昂贵的缺点，具有研究和临床诊断的良好价值。     

锌缺乏导致小鼠睾丸游离锌离子和附睾精子数量减少

目的研究锌缺乏对小鼠睾丸游离锌离子和附睾精子数量的影响。方法锌缺乏饲养小鼠5周后,应用ZnSe金属自显影技术对小鼠睾丸和附睾进行染色,观察睾丸和附睾锌离子的分布,同时计数附睾精子数量,并与对照组小鼠的精子数量做对比。结果缺锌喂养后的小鼠睾丸游离锌离子明显减少,并且睾丸生精小管管腔狭小,生殖细胞层厚度增加,此外,附睾精子数量也明显低于对照组。结论睾丸和附睾内游离锌离子减少是锌缺乏小鼠精子数量下降的原因,本结果有助于改善男性生殖健康的状况。     

层黏连蛋白受体在小鼠睾丸和附睾中的表达

目的 探讨层黏连蛋白受体(LAMR1)在小鼠睾丸和附睾中的表达.方法收集3只正常成年昆明小鼠睾丸和附睾.采用原位杂交和免疫组织化学方法,检测LAMR1 mRNA及蛋白在成年小鼠睾丸和附睾中的分布.结果 LAMR1 mRNA在附睾头和附睾尾表达最强;在睾丸生精细胞中也有表达.免疫组织化学结果显示,LAMR1蛋白从附睾头到...     

Purification of GP-83, a glycoprotein secreted by the human epididymis and conjugated to mature spermatozoa

Epididymal secretions are critical for mammalian spermatozoa to acquire both forward motility and an ability to recognize and penetrate oocytes. Previous studies identified two glycoproteins, GP-83 and GP-39, which were secreted by the human epididymis and may be related to maturation of sperm function. In this study, GP-83 was purified from human seminal fluid by DEAE-ion exchange, gel filtration chromatography and preparative gel elution. The isoelectric point (pI) of purified GP-83 was 6.57. Monospecific antiserum to GP-83 was induced in male New Zealand rabbits and confirmed on immunoblots. GP-83 was found in fluid, tissue and sperm extracts of corpus and cauda epididymis, but not in the caput. Immunohistochemical localization identified GP-83 in the luminal contents and in the supranuclear region and cell membrane of principal cells of the corpus and cauda epididymis. GP-83 was found on the anterior acrosome in ejaculated spermatozoa, and shifted to the equatorial region after capacitation and the acrosome reaction.     

牛精子膜SA—30在获能与顶体反应后的定位变化

为了认识精子膜抗原在受精中的作用，利用ＣｏｎＡ－Ｓｅｐｈａｒｏｓｅ４Ｂ亲合层析，结合ＳｅｐｈａｄｅｘＧ－１００凝胶过滤的方法，分离出牛精子膜与与Ｃdisplay status结合的一种分子量约３０ＫＤ的糖蛋白称为ＳＡ－３０。     

Morphometric analysis of intact sperm heads and of sperm nuclei in the mouse

A morphometric analysis of mouse sperm and of their nuclei was undertaken to investigate their respective post-testicular maturation. Sperm were collected from the testis, caput and cauda epididymidis, and their corresponding nuclei were isolated. Results indicate that the post-testicular maturation of sperm is distinct from that of nuclei. The size of intact sperm heads increases in the caput followed by a subsequent decrease in the cauda. In contrast, sperm nuclei decrease progressively in size. In general, a greater magnitude and number of alterations in intact heads and nuclei occur while in transit from the testis to the caput than during passage to the cauda epididymis. These results suggest that the period immediately following their release from the testis is crucial to the complete morphological maturation of sperm heads and nuclei. © 1993 Wiley-Liss, Inc.     

Maturation antigen of the mouse sperm flagellum: II. Origin from holocrine cells of the distal caput epididymis

During epididymal transit, the mouse sperm flagellum acquires a surface glycoprotein (SMA4) from epididymal fluid that functions as a sperm antiagglutinin. To determine the origin of this molecule, testes and epididymides of male mice were sectioned for light microscopy and stained with wheat germ agglutinin (WGA)-peroxidase, a probe that has been used previously to examine the biology of SMA4. WGA reactivity was localized to the cytoplasm in a small population of cells in the distal caput epididymis. Testis cells and principle cells of the caput were nonreactive with WGA, while stereocilia were stained on principle cells in the corpus and cauda. The WGA-positive cells in the distal caput were identified as holocrine cells on the basis of morphology, distribution, and PAS + reaction. At high magnification, intense WGA reactivity was due to the presence of numerous apical granules in the cytoplasm. The location of the cells in distal caput coincided exactly with the region of tubule in which sperm first acquired SMA4 on their flagellae. These data suggest that holocrine cells near the junction of caput and corpus epididymis are the source of the sperm antiagglutinin SMA4.     

Andrology: Evaluation of motility, fertilizing ability and embryonic development of murine epididymal sperm after coculture with epididymal epithelium

Murine sperm from the caput, corpus and cauda epididymis werecocultured with epididymal epithelial cells of their own regionor more distal regions, in the presence and absence of androgens(testosterone and dihydrotestosterone). Epitheial cell cultureswere used 3 or 10 days after preparation in a complex tissueculture medium (Chang's) as plated tubules. The coculture studiesinvolving spermatozoa and oocytes with epithelial cells werecarried out in T6 medium. Motility of caput spermatozoa wasmaintained for 24 h in the presence of day 3 corpus and caudaepithelial cells and hormones but not under other conditions.Likewise, the motility of corpus spermatozoa was maintainedfor 24 h in the presence of day 3 cauda epithelial cells andhormones but not other conditions. Fertilization of zonaintactoocytes by epididymal spermatozoa was not affected by theircoculture for 24 h with epithelial cells but fertilization ratesfor zona-free oocytes were increased for caput spermatozoa coculturedwith more distal epithelial cells. Fertilization rates for bothzona-intact and zona-free oocytes were increased for corpusspermatozoa cocultured with more distal cauda epithelial cells.The developmental capacity of embryos derived from caput spermatozoawas not significantly increased by coculture with epithelialcells but those derived from corpus spermatozoa cocultured withcauda epithelial cells were signilicantly increased. We concludethat the presence of more distal epithelial cells of the mouseepididymis maintains motility in culture, increases the abilityof caput and corpus spermatozoa to fertilize zona-free oocytesand increases the developmental capacity of embryos formed fromcorpus spermatozoa. These observations demonstrate the functionof epididymal regions in the maturation of murine spermatozoafor fertilization and embryo development.     

SA-30诱导的小鼠子宫内诱导型一氧化氮合酶的变化

目的　检测精子膜抗原 SA- 30对小鼠子宫内诱导型一氧化氮合酶 (i NOS)含量及分布的影响 ,从而了解 SA - 30的抗生育机制。　方法　 SA- 30免疫雌性昆明小鼠后 ,应用免疫组织化学方法 ,对子宫内的 i NOS进行了检测。结果　 SA - 30免疫后对间情期小鼠子宫内的 i NOS没有明显影响 ,无论是免疫组还是对照组 ,在子宫内膜基质中均有大量阳性细胞分布 ,两者无明显差异 ;在妊娠第 3d时 ,对照组的小鼠子宫中有较多 i NOS表达 ,阳性标记主要集中在内膜上皮及固有膜中的子宫腺上 ,经 SA- 30免疫的小鼠子宫中 ,i NOS表达则显著减少 ,整个内膜标记极弱 ,只在子宫肌层内有少量 i NOS阳性细胞分布。　结论　 SA - 30可能通过使妊娠早期小鼠子宫内 i NOS表达显著减少 ,而影响小鼠胚胎植入及早期胚胎的发育     

Decline in fertility of mouse sperm with abnormal chromatin during epididymal passage as revealed by ICSI

BACKGROUND: Recent studies showed that ICSI with cauda epididymal or ejaculated sperm of infertile mice or men, respectively, was less effective in fertilization and normal embryo development than ICSI using sperm from the testes. These studies suggested that sperm nuclear quality declined after release from the testis, but the site where this loss of fertility occurs has not been localized. METHODS: We performed ICSI with testicular, caput, and cauda epididymal sperm from infertile Tnp1-/-Tnp2+/- mutant mice, which have a minimal level of transition nuclear proteins and are sterile by natural mating. RESULTS: When the heads of motile sperm from the testis or caput epididymis of Tnp1-/-Tnp2+/- males were injected into enucleated mouse oocytes, sperm chromosomes showed no difference from those of wild-type mice, but the chromosomes from sperm taken from the cauda epididymis of mutant males showed increased abnormalities. Injection of testicular or caput epididymal sperm from Tnp1-/-Tnp2+/- males into intact oocytes resulted in normal embryonic and fetal development and yields of liveborn equivalent to wild-type, but cauda sperm from Tnp1-/-Tnp2-/- mice produced lower implantation rates and yields of liveborn than did those from wild-type mice. CONCLUSIONS: These results demonstrate that in mice with sperm chromatin abnormalities, the decline in fertility of sperm with ICSI occurs after the caput epididymis. The advantage of using caput epididymal sperm for ICSI in certain situations may be considered as an approach to be tested in human assisted reproduction.