水通道蛋白3在人胎盘和胎膜中的表达及分布

目的 研究正常妊娠晚期人胎盘和胎膜水通道蛋白3(AQP3)的表达及分布.方法 收集5例正常足月妊娠剖宫分娩的胎盘和胎膜样本,用RT-PCR测定AQP3 mRNA在胎盘和胎膜组织中的表达;用Western印迹和免疫组织化学方法检测AQP3蛋白质水平在胎盘和胎膜的表达.结果 RT-PCR显示AQP3 mRNA在胎盘和胎膜组织均有表达.Western印迹结果显示胎盘组织在29 000左右有一特异性条带.免疫组织化学结果显示AQP3表达于合体滋养细胞,而在羊膜上皮细胞未见表达.结论 AQP3在胎盘母儿液体平衡中可能发挥重要作用.     

水通道蛋白3在人胎盘和胎膜中的表达及分布

目的研究正常妊娠晚期人胎盘和胎膜水通道蛋白3(AQP3)的表达及分布。方法收集5例正常足月妊娠剖宫分娩的胎盘和胎膜样本,用RT-PCR测定AQP3 mRNA在胎盘和胎膜组织中的表达;用Western印迹和免疫组织化学方法检测AQP3蛋白质水平在胎盘和胎膜的表达。结果RT-PCR显示AQP3 mRNA在胎盘和胎膜组织均有表达。Western印迹结果显示胎盘组织在29 000左右有一特异性条带。免疫组织化学结果显示AQP3表达于合体滋养细胞,而在羊膜上皮细胞未见表达。结论AQP3在胎盘母儿液体平衡中可能发挥重要作用。     

水通道蛋白3在人前列腺组织中的表达及意义

目的： 揭示水通道蛋白3（AQP3）在人前列腺移行区和外周区的表达及意义,为深入探讨AQP在人前列腺及疾病状态下的表达及生理作用奠定基础。方法: 取前列腺组织标本5例,采用RT-PCR方法检测AQP3 mRNA在前列腺组织的表达。同时应用Western blotting和免疫染色技术研究AQP3蛋白在前列腺组织的定位表达。结果: RT-PCR结果显示在前列腺的移行区和外周区均存在AQP3 mRNA表达。Western blotting结果显示人前列腺的移行区和外周区均特异表达AQP3蛋白,且移行区表达强于外周区。免疫组织化学和免疫荧光染色结果表明AQP3表达在前列腺移行区分泌细胞的细胞膜。结论: 前列腺移行区分泌细胞表达AQP3提示, AQP3在前列腺液分泌过程中可能发挥重要的生理作用。     

水通道蛋白-4在大鼠甲状腺中的表达

目的:观察水通道蛋白-4(AQP4)在甲状腺滤泡细胞的表达和分布特点,为研究甲状腺内分泌及调节机制提供形态学基础。方法:利用免疫组织化学、免疫荧光组织化学和原位杂交组织化学技术,检测大鼠AQP4及其mRNA在甲状腺滤泡上皮细胞的表达。结果:免疫组织化学和免疫荧光组织化学显示,AQP4在甲状腺滤泡上皮细胞膜上有表达,而在上皮细胞基底面和管腔面表达更为明显;原位杂交组织化学显示AQP4 mRNA在上皮细胞中表达明显。结论:AQP4在甲状腺的表达和分布特点,提示其参与了甲状腺素的合成和分泌过程中渗透压的精细调节作用。     

肝细胞核因子6在小鼠肝内胆管发育过程中的表达

目的 探讨肝细胞核因子6(HNF6)在肝内胆管发育过程中的作用. 方法 用RT-PCR及免疫组织化学等方法 检测HNF6在小鼠胚胎发育的各个阶段以及成年肝中的表达. 结果 RT-PCR结果 显示,HNF6 mRNA的表达在E9d开始出现,与肝芽形成的时间吻合,E13d HNF6 mRNA的表达消失,E15d又重新出现,并一直维持到出生.免疫组织化学显示,CK19免疫反应在E13d开始出现,此时免疫反应阳性细胞在肝索内散在分布.E15d时在近肝门处的门管区丌始出现由单层的、CK19阳性细胞组成的胆管板,之后,阳性细胞反应主要分布于胆管板和小叶间胆管.E9～11d,多数肝索细胞呈HNF6阳性反应,E13d的肝索中未观察剑HNF6的表达.E15～17d,HNF6阳性反应的分布与CK19相似,成年小鼠肝的胆管上皮细胞仍呈HNF6阳性反应. 结论 HNF6可能与肝干细胞的特化关系不大,而与肝发育的启动、肝干细胞向胆管上皮细胞的分化及其分化状态的维持有关.     

水通道蛋白1在小鼠口腔粘膜表达与分布

目的观察水通道蛋白1(AQP1)在小鼠口腔粘膜的表达与分布,为研究AQP1在口腔水代谢和感觉传递中的作用提供形态学依据。方法制备成年小鼠口腔各部粘膜石蜡切片,进行AQP1免疫组织化学染色。结果 AQP1免疫阳性产物在唇、颊、硬腭、软腭和舌粘膜下层的毛细血管和神经纤维上广泛表达。另外在上述部位的腺泡和腺管周围的肌上皮细胞以及舌的味蕾和丝状乳头上也有AQP1免疫阳性信号。结论 AQP1在口腔广泛分布,可能参与了唾液分泌和口腔感觉传递等生理过程。     

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。     

β-catenin在四氯化碳诱导的小鼠肝纤维化过程中的定位、表达及意义

目的 探讨肝纤维化发生过程中β-连环蛋白(β-catenin)定位、表达及意义.方法 健康雄性昆明小鼠(n=45)随机分为对照组(n=15)与实验组(n=30).实验组小鼠皮下注射50% 四氯化碳(CCl4)-粟米油混合液(6ml/kg),2次∕周,对照组皮下注射同等剂量的粟米油.各组分别于造模1周、4周和8周后取小鼠肝脏,常规制作石蜡切片,Masson染色及Desmin免疫组织化学法观察比较不同组别小鼠肝纤维化病理变化,RT-PCR及免疫组织化学方法检测不同时间点各组小鼠肝组织内β-catenin的表达及定位.结果 Masson染色结果显示,对照组肝汇管区结缔组织内及血管壁有少量细小纤维,实验组小鼠肝脏汇管区和中央静脉及其周围胶原纤维增多;随损伤时间延长纤维增生愈加明显.Desmin免疫组织化学结果显示,各组别均有阳性表达,但损伤组各时间点desmin阳性表达细胞数明显高于对照组(P<0.05 或P<0.001).RT-PCR结果显示,CCl4损伤1周后肝内β-catenin mRNA水平与对照组相比无明显差别(P＞0.05),损伤4周及8周β-catenin mRNA水平则明显下降,与对照组相比差异均有显著性(P<0.01).免疫组织化学结果显示,对照组β-catenin弱表达于肝细胞膜及胆管上皮细胞膜和胞质,而损伤组β-catenin阳性反应主要定位于肝内增生的细胞团及新生胆管上皮细胞质,各组间积分吸光度值差别有显著性(P<0.01).结论 CCl4诱导的肝纤维化过程中β-catenin mRNA表达与蛋白表达不同步,阳性表达细胞主要为新生的细胞团及胆管上皮细胞.     

脾气虚胃溃疡大鼠胃黏膜生长抑素相关信号转导分子的变化

目的 探讨生长抑素及其信号传递分子与脾气虚胃溃疡的发生及转归的关系. 方法 选用成年Wistar大鼠,雌、雄性各35只,建立脾气虚胃溃疡大鼠模型;运用HE染色、免疫组织化学分析、RT-PCR及免疫印迹法,分别观察胃黏膜的组织结构、生长抑素含量、2型生长抑素受体mRNA、细胞外信号传递激酶2(ERK2)表达的变化. 结果 脾气虚胃溃疡大鼠胃黏膜中,2型生长抑素的蛋白含量增加,生长抑素受体mRNA表达上调,(ERK2)表达下降. 结论 生长抑素信号传递途径中相关信号分子的变化可能足引起脾气虚胃溃疡发生的原因之一.     

肺泡Ⅱ型上皮细胞染色体脆性位点基因FHIT与肺间质纤维化的关系

目的:研究肺间质纤维化模型大鼠肺泡Ⅱ型上皮细胞染色体脆性位点基因FHIT表达的改变.方法:SD雄性大鼠随机分为假手术组、模型组及川芎嗪用药组,分别于给药7、14、28 d后提取肺泡Ⅱ型上皮细胞行体外培养,RT-PCR法检测FHIT基因mRNA表达的改变,免疫组织化学检测FHIT蛋白表达.结果:与假手术组比较,造模7d后模型组大鼠肺泡Ⅱ型上皮细胞染色体脆性位点基因FHIT mRNA及蛋白表达无变化.用药14、28 d后,FHIT基因mRNA及蛋白表达则明显下降.而川芎嗪组比模型组有显著上调.结论:在肺间质纤维化过程中存在染色体脆性位点基因FHIT的改变,川芎嗪可以一定程度上抑制这种改变从而起到防治作用.     

Spontaneous mutation in mice provides new insight into the genetic mechanisms that pattern the seminal vesicles and prostate gland.

The seminal vesicles and prostate gland are anatomically adjacent male sex-accessory glands. Although they arise from different embryonic precursor structures and express distinct sets of secretory proteins, these organs share common features in their developmental biology. A key shared developmental feature is the elaboration of complex secretory epithelia with tremendous surface area from simple precursor structures with juxtaposed epithelial and mesenchymal cells. In this study, new insight into the nature of the biological processes that underlie glandular morphogenesis is achieved by analyzing the phenotypes present in mice that harbor a spontaneous mutation, seminal vesicle shape (svs), previously identified for causing altered seminal vesicle morphology in adults. An examination of seminal vesicle development in svs mice provides the first evidence that the concurrent processes of epithelial branching and epithelial infolding are distinct processes under separate genetic control. It also provides the first direct evidence that the thickness and topology of the smooth muscle layer in the seminal vesicles are determined by interaction with the glandular epithelium during the branching process. In addition, the seminal vesicle phenotype in svs mice is shown to phenocopy the morphologic form present in certain other mammals such as the guinea pig, raising the possibility that the svs mutation is the sort of variant that arises during evolution. By also including an investigation of the prostate gland, this study also identifies previously unrecognized phenotypes in svs prostates, including increased gland size and dramatically reduced levels of branching morphogenesis. Finally, this study advances the goal of identifying the svs gene by mapping the svs mutation relative to known molecular markers and testing Fgfr2 as a candidate gene. The finding that the svs mutation maps to a genomic region syntenic to a region frequently deleted in human prostate tumors, together with the prostatic phenotype present in svs mice, further raises the interesting possibility that the svs mutation will identify a candidate prostate tumor suppressor gene.     

The distribution of plasminogen activator in the male genital tract

The distribution of fibrinolytic activity in the tissues of the male genital tract was studied by a histological technique. Preparations made from testis, epididymis, vas deferens, seminal vesicle, prostate, bulbo-urethral gland, and urethra showed that most activity was related to the blood vessels. However, inconsistent fibrinolytic activity related to epithelium was found in all parts of the genital tract. This epithelial activity was least in the testis, greater in the seminal vesicle and prostate gland, and was greatest in the bulbo-urethral gland and terminal urethra. No fibrinolytic activity could be demonstrated in relation to spermatozoa.     

Mucin gene expression in human male urogenital tract epithelia

BACKGROUND: Mucins are large, hydrophilic glycoproteins that protect wet-surfaced epithelia from pathogen invasion as well as provide lubrication. At least 17 mucin genes have been cloned to date. This study sought to determine the mucin gene expression profile of the human male urogenital tract epithelia, to determine if mucins are present in seminal fluid and to assess the effect of androgens on mucin expression. METHODS AND RESULTS: Testis, epididymis, vas deferens, seminal vesicle, prostate, bladder, urethra and foreskin were assessed for mucin expression by RT-PCR (for 14 mucin genes) and immunohistochemistry (nine antibodies for five mucins). Epithelia of the vas deferens, prostate and urethra expressed the greatest number of mucins, each with mRNA for between 5 and 8 mucins. Except for MUC20 in epididymis, mRNA for MUC1 and MUC20, both membrane-associated mucins, was detected in all tissues analysed. By comparison, MUC6 was more restricted in expression, being primarily detected in seminal vesicle. MUC1, MUC5B and MUC6 were detected in seminal fluid samples by immunoblot analysis. Androgens had no effect on mucin expression in cultured human prostatic epithelial cells. CONCLUSIONS: Each region of urogenital tract epithelium expressed a unique mucin gene repertoire. Secretory mucins are present in seminal fluid, and androgens do not appear to regulate mucin gene expression in prostatic epithelial cells in culture.     

Follistatin and activin A production by the male reproductive tract

Follistatin is a binding protein for the activin and inhibin family of hormones, regulating their biological activity. In the male reproductive tract, the interaction of these factors is likely to be involved in the regulation of the proliferation of several cell types. We have investigated the presence of follistatin and activin A in seminal plasma using specific immunoassays and have localized follistatin and activin/inhibin subunits in the adult human testis, prostate and seminal vesicle to establish their likely sources. High concentrations of immunoreactive follistatin were present in seminal plasma in normal men (mean 97.9 ng/ml; 1.43 ng/ml in peripheral plasma) and were similar in men with oligo/azoospermia and following vasectomy. Follistatin immunoreactivity was localized to both Leydig and Sertoli cells of the testis, and to epithelial cells of the prostate gland and seminal vesicle, which are likely to be the predominant sources of the hormone in seminal plasma. Activin A was also present in seminal plasma in normal men but was undetectable following vasectomy, thus deriving from the testis. Consistent with this finding, the betaA-subunit was immunolocalized in Sertoli and Leydig cells but was not present in seminal vesicle or prostate gland. The functional significance of the high concentrations of follistatin secreted into seminal plasma by the prostate gland and/or seminal vesicle is uncertain, but they may regulate the biological activity of testis-derived activin A and inhibin B.      

Localization of certain phosphatases in the male sex accessory glands of Suncus murinus sindensis, Anderson, the common shrew.

Histochemical localization of various phosphatases, alkaline phosphatase, acid phosphatase, adenosine-tri-phosphatase and glucose-6-phosphatase, have been carried out in the male sex accessory glands of Suncus murinus sindensis, ANDERSON. The seminal vesicle and the COWPER'S gland in Suncus display strong phosphatases activities in the epithelium, except the alkaline phosphatase in the in the COWPER'S gland which is more pronounced in the stroma. The possible role of these phosphatases in the secretory activities of the organ where they are localized have been discussed. In the prostate gland, no phosphatase activity could be revealed in the epithelium and the secretions.     

The influence of progestin and androgen on the fine structure of the male reproductive tract of the rat. II. Epididymis and sex accessory glands

Young adult male rats were administered medroxyprogesterone (Provera, Upjohn) alone and in combination with testosterone, as has been done to inhibit male fertility. The histology and fine structure of several segments of the epididymis, the ventral prostate, and the seminal vesicle were studied at intervals after treatment for up to 16 weeks. The epididymides of treated animals weighed less than those of control rats. Microscopic alterations in the epididymis were similar in rats treated with Provera alone and in those animals that received Provera and testosterone, but the changes varied with the segment of the epididymis. In the middle segment in the caput epididymidis, the normally abundant luminal sperm were absent but the epithelium retained its normal ultrastructural features. In the terminal segment in the cauda epididymidis, different changes were observed in the proximal and distal portions. In the proximal cauda epididymidis, the lumen was small, irregular in outline, and virtually devoid of sperm. The light cells of the epididymal epithelium in the proximal cauda contained extremely large numbers of dense bodies resembling lysosomes, which occupied most of the supranuclear and basal cytoplasm. In contrast, in the distal part of the cauda epididymidis, the epithelium had a normal appearance but the lumen was filled with debris, sperm, and spherical masses of cytoplasm that were apparently derived from germ cells. It is suggested that the clearing of the lumen of the proximal cauda epididymidis may reflect the greater activity of light cells of the epididymal epithelium in that region. Although alterations in spermatogenesis may be most important in the antifertility effect of progestin and androgen, these alterations in epididymal sperm and epithelium may also play a role. The weights of the prostate and seminal vesicles of rats treated with Provera (1 mg/100 g/day) were greatly reduced compared to those of control rats. Although there was considerable variation, in many specimens treated with Provera alone the epithelium of the prostate showed a change from a columnar to a cuboidal or squamous shape, and there was a reduction in the size and abundance of organelles involved in the formation of secretions. The microscopic structure of the seminal vesicle of rats treated with Provera was less severely affected than the prostate. Although the seminal vesicle epithelium of Provera-treated rats was generally not as tall as in control animals, the cells possessed parallel cisternae of rough endoplasmic reticulum, secretory vacuoles, and an active-appearing Golgi apparatus, suggesting that they continued to be able to form secretions in the presence of Provera. The weights of the sex accessory glands were maintained at control levels by the administration of testosterone, 100 μg/100 g/day, along with the Provera. A normal fine structure was present in the epithelium of both the prostate and seminal vesicle of rats administered this amount of testosterone in addition to Provera. Lower doses of testosterone (15 or 30 μg/100 g/day) were insufficient to maintain normal weight or ultrastructure of the sex accessory glands in the presence of Provera.