子宫内膜癌组织KIF20A、LAPTM4B-35表达与临床病理特征及预后的关系研究

摘要 目的：探讨子宫内膜癌组织驱动蛋白家族成员20A(KIF20A)、溶酶体相关4次跨膜蛋白B-35(LAPTM4B-35)表达与临床病理特征及预后的关系。方法：选择2012年4月至2015年8月期间于我院行手术治疗的80例子宫内膜癌患者作为研究对象。检测子宫内膜癌组织以及距离肿瘤边缘2 cm以上癌旁组织中KIF20A、LAPTM4B-35 mRNA表达水平。分析子宫内膜癌组织中KIF20A、LAPTM4B-35 mRNA表达与临床病理特征的关系。分析不同KIF20A、LAPTM4B-35 mRNA表达患者5年总生存率的差异。分析子宫内膜癌患者预后的影响因素。结果：与癌旁组织相比,子宫内膜癌组织中KIF20A、LAPTM4B-35 mRNA表达水平明显升高,差异有统计学意义（P<0.05）。有淋巴结转移以及FIGO分期Ⅲ期患者的癌组织KIF20A、LAPTM4B-35 mRNA表达水平明显高于无淋巴结转移以及FIGO分期I～II期患者的癌组织,差异有统计学意义（P<0.05）。KIF20A低表达组患者5年总生存率明显高于KIF20A高表达组;LAPTM4B-35低表达组患者5年总生存率明显高于LAPTM4B-35高表达组患者,差异有统计学意义（P<0.05）。Cox回归分析结果显示：FIGO分期Ⅲ期、有淋巴结转移、KIF20A mRNA高表达和LAPTM4B-35 mRNA高表达是子宫内膜癌患者预后的影响因素（P<0.05）。结论：在子宫内膜癌组织中KIF20A、LAPTM4B-35 mRNA表达水平升高,有淋巴结转移、FIGO分期较高患者癌组织KIF20A、LAPTM4B-35 mRNA表达水平上调。KIF20A 、LAPTM4B-35高表达患者5年总生存率下降。     

纤溶酶原激活因子与猕猴精子运动力的关系及其在附睾和附性腺中的表达调节

用11酸睾酮诱导猕猴少精子症和弱精子症及单侧隐睾手术诱导单侧少精子症和弱精子症模型,观察对附睾头、附睾体、附睾尾、前列腺和精囊组织型PA（tPA）、尿激酶型PA（uPA）及抑制因子-1（PAI-1）mRNA表达的影响。原位杂交的结果表明11酸睾酮诱导少精子症和弱精子症,tPA mRNA的表达在附睾头、精囊及前列腺减少,而在附睾体升高,附睾尾表达基本无变化;uPA mRNA的表达在附睾头、附睾体、前列腺减少,而在精囊升高,附睾尾表达基本无变化;PAI-1 mRNA的表达在附睾头、附睾体、精囊下降,而在前列腺升高,附睾尾表达无显著变化。单侧隐睾手术不影响tPA、uPA和PAI-1 mRNA的表达。这些结果提示附睾头和附睾体分泌的uPA可能与精子前向运动能力的获得相关。tPA、uPA和PAI-1 mRNA在猕猴附睾头部和体部、前列腺和精囊中的表达可能受睾酮的调节,但不受睾丸分泌因子及温度的影响,且在不同部位睾酮的调节具不同的特征,而附睾尾tPA、uPA和PAI-1的表达则可能是组成性表达。     

染色域Y样蛋白介导的组蛋白巴豆酰化与抑郁小鼠模型中NGF和炎症因子水平及神经功能紊乱的关系

摘要 目的：探究染色域Y样蛋白介导的组蛋白巴豆酰化与抑郁小鼠模型中神经生长因子（Nerve growth factor,NGF）和炎症因子水平及神经功能紊乱的关系。方法：32只成年雄性C57BL/6小鼠分为4组：对照组、模型组、CDYL过表达组、CDYL敲低组,各8只。通过慢性束缚应激诱导抑郁模型,通过旷场试验记录中心时间和中心距离,通过强迫游泳测试记录不动时间。通过RT-qPCR和蛋白质免疫印迹检测CDYL mRNA和蛋白质表达水平。通过ChIP-seq检测组蛋白赖氨酸巴豆酰化。通过ELISA检测TNF-α,IL-1β和IL-6表达水平。通过RT-qPCR检测检测海马组织5-HT和IDO mRNA表达水平。通过蛋免疫组织化学染色检测检测海马组织NGF、BDNF和突触素（Synaptophysin,SYP）表达水平。通过BrdU免疫荧光染色检测神经系统的发育以及识别大脑中的神经发生。结果：模型组中心时间和中心距离较对照组降低,不动时间较对照组升高（P<0.05）。CDYL过表达组中心时间和中心距离较模型组降低,不动时间较模型组升高（P<0.05）。CDYL敲低组中心时间和中心距离较模型组升高,不动时间较模型组降低（P<0.05）。模型组CDYL mRNA和蛋白质表达水平较对照组升高（P<0.05）。CDYL过表达组CDYL mRNA和蛋白质表达水平较模型组升高（P<0.05）。CDYL敲低组TCDYL mRNA和蛋白质表达水平较模型组降低（P<0.05）。模型组组蛋白巴豆酰化水平较对照组降低（P<0.05）。CDYL过表达组组蛋白巴豆酰化水平较模型组降低（P<0.05）。CDYL敲低组组蛋白巴豆酰化较模型组升高（P<0.05）。模型组TNF-α,IL-1β和IL-6表达水平较对照组升高（P<0.05）。CDYL过表达组TNF-α,IL-1β和IL-6表达水平较模型组升高（P<0.05）。CDYL敲低组TNF-α,IL-1β和IL-6表达水平较模型组降低（P<0.05）。模型组5-HT mRNA表达水平较对照组降低,IDO mRNA表达水平较对照组升高（P<0.05）。CDYL过表达组5-HT mRNA表达水平较模型组降低,IDO mRNA表达水平较模型组升高（P<0.05）。CDYL敲低组5-HT mRNA表达水平较模型组升高,IDO mRNA表达水平较模型组降低（P<0.05）。模型组NGF,BDNF和SYP表达水平较对照组降低（P<0.05）。CDYL过表达组NGF、BDNF和SYP表达水平较模型组降低。CDYL敲低组NGF,BDNF和SYP表达水平较模型组升高（P<0.05）。结论：慢性束缚应激诱导抑郁模型组蛋白巴豆酰化降低、炎症因子水平升高、NGF、BDNF和SYP水平降低以及神经功能紊乱,CDYL过表达组较慢性束缚应模型激进一步加剧炎症反应和神经功能紊乱,而CDYL敲低对慢性束缚应激引起的新生细胞和未成熟神经元损伤具有保护作用。     

NOD1和NOD2基因在牦牛雄性生殖器官中的表达检测

为了解NOD1和NOD2基因在牦牛生殖系统中的表达情况,从脾脏组织总RNA中RT-PCR扩增牦牛NOD1和NOD2基因,半定量RT-PCR分析NOD1和NOD2 mRNA在牦牛雄性生殖组织的表达水平。结果显示,NOD1在脾脏、睾丸、附睾头、附睾体、附睾尾、输精管和阴茎中均相对低表达,而NOD2在脾脏、睾丸、附睾头、附睾体、附睾尾、输精管和阴茎中均相对高表达。结果表明,NOD1和NOD2在牦牛雄性生殖组织中广泛表达,提示其在牦牛生殖过程中可能发挥一定的作用。     

肌层浸润性膀胱癌组织KMT2D、MCM6与临床病理特征和预后的关系

摘要 目的：探讨肌层浸润性膀胱癌（MIBC）组织赖氨酸甲基转移酶2D（KMT2D）、微小染色体维持蛋白6（MCM6）与临床病理特征和预后的关系。方法：选择2017年5月至2019年10月长治医学院附属和平医院行手术治疗的MIBC患者96例,应用逆转录-实时定量聚合酶链式反应（qRT-PCR）检测癌组织及癌旁正常组织KMT2D 信使核糖核酸（mRNA）、MCM6 mRNA表达,分析KMT2D mRNA、MCM6 mRNA表达与MIBC患者临床病理特征的关系,应用Pearson相关分析法分析MIBC患者癌组织KMT2D mRNA及MCM6 mRNA表达的相关性。随访3年,比较死亡MIBC患者与存活MIBC患者癌组织KMT2D mRNA、MCM6 mRNA表达,并应用Kaplan-Meier生存曲线分析不同KMT2D mRNA、MCM6 mRNA表达分组MIBC患者预后情况。结果：MIBC癌组织中KMT2D mRNA表达水平显著低于癌旁组织,MCM6 mRNA表达水平显著高于癌旁组织（P<0.05）。MIBC患者癌组织中KMT2D mRNA、MCM6 mRNA表达与侵犯输尿管、淋巴结转移、TNM分期显著相关（P<0.05）,且两者表达呈负相关（P<0.05）。至随访截止,MIBC患者死亡45例,死亡组患者癌组织中KMT2D mRNA表达水平显著低于存活组、MCM6 mRNA表达水平显著高于存活组（P<0.05）。生存曲线结果显示KMT2D mRNA高表达组3年生存率显著高于KMT2D mRNA低表达组;MCM6 mRNA低表达组3年生存率显著高于MCM6 mRNA高表达组（P<0.05）。结论：MIBC癌组织中KMT2D低表达、MCM6高表达,与MIBC侵犯输尿管、淋巴结转移、TNM分期及患者预后不良有关。     

肝细胞肝癌组织GPSM2、GFPT2、SNORA51 mRNA表达与临床病理特征的关系及对预后的影响研究

摘要 目的：探讨肝细胞肝癌（HCC）组织G蛋白信号调节蛋白2（GPSM2）、谷氨酰胺果糖-6-磷酸转氨酶2（GFPT2）、核仁小RNA 51（SNORA51） mRNA表达与临床病理特征的关系及对预后的影响。方法：选择2017年1月～2018年12月于内蒙古医科大学附属医院诊断并经手术切除治疗的HCC患者60例。采用实时荧光定量聚合酶链式反应（RT-qPCR）检测HCC组织、癌旁组织中GPSM2 、GFPT2 、SNORA51 mRNA表达情况,分析GPSM2、GFPT2 、SNORA51的mRNA表达与HCC患者临床病理特征的关系。随访3年,应用Kaplan-Meier生存曲线分析不同分组HCC患者预后情况,并应用单因素和多因素Logistic回归分析HCC患者预后的影响因素。结果：HCC组织中GPSM2、GFPT2 、SNORA51 mRNA表达水平显著高于癌旁组织（P<0.05）。HCC组织中GPSM2 mRNA高表达组、GFPT2 mRNA高表达组、SNORA51mRNA高表达组血管侵犯、TNM分期Ⅲ期比例显著高于GPSM2 mRNA低表达组、GFPT2 mRNA低表达组、SNORA51mRNA低表达组（P<0.05）。Kaplan-Meier法分析显示GPSM2 mRNA低表达组、GFPT2 mRNA低表达组、SNORA51mRNA低表达组3年生存率显著高于GPSM2 mRNA高表达组、GFPT2 mRNA高表达组、SNORA51mRNA高表达组（P<0.05）。多因素Logistic回归分析模型结果显示,血管侵犯、TNM分期Ⅲ期、GPSM2 mRNA高表达、GFPT2 mRNA高表达、SNORA51 mRNA高表达是HCC患者死亡的危险因素（P＜0.05）。结论：GPSM2 、GFPT2、SNORA51在HCC组织中异常高表达,且与血管侵犯、TNM分期等临床病理特征有关,GPSM2 、GFPT2 、SNORA51高表达是HCC患者死亡的危险因素。     

膀胱癌组织CALD1 mRNA表达水平与临床病理特征和预后的关系分析

摘要 目的：探究膀胱癌组织钙调蛋白结合蛋白（CALD1）信使RNA（mRNA）表达水平与临床病理特征和预后的关系。方法：选取2015年7月-2017年7月于我院收治并确诊的111例膀胱癌患者为研究对象,收集癌组织及癌旁正常组织,采用逆转录聚合酶链式反应（RT-PCR）法检测CALD1 mRNA在膀胱癌组织与癌旁正常组织中的表达差异,分析CALD1 mRNA相对表达量与临床病理特征的关系。对患者进行5年随访,分析CALD1 mRNA相对表达量与预后的关系。结果：CALD1 mRNA在膀胱癌组织的相对表达量显著高于癌旁正常组织（P<0.05）;CALD1mRNA相对表达量与膀胱癌患者组织分化程度、TNM分期、淋巴结转移相关（P<0.05）;111例患者出院后随访5年,4例失访,107例患者获得随访,CALD1低表达组和高表达组的5年总生存率分别为72.34%（34/47）、50.00%（30/60）（P＜0.05）,CALD1低表达组5年总生存率高于高表达组;Cox比例风险回归模型分析显示,组织低分化程度、TNM高分期、淋巴结转移、CALD1mRNA高表达是影响膀胱癌患者预后的危险因素（P<0.05）。结论：CALD1在膀胱癌组织中呈高表达,CALD1mRNA相对表达量与患者组织分化程度、TNM分期、淋巴结转移及预后相关。     

慢性鼻病中乏氧诱导因子与5-脂加氧酶表达变化及其与疾病严重程度关系研究

摘要 目的：探究慢性鼻病中乏氧诱导因子（HIF-1α）与5-脂加氧酶（5-LOX）表达变化及其与疾病严重程度关系。方法：选取我院2019年7月-2020年11月期间收治的75例慢性鼻病患者作为研究对象;其中鼻息肉25例,变态反应性鼻炎25例,鼻窦炎25例;同期选择30例经鼻中隔偏曲矫正下鼻甲成形术患者的下鼻甲黏膜组织设为对照组。比较四组患者乏氧诱导因子与5-脂加氧酶蛋白及mRNA水平;并分析在不同鼻部疾病中,HIF-1α、5-LOX的相关性。结果：鼻息肉组、变态反应性鼻炎组、鼻窦炎组患者HIF-1α、5-LOX表达水平高于对照组（P<0.05）;鼻息肉组、变态反应性鼻炎组、鼻窦炎组患者HIF-1α、5-LOX表达水平比较无统计学意义（P>0.05）。鼻息肉组、变态反应性鼻炎组、鼻窦炎组患者HIF-1α、5-LOX mRNA表达水平高于对照组（P<0.05）;鼻息肉组、变态反应性鼻炎组、鼻窦炎组患者HIF-1α、5-LOX mRNA表达水平比较无统计学意义（P>0.05）。Parman相关分析,在不同鼻部疾病中,HIF-1α蛋白表达与5-LOX蛋白表达呈正相关（P<0.05）。结论：在慢性鼻病（鼻息肉、变态反应性鼻炎、鼻窦炎）中,HIF-1α、5-LOX表达水平显著升高,且在慢性疾病发展中可能相互促进、互相影响。     

Sirt3在不同热量饮食下的大鼠脂肪组织中的表达

Sirt3是一种NAD+依赖性组蛋白去乙酰化酶,在肾脏、棕色脂肪、心脏和其它代谢活跃的组织中高表达. 多项研究表明,Sirt3在细胞能量代谢、衰老、肿瘤发生等方面起着重要的作用.为探讨Sirt3基因在不同热量饮食下的SD大鼠脂肪组织中的表达差异和意义,将60只3周龄,重量(60±5) g的断乳SD雄性大鼠,随机分为热量限制组、自由摄食组、高热量组,其中高热量组用于构建肥胖SD大鼠模型,喂养16周后处死,分别用Western印迹和real-time RT-PCR方法检测各组大鼠肾周脂肪组织中Sirt3基因的蛋白和mRNA的表达量.Western印迹检测与real-time RT-PCR检测结果一致: Sirt3基因的蛋白和mRNA在肥胖大鼠脂肪组织中的表达低于自由摄食组（P<0.05）,在热量限制组大鼠脂肪中的表达高于自由摄食组(P<0.05).研究结果说明,Sirt3基因在不同热量饮食下的SD大鼠脂肪组织中的表达有差异,可能与能量代谢及肥胖有着密切关系.     

IL-4 和IL-12 在宫颈癌中的表达及对紫杉醇过敏的影响

目的:研究IL-4,IL-12在宫颈癌组织中的表达,探讨其对宫颈癌发生及术后对紫杉醇过敏的影响。方法:应用半定量逆反应-聚合酶链反应(RT-PCR)技术检测IL-4mRNA,IL-12p35以及IL-12p40 mRNA在正常宫颈组和宫颈癌组中的表达,并分析两者之间的相关性以对紫杉醇过敏的影响。结果:1.宫颈癌组中IL-4mRNA表达水平高于正常宫颈组,而IL-12p35和IL-12p40mRNA表达低于正常宫颈组,差异有统计学意义(P<0.05);2.在术后给予紫杉醇治疗的宫颈癌患者中,过敏组中IL-4mRNA的表达高于不过敏组;IL-12p35和IL-12p40mRNA则低于后者,差异有统计学意义(P<0.05)。结论:体内IL-12降低和(或)IL-4升高可促进宫颈癌的发生发展增加紫杉醇过敏的发生率。     

Different expression of lipocalin-type prostaglandin D synthase in rat epididymidis

This study was designed to explore the different expression of L-PGDS (lipocalin-type prostaglandin D synthase) in rat epididymidis and to gain further insight into the potential function of L-PGDS in male reproduction. The expression of L-PGDS in rat epididymidis was assessed using real-time quantitative PCR and immunoblotting. The distribution of L-PGDS in rat epididymidis was explored by immunohistochemical methods. The result of immunohistochemistry displayed that L-PGDS was mainly distributed in epididymidis and localized within the cytoplasm and the cilia of the epithelial cells. Real-time quantitative PCR and immunoblotting showed that L-PGDS was strikingly expressed in the caput epididymidis, while a moderate to weak expression was observed in the corpus and cauda epididymidis, the level of mRNA was 0.52+/-0.02 in the caput, 0.48+/-0.03 in the corpus and 0.32+/-0.01 in the cauda epididymidis, the level of protein expression in caput, corpus and the cauda groups was 1, 0.89+/-0.03 and 0.62+/-0.01, which suggested that L-PGDS may play certain kind of role during the process of the spermatozoa maturation.     

Les “Liver X receptors” (LXRs) et le métabolisme lipidique dans l’épididyme murin

This study investigated the role of Liver × Receptors (LXRs) in the lipid composition and gene expression regulation in mouse caput epididymidis. LXRs are nuclear receptors for oxysterols, molecules derived from cholesterol metabolism, which are present in mammals in two isoforms: LXRα, which is more specifically expressed in lipid metabolising tissues such as liver, adipose and steroidogenic tissues, while LXRβ is ubiquitous. Their importance in reproductive physiology has been sustained by the fact that male knockout mice for both LXRs have impaired fertility from the age of 5 months, leading to complete sterility by the age of 9 months. These disorders are associated with epididymal epithelium degeneration incaput epididymidis segments one and two, and with sperm midpiece fragility, leading to the presence of isolated heads and flagellae when spermatozoa are recovered from thecauda epididymidis. To further the phenotypic characterization of LXR knockout mice, the lipid composition ofcaput epididymides from wild-type and LXR knockout mice was assessed using oil red O staining on tissue cryosections, lipid extraction followed by high performance liquid chromatography or gas chromatography. Gene expression was determined by quantitative real-time PCR. We showed an accumulation of cholesteryl esters incaput epididymides fromlxrβ -/- andlxrαβ -/- mice. This accumulation was not associated with modifications in the fatty acid profiles, which are similar in all four genotypes. Changes in the expression levels of several genes are discussed in this physiological context, but cellular cholesterol efflux pathways appear to be altered in an LXRβ-dependent fashion. Altogether, these results show that LXRs are important regulators of epididymal functions, and could therefore play a key role in lipid maturation processes occurring during sperm epididymal maturation.     

Distribution of 5 alpha-reductase in the epididymis of the tammar wallaby (Macropus eugenii) and dependence of the epididymis on systemic testosterone and luminal fluids from the testis

The activity of 5 alpha-reductase was much higher in the caput and corpus epididymidis than in the cauda epididymidis. Orchidectomy caused a reduction in 5 alpha-reductase activity in the caput and corpus epididymidis, and regression of the epithelium and reduction in mass of all regions of the epididymis. Subsequent testosterone therapy caused a substantial increase in amount of epithelium and overall mass of the cauda epididymidis but showed little or no increase in any of the responses measured in the caput and corpus epididymidis. We concluded that the caput and corpus epididymidis of the tammar respond to factors other than testosterone, probably some constituent in the luminal fluid, and therefore are homologous with the initial segments of the epididymis in eutherians.     

Comparison of glutathione peroxidase 1 and iodothyronine deiodinase 1 mRNA expression in murine liver after feeding selenite or selenized yeast

The experiment was conducted to compare the effect of different selenium sources on the expression of glutathione peroxidase 1 (GPx1) and iodothyronine deiodinase 1 (Dio1) mRNA in mice by quantitative real-time PCR. A total of 60 male Kunming mice at average body weight of 20 g were allotted to three groups in a randomized complete block design, namely two treatments and one control. Mice in Group 1 were fed a basal diet as control, while mice in Groups 2 and 3 were fed the basal diet supplemented with 0.1 mg/kg selenium as sodium selenite or selenized yeast, respectively. Whole feeding experiment lasted for 30 d. At the end of the feeding trial, liver mRNA levels of GPx1 and Dio1 were determined by quantitative real-time PCR, as well as growth performance, body composition, blood and GPx activity were determined. The results showed that no significant differences in overall growth performance and body composition, including body weight, body length, heart weight, kidney weight and liver weight, were found between the experimental groups (P>0.05). Blood GPx activity increased in all of the selenium supplemented groups compared with control group (P<0.01). However, blood GPx activity in selenized yeast group was higher than that in sodium selenite group (P<0.05). Liver mRNA levels of GPx1 and Dio1 also increased in the two selenium supplemented groups compared with the control group (P<0.05), while there was no significant difference between the sodium selenite and selenized yeast groups (P>0.05). In conclusion, selenium increased the mRNA expression of GPx1 and Dio1 genes in murine liver, and there was no significant difference between the organic or inorganic form of selenium used.     

Epididymal specificity and androgen regulation of rat EP2

In primates, expression of the EP2 gene is androgen-dependent and epididymis-specific. EP2 mRNA expression was investigated in caput, corpus, and cauda regions of rat epididymis and in 15 other rat tissues. Polymerase chain reaction and Northern analyses showed that rat EP2 is expressed predominantly in the proximal caput epididymidis. EP2 mRNA expression was determined in proximal epididymides from castrated, sham-operated, and efferent duct-ligated rats. In castrated rats, EP2 mRNA decreased to <10% of that in sham-operated rats between Days 3 and 4 postcastration, demonstrating the androgen dependence of EP2 expression. In epididymides ligated unilaterally at the efferent ducts, EP2 mRNA levels were approximately equal to those in the unligated contralateral epididymides or in sham-operated rats, indicating that EP2 expression does not depend on testicular factors. In bilaterally castrated rats, immediate and delayed testosterone replacement showed the dependence of EP2 expression on circulating androgens. Injection of testosterone propionate (TP) on Days 0, 1, 2, and 3 postcastration maintained EP2 mRNA levels approximately equal to those in sham-operated rats. Starting at Day 4 postcastration, daily injection of TP for 7 days restored EP2 mRNA to approximately normal levels. These data indicate for the rat that EP2 is expressed specifically in the proximal caput epididymidis and that its expression depends on circulating androgens but not on testicular factors.     

Ultrastructural localization of PGP 9.5 and ubiquitin immunoreactivities in rat ductus epididymidis epithelium

Summary The distribution of protein gene product 9.5 (PGP) and ubiquitin in the spermatozoa and epithelial cells in the different regions of the rat duetus epididymidis (proximal caput, distal caput, corpus and cauda) was studied by Western blotting analyses and electron microscopical immunogold labelling. Western blotting analyses showed that the PGP immunoreactive band was very intense in the caput and cauda epididymidis and almost irrelevant in the corpus, while the ubiquitin immunoreactive band was intense in the distal caput and cauda. No ubiquitin immunoreactive band was observed in the proximal caput and only a very weak band was seen in the corpus. The results of electron microscopical immunogold labelling varied from one epididymal region to another. The proximal caput epididymidis presented immunoreaction to PGP in the rough endoplasmic reticulum, cytosol, mitochondria and microvilli of most principal cells, and in the cytosol, rough endoplasmic reticulum and mitochondria of most basal cells. No ubiquitin immunoreaction was observed in this epididymal region. In the distal caput epididymidis, PGP immunoreactivity was detected in some principal and basal cells in the same intracellular locations as described in the proximal caput. In this region, ubiquitin immunoreactivity appears in the apical cytosol and mitochondria of principal cells. The corpus epididymidis showed no immunoreaction to PGP or ubiquitin. In the cauda epididymidis, immunostaining to PGP was observed in most clear cells and in isolated principal cells. The intracellular location of PGP in both cell types was the cytosol, mitochondria and microvilli. Ubiquitin immunoreactivity was detected in the perinuclear cytosol and mitochondria — but not in the digestive vacuoles — of some clear cells. Scanty ubiquitin immunolabelling was also found in the microvilli, cytosol and mitochondria of some principal cells. The head of the spermatozoa present in the ductal lumen in all epididymal regions immunoreacted intensely to PGP. Ubiquitin was detected in the intermediate piece and residual cytoplasm of intraluminal spermatozoa present in the corpus and cauda epididymidis. These findings suggest that a non-ubiquitinated PGP irnrnunoreactive protein is secreted by the principal cells in caput epididymidis and binds the spermatozoon heads. It is possible that the clear cells of the cauda epididymidis secrete the ubiquitin that binds to spermatozoon tail.     

A 5-kilobase pair promoter fragment of the murine epididymal retinoic acid-binding protein gene drives the tissue-specific, cell-specific, and androgen-regulated expression of a foreign gene in the epididymis of transgenic mice

The murine epididymis synthesizes and secretes a retinoic acid-binding protein (mE-RABP) that belongs to the lipocalin superfamily. The gene encoding mE-RABP is specifically expressed in the mouse mid/distal caput epididymidis under androgen control. In transgenic mice, a 5-kilobase pair (kb) promoter fragment, but not a 0.6-kb fragment, of the mE-RABP gene driving the chloramphenicol acetyltransferase (CAT) reporter gene restricted high level of transgene expression to the caput epididymidis. No transgene expression was detected in any other male or female tissues. Immunolocalization of the CAT protein and in situ hybridization of the corresponding CAT mRNA indicated that transgene expression occurred in the principal cells of the mid/distal caput epididymidis, thereby mimicking the spatial endogenous mE-RABP gene expression. Transgene and mE-RABP gene expression was detected from 30 days and progressively increased until 60 days of age. Castration, efferent duct ligation, and hormone replacement studies demonstrated that transgene expression was specifically regulated by androgen but not by any other testicular factors. Altogether, our results demonstrate that the 5-kb promoter fragment of the mE-RABP gene contains all of the information required for the hormonal regulation and the spatial and temporal expression of the mE-RABP gene in the epididymis.     

Effects of caput ligation on rat sperm and epididymis: protein thiols and fertilizing ability.

Mammalian spermatozoa mature while passing through the epididymis. Maturation is accompanied by thiol oxidation to disulfides. In rats, sperm become motile and fertile in the cauda. We have previously demonstrated that rat caput sperm contain mostly thiols and that upon passage from the corpus to the cauda epididymidis, sperm protein thiols are oxidized. The present work was undertaken to study the role of the regions of the epididymis in sperm maturation as reflected in the thiol status, fertility, and motility of the spermatozoa. The distal caput epididymidis of mature albino rats was ligated on one side. After 5 days, sperm were isolated from the ligated caput and from caput and cauda of the control side. Thiol groups in sperm, epididymal luminal fluid (EF), and epididymal tissue were labeled using the fluorescent thiol-labeling agent monobromobimane. After ligation, changes were observed in a) sperm proteins, sperm nuclear proteins, and epididymal fluid by electrophoresis; b) epididymal tissues by histochemistry; c) progressive motility by phase microscopy; and d) fertilizing ability after insemination into uteri of immature females. We found that after ligation, caput sperm thiols, especially protamine thiols, are oxidized, rendering them similar to mature sperm isolated from the cauda epididymidis. Spermatozoa from ligated caput epididymidis gain progressive motility and partial fertilizing ability. Morphology of epithelial cells of ligated caput is similar to that of cauda cells. However, other changes in caput EF and epithelium induced by ligation render the ligated caput epididymidis different from either control caput or cauda. Hence, sperm thiol oxidation, along with the development of fertilizing ability, can occur in sperm without necessity for sperm transit through the corpus and cauda epididymidis.