Effects of 6-Chloro-6-deoxyglucose on Electrolyte and Water Transport in the Epididymis and Fertility of Male Rats

The effects of 6-chloro-6-deoxyglucose (6 CDG) on the transport of electrolytes and water in the cauda epididymidis and fertility of male rats were studied. Injection of 6 CDG into male rats at a dose rate of 120 μM/kg/day for 7–14 days induced sterility and inhibited sodium and water reabsorption in the perfused cauda epididymidis by about 60%. The rates of potassium and protein secretion were unaffected. When these rats were allowed to recover for 10 weeks, both fertility and the Na and water reabsorption of the cauda were restored. It is proposed that the chlorinated sugar may affect cpididymal sperm metabolism through an effect on the transport function of the epididymis.     

Increase in Sperm Type Hexokinase Activity of Rat Spermatozoa During Maturation

Rat hexokinases from caput sperm (immature) and caudal sperm (mature) were investigated. The hexokinases from both sources were studied by DEAE-cellulose column chromatography and by cellulose acetate electrophoresis. The specific activity of caput sperm hexokinase was not significantly different from that of the caudal sperm enzyme. Spermatozoa possess two isozymes of hexokinase. Type I hexokinase was the predominant type in caput sperm whereas the sperm type hexokinase was predominant in caudal sperm. Hexokinase types II and III were absent in both extracts.     

增精颗粒对大鼠附睾精子质量的影响

目的 :观察中药增精颗粒 (ZJG)对不育大鼠附睾精子质量的影响 ,探讨ZJG改善附睾精子质量的机制。　方法 :将 4 0只雷公藤多甙 (GTW )实验性SD雄性大鼠不育模型随机分成 4组 ,每组 10只 ,包括不育组 [GTW 2mg/(ml·10 0g) ]和ZJG高、中、低剂量组 [高 :ZJG0 .6 7g/(ml·10 0g) +GTW 2mg/(ml·10 0 g) ,中 :ZJG0 .33g/(ml·10 0g) +GTW 2mg/(ml·10 0 g) ,低 :ZJG0 .17g/(ml·10 0 g) +GTW 2mg/(ml·10 0g) ],另设正常组 (对照 ) 10只(等量的 1%羧甲基纤维素钠溶液 )。整个实验周期 6周 ,由 3周造模期和 3周用药期组成。ZJG各组按指定的剂量用药 3周 (实验第 4 3d)观察大鼠附睾精子质量、附睾腺管壁厚度及主要性腺器官系数的变化。　结果 :ZJG高、中、低剂量组附睾精子密度分别为 (5 9.6± 3.72 )、(6 3.3± 5 .70 )、(6 9.7± 6 .91)× 10 6/ml,精子活率分别为 (6 5 .4±6 .33) %、(6 9.3± 10 .96 ) %和 (72 .6± 9.6 1) % ,畸形率分别为 (5 2 .3± 7.4 7) %、(4 6 .2± 7.73) %和 (33.2± 7.97) % ,与不育组 [精子密度 (13.1± 6 .81)× 10 6/ml,精子活率 (7.6± 5 .87) % ,畸形率 (77.2± 8.75 ) % ]相比 ,ZJG各组差异均有显著性 (P <0 .0 5 ) ,与正常组 [精子密度 (75 .6± 10 .82 )× 10 6/ml,精子?     

Physico-Chemical Characterization of the NADPH Dependent Soluble 3α-Hydroxysteroid Oxidoreductase in the Rat Epididymis

The present paper describes some important physico-chemical characteristics of the NADPH dependent, 3α-hydroxysteroid oxidoreductase (3α-oxidoreductase) in the rat epididymis. This enzyme activity is stable in the presence of reducing agents (1–10 mM 2-mercaptoethanol), EDTA (1 mM), and glycerol (10% v/v). The pH optium is in the physiological range (pH 6.0–8.0).
Chromatography of epididymal cytosol fractions on calibrated G-200 columns indicated a molecular weight of 34 000, and sucrose gradient centrifugation a sedimentation coefficient of 3.3 S20,w. The enzyme has a Stokes radius of approximately 25 Å. Calculation of molecular weight using both the sedimentation rate and the Stokes radius indicated a molecular weight of 34 400 and a frictional ratio (f/f₀) of 1.17. The enzyme migrated with an R_x of 0.37 (relative to bromophenol blue) in 6.5% polyacrylamide gels and the iso-electric point (pI) determined by iso-electric focussing in 3.5 % polyacrylamide gels was 5.9. Further indication that the enzyme is negatively charged at neutral pH was obtained by DEAE-cellulose ion exchange chromatography, from which the enzyme activity was eluted between 0.05 and 0.08M KCl.
The epididymal 3α-oxidoreductase is temperature sensitive, showing a greatly reduced activity after preincubation at 50°C for 30 min. Preincubation at 60°C caused a complete loss of enzyme activity. Interestingly, preincubation at 25°C for 30 min seemed to cause a significant increase in enzyme activity. Whether this is due to "temperature activation", "cold inactivation" or to a temperature dependent disappearance of inhibitors is not known.     

Effects of Low Doses of Alpha Chlorohydrin on the Dehydrogenases and Oxidases of Rat Epididymal Epithelium and Sperms: A Correlative Histochemical and Biochemical Study

Histochemical studies have been made of the isocitrate dehydrogenase, succinic dehydrogenase, malate dehydrogenase, glutamate dehydrogenase, DPN diaphorase, TPN diaphorase, delta 5-3 beta-hydroxysteroid dehydrogenase and monoamine oxidase in the caput, corpus and cauda epididymides of normal and alpha chlorohydrin (6.5 mg/kg/9 days) treated rats. Administration of alpha chlorohydrin in a low dose caused a conspicuous decrease in all these enzymes except delta 5-3 beta-HSD, in various cell types of epididymal epithelium and sperms. Biochemical estimations of isocitrate dehydrogenase, succinic dehydrogenase, malate dehydrogenase and delta 5-3 beta-HSD have further supported and confirmed these histochemical observations. These changes in enzyme activities after treatment with low dose of alpha chlorohydrin strongly suggest that TCA cycle and amino acid metabolism of epididymis become defective, much earlier before any histological damage to the epididymis becomes visible.     

年龄相关分子c-kit、HIWI和波形蛋白在不同月龄大鼠睾丸和附睾中的表达

目的：检查c-kit、HIWI和波形蛋白在不同月龄大鼠睾丸和附睾组织中的表达差异,研究睾丸和附睾在成熟和衰老过程中特征性分子表达的规律性改变,为研究男性生殖系统衰老提供实验依据。方法：运用免疫组织化学方法检查6、12、24月龄SD大鼠睾丸和附睾组织中c-kit、HIWI和波形蛋白抗体免疫反应强度,并对其进行比较分析。结果：c-kit特异性免疫反应阳性主要表达于精原细胞,其他细胞表达水平甚弱,不同月龄差异不显著。附睾上皮、附睾管腔内成分亦有c-kit阳性表达。HIWI在大鼠睾丸和附睾组织中表达也很丰富,6月龄和12月龄大鼠中各级生精细胞,尤其是精原细胞和初级精母细胞为免疫反应强阳性细胞;支持细胞、间质细胞、类肌细胞和血管内皮细胞亦为HIWI阳性细胞。附睾上皮有HIWI阳性表达。但是24月龄大鼠睾丸和附睾组织中HIWI表达下调。波形蛋白在睾丸和附睾组织中表达模式为6月龄大鼠睾丸和附睾组织中低表达,24月龄大鼠睾丸和附睾组织表达明显上调（P〈0.01）。结论：c-kit、HIWI在24月龄动物睾丸和附睾组织中表达下调,波形蛋白表达随增龄而明显增强。结果提示,睾丸和附睾组织的衰老与细胞和细胞信号转导异常密切相关。     

Histochemical Changes of the Rat Testis and Epididymis after Treatment of α-Chlorohydrin-Effects of a Single Low Dose

Histochemische Veränderungen des Rattenhodens und -Nebenhodens nach einer Behandlung mit α-Chlorhydrin. Effekt einer kleinen Einzeldosis
In einer histochemischen Studie wurden die Lokalisation und die Veränderungen der Lipide, Kohlenhydrate, ATPase und 5'-Nukleotidase in frischen und fixierten Präparaten des Hodens und Nebenhodens bei normalen und α-chlorhydrinbehandelten Ratten untersucht. Nach Gabe einer niedrigeren Einzeldosis von α-Chlorhydrin zeigte sich ein Abfall der Phospholipide bei correspondierendem Anstieg der Triglyceride sowohl im Hoden als auch im Nebenhoden. Glykogen, ATPase und 5'-Nukleotidase zeig- ten ebenfalls einen Rückgang nach dieser Therapie. Die physiologische Bedeutung die-ser histochemischen Veränderungen werden diskutiert.     

Purification and Localization of Acidic Epididymal Glycoprotein (AEG): A Sperm Coating Protein Secreted by the Rat Epididymis

A protein designated acidic epididymal glycoprotein (AEG) was purified from rat epididymis using ion exchange chromatography on DEAE-Sephadex, gel filtration in Sephadex G-75 and affinity chromatography on Concanavalin-A Sepharose. AEG is a major secretory product of the epididymis making up 2–3 per cent of total soluble protein. Antibody to AEG was raised in rabbits and purified by affinity chromatography on AEG-Sepharose. Quantitation of AEG in cytosol using "rocket" immunoelectrophoresis showed AEG to increase in epididymal segments from caput to cauda. Ligation of the midcorpus decreased AEG in the cauda. Localization of AEG using an immunoperoxidase method revealed that it is secreted largely by the epithelium of caput and corpus beginning with the region distal to the initial segment. It appears to be secreted by a specific cell type, probably the so-called "principal" cell. Specific staining of AEG was also noted in "clear" cells in the cauda. Spermatozoa become coated with AEG as they leave the initial segment and remain so during passage through the cauda.     

Distribution of Lectin Binding in Rat Testis and Epididymis

Summary: Seven lectins (PNA, RCA I, SBA, Con A, WGA, UEA I, DBA) conjugated with rhodamine were employed to analyse the staining pattern of glycoproteins with varying sugar residues in the testis and epididymis of adult Wistar rats. Some lectins (UEA I, SBA, DBA) gave rather specific staining of the mature acrosome, while others (PNA, RCA I) showed affinity for the early stages of acrosome formation or had a wide affinity for germinal and non-germinal cells and structures (Con A, WGA). In the epididymis the sperm mass had a homogenous staining reaction with some lectins (PNA, RCA I, Con A, WGA, DBA) which also showed a rather strong reaction on the epithelial surface. It was concluded that this reaction is at least partially due to the secretory products synthetized by principal, apical, narrow and light cells of the epididymal epithelium. Some differences in the staining pattern of these cells were recorded indicating specialization of the cells for the production of distinct glycoproteins. The staining pattern of the interstitial and intertubular compartment of the testis and epididymis was also recorded. Zusammenfassung: Die Verteilung der Lektin-Bindung im Hoden und Nebenhoden der Ratte Sieben Lektine (PNA, RCA I, SBA, Con A, WGA, UEA I, DBA) wurden verbunden mit Rhodamin verwendet, um das Darstellungsmuster von Glykoproteinen mit verschiedenen Zuckerresten im Hoden und Nebenhoden erwachsener Wistar-Ratten zu analysieren. Einige Lektine (UEA I, SBA, DBA) gaben eine recht spezifische Darstellung des reifen Akrosoms, wohingegen andere (PNA, RCA I) eine Affinität zu den frühen Stadien der Akrosomformation zeigten oder eine breite Affinität zu Germinal- und Nichtgerminalzellen und -strukturen aufwiesen (Con A, WGA). Im Nebenhoden zeigte die Spermatozoenmasse ein homogenes Darstellungsmuster mit einigen Lektinen (PNA, RCA I, Con A, WGA, DBA), welche auf der epithelialen Oberfläche auch eine recht starke Reaktion aufwiesen. Es wurde daraus geschlossen, daβ diese Reaktion zumindest teilweise auf die sekretorischen Produkte, die in den Haupt-, Apikal-, Schmal- und Hellzellen des Nebenhodens synthetisiert werden, zurückzuführen ist. Einige Unterschiede im Darstellungsmuster dieser Zellen wurden aufgezeichnet, die die Spezialisierung der Zellen für die Produktion bestimmter Glykoproteine anzeigen. Das Darstellungsmuster des interstitiellen und intertubulären Anteils des Hodens und Nebenhodens wurde ebenfalls aufgezeichnet.     

有机阳离子转运子2在人类附睾中的表达及其意义

目的:研究人类附睾有机阳离子转运子2(OCTN2)mRNA的表达,探讨附睾肉碱转运机制,为探索男性避孕节育新技术提供理论依据。方法:应用RT-PCR方法检测人类附睾头、体、尾组织中OCTN2 mRNA的表达。结果:人类附睾头、体、尾组织中都存在OCTN2 mRNA表达。结论:人类附睾可能依赖OCTN2转运肉碱进入附睾管,为精子提供能量,促进精子成熟。对人类附睾OCTN2的进一步研究,将成为男性节育研究中新的分子靶标。     

附睾分泌蛋白2β1在青春期雄性大鼠睾丸和附睾中的表达

目的:探讨附睾分泌蛋白2(HE2/EP2)的一种异构体———HE2β1在青春期雄性大鼠睾丸和附睾中的表达及其意义。方法:应用免疫组化SP法检测15只青春期SD大鼠睾丸和附睾组织中HE2β1的定位及其表达情况。结果:HE2β1在青春期大鼠睾丸和附睾组织中均有表达。在附睾中,HE2β1主要表达于附睾管上皮主细胞胞质内,而在亮细胞、晕细胞及基细胞内未见阳性表达;其表达水平在附睾头部远段较弱,在体部近、中段及尾部较强,而在附睾始段未见阳性表达。在睾丸生精小管中,部分精原细胞核及支持细胞核均可见明显的棕褐色的阳性颗粒,其他生精细胞以及间质细胞均为阴性。结论:HE2β1在青春期雄性大鼠的睾丸和附睾上皮中均有表达,其定位及表达水平具有区域特异性和细胞特异性,提示其在大鼠精子发生、成熟及附睾上皮天然抗感染机制中发挥重要的作用。     

大鼠精索静脉曲张后附睾上皮细胞凋亡及管腔α-1,4-葡糖苷酶、唾液酸含量观察

目的:探讨大鼠左侧精索静脉曲张后同侧附睾上皮细胞凋亡与附睾管腔中-α1,4-葡糖苷酶和唾液酸含量改变的关系及意义。方法:16只雄性SD大鼠建立左侧精索静脉曲张模型,分为对照组和实验组,每组8只。缺口末端转移酶标记技术检测附睾上皮细胞凋亡;硝基酚法和5-甲基苯二酚法分别检测附睾管腔液-α1,4-葡糖苷酶和唾液酸含量。结果:大鼠左侧精索静脉曲张模型建立后的第7 d,实验组同侧附睾上皮细胞凋亡指数显著高于对照组(P<0.001);实验组附睾管腔液-α1,4-葡糖苷酶、唾液酸含量显著低于对照组(P<0.05,P<0.005)。结论:大鼠左侧精索静脉曲张可导致同侧附睾上皮细胞凋亡增加,进而影响该侧附睾上皮细胞的合成和分泌功能。     

L-肉碱在奥硝唑所致大鼠睾丸和附睾损伤中的保护作用

目的:探讨L-肉碱(LC)在奥硝唑(ORN)所致大鼠附睾和睾丸损伤中的的保护作用。方法:40只雄性SD大鼠(200～230g)随机均分为5组:①A组:给予0.5%的羧甲基纤维素钠(溶剂)灌胃;②B组:每天给予400mg/kgORN灌胃;③C组:每天给予800mg/kgORN灌胃;④D组:每天给予[ORN(400mg/kg)+LC(100mg/kg)]灌胃;⑤E组:每天给予[ORN(800mg/kg)+LC(100mg/kg)]灌胃。上述各组均连续灌胃20d,末次给药24h后,所有大鼠麻醉后处死,分别取睾丸、附睾,进行称重和HE染色,计算睾丸、附睾系数并观察睾丸和附睾病理组织学改变。结果:①与A组相比,B组睾丸、附睾系数明显降低(P<0.05);而C组睾丸、附睾系数为极显著性降低(P<0.01);D组与A组相比无差异,E组与A组相比有极显著性差异(P<0.01);②HE染色显示,与A组相比,B组睾丸生精小管内各级生精细胞排列基本整齐,部分生精小管管腔内有脱落的生精细胞,附睾管腔中精子数目下降,有时可见散在的生精细胞;C组大鼠睾丸生精小管管腔内均可见坏死脱落的生精细胞,附睾管腔中精子数目明显减少,且有较多的非精子细胞成分。D组睾丸生精小管无明显改变,附睾管腔中精子数目也未见明显下降;E组睾丸生精小管管腔内精子数目减少,可见坏死脱落的生精细胞,附睾腔中精子数目明显减少,并伴有较多的非精子细胞成分。结论:奥硝唑(ORN)可导致雄性大鼠附睾和睾丸病理组织学改变,LC对ORN引起大鼠附睾和睾丸损伤具有一定的保护作用。     

一氧化氮合酶在食蟹猴睾丸及附睾中的表达及意义

目的:探索一氧化氮合酶(NOS)在食蟹猴睾丸及附睾中的表达及意义。方法:运用免疫组化染色法观察NOS在8只猴龄为8岁左右性成熟食蟹猴睾丸及附睾中的分布。结果:①神经元型NOS(nNOS)免疫反应阳性见于睾丸生精小管上皮内各级生精细胞、腔内精子、附睾输出小管上皮、血管内皮细胞。②诱导型NOS(iNOS)免疫反应阳性见于附睾输出小管上皮、腔内精子、管周类肌细胞、血管内皮细胞。③内皮型NOS(eNOS)免疫反应阳性见于睾丸间质细胞、附睾输出小管上皮、腔内精子、管周类肌细胞、血管内皮细胞。结论:NOS广泛表达于性成熟食蟹猴睾丸及附睾组织细胞中,推测其在参与精子发生、成熟及睾丸激素分泌等过程中起到重要作用。     

Age Related Development of Angiotensin Converting Enzyme in Testes and Epididymis of Rat

Altersabhängige Entwicklung von Angiotensin-Converting-Enzym (ACE) in den Hoden und Nebenhoden der Ratte
ACE wurde in den Hoden und in verschiedenen Bereichen des Nebenhodens der Ratte unterschiedlichen Alters untersucht. Die ACE-Aktivität erreichte ihren Gipfel, wenn die Tubuli seminiferi die Spermatozoenproduktion des Erwachsenenstadiums aufwiesen; sie pendelte sich auf dieser Höhe danach ein. Die Spitzenkonzentration im Nebenhoden wurde später beobachtet als im Hoden. Der Konzentrationsgrad im Nebenhoden war proportional zum Grad der Spermatozoenreifung und zur Fertilität; die höchste ACE-Aktivität im Nebenhoden fand sich im Kopfbereich.     

血管内皮生长因子和其受体Flt-1在青春期大鼠睾丸、附睾及附睾内精子上的表达研究

目的:通过对血管内皮生长因子(VEGF)及其受体Flt-1在大鼠睾丸、附睾及附睾内精子上表达的研究,探讨其在雄性生殖系统中的作用。方法:免疫组化SP法和免疫荧光法检测20只青春期SD大鼠睾丸、附睾及精子上VEGF和Flt-1蛋白的表达情况。结果:VEGF和Flt-1在大鼠睾丸和附睾组织及精子上均有特征性表达。睾丸内VEGF蛋白表达于生精细胞、精子细胞发育中的顶体、Sertoli和Leydig细胞胞质内;Flt-1只见于精子细胞发育中的顶体及Leydig细胞胞质中。附睾中VEGF表达于各段上皮主细胞胞质内,而Flt-1表达于头、尾段上皮主细胞胞质内,体部免疫染色阴性;两者在附睾上皮亮细胞、晕细胞和基细胞中均为阴性表达。免疫荧光染色显示,VEGF和Flt-1共同定位于附睾内精子头部的顶体,尾部的颈、中和主段。结论:VEGF和Flt-1蛋白在大鼠睾丸、附睾及精子中的特异性表达提示,他们可由不同生精上皮细胞、间质细胞和附睾主细胞产生,可能以自分泌或旁分泌的形式单独或共同作用于睾丸和附睾的生殖细胞或Leydig细胞,直接或间接地影响精子的发生、发育和成熟过程,并与精子的活动和受精能力有关。     

The General Importance of Proteins and other Factors in the Transfer of Steroids into the Rat Epididymis

The entry of total radioactivity into the perfused tubule of the cauda epididymidis was measured during intravenous infusion of labelled steroids into the anaesthetised rat. The effect of altering the composition of the perfusate on this entry and the nature of radiometabolites was also studied. No radioactivity entered the epididymal lumen during infusions of cholesterol, and in the absence of luminal proteins the entry of isotope was generally low (< 14% plasma levels) for all steroids except dehydroepi-androsterone. C_21/22-steroids exhibited a delay of about 30 min before a slow rise in luminal activity reached between 3% and 14% plasma levels. For C₁₉-steroids a low plateau level of radioactivity was reached (at 6%-9% plasma levels) with dihydrotestosterone and androstenedione. whereas luminal activity continued to rise with respect to blood during infusions of dehydroepiandrosterone. When protein was added to the perfusing solution the delay in entry of radioactivity was decreased and plateau levels established for C_21/22-steroids (at 14%-38% plasma levels). The extents of entry of dihydrotestosterone and androstenedione were raised (34%-47%) and radioactivity continued to rise in the lumen relative to blood, but at a faster rate during infusions of dehydroepiandrosterone and perfusion with protein. Testicular fluid protein had little effect on the entry of activity for any steroid. There was no correlation between the extent of entry of tracer with binding of the steroids to plasma or perfusates, or partition into lipids. Tentative identification of metabolites indicated an entry of both dihydrotestosterone and dehydroepiandrosterone during their respective infusions. Androstenedione and andro-stanediols were excluded whereas 5u-reduced metabolites appeared during of androstenedione and dehydroepiandrosterone.     

In vitro Metabolism of (3H)-Androstenedione in the Rat Epididymis and Vas Deferens

The in vitro metabolism of (3H)-androstenedione in the epididymis and vas deferens of intact and castrated rats was investigated and the metabolites formed were identified by radio gas chromatography. Incubation of slices of caput epididymidis for 2 hr at 34 degrees C metabolised 90% androstenedione. Similar incubations of tissue samples from cauda epididymidis and vas deferens metabolized 60 and 25% of androstenedione respectively. The major metabolites formed in the epididymis were androstanedione (caput: 48%; cauda: 33%) and androsterone (caput: 35%; cauda: 13%). These metabolites appeared in much less concentration in the incubations with vas deferens (about 8% each). In general, conversion to testosterone and dihydrotesterone was low in all the three organs examined. Castration did not significantly alter the metabolic pattern in the caput epididymidis and vas deferens but promoted the formation of androsterone (38%) in the cauda epididymidis. The conversion of androstenedione, a weak androgen to testosterone, dihydrotestosterone and 3 alpha/3 beta-diols in the epididymidis and vas deferens of castrated rats may be of physiological significance. In addition, androsterone appears to be an important androgenic metabolite in the epididymis.     

Acid Phosphatase of the Rat Epididymis. III Histochemical and biochemical responses in experimental conditions

Die spezifischen Aktivitäten der drei sauren Phosphatasen des Nebenhodens wurden bei heranwachsenden Ratten untersucht. Alle Aktivitäten waren konstant erhöht, aber sie offenbarten zwei steilere Anstiege wahrscheinlich korrespondierend mit dem Anstieg der Androgensekretion und dem Auftreten der Spermatozoen im Nebenhoden. Gleichzeitig wurde ein Aktivitätsanstieg bei den histochemischen Färbungen in allen Nebenhodensegmenten beobachtet.
Sowohl die biochemischen als auch die histochemischen Untersuchungen zeigen, daß die Kastration die Aktivitäten der sauren Phosphatasen reduziert; dieser Effekt ließ sich wiederaufheben mit Testosteron-östradiol-Behandlung. Östradiol hat keinen Einfluß auf die Aktivität bei kastrierten Tieren; bei Normaltieren ergab sich hierunter eine Erholung der Enzym-I-Aktivität. In der histochemischen Studie scheint Östradiol die Aktivität der sauren Phosphatase wiederherzustellen — Enzym I reagiert langsamer als die anderen gegenüber Kastration und Hormontherapie. Eine erhöhte Eliminierung durch Ligatur des Nebenhodens verursachte eine Reduzierung der Aktivität aller Enzyme innerhalb von 4 Tagen; das war vor allem im Korpusbereich des Nebenhodens der Fall. Die vorliegende Untersuchung bringt keine weitere Erklärung für die Annahme, daß die sauren Phosphatasen an der Eliminierung von überzähligen und defekten Spermatozoen teilhaben.