Inhibition of Glyceraldehyde 3-Phosphate Dehydrogenase by Adenine Nucleotides in Pachytene Primary Spermatocytes from Rat Testes

The effect of adenine nucleotides on the catalytic activity of spermatocyte glyceraldehyde 3-phosphate dehydrogenase (GA3PDH) was studied for its significance to metabolic regulation. In the presence of glucose (10 mM), the ATP level declined whereas the ADP and AMP levels remained unchanged. During incubation with glucose, fructose 1,6-bisphosphate (fruc 1,6-P2) and dihydroxyacetone phosphate (DHAP) were accumulated markedly. GA3PDH was inhibited by ATP (Ki = 2.27 mM), ADP (Ki = 1.21 mM) and AMP (Ki = 0.73 mM) competitively with NAD (Km = 0.24 mM). The results suggest that glycolysis in spermatocytes is regulated by GA3PDH.     

Effect of Hypophysectomy on Ribonucleic Acid Synthesis in Primary Spermatocytes Isolated from Rat Testes

RNA synthesis was examined in pachytene spermatocytes isolated from intact immature rats and from hypophysectomized immature rats (2–4 days after hypophysectomy). Hypophysectomy did not result in changes in the synthesis and turnover of "heterogeneous" RNA as well as the synthesis and processing of rRNA from spermatocytes. The uptake and incorporation of ³H-uridine in the spermatocyte population isolated from hypophysectomized rats, however, was decreased. These contradictory results were explained by radioautographic experiments which showed that 25 ± 6% of the pachytene spermatocytes isolated from hypophysectomized rats were unlabelled after incubation with ³H-uridine, whereas only 3 ± 2% of the pachytene spermatocytes from intact rats were unlabelled. It appeared therefore that in the absence of gonadotropic hormones from the pituitary gland, RNA synthesis in many spermatocytes remained unchanged for at least several days, although an increased number of the spermatocytes became inactive.     

Histopathological Changes in Testes of Adult Inbred Rats Immunized Against Pachytene Spermatocytes or Sertoli Cells

Adult inbred Lewis/Wistar rats, immunized against pachytene spermatocytes or Sertoli cells prepared from maturing rats of the same strain, develop histopathological changes in the testes comparable in several respects to those reported by others in classical experimental allergic orchitis. Early changes in the structure of pachytene spermatocytes are followed by germ cell degeneration, and defoliation of germinal cells into the lumen. Concomitantly, abnormalities develop in Sertoli cell structure, and the lymph-testis barrier becomes increasingly permeable. Sertoli cells are the only testicular somatic cells which appear to be directly sensitive during the immunologic response. Disturbances in Sertoli cell functions are postulated to be crucial in the etiology of aspermatogenesis. The data are discussed in relation to the possible roles in spermatogenesis of specific antigenic determinants present on the surfaces of mature Sertoli cells and advanced germinal cells located within the adluminal compartment of the seminiferous tubule.     

Ocucrrence of Primary Spermatocytes in the Infant and Child Testis

Summary:  Light microscope and ultrastructural studies of normal infants and children testes revealed the presence of primary spermatocytes and occasional spermatids at 4, 8, 9, 11, 12 and 13 years of age, but not in all the boys of these ages. When they were found they appeared in both testes, but only in a 5–25% of the seminiferous tubules. These spermatocytes undergo degeneration or progress to abnormal spermatids which degenerate in turn. Spermatozoa were never found.
Zusammenfassung:  Das Auftreten primarer Spermatozyten in den Hoden von Kindern und Kleinkindern
Lichtmikroskopische und ultrastrukturelle Untersuchungen der Hoden normaler Kinder und Kleinkinder wiesen das Vorhandensein primärer Spermatozyten und gelegentlich Spermatiden im Alter von 4, 8, 9, 11, 12 und 13 Jahren nach, jedoch nicht bei alien Jungen der genannten Altersgruppen. Wenn diese Zellen nachgewiesen werden konnten, waren sie in beiden Hoden vorhanden, aber nur in 5–25% der Tubuli seminiferi. Diese Spermatozyten unterliegen einer Rückbildung oder einer Umwandlung in abnorme Spermatiden, die sich in der Folge zurückbilden. Spermatozoen warden nie gefunden.     

Prostatic Origin of Pyruvate Kinase Activity in Human Seminal Plasma

Pyruvat-Kinase-Aktivität aus der Prostata im Spermaplasma des Menschen
Die Pyruvat-Kinase-Aktivität findet man im Sperma-Plasma bei Menschen mit Normozoospermie, Oligozoospermie, Azoospermie und Vasektomie. Um die Herkunft dieses Enzyms herauszufinden, wurde die fraktionierte Ejakulat-Methode angewandt. Spermato zoenzahl, Aktivität der sauren Phosphatase und Fruktosekonzentration waren die Kontrollparameter, um genaue Ergebnisse zu sichern. Der Beweis wird erbracht, daß die Prostata die Hauptquelle der Pyruvat-Kinase im Spermaplasma des Menschen darstellt.     

重组人睾丸精子结合蛋白对人精子运动参数的影响

目的:探讨重组人睾丸精子结合蛋白(TSBP)对体外培养人精子运动参数的影响。方法:将22例生育男性的精液经Percoll密度梯度离心后,分别与0.01mg/ml及0.1mg/ml的重组His6-TSBP在体外共同孵育1h或3h,同时设立对照组,Western印迹检测重组His6-TSBP与精子膜的结合情况,计算机辅助精液分析(CASA)系统测定重组His6-TSBP对精子运动参数的影响。将12例弱精子症患者的精液按同样方法处理,检测重组His6-TSBP对弱精子症患者精子运动参数的影响。结果:0.1mg/ml重组His6-TSBP与生育男性精子作用1h可以提高体外培养精子的前向运动百分率(a+b级精子百分率),培养3h后前向运动百分率和活率均有所提高,差异具有显著性(P<0.05);0.01mg/ml重组His6-TSBP对检测各指标均无显著性影响。0.1mg/ml重组His6-TSBP与弱精子症患者精子作用3h可以提高精子前向运动百分率(P<0.05),但对活率无显著影响。结论:0.1mg/ml的重组His6-TSBP在体外可以提高生育男性精子的前向运动百分率和活率及弱精子症患者精子的前向运动百分率。     

Protein Synthesis in the Rat Seminiferous Epithelium During Onset of Spermatogenesis — an Autoradiographic Study

The pattern of protein synthesis in Sertoli and germ cells during the ontogeny of spermatogenesis in the rat was followed by autoradiography using (3H)-Leucine as precursor. The results suggest that the protein synthetic potential of Sertoli cells is greater than that of germ cells at any stage of differentiation. Significantly enough, the ontogeny-related protein synthetic capabilities of the germ cell compartment find a parallel in those reported for the adult rodent. Furthermore, the data support the suggestion that proteins may be transported from Sertoli cells to germ cells.     

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。     

Transformed Spermatocytes Constituting the Ejaculate of an Infertile Man/Strukturveränderte Spermatozyten im Ejakulat eines infertilen Mannes

In the ejaculate of an infertile man exclusively free immature germ cells were found. Electron microscopic examination of their cytology revealed some characteristics of primary spermatocytes. After having been released from the germinal epithelium, these cells, however, have developed into totally new cell forms, which finally degenerate and disintegrate.     

Synthesis and secretion of proteins in vitro by isolated epithelial strips from the proximal and distal vas deferens of the rat

Epithelial strips of rat vas deferens were isolated by a new technique and used to study differences in protein synthesis and secretion between morphologically defined segments of the vas deferens. The isolated strips were viable as judged by linear oxygen uptake over the incubation period and by preservation of structure. Epithelium from the proximal vas deferens incorporated more labelled amino acids into cytosolic (P less than 0.02) and incubation medium (P less than 0.01) proteins than did epithelium from distal vas deferens; this incorporation was inhibited by cycloheximide. Although some of the incubation medium proteins arose by leakage from damaged cells, specific protein secretion was indicated by differences in SDS-PAGE autoradiogram banding patterns and by differences in glycosylation between proteins in the incubation medium and those in the cytosol. Thus, the former contained more label from [14C]galactose incorporation than did cytosolic proteins (proximal: P less than 0.05; distal: P less than 0.005).     

人精子表面蛋白P34H的基因克隆及其在睾丸和附睾中表达分析

目的 :克隆精子表面蛋白P34H基因的编码区 ,为进一步体外表达P34H蛋白做准备。　方法 :提取人附睾体部总RNA ,并以此为模板 ,进行反转录PCR获得编码P34H蛋白的基因片段。应用T/A克隆策略 ,将扩增的P34H基因编码区克隆入T载体 ,并通过双酶切和DNA测序进行鉴定。同时 ,以 β actin为内参照物 ,进行反转录PCR半定量分析 ,比较P34H在附睾头部、体部和尾部及睾丸组织中的表达量。　结果 :成功地克隆了P34H基因。将P34H的cDNA序列登录GenBank ,登录号为AF5 15 6 2 5。反转录PCR半定量分析表明P34H主要在附睾体部表达。　结论 :克隆的P34H基因可用于构建表达载体     

Molecular Cloning of Human Sperm Surface Protein P34H Gene and Semi-quantitative Analysis of Its Expression in Testis and Epididymidis

Molecular Cloning of Human Sperm Surface Protein P34H Gene and Semi-quantitative Analysis of Its Expression in Testis and Epididymidis     

大鼠睾丸间质tau蛋白和微管结合蛋白表达的变化

目的 :检查老龄和青年大鼠睾丸间质组织中tau蛋白和微管结合蛋白的表达情况 ,籍此评价老龄大鼠睾丸衰退的发生机制。　方法 :应用二步法免疫组化技术检查 6月龄SD雄性大鼠 (10只 ,青年组 )和 2 8月龄同品系大鼠 (10只 ,老龄组 )睾丸组织抗Tau蛋白和抗微管结合蛋白的抗体免疫反应情况。　结果 :老龄大鼠睾丸间质组织中tau蛋白免疫反应阳性细胞数明显高于青年组 (P <0 .0 0 1) ,而青年组大鼠睾丸间质组织中微管结合蛋白免疫反应阳性细胞数明显高于老龄组 (P <0 .0 1)。　结论 :tau蛋白和微管结合蛋白免疫反应阳性细胞数量的变化可能与睾丸衰退有密切关系     

实验性精索静脉曲张对青春期大鼠睾丸、附睾中CRES蛋白表达的影响

目的:研究青春期大鼠实验性左侧精索静脉曲张(ELV)对睾丸和附睾中胱蛋白酶抑制剂相关的附睾精子发生(CRES)蛋白表达的影响,探索精索静脉曲张导致不育的机制。方法:建立青春期雄性SD大鼠ELV模型,采用免疫组化及蛋白印迹方法检测CRES蛋白在ELV 2周组(n=8)、4周组(n=8)及其相应的对照组(n均=5)大鼠睾丸和附睾头、体、尾中的表达变化。结果:免疫组化显示,CRES蛋白在ELV实验组和对照组大鼠睾丸和附睾中均有表达。在睾丸中,CRES蛋白主要定位于圆形和长形精子细胞的胞质、精子的顶体以及残余体中,其表达与生精周期密切相关,Ⅰ~Ⅲ和Ⅸ~ⅩⅣ期表达最强,Ⅶ~Ⅷ期表达次之,Ⅳ~Ⅵ期表达减弱。在附睾中,CRES蛋白主要表达于从附睾起始段到尾部的上皮主细胞的胞质中,且以体、尾部表达最强,头部次之,管腔分泌物也呈阳性表达。实验组比对照组CRES蛋白表达增强。W estern印迹显示:实验组和对照组大鼠睾丸和附睾在相对分子质量(Mr)为19 000和14 000处均可见CRES蛋白条带,其中以Mr19 000处表达更为明显。实验组CRES蛋白的表达比对照组明显增强。对免疫组化和W estern印迹结果进行灰度值和积分吸光度值(IA)测定,并经统计学分析显示:ELV 2周组和4周组比其相对应的对照组表达增强且差异有显著性(P<0.05或P<0.01),而ELV各组间未见CRES蛋白表达有明显差异(P>0.05)。结论:CRES蛋白在大鼠睾丸和附睾中均有表达,睾丸中表达呈现生精周期特异性和细胞特异性,附睾中表达呈现附睾上皮区段和细胞特异性;CRES蛋白在青春期ELV大鼠睾丸和附睾中的表达比对照组明显增强。这些结果提示CRES蛋白在精子发生和精子成熟过程中可能起着重要的作用,并且可能与ELV引起的不育或生育力低下有关。     

Effect of Chronic Chloroquine Treatment on Prostaglandin and Human Chorionic Gonadotropin (hCG) Stimulation of Testosterone Secretion by the Rat Testis

The effect of chronic chloroquine administration on prostaglandin E1 (PGE1) and human chorionic gonadotropin (hCG) stimulation of testosterone secretion by the rat Leydig cells was studied. Adult albino (Wistar strain) rats were injected with 4 mg/kg body weight of chloroquine phosphate every other day for 30 days. At the end of the period of treatment, the rats were sacrificed and the Leydig cells incubated with hCG and/or PGE1 for 3 hrs. The amount of testosterone secreted was estimated by radioimmunoassay. Chronic chloroquine treatment did not affect the activity of the Leydig cells and combined PGE1 and hCG did not show any additive effects.     

Involvement of Lysosomal Enzymes in Flutamide-Induced Stimulation of Rat Testis

Beteiligung lysosomaler Enzyme bei Flutamid-induzierter Stimulation des Rattenhodens
Das Antiandrogen Flutamid läßt den Plasmaspiegel von LH ansteigen, stimuliert die Steroidbiosynthese im Hoden und hebt die Wirkung von Androgen auf die Zielorgane auf.
Flutamid wurde Ratten verabreicht, um die intrazellulären Mechanismen im Hoden zu beobachten, wo die Steroidsyntheserate in Gegenwart des Antiandrogens erhöht war.
Das Gewicht der ventralen Prostata sowie der Bläschendrüsen zeigte die antiandrogenen Auswirkungen von Flutamid. Jedoch spiegelte sich das Vorhandensein von Testosteron in der erhöhten Aktivität der β-Glucuronidase der Niere wieder. Das Gewicht der Hoden und die Inkorporation von ³²P in die Nukleinsäurefraktion waren nach 45 Minuten erhöht. Die Konzentration der DNA des Hoden und der Ascorbinsäure hatte abgenommen. Die Aktivität der lysosomalen Fazyme, sauren Phosphatase und β-Glucuronidase war erhöht.
Unter Flutamid kam es demnach zu einem Zustand, in dem ein Wechsel der lysosomalen Stabilität in Verbindung mit einer Veränderung der Hodenfunktion gezeigt wurde.     

Synthesis of alpha 1-acid glycoprotein by the human prostate

In order better to define the extent of protein synthesis capacity of the human prostate, we have studied the translation of selected serum proteins using isolated poly(A)+ RNA preparations and the rabbit reticulocyte lysate system. The translation of alpha 1-acid glycoprotein could be conclusively demonstrated but there was no apparent translation of albumin and plasmatic transferrin. Labeled alpha 1-acid glycoprotein was identified by specific immunoprecipitation with a commercial anti alpha 1-acid glycoprotein antiserum and correct processing by canine pancreatic microsomal membranes. Furthermore, we have shown by the immunoperoxidase technique that alpha 1-acid glycoprotein was indeed localized mainly in prostatic epithelial cells in 2 out of 2 patients with benign prostatic hypertrophy and in 3 out of 11 patients with prostatic adenocarcinoma. The significance of the synthesis and secretion of alpha 1-acid glycoprotein by prostatic cell themselves is presently unknown. However, we think that it could represent an interesting subject to explore further in relation with prostatic inflammation.     

Selective uptake by the Sertoli cells of cytoplasm from normal spermatogonia in the rat testis

Electron microscopical examination of germ cells during their development from early type A spermatogonia to late pachytene spermatocytes showed that small, spherical pseudopodia emerged from type B spermatogonia and, to a lesser degree, from intermediate spermatogonia and early spermatocytes. Serial sections showed that the pseudopodia pinched off from the type B spermatogonia and were engulfed by the adjacent Sertoli cells. Groups of dense bodies were found in the Sertoli cells adjacent to the engulfed islands of germ cell cytoplasm. At a few instances islands of germ cell cytoplasm were seen to fuse with dense bodies in the Sertoli cells. The fate of the cytoplasmic islands is unknown, but phagocytosis by the Sertoli cells may be suggested. The findings indicate a new type of interaction between Sertoli cells and certain classes of spermatogonia.     

Effects of Oestrogens and FSH on LH Stimulation of Steroid Production by Testis Leydig Cells from Immature Rats

Hypophysectomy of immature male rats results after 5 days in a decreased production of testosterone by isolated testis Leydig cells in response to LH. The LH responsiveness of the Leydig cells can be partly restored by treatment of the hypophysectomized rats with FSH. In continuation of previous reports on this subject ( Steroids 28 (1976) 847; and 30 (1978) the following conclusions were derived from the results in the present paper:
1. After hypophysectomy of immature male rats the production of testosterone (T) as well as of 5-pregnenolone (Δ5P) by isolated Leydig cells in response to LH is reduced.
2. Daily administration of FSH after hypophysectomy restores the Δ5P production in response to LH almost completely, but has a much smaller effect on the restoration of T production.
3. Administration of oestradiol benzoate (E₂B) together with FSH has no effect on the restoration of LH-stimulated Δ5P production, but causes a reduction of T production, when compared with Leydig cells from animals treated with FSH only.
4. Treatment of intact immature rats with E₂B results in a decreased production of T and an increased production of Δ5P in isolated Leydig cells.
5. From experiments with labelled pregnenolone it appears that E₂B and diethylstilboestrol (DES) inhibit the 17α-hydroxylase activity of Leydig cells from intact as well as from hypophysectomized rats. This results in a reduced conversion of pregnenolone to C1:)-steroids and in increased production of 3α-hydroxy-5α-pregnan-20-one from δ5P.
6. The observed effects of FSH and E, were similar within a dose range of 100–10000 ng LH per 106 Leydig cells.