OBJECTIVESTo study the expression and the significance of telomerase gene hTERT in testes of infertile male.

METHODSBy using in situ hybridization(ISH) techniques, the expression of telomerase gene hTERT mRNA in testes of 47 infertile male and 10 normal testicular tissues were observed.

RESULTSIn male testes, there was a positive correlation between the expression of hTERT and the quantity and density of germ cells(spermatogonia, spermatocyte, spermatid). The expression of hTERT in some germinal cell of maturation arrest patients were not significantly different with those of normal.

CONCLUSIONSOur results suggest that the deficiency of telomerase might be a factor for germinal cell maturation arrest and there might be some other etiological factors in these patients. Our study provides experimental groundwork for the gene therapy of male infertility.

Humans ; Infertility, Male ; enzymology ; Male ; Spermatids ; Spermatocytes ; Spermatogenesis ; Spermatogonia ; Telomerase ; deficiency ; genetics ; metabolism ; Testis ; enzymology ; physiology

OBJECTIVETo investigate the expression of cyclooxygenase-2 (cox-2) in the testes and epididymides of adult male rats and its significance.

METHODSImmunohistochemical staining was used to detect the expression and localization of cox-2 in the testicle and epididymal tissues of 40 adult male SD rats.

RESULTSStrong cox-2 immunoreactivity was detected in the epididymides and testes of the rats. In the caput epididymides, cox-2 expressed mainly in the epithelial nuclei and partly in the cytoplasm. Cox-2 was also found positive in the testis nuclei and cytoplasm.

CONCLUSIONImmunohistochemical staining is a fairly sensitive method for detecting cox-2 expression in the testes and epididymides of adult male rats.

Animals ; Cyclooxygenase 2 ; biosynthesis ; Epididymis ; enzymology ; Immunohistochemistry ; Male ; Rats ; Rats, Sprague-Dawley ; Sensitivity and Specificity ; Testis ; enzymology

OBJECTIVESTo study the changes of sexual gland 5 alpha-reductase type II activity in pubertal and adult rats with diabetes.

METHODSWe selected 40 and 90 days old male Wistar rats as pubertal and adult animal model respectively, 30 rats in each group. The rats were randomly divided into three groups: control group (C), diabetic group (D) and diabetes with insulin replacement group (ID). The activity of 5 alpha-reductase type II was measured with thin layer chromatography in the epididymis, prostate and testis.

RESULTS1. In all sexual glands of pubertal rats, the activity of 5 alpha-reductase type II in D group is significantly lower than that in C and ID groups. 2. In all sexual glands of adult rats. there is no difference in the activity of 5 alpha-reductase type II among these groups.

CONCLUSIONSThe activity of 5 alpha-reductase type II is likely to be influenced by metabolic environment, hormonal levels and local specific factors in pubertal rats, but it is relatively stable in adult rats.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase ; metabolism ; Animals ; Diabetes Mellitus, Experimental ; enzymology ; Epididymis ; enzymology ; Male ; Prostate ; enzymology ; Rats ; Rats, Wistar ; Testis ; enzymology

OBJECTIVETo investigate the location of heme oxygenase (HO) enzyme in the human testis, and explore the correlation of the expression of HO enzyme with azoospermia by analyzing its different expression levels in the testes of nonobstructive azoospermia, obstructive azoospermia and normal men.

METHODSWe detected the location of the cells expressing HO enzyme in the human testis tissue using immunohistochemistry, determined the mRNA and protein expression levels of HO-1 and HO-2 in the testes of azoospermia patients and normal healthy men by RT-fluorescence quantitative PCR (RT-FQ-PCR) and Western blot, and explored the correlation of HO expressions with the pathogenesis of azoospermia.

RESULTSHO-1 enzyme was expressed mainly in the Sertoli cells and HO-2 enzyme chiefly in the germ cells of the testis tissue. RT-FQ-PCR showed that the expression of HO-1 in the testis tissue was significantly lower in the nonobstructive azoospermia than in the normal and obstructive azoospermia groups (P < 0.05), with no significant difference between the latter two. Western blot revealed no obvious difference between the expression level of HO-1 protein and that of HO-1 mRNA. There were no differences in the expression level of HO-2 protein among the three groups.

CONCLUSIONThe expression level of HO enzyme is significantly decreased in the testis tissue of nonobstructive azoospermia patients, and the expression of HO-1 protein is consistent with that of HO-1 mRNA. As HO-1 protects the testis tissue against various stress injuries through its antioxidant, anti-inflammatory and anti-apoptotic effects, its decreased expression level may be correlated with spermatogenic dysfunction, and therefore considered as a possible mechanism of nonobstructive azoospermia.

Azoospermia ; enzymology ; metabolism ; Case-Control Studies ; Heme Oxygenase (Decyclizing) ; metabolism ; Heme Oxygenase-1 ; metabolism ; Humans ; Male ; Spermatogenesis ; Testis ; enzymology ; metabolism

OBJECTIVESTo determine the expression of nitric oxide synthase (NOS) in testis and to investigate the effects of NO on the reproductive function of testis.

METHODSTestes of adult male Sprague-Dawley rats were fixed in 4% paraformaldehyde. The paraffin sections were made as routine. Immunohistochemical ABC method was used to observe the localization of NOS.

RESULTSEndothelia NOS (eNOS), neuronal NOS (nNOS) and inductive NOS (iNOS) were all expressed in Leydig cells. Only eNOS was expressed in peritubular myoid cells, endothelial and smooth muscle cells of blood vessel, while only nNOS expressed in tunica adventitia of testicular blood vessels. The reactive substance distributes in cytoplasm with negative nuclei. Immunoreactivity for eNOS, nNOS and iNOS in all spermatogenic cells was negative.

CONCLUSIONSThree kinds of NOS were all expressed in testis and the distribution of different NOS had a little difference.

Animals ; Immunohistochemistry ; Male ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase ; analysis ; Rats ; Rats, Sprague-Dawley ; Testis ; enzymology

The serine/threonine phosphatase (PP1) isoform PP1 gamma 2, predominantly expressed in the testis, is a key enzyme in spermatozoa. High PP1 gamma 2 catalytic activity holds motility in check in immature spermatozoa. Inhibition of PP1 gamma 2 causes motility initiation in immature spermatozoa and motility stimulation and changes in flagellar beat parameters in mature spermatozoa. The PP1 gamma 2 isoform is present in all mammalian spermatozoa studied: mouse, rat, hamster, bovine, non-human primate and man. We have now identified at least four of its regulatory proteins that regulate distinct pools of PP1 gamma 2 within spermatozoa. Our studies provide new insights into biochemical mechanisms underlying development and regulation of sperm motility. We hypothesize that changes in sperm PP1 gamma 2 activity as a result of phosphorylation and reversible binding of the regulatory proteins to the catalytic subunit are critical in the development and regulation of motility and the ability of sperm to fertilize eggs. Targeted disruption of the Ppp1cc gene, which encodes the PP1 gamma 1 or PP1 gamma 2 isoforms, causes male infertility in mice as a result of impaired spermiogenesis. Our observations suggest that, in addition to motility, the protein phosphatase PP1 gamma 2 might play an isoform-specific function in the development of specialized flagellar structures of mammalian spermatozoa.
Animals ; Cattle ; Cricetinae ; Epididymis ; enzymology ; physiology ; Homeostasis ; Humans ; Male ; Mice ; Phosphoprotein Phosphatases ; genetics ; metabolism ; Sperm Motility ; physiology ; Spermatozoa ; enzymology ; physiology ; Testis ; enzymology

To study the expression of mTERT gene in the testis of SD rats and its significance, in situ hybridization (ISH) techniques were used to detect the expression of telomerase gene mTERT mRNA in the testis of SD rats. The expression of mTERT was detectable in different-age male SD rats testis. There was a positive correlation between the expression of mTERT and the location of germ cells (spermatogonia, spermatocyte, spermatid). In Sertoli cells, leydig cell and spermatozoa, telomerase mTERT was not detected. Type A spermatogonia expressed the highest level of telomerase mTERT mRNA. Our results suggest that the expression of mTERT gene in the testis of SD rats is of lifetime and coincide with the telomerase activity.
Animals ; Catalysis ; Cell Differentiation ; DNA-Binding Proteins ; Gene Expression Regulation, Developmental ; Male ; RNA ; Rats ; Rats, Sprague-Dawley ; Spermatids ; enzymology ; ultrastructure ; Spermatogenesis ; genetics ; Spermatogonia ; enzymology ; ultrastructure ; Spermatozoa ; enzymology ; ultrastructure ; Telomerase ; biosynthesis ; genetics ; metabolism ; Testis ; enzymology ; growth & development

Angiotensin-converting enzyme functions in the male reproductive system, but the extent of its function in reproduction is not fully understood. The primary objective of this work was to investigate the relationship between the testicular isoform of angiotensin-converting enzyme present in human spermatozoa and semen parameters, human embryo quality, and assisted reproduction success. A total of 81 semen samples and 635 embryos from couples undergoing oocyte donation cycles at the IVI Bilbao Clinic were analyzed. Semen parameters, embryos quality, and blastocyst development were examined according to the World Health Organization standards and the Spanish Association of Reproduction Biology Studies criteria. The percentage of testicular angiotensin-converting enzyme-positive spermatozoa and the number of molecules per spermatozoon were analyzed by flow cytometry. Both parameters were inversely correlated with human sperm motility. Higher percentages of testicular angiotensin-converting enzyme-positive spermatozoa together with fewer enzyme molecules per spermatozoon were positively correlated with better embryo quality and development. Our results suggest that embryos with a higher implantation potential come from semen samples with higher percentages of testicular angiotensin-converting enzyme-positive cells and fewer enzyme molecules per spermatozoon. Based on these findings, we propose that testicular angiotensin-converting enzyme could be used to aid embryologists in selecting better semen samples for obtaining high-quality blastocysts during in vitro fertilization procedures.
Adult ; Embryo Implantation/physiology* ; Embryo Transfer ; Embryonic Development/physiology* ; Fertility/physiology* ; Fertilization in Vitro ; Humans ; Male ; Middle Aged ; Peptidyl-Dipeptidase A/metabolism* ; Sperm Motility/physiology* ; Spermatozoa/enzymology* ; Testis/enzymology*

AIMTo investigate wether the corresponding protein of mono-ADP-ribosyltransferase 3 (ART3) mRNA is expressed in human testes and, if so, whether the expression is cell type-specific.

METHODSART3 mRNA was determined in human testes and sperm by reverse transcription-polymerase chain reaction (RT-PCR). The glycosyl-phosphatidylinositol linkage of ART3 was shown by treating ART3-transfected HEK-293-T cells with phospholipase C. Fluorescent activated cell sorter (FACS)-analyses were used to detect ART3 on mature spermatozoa and immunohistological studies to detect the protein in testes.

RESULTSART3 protein was shown to be present in testes. It was found on spermatocytes only. It was absent from spermatogonia, spermatids and spermatozoa. The absence of ART3 from spermatozoa was confirmed by FACS-analysis. ART3 protein was detected neither within a seminoma nor on Leydig cells.

CONCLUSIONHere we show for the first time that ART3 protein is expressed in testes in particular on spermatocytes, indicating that ART3 exerts a specific function only required at a particular stage of spermatogenesis.

ADP Ribose Transferases ; genetics ; Cell Line ; Flow Cytometry ; GPI-Linked Proteins ; Humans ; Male ; Membrane Proteins ; genetics ; Organ Specificity ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Spermatocytes ; enzymology ; Spermatozoa ; enzymology ; Testis ; enzymology ; Transfection

OBJECTIVETo investigate the effect of immunological orchitis on spermatic specific enzyme and fertility.

METHODSExperimental allergic orchitis (EAO) model of guinea pigs was duplicated. The histological and morphological changes of spermatic acrosomal protease and hyaluronidase, lactate dehydrogenase, sperm in epididymis and testes were observed by means of enzyme kinetical spectrophotometry and gelatin fixation of substrate thin membrane.

RESULTSThe activity of acrosomal protease, hyaluronidase and spermatic cytoplasmic lactic dehydrogenase in the epididymis acrosomal enzyme system became low, and so did the quality of sperm in epididymis. Remarkable morphological changes of spermatogenic cells developed in the convoluted seminiferous tubules.

CONCLUSIONSEAO remarkably affects the fertility of male guinea pigs. The orchis and epididymal sperms might be the sites of action.

Acrosin ; metabolism ; Animals ; Autoimmune Diseases ; enzymology ; pathology ; physiopathology ; Disease Models, Animal ; Fertility ; physiology ; Guinea Pigs ; L-Lactate Dehydrogenase ; metabolism ; Male ; Orchitis ; enzymology ; pathology ; physiopathology ; Spermatozoa ; enzymology ; pathology ; Testis ; pathology