We have performed bilateral testicular biopsies on 36 patients with varicocele and discussed the relation between histologic findings and semenogram. The results obtained in the study are as follows. 1. The volume of the semen increased more than 4.0 ml in our series by 19. 5%, and decreased below 1.5ml by 8.3%. The sperm count, motility and morphology were seen to depend on the size of varicocele 2. Testicular histology showed germinal cell hypoplasia and premature sloughing of immature cells into the lumen of the tubules, which seems to be impaired spermatogenesis. These cells are similar to those seen in the ejaculate, including tapering forms and spermatids. In most of the cases the tubular thickening was found in variable degree, and this is considered as a possible prognostic factor. Our study showed on preferential influence on testicular histology in relation to right or left side, except a few cases. The size of varicocele is no concerned with the histologic change of the testicle. 3. In comparing the histologic change of the left testicle with the semenogram, the sperm morphology was thought to be correlated with the histologic change but the count and motility were not so.
Biopsy* ; Humans ; Semen ; Sperm Count ; Spermatids ; Spermatogenesis ; Spermatozoa ; Testis ; Varicocele*

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

OBJECTIVESTo develop a simple and effective method by which spermatids can be isolated from mouse testis.

METHODSCombination of enzymatic digestion was used to prepare suspension of spermatogenic cells from adult mouse testis, and then a modified discontinuous Percoll gradient (15%, 22%, 30%, 40%, 50%, 60%) centrifugation method was introduced to isolate spermatids from the cellular suspension. The content of spermatids in each isolated fraction by Percoll method was determined by morphology (Wright-Giemsa staining) and flow cytometry analysis, and the viability of spermatogenic cells was assessed using Eosin Y exclusion test.

RESULTSMore than 97% of the testicular cells remained their viability after enzymatic digestion. After Percoll centrifuged, six fractions were formed. In each isolated fraction, the 22% fraction contained mostly spermatids(mean 86.7%) and cell viability was more than 85.5%. While in the 30% fraction, immature spermatogenic cells were present, and more than 92% of the cells remained their viability.

CONCLUSIONSA large of relatively purified spermatids can be isolated from mouse testis by enzymatic digestion combined discontinuous Percoll gradient centrifugation method.

Animals ; Cell Separation ; methods ; Centrifugation, Density Gradient ; methods ; Male ; Mice ; Spermatids ; cytology ; Testis ; cytology

It would appear that unilateral rupture of the testis causes the breakdown of sequestration mechanisms which normally prevent presentation of antigens within the genital tract to the immune system. We performed this experiment to evaluate the histological changes of contralateral testis following rupture of unilateral testis in rats. Left testicular rupture was induced in Sprague- Dawley rats by incision of tunica albuginea and partial resection of seminiferous tubules. After 1 and 7 days, the ruptured testes were surgically removed and all rats were fed for 5 months. And there was algo an experimental group that the ruptured tested were not removed and were fed for 5 months. Right testes were obtained just after being sacrificed in each group and histological changes were compared with the control group. The size of seminiferous tubule was markedly decreased and the basement membrane was thickened in groups that the ruptured tested were removed after 7 days and not removed for 5 months. Also, spermatid was not found and only sertoli cells were lined in these groups. However, these changes were not noticed in control group and the group that ruptured testes were removed after 1 day. These results imply that unilateral testicular rupture may cause histological changes of contralateral testis and early exploration may reduce contralateral testicular damage.
Animals ; Basement Membrane ; Immune System ; Infertility ; Rats* ; Rupture* ; Seminiferous Tubules ; Sertoli Cells ; Spermatids ; Testis*

Glucose is essential for testicular function; the uptake of carbohydrate-derived glucose by cells is mediated by glucose transporters (GLUTs). In the present study, we investigated the activity of GLUT1 and GLUT3, the two main isoforms of GLUTs found in testes, in the left scrotal and right abdominal testes of a German Shepherd dog. Immunohistochemical analysis showed that GLUT1 immunoreactivity was absent in the scrotal and abdominal testes. In contrast, weak to moderate GLUT3 immunoreactivity was observed in mature spermatocytes as well as spermatids in the scrotal testis. In the abdominal testis, relatively strong GLUT3 immunoreactivity was detected in Leydig cells only and was absent in mature spermatocytes and spermatids. GLUT3 immunoreactivity was significantly decreased in the tubular region of abdominal testis and significantly increased in the extra-tubular (interstitial) region of abdominal testis compared to observations in the each region of scrotal testis, respectively. These results suggest that GLUT3 is the major glucose transporter in the testes and that abdominal testes may increase the uptake of glucose into interstitial areas, leading to an increased risk of developing cancer.
Animals ; Cryptorchidism ; Dogs* ; Glucose Transport Proteins, Facilitative* ; Glucose* ; Leydig Cells ; Male ; Protein Isoforms ; Spermatids ; Spermatocytes ; Testis*

Number of germ cells in the seminiferous epithelium was analyzed quantitatively in testicular biopsy specimens of 23 patients without ductal obstruction and of 4 patients with ductal obstruction. Roth number of mature spermatids within each cross-section of seminiferous tubule and number of atrophic tubule were counted in biopsy specimens. Results were expressed as cell number of mature spermatids per seminiferous tubule and percentage of atrophic tubules. A significant correlation was demonstrated between sperm density and mature spermatid counts. Patients with sperm counts of less than 40 x l0(6)/ml had mature spermatids counts of less than 25 per seminiferous tubule. Coefficients of correlation between mature spermatid count and percentage of atrophic tubules were higher than those of correlation between sperm counts and percentage of atrophic tubules. In asoospermrc patients with epididymal obstruction, sperm count after corrective surgery could be predicted correctly by this quantitative analysis technique of testicular biopsy specimens and partial obstruction of anastomotic site of seminal tract could be proved in oligozoospermic patients after corrective surgery.
Biopsy ; Cell Count ; Germ Cells ; Humans* ; Seminiferous Epithelium* ; Seminiferous Tubules ; Sperm Count ; Spermatids ; Spermatozoa ; Testis*

The relationship between Sertoli cells and germ cells in varicocele remains controversial. To study this relationship in varicocele, seminiferous tubular changes were observed in pubertal rats according to the length of time after induction of the varicocele and the interval between induction and repair of the varicocele. As the length of time of the varicocele increased, accumulation of lipid inclusions within the Sertoli cell cytoplasm appeared first and then premature sloughing of the early spermatids appeared. Lastly, decrease in testicular weight and mean seminiferous tubular diameter (MSTD) together with decrease in the number of late spermatids were observed. Inter-Sertoli cell junctions were preserved unrelated to the duration of the varicocele. When Sertoli cell changes were reversed after varicocele repair, premature sloughing of the early spermatids was not observed. The testicular weight, MSTD and number of late spermatids were significantly increased compared to controls. When Sertoli cell changes were not fully reversed after varicocele repair, premature sloughing of the early spermatids was still observed. The testicular weight, MSTD and number of late spermatids were not significantly increased compared to controls. These results suggest that the blood-testis barrier remains intact in varicocele. The Sertoli cell is the primary intratubular site of alteration leading secondarily to spermatogenic disruption in varicocele. Changes in the Sertoli cell cause premature sloughing of the early spermatids and affect maximally the spermatid Stage.
Animals ; Blood-Testis Barrier ; Cytoplasm ; Germ Cells* ; Intercellular Junctions ; Rats* ; Sertoli Cells ; Spermatids ; Varicocele*

OBJECTIVETo establish an effective method for haploid spermatid enrichment by Hoechst 33342 staining and flow sorting.

METHODSMouse testicular monoplast suspension was prepared by two-step enzyme digestion, and the cells were incubated in the medium containing Hoechst 33342 and Verapamil. Haploid spermatids were separated and enriched according to their DNA content by flow sorting. The gene expressions in the spermatids of several histone-modified enzymes, including the histone acetylases (HAT) and histone deacetylases (HDAC), were examined by RT-PCR and compared with that in the HAT-inhibitor curcumin-treated counterparts.

RESULTSWe successfully enriched the haploid spermatids with high purity and further purified the round and elongated spermatids. RT-PCR results indicated the specificity of the expression of the HAT gene in the spermatids, and that it was influenced by curcumin.

CONCLUSIONFlow sorting can efficiently improve the purity of haploid spermatid enrichment, which helps a lot to elucidate the mechanisms of spermiogenesis.

Animals ; Cell Separation ; methods ; Flow Cytometry ; methods ; Haploidy ; Male ; Mice ; Mice, Inbred ICR ; Spermatids ; cytology

PURPOSE: We determined the usefulness of in vitro germ cell culture in nonobstructive azoospermic patients diagnosed with Sertoli cell only syndrome, no sperm in testicular sperm extraction. MATERIALS AND METHODS: This study included 44 patients (45 testicular tissues) with nonobstructive azoospermia who were diagnosed with Sertoli cell only syndrome and were found to have no sperm in testicular sperm extraction between January 2006 and July 2008. Among the 45 testicular tissues, 22 tissues were processed for culture. In the in vitro cultures, the testicular tissues were dissociated and plated on gelatin-coated dishes. Patients were divided into 2 groups according to culture success: group I, culture positive (+; n=10); and group II, culture negative (-; n=12). RESULTS: The mean patient ages were 31.73 and 31.68 years for groups I and II, respectively. The mean testicular sizes were 10.19 and 10.42 cc, respectively; the semen volumes were 2.86 and 3.04 cc, respectively; and the mean FSH, LH, and testosterone levels were 18.86 mIU/ml, 5.99 mIU/ml, and 4.46 ng/ml vs. 21.02 mIU/ml, 6.29 mIU/ml, and 4.32 ng/ml for groups I and II, respectively, with no significant differences between the groups (p>0.05). The culture rate of nonobstructive azoospermic patients diagnosed with Sertoli cell only syndrome was 45.5% (10/22). Round spermatid injection was done in 2 patients with consent of the patients, but implantation failed. Among the 45 tissues, germ cells were found in 8 tissues after pathologic reexamination. CONCLUSIONS: The in vitro culture of germ cells would be useful in the advanced treatment of nonobstructive azoospermic patients.
Azoospermia ; Germ Cells ; Humans ; Semen ; Sertoli Cell-Only Syndrome ; Spermatids ; Spermatozoa ; Testosterone

Descending of the testes is an important process for spermatogenesis and cryptorchidism is one of the most relevant genital defects in dogs. In a previous study, we observed abnormal morphology and proliferation of Sertoli cells in a cryptorchid testis. In the present study, we investigated the expression of estrogen and progesterone receptors in the normal and cryptorchid testis of a dog. Elective orchidectomy was performed on the dog's abdominal right testis (undescended, cryptorchid) and scrotal left testis (descended, normal). In the normal testis, estrogen receptor α immunoreactivity was detected in Leydig cells alone, while estrogen receptor α immunoreactivity in the cryptorchid testis was significantly prominent in the Sertoli cells as well. In addition, progesterone receptor immunoreactivity in the control testis was detected in the spermatids, but was not detected in the cryptorchid testis. This result suggests that unilateral cryptorchidism causes increases of estrogen receptor α expression in Sertoli cells.
Animals ; Cryptorchidism ; Dogs* ; Estrogens* ; Leydig Cells ; Male ; Orchiectomy ; Progesterone* ; Receptors, Progesterone* ; Sertoli Cells ; Spermatids ; Spermatogenesis ; Testis*