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*

Evidence of the antigenicity of testis and semen was first presented at the end of the last century. Landsteiner (1899), Metchnikoff (1900), and Metalnikoff (1900) demonstrated the induction of a spermotoxic antibody in animals sensitized with testicular homogenate or semen; this antibody was capable of immobilizing sperm cells. The earliest manifestation of homologous type of antisperm sensitization (Kennedy, 1924) was the immobilization of spermatozoa, and in some cases atrophy of germinal epithelium, following repeated injection of testicular homogenate or epidydimal sperm. Ryoo and Kim (1982) reported that spermatogenesis was adversely affected with degeneration and sloughing of germinal cells of the seminiferous tubules in the mice which were immunized with testis homogenate plus complete Freund's adjuvant. The purpose of this study was to observe the effect of antitesticular rabbit serum produced against rat testis on spermatogenesis in rat. The results were as follows: 1.Theseminiferous tubules showed mild to moderate impairment of spermatogenesis such as degeneration and exfoliation of germinal epithelium in all experimental groups. Intraluminal spermatozoa of seminiferous tubules were decreased in number. Interspaces of seminiferous tubules were wider than normal and were infiltrated with mononuclear cells with some hemorrhage. 2. Intraluminal spermatozoa of the epididymides were markedly decreased in number but immature sperm cells were observed much more often than in normal control group.
Animals ; Atrophy ; Epithelium ; Freund's Adjuvant ; Hemorrhage ; Immobilization ; Mice ; Rats* ; Semen ; Seminiferous Tubules ; Spermatogenesis ; Spermatozoa ; Testis*

OBJECTIVE: The aim of this study was to investigate the morphological aspects of testicular tissue before and after freezing-thawing by light and transmission electron microscopy. METHODS: Tissue biopsies were carried out on mouse testis for freezing. Samples in medium containing 20% glycerol were frozen by computer-controlled freezing program. The effect of freezing-thawing on the structural change of testicular tissues were examined by light and electron microscopy. RESULTS: The freezing-thawing procedure had no significant effect on tubular diameter. However, it caused folding of the lamina propria, and notable damage to Sertoli cells, spermatogonia and spermatocytes. The cells were detached, desquamated from the basal lamina and had increased vacuolization. Round spermatids, elongated spermatids and spermatozoa were less affected, and most of them maintained their normal structure. CONCLUSIONS: The structure of spermatogonia, spermatocyte and basal compartments in seminiferous epithelium was significantly altered by freezing-thawing procedure of mouse testicular tissues. Thus, we need to develop a more reliable method for the cryopreservation of testicular tissues.
Animals ; Basement Membrane ; Biopsy ; Cryopreservation ; Freezing ; Glycerol ; Mice* ; Microscopy, Electron ; Microscopy, Electron, Transmission ; Mucous Membrane ; Seminiferous Epithelium ; Seminiferous Tubules ; Sertoli Cells ; Spermatids ; Spermatocytes ; Spermatogonia ; Spermatozoa ; Testis

OBJECTIVE: The aim of this study was to investigate the morphological aspects of testicular tissue before and after freezing-thawing by light and transmission electron microscopy. METHODS: Tissue biopsies were carried out on mouse testis for freezing. Samples in medium containing 20% glycerol were frozen by computer-controlled freezing program. The effect of freezing-thawing on the structural change of testicular tissues were examined by light and electron microscopy. RESULTS: The freezing-thawing procedure had no significant effect on tubular diameter. However, it caused folding of the lamina propria, and notable damage to Sertoli cells, spermatogonia and spermatocytes. The cells were detached, desquamated from the basal lamina and had increased vacuolization. Round spermatids, elongated spermatids and spermatozoa were less affected, and most of them maintained their normal structure. CONCLUSIONS: The structure of spermatogonia, spermatocyte and basal compartments in seminiferous epithelium was significantly altered by freezing-thawing procedure of mouse testicular tissues. Thus, we need to develop a more reliable method for the cryopreservation of testicular tissues.
Animals ; Basement Membrane ; Biopsy ; Cryopreservation ; Freezing ; Glycerol ; Mice* ; Microscopy, Electron ; Microscopy, Electron, Transmission ; Mucous Membrane ; Seminiferous Epithelium ; Seminiferous Tubules ; Sertoli Cells ; Spermatids ; Spermatocytes ; Spermatogonia ; Spermatozoa ; Testis

Cimetidine is an H2 receptor antagonist that has an antiandrogenic effect. It intervenes with the conversion of testosterone into estrogen in the Sertoli cells with accompanying testicular structural changes. In the present study, the microscopic and the ultrastructural changes induced by cimetidine and the effect of vitamin B12 as a protective agent on rat testes were studied. Immunoexpression of estrogen receptor β (ERβ) in testes was evaluated. Twenty-four adult male rats were divided into four groups: control, cimetidine-treated, vitamin B12 treated, and combined cimetidine and vitamin B12 treated. The experimental rats were administered with cimetidine and/or vitamin B12 for 52 days. Group II rats showed marked atrophy of the seminiferous tubules with a significant increase in tubular diameter and decrease in the tubular luminal and epithelial areas. Ultrastructure of this group showed irregular Sertoli cells with basal cytoplasmic vacuolation and significantly thickened basement membrane. ERβ immunoexpression was similar to controls. Group III rats showed near normal seminiferous tubular structures with minimal cellular alterations and the immunoreactivity of the testicular sections was very close to normal. However, group IV rats showed markedly immunopositive detached cells, spermatids, and primary spermatocytes. Cimetidine interferes with the control of spermatogenesis as evidenced by microscopic and ultrastructural studies and affection of ERβ receptors and vitamin B12 has a protective action against this harmful effect.
Adult ; Animals ; Basement Membrane ; Cimetidine ; Cytoplasm ; Estrogens ; Humans ; Male ; Phenobarbital ; Rats* ; Seminiferous Epithelium* ; Seminiferous Tubules ; Sertoli Cells ; Spermatids ; Spermatocytes ; Spermatogenesis ; Testis ; Testosterone ; Vitamin B 12* ; Vitamins*

Spermatogenesis is a particularly difficult process to study the unique multiple cellular associations within the seminiferous epithelium. Laser capture microdissection (LCM) is a recently developed technique that enables the isolation of individual cell populations from complex tissues. The superoxide dismutase (SOD) is the first and most important enzyme of antioxidant defense systems against superoxide anion. The aim of this study was to investigate the quantitative changes of SOD gene expression according to the spermatogenic cycle in mouse testes using LCM and real-time polymerase chain reaction (PCR) techniques. Frozen sections (10 micrometer) were obtained from the testes of 8-weeks-old ICR mice. LCM was used to capture all cells in cross-sectioned seminiferous tubules which were grouped into stages I-V, VII-VIII, and IX-XI. The expression level of cytoplasmic Cu, Zn-SOD (SOD1) mRNA was remarkably higher than those of mitochondrial Mn-SOD (SOD2) and extracellular Cu, Zn-SOD (SOD3) mRNAs in mouse testes. During spermatogenesis, the expressions of SOD1 and SOD2 mRNAs were highest on stages I-V, began to decrease after stage VII, and showed a lowest level on stage IX-XI. However, the expression of SOD3 mRNA was highest on stages VII-VIII. These findings suggest that the subtypes of SOD are expressed differentially in mouse testes during spermatogenesis.
Animals ; Cytoplasm ; Frozen Sections ; Gene Expression ; Laser Capture Microdissection ; Mice ; Mice, Inbred ICR ; Real-Time Polymerase Chain Reaction ; RNA, Messenger ; Seminiferous Epithelium ; Seminiferous Tubules ; Spermatogenesis ; Superoxide Dismutase ; Superoxides ; Testis

AIMTo evaluate the effect of a protein synthesis inhibitor cycloheximide on arresting activity in spermatogenesis and sperm count in male rats.

METHODSThe study used seminiferous tubule (ST) segments from adult rats cultured in vitro with or without cycloheximide to condition culture media, which have been concentrated, size fractioned (30-50 kDa) and administered 7 days to adult rats by intraperitoneal injections. The effects on testicular and epididymal weights, spermatogenesis and epididymal sperm count were determined.

RESULTSThe fraction (30-50 kDa), named arresting, obtained from the culture without cycloheximide decreased testicular and epididymal weights (P<0.01) and reduced the epididymal sperm count significantly. Study of the spermatogenic cycle by transillumination showed spermatogenic arrest at stage VII in rats treated with arresting compared to that observed in controls. The length of stage VII in the group receiving the seminiferous tubules culture media with cycloheximide (30-50 KDa CHX-STCM fraction) was similar to control.

CONCLUSIONThe difference in the effect may be the result of the presence or absence of arresting, a protein secreted by the tubules.

Animals ; Arrestin ; biosynthesis ; Culture Media, Conditioned ; Cycloheximide ; pharmacology ; Epididymis ; cytology ; drug effects ; growth & development ; Male ; Protein Synthesis Inhibitors ; pharmacology ; Rats ; Seminiferous Epithelium ; cytology ; drug effects ; physiology ; Seminiferous Tubules ; metabolism ; Sperm Count ; Testis ; drug effects ; growth & development

A number of tissues have been studied in the past with respect to their organ-specific antigens. In many instances it has been possible to produce autoantibodies against characteristic components. The testis, epididymis, and seminal plasma have been largely explored from this angle. Interest in the field of accessory glands began many years ago, when the first cross-reactions between extracts of prostate, seminal vesicles and seminal plasma were demonstrated. As a consequence, the possibility that some seminal plasma antigens might be present in the accessory glands before being secreted into the genital tract opened up a new approach to possible autoimmunologic damage of these glands and of seminal spermatozoa as well. The purpose of this study is made to observe the effect of homologous epididymal extracts on the spermatogenesis in mouse. Isoimmunization with extracts of mouse epididymis, administered with complete Freund`s adjuvant, has been performed in this study.The results were as follows: 1. The histological observations revealed that spermatogenesis was adversely affected by the immunization with homologous epididymal extract added with an equal amount of complete Freund's adjuvant for 6 weeks. It was observed that spermatogenesis was remarkably impaired in the experimental group whereas it was unaffected in the control group of male mouse. The results further indicated that the degeneration and exfoliation were found in the germinal cell of seminiferous tubules and in the epithelium of the epididymal ducts besides intercanalicular infiltration of m0nonuclear round cells. 2. The cross-reactions between extracts of epididymis and testicular tissues were demonstrated in mouse. 3. The immunological examination such as immune diffusion test and sperm agglutination test showed negative reaction on all of the experimental animals in this study. Therefore. the immunological change in this experiments seems to be caused by cell mediated immunity.
Animals ; Autoantibodies ; Diffusion ; Epididymis ; Epithelium ; Freund's Adjuvant ; Humans ; Immunity, Cellular ; Immunization ; Male ; Mice* ; Prostate ; Semen ; Seminal Vesicles ; Seminiferous Tubules ; Sperm Agglutination ; Spermatogenesis* ; Spermatozoa ; Testis

Evidence of antigenicity of testis and semen has been presented since Landsteiner (1899), Metchinikoff (1900) and Metalnikoff (1900) first demonstrated the induction of a spermatoxic antibody in animals sensitized with testicular homogenates or semen. Interest in the field of male accessory sex gland began longtime ago, when the first cross-reaction between extracts of prostate, seminal plasma were demonstrated. Saline extracts of prostatic secretion from bulls, tested by double agar diffusion technique showed four antigens common to serum proteins and spermatozoa. The seminal vesicle have been found to have three to five antigens, also with common reactivity to spermatozoa. Attempts have been made to induce cross-immunologic damage in the testes by repeated immunization of mice with epididymal extracts (free of sperm) plus adjuvant and it was claimed that spermatogenesis was adversely effected and fertility of females was markedify reduced following mating with immunized male (Shethye and Rao, 1968; Kim and Kim, 1982). Rabit antiserum produced against the tissue protein of rat epididymis and seminal vesicle was capable of immobilizing and agfflutinating the sperm of both animals and the rabbit antiserum against complex antigen of epididymal tissue protein and seminal vesicle tissue protein of rat was most potent on sperm immobilization and agglutination of both animals (Cha and Kim, 1975). The purpose of this study is to observe the effect of rabbit anti-rat epididymal serum on epididymis and spermatogenesis in rat. The results were as follow; 1. The intraluminalspermatozoa of epididymis were decreased in number but immature sperm cells were much more noted than normal control group. The interspaces of epididymal ducts were widened and infiltrated with mononuclear cells and congestion in some places. There was no definite degenerative changes on epididymal epithelium. 2. Spermatogenesis was mildly to moderately impaired in the experimental group whereasit was unaffected in the control group. Degeneration and exfoliation were found in the germinal cells of seminiferous tubules. Intraluminal Spermatozoa of seminiferous tubules were decreased in number.
Agar ; Agglutination ; Animals ; Blood Proteins ; Diffusion ; Epididymis* ; Epithelium ; Estrogens, Conjugated (USP) ; Female ; Fertility ; Humans ; Immobilization ; Immunization ; Male ; Mice ; Prostate ; Rats* ; Semen ; Seminal Vesicles ; Seminiferous Tubules ; Spermatogenesis ; Spermatozoa ; Testis

It is highly desirable to achieve optimal reproductive performance, reliable morphological and physiological basic data of the reproductive organs. Therefore, seasonal changes in serum testosterone, LH, and FSH concentrations, and morphological changes in testicular epithelial cells were studied in the Korean native pheasant throughout the annual cycle. Mature male pheasants[14-16 months after hatching] were used in this study. The general morphological changes of the epithelia of the seminiferous tubules were observed by dibasic stain, and semithin section from Epon blocks with a phase contrast microscopy. The ultrastructural changes of the the epithelia of the seminiferous tubules were investigated by ultrathin section with transmission electron microscope. The changes in the profiles of the serum FSH, LH, and testosterone concentratioins were measured by RIA[radioimmunoassay]. The results obtained are summarized as follows : 1. There was little variation in the average diameter of the seminiferous tubules from autumn[67.13+/-5.95micrometer] to winter[68.59+/-6.07micrometer], but the highest levels were reached in spring[192.78+/-41.58micrometer]. Thereafter, the diameter decreased slowly in summer[146.57+/-43.68micrometer], then decreased significantly in autumn[67.13+/-5.95micrometer]. 2. Serum testosterne concentration was low from autumn[13.+/-7.21ng/100ml] to winter[17.39+/-13.75ng/100ml], but the highest levels were reached in spring[127.72+/-66.47 ng/100ml]. Thereafter, the concentration was lowest in autumn[13.+/-7.21ng/100ml]. 3. Serum LH concentration increased slowly and linealy from autumn[5.04+/-1.04ng/100ml] to winter[6.23+/-1.08ng/100ml], but the highest levels were reached in spring[11.3+/-3.6 ng/100ml]. Thereafter, the concentration reached the lowest level in autumn[5.04+/-1.04 ng/100ml]. 4. Serum FSH concentration was low from autumn[4.65+/-0.63ng/100ml] to winter[4.2+/-0.98ng/100ml], but the highest levels were reached in spring[17.41+/-8.35ng/100ml]. Thereafter, concentration was the lowest in autumn[4.65+/-0.63ng/100ml]. 5. The seminiferous tubules showed the onset of the spermatongenic cycle in spring but the seminiferous tubules collected in summer exhibited partially degenerative changes. 6. The cytoplasmic process of Sertoli cells of the seminiferous tubules of the pheasant were collected in summer. Many vesicles and degeneratiye changes were included but many number of spermatozoa were embedded partially in the multivesicular bodies in these processes. 7. The diameter of the seminiferous tubules of the pheasant narrowed markedly in autumn, and atrophied in winter. The spermatogonia and Sertoli cells were arranged in monolayer. 8. The myelin figures in the cytoplasmic process of Sertoli cells of the seminiferous tubules of the pheasant in autumn. The nucleus of the Sertoli cells were of a round configuration elongated and oriented perpendicularly to the basement membrane. The results obtained provide basic data for reproductive physiology and are useful for studying the male genital organs of the Korean native pheasant. Structural changes of the seminiferous epithelial cells significantly and postively correlated with serum FSH, LH. The correlation of changes in the hormonal status with alterations of Sertoli cell organells precedes the breeding season.
Basement Membrane ; Breeding ; Cytoplasm ; Epithelial Cells ; Epithelium* ; Genitalia, Male ; Humans ; Male ; Microscopy, Phase-Contrast ; Multivesicular Bodies ; Myelin Sheath ; Physiology ; Seasons* ; Seminiferous Tubules* ; Sertoli Cells ; Spermatogonia ; Spermatozoa ; Testosterone*