Spermatogenic cycle length and spermatogenic efficiency in the gerbil (Meriones unguiculatus)

The gerbil (Meriones unguiculatus) is a rodent native of the arid regions of Mongolia and China. Because the gerbil can be easily bred in laboratory conditions, this species has been largely used as an experimental model in biomedical research. However, there is still little information concerning the testis structure and function in the gerbil. In this regard, we performed a detailed morpho-functional analysis of the gerbil testis and estimated the spermatogenic cycle length utilizing 3H-thymidine as a marker for germ cell progression during their evolution through the spermatogenic process. The stage frequencies of the XII stages characterized according to the acrosome formation and development were (I-XII) 13.8, 10.1, 8.1, 7.8, 4.0, 11.2, 7.5, 7.1, 5.9, 7.6, 8.1, and 8.9. The mean duration of each seminiferous epithelium cycle was determined to be 10.6 +/- 1.0 days and the total duration of spermatogenesis, based on 4.5 cycles, was approximately 47.5 days. The volume density of tubular and interstitial compartments was approximately 92% and 8%, respectively. Based on the volume occupied by seminiferous tubules in the testis and the tubular diameter, about 9 and 18 m of seminiferous tubules were found per testis and per gram of testis, respectively. Twelve primary spermatocytes were formed from each type A1 spermatogonia. The meiotic index was 2.8, indicating that 30% of cell loss occurs during meiosis. The number of Leydig and Sertoli cells per gram of the testis was 28 million and each Sertoli cell was able to support approximately 13 spermatids. The daily sperm production per gram of testis (spermatogenic efficiency) was 33 million. Taken together, these data indicate that, mainly due to the high seminiferous tubule volume density and Sertoli cell support capacity for germ cells, the gerbil presents high spermatogenic efficiency compared with other mammalian species already investigated. The data obtained in the present study might provide the basis for future research involving the reproductive biology in this species.     

Quantitative testicular biopsy in congenital and acquired genital obstruction

To determine if congenital obstruction of the genital tract is associated with significant testicular histopathological conditions compared to acquired forms of obstruction we performed testicular biopsy in 8 vasectomized men and 5 men with vasal agenesis. Quantitative analysis of the seminiferous tubular and epithelial parameters demonstrated a statistically significant increase in tubular wall thickness in the vasectomized group. There was no significant difference among the groups with reference to the mean number of late spermatids per seminiferous tubules, mean number of Sertoli cells per seminiferous tubules, mean number of seminiferous tubules per field (100 times) or mean seminiferous tubular diameter. We conclude that despite a lifelong duration of obstruction, men with vasal agenesis demonstrate a more favorable testicular histological status compared to men after vasectomy. This finding may have therapeutic implications when considering assisted pregnancy techniques as a method of treatment of male genital tract atresia.     

BCG-induced orchitis: Structural changes during the degeneration of seminiferous tubules of rats and rabbits

Summary The effect of BCG-induced orchitis on the structure of the seminiferous tubules in rats and rabbits was investigated by light and electron microscopy. The formation of cavities between Sertoli cells and the displacement of the cells of the spermatogenic cycle are the earliest changes to be observed. Individual Sertoli cells degenerate and separate from spermatocytes and spermatids. The latter form multinuclear complexes by a broadening of the intercellular bridges. The nuclei of spermatids undergo ring-like chromatin condensation in the rat and swelling in the rabbit. After the loss of spermatocytes and spermatids from the germinal epithelium, the remaining Sertoli cells have a very irregular shape and contain many residual bodies, which are probably derived from previously phagocytosed spermatids. They often contain crystalline inclusions. The nuclei of Sertoli cells show small chromatin condensations. In the rabbit, the tubular wall increases considerably in diameter. In the vicinity of a granuloma in the interstitium caused by BCG inflammatory cells accumulate around the wall of the seminiferous tubules. Although the basal lamina seems to be an obstacle, penetration of macrophages into the tubular lumen could be observed. However, this occurred only after the degeneration of the germinal epithelium.     

On the Postnatal Development of the Terminal Segment of the Seminiferous Tubules in Normal and Germ Cell Depleted Rat Testes

The development of the terminal segment of the seminiferous tubules was studied in 5 to 50 days old normal rats. At the age of 5, 10, and 15 days the terminal segment contained fewer gonocytes or spermatogonia than did the corresponding seminiferous tubule. The differentiation of the terminal segment was obvious at 20 days of age due to the high number of germ cells in the seminiferous tubules, where the epithelium became stratified at this stage. The blood-testis barrier in the terminal segment was chiefly established between 15 and 20 days of age as revealed by the lanthanum tracer technique.
To study the effect of the germ cells on the differentiation, the germ cell depleted testes of prenatally irradiated rats were also studied. The modified Sertoli cells of the terminal segment were more vacoulated and had fewer lipid droplets and inter-Sertoli cell junctions than did the Sertoli cells of the seminiferous tubules. The ultrastructure of the modified Sertoli cells of the terminal segment was similar in adult normal and adult SCO (Sertoli cell only) rats. The amount of lipid droplets in the Sertoli cells of SCO rats showed considerable variation among different tubular cross-sections within one testis.     

Tubular Hyalinization in Human Testis

Tubular hyalinization was studied both light and electron microscopically in 22 cases. 8 of them were found in undescended testes and 5 cases represented possible intermediary forms with only partial hyalinization of the tubules. In these testes spermatogenic arrest, hypospermatogenesis, only Sertoli cells and areas with normal spermatogenesis were found. The only remaining cell type in the tubular wall as well as, in some cases, also within the tubules was the fibroblast. Ultrastructurally the tubules were filled with collagen fibers. Based on these findings it is suggested that tubular hyalinization may be the end stage of a variety of pathologic conditions causing damage to the seminiferous tubules.     

Testicular focal atrophy – tubular blockade as partial degeneration of the tubule

This study investigated whether focal atrophy is a degenerative process of a whole tubule or tubular blockade as partial intratesticular degeneration. Serial section analyses of testicular tissue from 19 men with different andrologic diseases were examined. From every fifth section of any series, defined areas were viewed under a light microscope, and tubule sections were drawn. Three-dimensional reconstructions were made from these materials. Reconstructions of the seminiferous tubules showed tubular blockade as partial degeneration of the tubules. Transition from an intact portion to a blockade was accompanied by an increase in the thickness of the lamina propria. The blockade was a cell cord that contained Sertoli cells and, at most, spermatogonia, or it was a completely atrophied tubule, a so-called tubular scar. In a given tubule, defined areas of atrophy and areas of spermatogenic activity were both found. Occurrence of tubular blockade increased with age. Serial section analyses of testicular tissue showed tubular blockade as the partial degeneration of seminiferous tubules. In focal atrophy, a given tubule can have defined areas of atrophy and areas of spermatogenic activity. Received: 25 August 1999 / Accepted: 21 April 2000     

Local regulation of Leydig cells by the seminiferous tubules. Effect of short-term cryptorchidism

Unilateral cryptorchism was induced in adult rats for 24 h, and its effect on testicular morphology and intratesticular testosterone concentration after hCG-stimulation were studied. In seminiferous, tubules from abdominal testes an increased number of degenerating germ cells was noted in stages XIV-III of the spermatogenic cycle and Sertoli cells contained an increased amount of lipid droplets in stages XIV-VIII. However, germ cells and Sertoli cells from tubules at other stages of the cycle appeared unaffected. In scrotal testes the size of peritubular Leydig cells varied in phase with the spermatogenic cycle. The largest cells were found adjacent to stage VII-VIII and the smallest adjacent to stage XI-XII. In abdominal testes no stage-dependent variation in the size of peritubular Leydig cells was seen. Perivascular Leydig cells were of equal size in abdominal and scrotal testes. The testicular testosterone concentration following stimulation with a low dose of hCG was significantly lower in abdominal testes. It is suggested that the seminiferous tubules locally modulate Leydig cell function and that the stage specific stimulatory influence from stage VII-VIII is rapidly lost during experimental cryptorchidism.     

抑制素B βB亚单位在不同病理分型的无精子症患者睾丸组织中的表达

目的:探讨抑制素B(INH B)βB亚单位在不同生精功能状态的人睾丸组织中的表达情况。方法:对83例无精子症患者进行睾丸组织病理检查诊断,根据病理形态的不同分为:唯支持细胞综合征型(n=21);生精功能低下型(n=20);生精阻滞型(n=24);生精功能基本正常型(n=18)。选择上述各型结构完整的睾丸组织,分别应用免疫组化法(SP)对血清INH B βB亚单位在不同生精功能状态的睾丸组织,进行定位研究。结果:各型睾丸组织内均存在血清INH B βB的表达,其分布特点为:间质细胞(Leydig cell)和早期生精细胞多为强阳性表达,呈深棕黄色;支持细胞(Sertoli cell)多为阳性表达;而晚期精子细胞和成熟精子未见表达;生精小管管周类肌细胞呈弱阳性表达。结论:INH B可能是睾丸Sertoli细胞和早期生精细胞的一个联合产物。     

睾丸细胞生物学研究进展

睾丸是男性生殖腺,由生精小管和间质构成。生精小管主要由生精细胞和支持细胞组成,是精子发生场所;间质中主要是间质细胞,间质细胞合成与分泌雄激素。本文介绍睾丸3种细胞的发育分化,以及成年期睾丸细胞的结构和生物学研究进展。     

Quantitative testicular biopsy in spinal cord injured men: comparison to fertile controls.

Spermatogenic abnormalities have been reported in the majority of spinal cord injured men on routine testicular biopsy. However, given the interim advances in their urological and rehabilitative care, a quantitative assessment of the germinal epithelium after spinal cord injury and comparison of these parameters to normal controls are warranted. Incisional testicular biopsy was performed in 14 spinal cord injured men. Quantitative micrometric techniques were applied to assess spermatogenesis and the results were compared to a normative data base of testicular biopsies previously obtained from a group of 15 fertile volunteers. From a minimum of 10 randomly selected round seminiferous tubules per subject the mean number of Sertoli cells, mature spermatids, tubular diameter and tubular wall thickness were determined in both groups and statistically analyzed. In the spinal cord injury group the mean number of spermatids per tubule was significantly lower and the mean number of Sertoli cells per tubule was significantly higher than in fertile controls (p less than 0.05). Moreover, the mean Sertoli cell-to-spermatid ratio per seminiferous tubule was significantly higher in the spinal cord injury group and discriminated between spinal cord injured men and controls, with a sensitivity of 93% and specificity of 100% (p less than 0.0001). Half of the spinal cord injury group showed a mean tubular spermatid density of less than 10. Compared to the fertile population, spinal cord injured men show significant differences in quantitative parameters of the germinal epithelium that may contribute to the reproductive dysfunction.     

大鼠严重烫伤后睾丸的病理变化

对大鼠严重体表烫伤后睾丸组织的病理变化进行了动态观察。大鼠体表３０％ＩＩ度烫伤后睾丸出现明显病变，其病理变化出现早、程度较重、形态变化多样，生精细胞、支持细胞、曲细精管界膜、间质细胞及血管内皮细胞均有不同程度的损伤性改变。生精细胞的病变以精母细胞及精细胞为重，精原细胞无明显损伤，伤后３０天生精上皮基本恢复正常。曲细精管界膜内纤维连接蛋白（Ｆｎ）于伤后早期减少。认为曲细精管界膜及支持细胞的损伤，特别是后者的损伤，改变了生精细胞生长、发育的微环境而可能导致或加重生精细胞的损伤。     

Pattern of Sertoli cell degeneration in cryptorchid prepubertal tests

Seventy-three testicular biopsies from 54 children (aged 2 months-14 years) with undescended testes were examined by light and electron microscopy. The biopsies included abdominal, inguinally fixed, inguinally moveable, and retractile testes. Alterations in Sertoli cell morphology were found in all biopsies. The alterations included dilated elements of rough endoplasmic reticulum, vacuolization of the cytoplasm, mitochondria with poorly preserved cristae, increase in electron density of the matrix, elongation of the nuclei, and irregularities of the nuclear membrane. According to the numerical appearance of these cells and to the extent of lesions in single Sertoli cells, seven phases in the continuous process of tubular alteration were distinguished. The most severe tubular damaged (phase VII) occurred when the seminiferous epithelium consisted exclusively of necrotic cells. All phases of tubular alterations were seen regularly in each of the biopsies investigated. Germ cells occurred only in phases I-IV and were never observed in tubules in phases V-VII. Significant differences became evident between inguinal and retractile testes by morphometric evaluation. It was demonstrated that the number of germ cells per cross-sectioned tubule (S/T value) correlated negatively with the percentage of tubules in phases V-VII. In contrast to inguinal testes, a complete absence of Sertoli cells and an S/T value less than 0.1 were never found in retractile testes and the percentage of tubules in phases V-VII was reduced significantly compared with inguinal testes. Our findings indicate that (i) maldescended testis in patients between 1 and 15 years-of-age is associated with a special pattern of Sertoli cell degeneration; (ii) Sertoli cell degeneration is a continuous process, which can lead eventually to complete dissolution of the seminiferous epithelium; (iii) total degeneration is not related to age but is dependent on testicular position; (iv) a defined phase of degeneration excludes germ cell development, and therefore enhanced Sertoli cell degeneration in cryptorchid testes must also account for the reduction in germ cell number.     

Immunohistochemical distribution of S-100 protein and subunits (S100-alpha and S100-beta) in the swamp-type water buffalo (Bubalus bubalis) testis

The distribution and localization of S-100 protein (S-100) and its subunits (S100-alpha and S100-beta) in the testis of swamp-type water buffalo were investigated using immunohistochemistry. S-100 was detected in the Sertoli cells in the convoluted seminiferous tubules, modified Sertoli cells lining the terminal segment of the seminiferous tubules and in the intratesticular excurrent ducts (straight tubules and rete testis). S100-beta showed the same distribution and localization with that of S-100. However, the cytoplasmic extension of the Sertoli cells in S100-beta staining showed less staining intensity compared with that of S-100. S100-alpha showed a positive staining only in the modified Sertoli cells of the terminal segment of the seminiferous tubule. Endothelial cells of blood vessels were also positive with the proteins while the Leydig and spermatogenic cells showed a negative reaction. The localization of S-100 in the testis of the water buffalo was in parallel with that of other artiodactyls which supports the hypothesis that this protein is a multifunctional protein. S100-beta in the Sertoli cells suggests that this protein is involved in establishing blood-testis barrier. Its presence in the modified Sertoli cells and in the epithelium of the excurrent ducts suggest secretory and absorptive function, respectively. Meanwhile, S100-alpha, which was detected only in the modified Sertoli cells, is involved in the secretory activity of these cells that are related to exocrine function.     

Development of stage-specific paracrine regulation of Leydig cells by the seminiferous tubules

The size of peritubular Leydig cells surrounding tubules in different stages of the spermatogenic cycle was determined in 43- and 47-day-old male rats. A stage-dependent variation in the size of peritubular Leydig cells was not present in 43-day-old rats, but by 47 days those Leydig cells closely adjacent to tubules at stages VII-VIII were larger than others. At 43 days of age spermatogenesis had developed up to step 18 spermatids in late stage VI tubules. At 47 days of age the first mature sperm had just been released from the seminiferous epithelium, and consequently the first wave of the spermatogenic cycle was completed. Tubules at stages VII-VIII therefore acquire the ability to influence surrounding Leydig cells when they contain step 19 spermatids. It remains to be shown whether this maturation step is due to inherent maturation of the Sertoli cells or if step 19 spermatids specifically modulate Sertoli cell function.     

Morphological evidence for two types of idiopathic ''Sertoli-cell-only'' syndrome

Biopsies of testicular specimens taken from 41 patients that were diagnosed as having idiopathic Sertoli-cell-only syndrome were classified into two types, A and B, on the basis of histological and immunohistochemical findings. Thirty eight specimens that were classified as type A exhibited seminiferous tubules of small diameter and with tubular wall hyalinization, but containing normal adult type Sertoli cells. The other three specimens that were classified as type B showed no seminiferous tubular wall hyalinization, and their Sertoli cells had vimentin distribution localized in the subnuclear cytoplasm and had a pseudostratified lining, features resembling the appearance of fetal Sertoli cells. In one patient with a seminoma, a comparative study of the same testis prior to and post-irradiation was undertaken. Judging from this, postpubertal depletion of the germ cell population was considered to be responsible for the tubular atrophy observed in type A. Type B testes, though small in number, were characterized by a morphology distinct from the type A, but their pathogenesis remains unknown.     

Expression of endothelial nitric oxide synthase in the Sertoli cells of men with infertility of various causes

OBJECTIVE: To investigate how endothelial nitric oxide (eNOS) expression in the seminiferous tubules might be related to spermatogenesis, by examining eNOS expression in testicular tissue of patients infertile from various causes. PATIENTS AND METHODS: The study included five fertile men with a normal sperm concentration, nine patients with obstructive azoospermia, 20 with varicocele testes and eight with idiopathic azoospermia (Sertoli cell-only syndrome). Testicular biopsy specimens were examined by immunohistochemistry for eNOS protein expression, in addition to a routine pathological assessment. eNOS protein was detected using an eNOS monoclonal antibody. A Sertoli cell staining index (SSI) was defined as the ratio of stained Sertoli cells per total number of Sertoli cells, and was compared among the groups. RESULTS: eNOS was localized to Sertoli cells in the seminiferous tubules and Leydig cells in the interstium; although some degenerating germ cells stained, normal germ cells did not. The SSI was significantly lower in patients with Sertoli cell-only syndrome than in either fertile men or patients with obstructive azoospermia or varicocele. However, the SSI did not correlate significantly with the Johnsen score. CONCLUSION: The expression of eNOS in Sertoli cells may depend on the existence of germ cells and be associated with germ cell development.     

Êxpression of inducible nitric oxide synthase (iNOS) in the azoospermic human testis

Azoospermia, which is the absence of spermatozoa in the ejaculate, is not a rare cause of male infertility. Inducible nitric oxide synthase (iNOS) is a calcium-independent NOS, which is present in the testis and involved in spermatogenesis, and apoptosis of Sertoli and germ cells. Twenty idiopathic infertile men presenting nonobstructive azoospermia were enrolled in this study, and testicular sperm extraction procedures were performed. Tissue extracts were dissected, and the fluid samples were investigated to determine the presence of spermatozoa. Histologic evaluation of the spermatozoa-present samples revealed that seminiferous tubules were normal and were lined by Sertoli cells and spermatogenic cells. However, in the spermatozoa-absent samples, the diameter of the seminiferous tubules was small, and Sertoli-cell-only syndrome was determined in most of the tubules. iNOS expression was very weak in Sertoli cells, germ cells, and in Leydig cells in the spermatozoa-present group. In the spermatozoa-absent group, the immunostaining was very intense in Sertoli and Leydig cells. Electron microscopy findings were supported the histologic results. In conclusion, complete germ cell loss and intense expression of iNOS in the Sertoli and Leydig cells in the spermatozoa-absent groups of azoospermic human testis suggest an essential role of iNOS in spermatogenesis.     

大鼠曲细精管生殖细胞体外共培养和精子发生过程的观察

目的建立体外长期共培养体系和观察方法,为精子发生过程的研究提供细胞模型。方法采用大鼠曲细精管生殖细胞及支持细胞共培养的方法,对睾丸生精细胞作显微镜观察。结果共培养的支持细胞和生精细胞在体外存活超过6个月。在共培养期间,观察到精母细胞、圆形精子细胞和长形精子细胞。结论在不添加任何细胞因子和生长因子的情况下,大鼠曲细精管生殖细胞长期增生分化,不断产生精子细胞。这一结果暗示组织块和共生的支持细胞可为生殖细胞的增生和分化提供必需的细胞因子。该方法为体外研究精子发生过程提供了实验依据。     

Torsion and the contralateral testicle

The histologic effect of unilateral torsion on the contralateral testicle in adult Noble rats was examined. Utilizing detorsion or orchiectomy at 3 hours and 24 hours after torsion, the effect of early and late treatment of these changes was reviewed. The histologic changes consisted of loss of tubular spermatozoa, clumping of chromatin within the spermatocytes, the presence of Sertoli-only cells, evacuolization of Sertoli cytoplasm and germinal epithelial sloughing. Depression of spermatogenesis and decrease in the mean seminiferous tubular size was seen in the "normal" testicle after unilateral torsion. This effect was negated by early treatment with either orchiectomy or detorsion. Late detorsion does not negate these effects and late orchiectomy only partially negates them. Despite the depressed tubular function, the presence of early spermatogenic elements seen in the majority of the tubules in the "normal" testicle implies the possible reversibility of these changes.     

Deletion of Y chromosome involving the DAZ (deleted in azoospermia) gene in XX males.

The testicular histology and the presence or absence of 32 Y DNA loci was investigated, with a focus on the long arm of Y chromosome (Yq) interval 6, by means of a polymerase chain reaction strategy in 2 XX males. Seminiferous tubules lined by only Sertoli cells and a slight thickening of tubular walls were observed. The men showed an absence of 32 Y DNA loci. These facts suggest that severe spermatogenic impairment is caused by deletions of Yq interval 6 in XX males.