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The cellularity of the human prepubertal testicular interstitium has not been well studied at the ultrastructural level. In this study, testicular biopsies were obtained from 35 boys aged three to nine years and examined by electron microscopy to clarify and quantitate the cell types present during the prepubertal period. The prepubertal testicular interstitium is found to consist of immature Leydig cells (9%), primitive fibroblastic cells (63%) (intertubular in location), and attenuated peritubular fibroblasts (28%). The primitive fibroblastic cells and peritubular fibroblasts appear closely related, being distinguished mainly by shape and location. The immature Leydig cell type contrasts with the fibroblastic cell types by exhibiting an irregular nucleus with relatively little heterochromatin. The most impressive cytoplasmic feature is the moderate to extensive development of smooth endoplasmic reticulum in the form of anastamosing tubules. In contrast, the rough endoplasmic reticulum is not well developed. Other cytoplasmic characteristics are the highly developed Golgi elements and occasional lipid droplets and lysosomes. Glycogen is also often present and is generally found in those cells that do not contain a well-developed smooth endoplasmic reticulum. The ultrastructure of the immature Leydig cell is compared with that of the mature fetal and adult Leydig cells. Although generally found in small clusters between tubules, these cells are often attenuated and closely associated with the seminiferous tubules. Occasional intermediate cell morphologies suggest a relationship between the primitive fibroblasts and immature Leydig cells. The presence of small cells exhibiting a steroid-producing morphology, classified as immature Leydig cells, in the prepubertal testicular interstitium is an interesting finding and is in accordance with earlier studies on nonhuman mammals. It is unknown whether these cells are remnants of the fetal Leydig cell population or have differentiated neonatally from the primitive fibroblastic cells. It is suggested that the immature Leydig cells are the progenitors of the adult Leydig cell population.
Anat Rec 1984 Jun
PMID:Ultrastructure of immature Leydig cells in the human prepubertal testis. 646 27

Stereological analysis was carried out on Leydig cells in perfusion-fixed testes of normal adult mice. In a decapsulated testis, the seminiferous tubules occupy 89.3% and the interstitial tissue makes up 10.7% of the volume of the testis parenchyma. The Leydig cells comprise 3.8% of testicular volume. There are 24.9 million Leydig cells per cm3 (or gm) of tissue. An average Leydig cell has a volume of 1,533 microns 3 and a surface area of 1150 microns 2. The smooth endoplasmic reticulum (SER) is the most prominent organelle in the Leydig cells, and has a membrane surface area of 2,428 cm2 per cm3 of fresh testis tissue, which is 8.5 times the surface area of the plasma membrane and constitutes 56.9% of the total membranes in Leydig cells. Mitochondria occupy 10.1% of the Leydig cell volume or 11.4% of cytoplasmic volume. The inner mitochondrial membrane (including tubular or vesicular cristae) provides a surface area of about 2855 microns 2/cell and is 2.26 times that of the outer membrane. There are approximately 712 cm2 of inner membranes per cm3 tissue. Mouse Leydig cells have numerous lipid droplets, which average 147 per cell and occupy 5.1% of the cell volume.
Anat Rec 1982 Dec
PMID:Morphometric analysis of testicular Leydig cells in normal adult mice. 718 Nov 38

Rapid and complete withdrawal of intratesticular testosterone was achieved via the destruction of all Leydig cells with the specific Leydig cell cytotoxin ethane dimethanesulphonate (EDS). Restoration of testosterone levels was accomplished by administration of a single dose (25 mg) of testosterone esters (T) known to reverse the antispermatogenic effects of androgen withdrawal. Quantitation of the degenerating germ cells in cross sections of seminiferous tubules (ST) at stages IV-V, VII, IX, and X-XI of the spermatogenic cycle was used as a sensitive biological index of the effects of testosterone withdrawal and restoration upon the function of the Sertoli cells. Compared to control testicular tissues, the mean numbers of pyknotic germ cells per ST cross section at stages VII, IX and X-XI increased significantly (P < 0.01-0.001) between 4 to 8 days post-EDS treatment, but only in stage VII tubules was this trend reversed significantly (P < 0.005) within 2 days by T supplementation. In EDS-treated rats, stages VII, VIII, IX, and X-XI also exhibited significant (P < 0.05-0.001) increases (compared to controls) in the volumetric proportions by which intraepithelial vacuoles appeared within the seminiferous tubules. Again, in EDS+T supplemented rats, the appearance of vacuoles was significantly (P < 0.001) suppressed in stage VII and VIII. In contrast to tubules at stages VII-XI, those at stages IV-V were completely unaffected by testosterone withdrawal or replacement. The results show that at selected time intervals after EDS treatment, testosterone supplementation is capable of preventing/reversing these morphological changes within 2 days in stage VII tubules. It is suggested that the induction and subsequent prevention of seminiferous epithelial damage will serve as an important in vivo and in vitro approach for studies on the androgen-mediated changes in Sertoli cell biology during phases of impairment and recovery of their function. Manipulation of adult Sertoli cell function as provided by our model should permit identification of androgen-regulated gene products together with an understanding of their role(s) in normal and abnormal spermatogenesis.
Anat Rec 1993 Apr
PMID:Stage-dependent changes in spermatogenesis and Sertoli cells in relation to the onset of spermatogenic failure following withdrawal of testosterone. 838 23

The objective of this study was to detect by immunohistochemical means nuclear accumulations of p53 and p21 proteins in testicular tumours of dogs. Intense p53 protein nuclear labelling was shown by each of seven seminomas and one intermediate collision tumour. Moderate or intense immunoreactivity was shown by three Sertoli cell tumours. All the tumours also showed p21 nuclear reactivity in parallel with the p53 reactivity. In contrast, four Leydig cell tumours showed no detectable immunoreactivity. The results suggested that high levels of p53 accumulation were associated with the expression of wild-type p53, which was able to activate the transcription of the p21 gene. In addition, weak p53- and p21-nuclear reactivities were detectable in primary spermatocytes within normal seminiferous tubules.
Vet Rec 2000 Mar 25
PMID:Immunohistochemical detection of p53 and p21 proteins in canine testicular tumours. 1080 82

Cyclosporine A (CsA) is known to have testicular toxicity, leading to male infertility. Stimulant and aphrodisiac properties have been attributed to the plant, Heteropterys aphrodisiaca. Thus, the present work was undertaken to evaluate the association of the drug and the medicinal herb in Wistar rats, applying testicular morphometry and ultrastructure. Twenty-four rats were used, divided into four groups: I, control; II, CsA; III, simultaneous use of CsA and H. aphrodisiaca; IV, H. aphrodisiaca. Daily administration by gavage was carried out, during 56 days, of water (sham), CsA in a dose of 15 mg/kg per day and/or H. aphrodisiaca in a dose of 0.5 ml of the infusion prepared with 25 g of roots/100 ml of boiling water. Increased body weight was observed for all groups, but the animals that received only CsA showed the smallest body weight gain. Morphometry showed increased connective tissue volumetric proportion and decreased Leydig cell volumetric proportion in CsA-treated rats. Using transmission electron microscopy, it was possible to ascertain that CsA caused seminiferous epithelium degeneration, resulting in Sertoli cell vacuolization, abnormal round and elongated spermatids and large accumulation of residual cytoplasm at the epithelium border next to the lumen. Expanded intercellular spaces between germ cells were still observed in H. aphrodisiaca-treated rat testes. The administration of H. aphrodisiaca infusion to CsA-treated rats diminished nearly all the CsA-induced damage to the testis ultrastructure, suggesting that H. aphrodisiaca infusion may be used combined with CsA to reduce CsA-induced injuries in the testis.
Anat Rec (Hoboken) 2008 Jul
PMID:Heteropterys aphrodisiaca infusion reduces the collateral effects of cyclosporine A on the testis. 1844 93


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