It was speculated that vasectomy might induce hyperplasia of Leydig cells associated with atrophy of seminiferous tubules, but estimates of the Leydig cell number were lacking in previous studies. This study aimed to test the speculation by determining the numerical change of Leydig cells and other interstitial cells after a vasectomy that induced spermatogenic damage. Twelve adult Japanese white rabbits and twelve mature Sprague-Dawley rats were subjected to a unilateral vasectomy away from the scrotum. Six months (rabbits) or thirty-seven days (rats) postoperation, testes on both sides were removed and methacrylate sections prepared. The total numbers (per testis) of all nuclei in the testicular interstitial tissue were estimated with a stereological technique – the optical disector. The results showed that marked spermatogenic damage associated seminiferous tubular atrophy on the vasectomized side occurred in 7 (rabbits) or 5 (rats) of the 12 animals. For the rabbit, the total numbers of myoid cells or leukocytes on the vasectomized side (compared with the contralateral nonvasectomized side) were unchanged but those of Leydig cells and other interstitial cells increased significantly by 41% and 52%, respectively, (a) with the increase in the testis with spermatogenic damage appearing to be larger than that in the testis without spermatogenic damage and (b) without significant increase in the total volume of the interstitium. For the rat, there were no significant differences in the total numbers of any cell type. In conclusion, hyperplasia or hyperproliferation of the interstitial cells, which might be a result of increased intra-testicular pressure for long, was possible postvasectomy, but it might not be indicative of a better function of the cells.

/pdf/a-stereological-quantitative-study-of-leydig-cell-4e2m7p2tk4.pdf

A Stereological (Quantitative) Study of Leydig Cell Hyperplasia in Vasectomized Rabbit and Rat Testes

The optical disector, a three-dimensional counting frame or probe in stereology, is often positioned in the middle (depth) of a thick section for unbiased nuclear counting. Using 30-40 μm thick methacrylate or paraffin sections for nuclear counting of neurons with the optical disector, however, some studies showed markedly higher nuclear densities at 10% of the section thickness near the top or bottom surface of the section, suggestive of deformation of section along its z axis and thus affecting the number estimation. To verify the findings, this study obtained two sets of 12-14 methacrylate sections (average thicknesses 21.7 and 29.4 μm) and two sets of 12 paraffin sections (average thicknesses 13.8 and 29.2 μm) from mature rat testes. Each section was used to count round spermatid nuclei in the seminiferous epithelium densely packed with the cells, using 3-4 consecutive disectors placed vertically (along the z axis of the section) from the top surface of the section, through the whole section thickness (two sets of methacrylate and paraffin sections) or in 80-83% of the thickness (other sections). The results demonstrated that, overall, there were no considerable nonuniform changes of the nuclear densities along the z axis of the sections.

Is there section deformation resulting in differential change of nuclear numerical densities along the z axis of thick methacrylate or paraffin sections

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Is there section deformation resulting in differential change of nuclear numerical densities along the z axis of thick methacrylate or paraffin sections