Excessive unilateral proliferation of spermatogonia in a patient with non-obstructive azoospermia – adverse effect of clomiphene citrate pre-treatment?

Abstract

Background: Clomiphene citrate has been proposed as pre-treatment for infertile men with non-obstructive, testicular azoospermia (NOA) before surgery for testicular sperm extraction (TESE), especially when serum testosterone is low. Case presentation: Here, we report on a 33-year old azoospermic patient with a previous history of repeated "fresh" TESE and clomiphene citrate therapy (50 mg/day over 6 months) before undergoing microscopically assisted, bilateral testicular biopsy. Comprehensive histological and immunohistochemical work-up revealed a heterogeneous spermatogenic arrest at the level of spermatogonia or primary spermatocytes, with focally preserved spermatogenesis up to elongated spermatids in the right testis. In the left testis, the majority of tubules (> 70%) showed no tubular lumen or regular seminiferous epithelium but a great number of spermatogonia-like cells. These cells proved to be normally differentiated spermatogonia (positive for melanoma associated antigen 4 (MAGEA4), negative for placental alkaline phosphatase (PlAP)) with increased proliferative activity (positive for proliferating cell nuclear antigen (PCNA)) and a slightly higher rate of apoptotic cells. When compared to a tissue control with normal spermatogenesis, expression of sex hormone receptors androgen receptor (AR), estrogen receptor (ER) alpha, and G-protein coupled estrogen receptor 1 (GPER1) was not altered in patient samples. Sertoli cells appeared to be mature (positive for vimentin, negative for cytokeratin 18), whereas the expression of zona occludens protein 1 (ZO-1), claudin 11, and connexin 43 was absent or dislocated in the tubules with abundance of spermatogonia. Conclusion: This result suggests that formation of the blood-testis barrier is disturbed in affected tubules. To our knowledge this is the first observation of excessive, non-malignant proliferation of spermatogonia in a NOA patient. Although underlying molecular mechanisms remain to be elucidated, we hypothesize that the unusual pathology was triggered by the high-dose clomiphene citrate treatment preceding testicular biopsy.

Co-administration of alcohol and combination antiretroviral therapy (cART) in male Sprague Dawley rats: A study on testicular morphology, oxidative and cytokines perturbations

“…Furthermore, the increased number of spermatogonia (proliferation of spermatogenesis) observed in testis of the alcohol and cART treated groups, could be an attempt to increase the germ cell pool so as to counteract the diminishing spermatogenesis processes which is in line with a previous clinical study that reported a massive increase in the number of normally differentiated spermatogonia and elevated spermatogonia proliferative activity in a biopsy of patient with non-obstructive azoospermia [43]. Additionally, the (LAF and diameter (LD) were both slightly increased in the A group but significantly increased in the A+cART group, suggesting varying degrees of germ cell layer reduction due to different treatments as well as seminiferous tubule shrinkage.…”

Research Progress of PCNA in Reproductive System Diseases

“…PCNA is also a useful marker for diagnosing germinal arrest, which is the result of deterioration in DNA synthesis [ 77 ]. In one study, clomiphene citrate was used to treat infertile men with NOA; subsequent testicular sperm extraction showed that most tubules (>70%) exhibited no tubular lumen or regular seminiferous epithelium, but there were a large number of spermatogonia-like cells that exhibited normally differentiated spermatogonia with enhanced proliferation activity (positive for PCNA) [ 78 ]. The mechanism for effective spermatogenesis involves germ cell type-specific posttranslational modifications of histones, and histone variants play an important role.…”