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1.
Pediatr Surg Int ; 35(11): 1309-1316, 2019 Nov.
Article in English | MEDLINE | ID: mdl-31435735

ABSTRACT

PURPOSE: Undescended testes (UDT) are subjected to heat stress, which can disturb gonocyte transformation as well as apoptosis. This study aims to describe the apoptosis pathway occurring during minipuberty of children with unilateral (UDT), and to investigate the role of inhibin-B. METHODS: Testicular biopsies at unilateral orchidopexy of 10 boys (6-9 months old) with normal inhibin-B (n = 5) or low inhibin-B (n = 5) were selected for immunohistochemistry and TUNEL (Terminal deoxynucleotidyl transferase dUTP nick end labelling) assay. Testicular tubules were labelled with antibodies against Anti-Müllerian hormone (AMH, Sertoli cell marker), mouse Vasa Homolog (MVH) and placental alkaline phosphatase (PLAP) (both germ cell markers), cleaved caspase3 (apoptotic marker), and followed by confocal imaging and cell counting with Fiji/ImageJ. Data were analyzed with GraphPad Prism. RESULTS: In males with low and normal inhibin-B, there was no statistical difference (p > 0.05) in the percentage of testicular tubules containing TUNEL + cells, number of cleaved caspase3 ± germ cells/tubule, total number of germ cells/tubule, and the percentage of fibrotic tubules or number of Sertoli cells/tubule. CONCLUSIONS: These results suggest that inhibin-B does not regulate cell death of gonocytes and further studies are required to uncover any role of inhibin-B in gonocyte transformation.


Subject(s)
Cell Differentiation , Cryptorchidism/pathology , Inhibins/blood , Seminiferous Tubules/cytology , Apoptosis , Caspase 3/metabolism , Cryptorchidism/surgery , Germ Cells/cytology , Germ Cells/metabolism , Humans , Immunohistochemistry , In Situ Nick-End Labeling , Infant , Male , Orchiopexy
2.
J Pediatr Surg ; 54(8): 1654-1659, 2019 Aug.
Article in English | MEDLINE | ID: mdl-30739749

ABSTRACT

INTRODUCTION: Neonatal testicular germ cells/gonocytes, transform into stem cells for spermatogenesis during 'minipuberty', driving change in timing of surgery. This study examined gonocyte transformation in cryptorchid testes in children ≤18 months of age with unilateral, bilateral undescended testes (UDT), complete or partial androgen insensitivity syndrome (CAIS, PAIS) [3,4]. MATERIAL AND METHODS: Testicular biopsies were taken from patients with unilateral or bilateral UDT, PAIS or CAIS, aged 10 days-18 months. These testicular sections underwent immunohistochemistry with antibodies (Oct4, Ki67, C-Kit, Sox9) followed by confocal imaging, cell counting and statistical analysis. RESULTS: Both Sertoli cells/tubule and germ cells (GC)/tubule decreased with age, and % empty tubules (no GC) increased with age but with no significant differences between patient groups. Oct4+ germ cells/tubule decreased with age. There are some GCs and Sertoli cells proliferating during the first year and most proliferating Oct4+ germ cells (Oct4+/Ki67+) were located off tubular basement membrane. CONCLUSION: Our study showed that Oct4 expression gradually decreased after minipuberty and transformation into spermatogonia. Germ cells and Sertoli cells undergo mitosis during the first 12 months although not abundantly. We propose that Oct4+ gonocyte transformation into spermatogonia via proliferation and migration to the basement membrane may be delayed in UDT.


Subject(s)
Androgen-Insensitivity Syndrome/pathology , Cryptorchidism/pathology , Spermatogonia/pathology , Testis/physiology , Androgen-Insensitivity Syndrome/physiopathology , Basement Membrane/pathology , Cell Count , Cell Differentiation , Cryptorchidism/physiopathology , Humans , Infant , Infant, Newborn , Male , Octamer Transcription Factor-3/metabolism , Sertoli Cells/pathology , Spermatogenesis , Spermatogonia/metabolism , Testis/pathology , Testis/physiopathology
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