Vanessa Wilson1,2, Jorgen Thorup3,4, Erick Clasen-Linde5, Dina Cortes4,6, John M Hutson1,2,7, Ruili Li8,9. 1. Douglas Stephens Surgical Research Group, Murdoch Children's Research Institute, Flemington Road, Parkville, VIC, 3052, Australia. 2. Department of Paediatrics, University of Melbourne, Melbourne, Australia. 3. Department of Paediatric Surgery, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark. 4. Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark. 5. Department of Pathology, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark. 6. Department of Pediatrics, Copenhagen University Hospital Hvidovre, Copenhagen, Denmark. 7. Department of Urology, The Royal Children's Hospital, Melbourne, Australia. 8. Douglas Stephens Surgical Research Group, Murdoch Children's Research Institute, Flemington Road, Parkville, VIC, 3052, Australia. Ruili.li@mcri.edu.au. 9. Department of Paediatrics, University of Melbourne, Melbourne, Australia. Ruili.li@mcri.edu.au.
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.
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), mouseVasa 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.
Authors: Francisco J Schneuer; Elizabeth Milne; Sarra E Jamieson; Gavin Pereira; Michele Hansen; Andrew Barker; Andrew J A Holland; Carol Bower; Natasha Nassar Journal: Lancet Child Adolesc Health Date: 2018-08-30