title: Immunofluorescent analysis of testicular biopsies with germ cell and Sertoli cell markers shows significant MVH negative germ cell depletion with older age at orchiopexy.
abstract: OBJECTIVE
  Undescended testis is the most common defect in male newborns. This condition is associated with increased risks of infertility and testicular malignancy due to abnormal germ cell development in the testes. Early surgery may limit such risks. We analyzed germ cell development vs age at orchiopexy using a germ cell marker and a Sertoli cell marker on testicular biopsies.
  
  
  METHODS
  A total of 22 testicular biopsies at orchiopexy in 20 patients 5 to 24.5 months old were fixed and embedded in paraffin. Sections were processed and labeled with AMH antibody for Sertoli cells and MVH antibody for germ cells for immunofluorescent histochemical analysis. Confocal images were counted using ImageJ (National Institutes of Health, Bethesda, Maryland) for germ cells and testicular tubules. The data were analyzed using linear regression.
  
  
  RESULTS
  Sertoli cells were clearly distinguished from MVH positive and negative germ cells located centrally or on basement membranes of tubules. Percentage of tubules with MVH negative germ cells significantly decreased with increasing age at orchiopexy (β = -0.03, p = 0.03). Total tubular numbers and "empty" tubules without germ cells significantly increased with age at orchiopexy (β = 1.15, p = 0.02 and β = 0.44, p = 0.04, respectively).
  
  
  CONCLUSIONS
  AMH antibody distinguished Sertoli cells from germ cells, and MVH antibody distinguished 2 types of germ cells at different developmental stages. Biopsy at orchiopexy in older patients showed significant germ cell depletion. These results lend support to early surgery to optimize germ cell number.
authors.affiliation: Surgical Research Laboratory, Murdoch Childrens Research Institute, Parkville, Victoria, Australia; Department of Pediatrics, University of Melbourne, Parkville, Victoria, Australia. Electronic address: ruili.li@mcri.edu.au.
authors.forename: Ruili
authors.initials: R
authors.lastname: Li
authors.affiliation: Department of Pediatric Surgery, Rigshospitalet, Copenhagen, Denmark; Faculty of Health Science, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Jorgen
authors.initials: J
authors.lastname: Thorup
authors.affiliation: Environmental and Genetic Epidemiology Research, Murdoch Childrens Research Institute, Parkville, Victoria, Australia; Department of Pediatrics, University of Melbourne, Parkville, Victoria, Australia.
authors.forename: Cong
authors.initials: C
authors.lastname: Sun
authors.affiliation: Faculty of Health Science, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Dina
authors.initials: D
authors.lastname: Cortes
authors.affiliation: Surgical Research Laboratory, Murdoch Childrens Research Institute, Parkville, Victoria, Australia; Department of Pediatrics, University of Melbourne, Parkville, Victoria, Australia.
authors.forename: Bridget
authors.initials: B
authors.lastname: Southwell
authors.affiliation: Surgical Research Laboratory, Murdoch Childrens Research Institute, Parkville, Victoria, Australia; Department of Pediatrics, University of Melbourne, Parkville, Victoria, Australia; Urology Department, Royal Children's Hospital, Parkville, Victoria, Australia. Electronic address: john.hutson@rch.org.au.
authors.forename: John
authors.initials: J
authors.lastname: Hutson
mesh_terms: D000367:Age Factors; D054304:Anti-Mullerian Hormone / Q000276:immunology; D001485:Basement Membrane / Q000473:pathology; D001706:Biopsy; D016923:Cell Death; D002675:Child, Preschool; D003456:Cryptorchidism / Q000473:pathology* / Q000601:surgery*; D053487:DEAD-box RNA Helicases / Q000276:immunology; D005355:Fibrosis; D005854:Germ Cells / Q000276:immunology / Q000473:pathology*; D006801:Humans; D007150:Immunohistochemistry; D007223:Infant; D008297:Male; D018613:Microscopy, Confocal; D056126:Orchiopexy; D012708:Sertoli Cells / Q000276:immunology / Q000473:pathology*; D013737:Testis / Q000473:pathology
chemical_list: D054304:Anti-Mullerian Hormone; C090864:Ddx4 protein, mouse; D053487:DEAD-box RNA Helicases
keywords: AD-S; AMH; GC; GCs/T; MVH; PLAP; Sertoli cells; UDT; adult dark spermatogonia; anti-Mullerian hormone; cryptorchidism; germ cell; germ cells; germ cells per tubule; mouse VASA homologue; orchiopexy; placental alkaline phosphatase; testis; undescended testes