title: ESX1 mRNA expression in seminal fluid is an indicator of residual spermatogenesis in non-obstructive azoospermic men.
abstract: OBJECTIVE
  Is the presence of ESX1 mRNA in seminal fluid (SF) an indicator of residual spermatogenesis in men with non-obstructive azoospermic (NOA)?
  
  
  CONCLUSIONS
  ESX1 mRNA in SF is a suitable molecular marker for predicting the presence of residual spermatogenesis in testis.
  
  
  BACKGROUND
  ESX1 is an X-linked homeobox gene whose expression in testis is restricted to germ cells. We previously reported, in the testicular biopsies from azoospermic men, a positive correlation between the presence of ESX1 mRNA and residual spermatogenesis.
  
  
  METHODS
  We investigated ESX1 mRNA expression in 70 testicular fragments (TF) and 56 (SF) of 70 NOA men. As controls, we analyzed 8 TF from men with obstructive azoospermic (OA) and 9 SF from normozoospermic men. For all patients we considered the histological classification of testis biopsies and the recovery of spermatozoa by surgical procedures.
  
  
  METHODS
  Relative ESX1 mRNA expression was evaluated by quantitative RT-PCR using the ΔΔCt method. The results were compared with the recovery of spermatozoa at surgery.
  
  
  RESULTS
  In TF from NOA patients we found that: (i) ESX1 mRNA level was significantly decreased as the severity of spermatogenic defects increased (P < 0.0001, one-way analysis of variance); (ii) the presence of ESX1 mRNA can predict the success of sperm retrieval (sensitivity: 80%). In SF from NOA patients we found that: (i) ESX1 mRNA was present in 78.5% of NOA men; (ii) the presence of ESX1 mRNA could predict the success of sperm retrieval (sensitivity: 84%).
  
  
  CONCLUSIONS
  Spermatozoa were recovered at surgery in 5 out of 12 patients whose SF was negative for ESX1 mRNA expression. We think that discrepancies between molecular and clinical results could be reduced by analyzing more than one ejaculate from each man.
  
  
  CONCLUSIONS
  The data confirm that the ESX1 transcript in the semen of men with NOA is a suitable molecular marker for predicting the presence of residual foci of spermatogenesis in the testis. The implication of these results is that some patients 'with azoospermia', although having a severe impairment of spermatogenesis, could still maintain residual foci of spermatogenesis in limited areas of the testes, not always recovered by surgery.
  
  
  BACKGROUND
  This work was supported by the Fondazione IRCCS Ca' Granda, Ospedale Maggiore Policlinico: Ricerca Corrente [grant number RC2014/519-02] to M.M. and from ASM onlus 2010-2011 to M.M. The authors declare that they have no conflict of interest.
authors.affiliation: Medical Genetics, Department of Health Sciences, Università degli Studi di Milano, 20142 Milano, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Pansa
authors.affiliation: Medical Genetics, Department of Health Sciences, Università degli Studi di Milano, 20142 Milano, Italy.
authors.forename: Silvia M
authors.initials: SM
authors.lastname: Sirchia
authors.affiliation: Urology II Unit - Andrology and Assisted Reproduction, San Paolo Hospital - Università degli Studi di Milano, 20142 Milano, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Melis
authors.affiliation: Urology II Unit - Andrology and Assisted Reproduction, San Paolo Hospital - Università degli Studi di Milano, 20142 Milano, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Giacchetta
authors.affiliation: Urology II Unit - Andrology and Assisted Reproduction, San Paolo Hospital - Università degli Studi di Milano, 20142 Milano, Italy.
authors.forename: Mirco
authors.initials: M
authors.lastname: Castiglioni
authors.affiliation: Medical Genetics, Department of Health Sciences, Università degli Studi di Milano, 20142 Milano, Italy.
authors.forename: Patrizia
authors.initials: P
authors.lastname: Colapietro
authors.affiliation: Division of Pathology, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, 20122 Milano, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Fiori
authors.affiliation: Division of Pathology, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, 20122 Milano, Italy.
authors.forename: Rossella
authors.initials: R
authors.lastname: Falcone
authors.affiliation: Division of Pathology, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, 20122 Milano, Italy.
authors.forename: Leda
authors.initials: L
authors.lastname: Paganini
authors.affiliation: Division of Pathology, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, 20122 Milano, Italy.
authors.forename: Eleonora
authors.initials: E
authors.lastname: Bonaparte
authors.affiliation: Urology II Unit - Andrology and Assisted Reproduction, San Paolo Hospital - Università degli Studi di Milano, 20142 Milano, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Colpi
authors.affiliation: Division of Pathology, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, 20122 Milano, Italy Molecular Pathology, Department of Pathophysiology and Transplantation, Università degli Studi di Milano, Milano 20122, Italy monica.miozzo@unimi.it.
authors.forename: Monica
authors.initials: M
authors.lastname: Miozzo
authors.affiliation: Molecular Pathology, Department of Pathophysiology and Transplantation, Università degli Studi di Milano, Milano 20122, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Tabano
mesh_terms: D053713:Azoospermia / Q000235:genetics* / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008297:Male; D012333:RNA, Messenger / Q000378:metabolism; D012661:Semen / Q000378:metabolism*; D055101:Semen Analysis; D053625:Sperm Retrieval; D013091:Spermatogenesis / Q000235:genetics*
chemical_list: D015415:Biomarkers; C432969:ESX1 protein, human; D018398:Homeodomain Proteins; D012333:RNA, Messenger
keywords: ESX1; azoospermia; infertility; sperm recovery