title: Supplementation of freezing and thawing media with brain-derived neurotrophic factor protects human sperm from freeze-thaw-induced damage.
abstract: OBJECTIVE
  To investigate the effects of brain-derived neurotrophic factor (BDNF) supplementation to freezing and thawing media on frozen-thawed human sperm parameters.
  
  
  METHODS
  Laboratory study.
  
  
  METHODS
  University hospital.
  
  
  METHODS
  Semen samples from 21 healthy fertile men.
  
  
  METHODS
  We measured reactive oxygen species (ROS) by flow cytometry using the probes dichlorofluorescin diacetate for intracellular hydrogen peroxide (H2O2) and dihydroethidium for intracellular superoxide anion (O2-•), sperm plasma membrane integrity by flow cytometry, caspase-3 activity using ELISA, and AKT phosphorylation status using Western blot in sperm that was cryopreserved and thawed in media either supplemented with BDNF or without BDNF supplementation (control).
  
  
  METHODS
  Sperm motility, viability, ROS levels, caspase-3 activity and AKT phosphorylation.
  
  
  RESULTS
  The percentage of motile and viable sperm cells was significantly higher in BDNF-supplemented groups as compared with the nonsupplemented (control) group. There was a significant difference in AKT phosphorylation status between BDNF-supplemented groups and the control group. Moreover, the levels of intracellular H2O2and caspase-3 activity were significantly lower in the sperm cells that were frozen and thawed in media supplemented with BDNF compared with in the control group.
  
  
  CONCLUSIONS
  BDNF supplementation to sperm freezing or thawing media has protective effects against oxidative stress and apoptosis in frozen-thawed human spermatozoa and could improve sperm function, probably through the activation of AKT.
authors.affiliation: Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Atefeh
authors.initials: A
authors.lastname: Najafi
authors.affiliation: Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Ebrahim
authors.initials: E
authors.lastname: Asadi
authors.affiliation: Department of Surgery, Urology Division, McGill University, Montreal, Quebec, Canada; Department of Theriogenology, Faculty of Veterinary Medicine, Cairo University, Giza, Egypt.
authors.forename: Adel R
authors.initials: AR
authors.lastname: Moawad
authors.affiliation: Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Saideh
authors.initials: S
authors.lastname: Mikaeili
authors.affiliation: Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Fardin
authors.initials: F
authors.lastname: Amidi
authors.affiliation: Department of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Adutwum
authors.affiliation: Department of Hematology, Faculty of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran.
authors.forename: Majid
authors.initials: M
authors.lastname: Safa
authors.affiliation: Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran. Electronic address: Aligholi.sobhani@yahoo.com.
authors.forename: Ali Gholi
authors.initials: AG
authors.lastname: Sobhani
mesh_terms: D000328:Adult; D019208:Brain-Derived Neurotrophic Factor / Q000494:pharmacology*; D053148:Caspase 3 / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D019610:Cytoprotection; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D008297:Male; D018384:Oxidative Stress / Q000187:drug effects; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D012662:Semen Preservation / Q000379:methods*; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D013481:Superoxides / Q000378:metabolism; D055815:Young Adult
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D003451:Cryoprotective Agents; D013481:Superoxides; D006861:Hydrogen Peroxide; D051057:Proto-Oncogene Proteins c-akt; C505407:CASP3 protein, human; D053148:Caspase 3
keywords: AKT; BDNF; ROS; Sperm cryopreservation; apoptosis; membrane integrity