title: Melatonin modulates the expression of BCL-xl and improve the development of vitrified embryos obtained by IVF in mice.
abstract: OBJECTIVE
  Antioxidant and anti-apoptotic effects of melatonin on development of in vitro fertilization (IVF)/vitrified two-cell mouse embryos were evaluated in this study.
  
  
  METHODS
  The IVF two-cell embryos were vitrified by cryotop, and were cultured in KSOM medium in different concentrations of melatonin (10(-6), 10(-9), 10(-12) M) and without melatonin. The blastocyst cell number, apoptotic cells and glutathione (GSH) level were evaluated by differential, TUNEL and cell tracker blue staining, respectively. The expression of Bax and Bcl-xl genes was evaluated by qPCR. The expression of melatonin receptors (Mtnr1a and Mtnr1b) in mouse 2-cell embryos and blastocysts was evaluated by RT-PCR.
  
  
  RESULTS
  Melatonin increased the rate of cleavage and blastulation at 10(-12) M concentration (p < 0.05). The number of trophectoderm and inner cell mass showed a significant increase (p < 0.05) in 10(-9) M melatonin. The 10(-9) M and 10(-12) M melatonin treatments significantly reduced (p < 0.05) the apoptotic index. The significant increase in the expression of Bcl-xl observed at 10(-9) M concentration however, reduced expression of Bax was not statistically significant. The levels of GSH in 10(-9) and 10(-12) M groups were significantly improved relative to the control group (p < 0.05). The Mtnr1a was expressed in 2-cell embryos and blastocysts in all groups, but the expression of Mntr1b was not detected.
  
  
  CONCLUSIONS
  Melatonin may have a special role against oxidative stress in protection of IVF/vitrified embryos.
authors.affiliation: Department of Transgenic Animal Science, Stem Cell Technology Research Center, Tehran, Iran.
authors.forename: Maryam
authors.initials: M
authors.lastname: Dehghani-Mohammadabadi
authors.forename: Mohammad
authors.initials: M
authors.lastname: Salehi
authors.forename: Fattaneh
authors.initials: F
authors.lastname: Farifteh
authors.forename: Sedigheh
authors.initials: S
authors.lastname: Nematollahi
authors.forename: Ehsan
authors.initials: E
authors.lastname: Arefian
authors.forename: Atena
authors.initials: A
authors.lastname: Hajjarizadeh
authors.forename: Kazem
authors.initials: K
authors.lastname: Parivar
authors.forename: Zahra
authors.initials: Z
authors.lastname: Nourmohammadi
mesh_terms: D000818:Animals; D046149:Embryo Culture Techniques; D047108:Embryonic Development / Q000187:drug effects; D005260:Female; D005307:Fertilization in Vitro; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D008550:Melatonin / Q000494:pharmacology; D051379:Mice; D018384:Oxidative Stress; D058989:Vitrification; D051020:bcl-X Protein / Q000235:genetics / Q000378:metabolism*
chemical_list: C494812:Bcl2l1 protein, mouse; D051020:bcl-X Protein; D008550:Melatonin