Exposing mouse oocytes to necrostatin 1 during in vitro maturation improves maturation, survival after vitrification, mitochondrial preservation, and developmental competence.
Necrostatin 1 (Nec1) is widely used in disease models to examine the contribution of receptor-interacting protein kinase 1 in cell death. The biological actions of Nec1 are blocking necrotic cell death. The purpose of this study was to investigate whether adding Nec1 into in vitro maturation (IVM) media, followed by vitrification procedures, could enhance the survival and developmental competency of oocytes. Germinal vesicle oocytes were matured in IVM medium containing 2 different doses of Nec1 (0.5 and 1 μmol/L). After IVM, the oocytes were vitrified using a 2-step exposure to equilibrium and vitrification solutions. After warming, the rates of survival, fertilization, embryonic development up to blastocyst in vitro, morphology of spindle and chromosome, membrane integrity, mitochondria integrity, and several gene expressions were evaluated. The survival and developmental competency of oocytes were higher in the 1 μmol/L Nec1-treated group than control. The proportion with intact spindles/chromosomes and stable membranes was similar in all the groups. The mitochondrial integrity of all Nec1-treated groups showed a higher score with strong staining. The 1 μmol/L Nec1 showed significantly increased expressions of Mad2, Gdf9, and Bcl2. The Cirp level had a tendency to be downregulated in the 0.5 µmol/L Nec1 but upregulated in the 1 μmol/L Nec1, compared with the control. The Mtgenome expressions were significantly decreased in both Nec1 groups. The supplementation of 1 μmol/L Nec1 into the IVM medium could be beneficial for the survival and development of immature oocytes after vitrification.
Department of Obstetrics and Gynecology, Seoul National University Bundang Hospital, Seongnam, Korea Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University, Seoul, Korea.
Jun Woo
JW
Jo
Department of Obstetrics and Gynecology, Seoul National University Bundang Hospital, Seongnam, Korea Department of Obstetrics and Gynecology, Seoul National University College of Medicine, Seoul, Korea.
Jung Ryeol
JR
Lee
Department of Obstetrics and Gynecology, Seoul National University Bundang Hospital, Seongnam, Korea Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University, Seoul, Korea Department of Obstetrics and Gynecology, Seoul National University College of Medicine, Seoul, Korea.
Byung Chul
BC
Jee
Department of Obstetrics and Gynecology, Seoul National University Bundang Hospital, Seongnam, Korea Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University, Seoul, Korea Department of Obstetrics and Gynecology, Seoul National University College of Medicine, Seoul, Korea suhcs@snu.ac.kr.
Chang Suk
CS
Suh
Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University, Seoul, Korea Department of Obstetrics and Gynecology, Seoul National University College of Medicine, Seoul, Korea Department of Obstetrics and Gynecology, Seoul National University Hospital, Seoul, Korea.
Seok Hyun
SH
Kim
D000818:Animals; D001755:Blastocyst / Q000187:drug effects* / Q000378:metabolism; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D015925:Cryopreservation; D005260:Female; D005307:Fertilization in Vitro; D018507:Gene Expression Regulation, Developmental; D007093:Imidazoles / Q000494:pharmacology*; D059471:In Vitro Oocyte Maturation Techniques / Q000379:methods*; D007211:Indoles / Q000494:pharmacology*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D051379:Mice; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D054315:Oocyte Retrieval; D009865:Oocytes / Q000187:drug effects* / Q000378:metabolism; D008941:Spindle Apparatus / Q000187:drug effects / Q000378:metabolism; D058989:Vitrification
D007093:Imidazoles; D007211:Indoles; C507699:necrostatin-1
in vitro maturation; mitochondria; necrostatin 1; oocyte; spindle; vitrification