Lysophosphatidic acid improves development of porcine somatic cell nuclear transfer embryos
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This study was conducted to investigate whether lysophosphatidic acid (LPA) could improve the development of porcine somatic cell nuclear transfer (SCNT) embryos. Porcine SCNT-derived embryos were cultured in chemically defined polyvinyl alcohol (PVA)-based porcine zygote medium (PZM)-4 without or with LPA, and the development, cell proliferation potential, apoptosis, and expression levels of pluripotent markers were evaluated. LPA significantly increased the rates of cleavage and blastocyst formation compared to those seen in the LPA un-treatment (control) group. The expression levels of embryonic development-related genes (IGF2R, PCNA and CDH1) were higher (p < 0.05) in the LPA treatment group than in the control group. LPA significantly increased the numbers of total, inner cell mass and EdU (5-ethynyl-2’-deoxyuridine)-positive cells in porcine SCNT blastocysts compared to those seen in the control group. TUNEL assay showed that LPA significantly reduced the apoptosis rate in porcine SCNT-derived embryos; this was confirmed by decreases (p < 0.05) in the expression levels of pro-apoptotic genes, BAX and CASP3, and an increase (p < 0.05) in the expression level of the anti-apoptotic gene, BCL2L1. In addition, LPA significantly increased Oct4 expression at the gene and protein levels. Together, our data suggest that LPA improves the quality and development of porcine SCNT-derived embryos by reducing apoptosis and enhancing cell proliferation and pluripotency.
Abstract
This study was conducted to investigate whether lysophosphatidic acid (LPA) could improve the development of porcine somatic cell nuclear transfer (SCNT) embryos. Porcine SCNT-derived embryos were cultured in chemically defined polyvinyl alcohol (PVA)-based porcine zygote medium (PZM)-4 without or with LPA, and the development, cell proliferation potential, apoptosis, and expression levels of pluripotent markers were evaluated. LPA significantly increased the rates of cleavage and blastocyst formation compared to those seen in the LPA un-treatment (control) group. The expression levels of embryonic development-related genes (IGF2R, PCNA and CDH1) were higher (p < 0.05) in the LPA treatment group than in the control group. LPA significantly increased the numbers of total, inner cell mass and EdU (5-ethynyl-2’-deoxyuridine)-positive cells in porcine SCNT blastocysts compared to those seen in the control group. TUNEL assay showed that LPA significantly reduced the apoptosis rate in porcine SCNT-derived embryos; this was confirmed by decreases (p < 0.05) in the expression levels of pro-apoptotic genes, BAX and CASP3, and an increase (p < 0.05) in the expression level of the anti-apoptotic gene, BCL2L1. In addition, LPA significantly increased Oct4 expression at the gene and protein levels. Together, our data suggest that LPA improves the quality and development of porcine SCNT-derived embryos by reducing apoptosis and enhancing cell proliferation and pluripotency.
참고문헌 (46)
- Rodriguez-Osorio N, "Reprogramming mammalian somatic cells", Theriogenology, 78, pp.1869~1886 (2012)[DOI]
- Wilmut I, "Viable offspring derived from fetal and adult mammalian cells", Nature, 385, pp.810~813 (1997)[DOI]
- Cibelli JB, "Cloned transgenic calves produced from nonquiescent fetal fibroblasts", Science, 280, pp.1256~1258 (1998)[DOI]
- Wakayama T, "Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei", Nature, 394, pp.369~374 (1998)[DOI]
- Polejaeva IA, "Cloned pigs produced by nuclear transfer from adult somatic cells", Nature, 407, pp.86~90 (2000)[DOI]
- Galli C, "Pregnancy: a cloned horse born to its dam twin", Nature, 424, pp.635~ (2003)[DOI]
- Lee BC, "Dogs cloned from adult somatic cells", Nature, 436, pp.641~ (2005)[DOI]
- Liu Z, "Cloning of macaque monkeys by somatic cell nuclear transfer", Cell, 172, pp.881~887.E7 (2018)[DOI]
- Vajta G, "Production of cloned pigs by handmade cloning", Methods Mol Biol, 2647, pp.183~195 (2023)
- Lai L, "Production of α-1, 3-galactosyltransferase knockout pigs by nuclear transfer cloning", Science, 295, pp.1089~1092 (2002)[DOI]
- Tanihara F, "Current status of the application of gene editing in pigs", J Reprod Dev, 67, pp.177~187 (2021)[DOI]
- Yang X, "Nuclear reprogramming of cloned embryos and its implications for therapeutic cloning", Nat Genet, 39, pp.295~302 (2007)[DOI]
- Terashita Y, "Improving the quality of miniature pig somatic cell nuclear transfer blastocysts : aggregation of SCNT embryos at the four-cell stage", Reprod Domest Anim, 46, pp.189~196 (2011)[DOI]
- Matoba S, "Somatic cell nuclear transfer reprogramming : mechanisms and applications", Cell Stem Cell, 23, pp.471~485 (2018)[DOI]
- Boruszewska D, "Studies on lysophosphatidic acid action during in vitro preimplantation embryo development", Domest Anim Endocrinol, 54, pp.15~29 (2016)[DOI]
- Beltrame JS, "Novel role for lysophosphatidic acid in vascular remodeling at the maternal–fetal interface", Reproduction, 159, pp.R55~R67 (2020)[DOI]
- Cheng HY, "Lysophosphatidic acid signaling protects pulmonary vasculature from hypoxia-induced remodeling", Arterioscler Thromb Vasc Biol, 32, pp.24~32 (2012)[DOI]
- Moolenaar WH, "Development of our current understanding of bioactive lysophospholipids", Ann NY Acad Sci, 905, pp.1~10 (2000)[DOI]
- Hashimoto T, "Lysophosphatidic acid enhances in vivo infiltration and activation of guinea pig eosinophils and neutrophils via a Rho", Rho-associated protein kinase-mediated pathway, 2003, pp.91, 8~14
- Zhang T, "The metabolic characteristic of decidual immune cells and their unique properties in pregnancy loss", Immunol Rev, 308, pp.168~186 (2022)[DOI]
- Hinokio K, "Lysophosphatidic acid stimulates nuclear and cytoplasmic maturation of golden hamster immature oocytes in vitro via cumulus cells", Life Sci, 70, pp.759~767 (2002)[DOI]
- Jo JW, "Addition of lysophosphatidic acid to mouse oocyte maturation media can enhance fertilization and developmental competence", Hum Reprod, 29, pp.234~241 (2014)[DOI]
- Shin MY, "Lysophosphatidic acid accelerates development of porcine embryos by activating formation of the blastocoel", Mol Reprod Dev, 85, pp.62~71 (2018)[DOI]
- Zhang JY, "Lysophosphatidic acid improves porcine oocyte maturation and embryo development in vitro", Mol Reprod Dev, 82, pp.66~77 (2015)[DOI]
- Boruszewska D, "The effect of lysophosphatidic acid during in vitro maturation of bovine cumulus–oocyte complexes:cumulus expansion, glucose metabolism and expression of genes involved in the ovulatory cascade, oocyte and blastocyst competence", Reprod Biol Endocrinol, 13, pp.44~ (2015)[DOI]
- Boruszewska D, "The effect of lysophosphatidic acid during in vitro maturation of bovine oocytes : embryonic development and mRNA abundances of genes involved in apoptosis and oocyte competence", Mediators Inflamm, 2014, pp.670670~ (2014)[DOI]
- Yu B, "Lysophosphatidic acid accelerates bovine in vitro-produced blastocyst formation through the hippo", YAP pathway, 2021, pp.22, 5915~
- Li Y, "Lysophosphatidic acid improves human sperm motility by enhancing glycolysis and activating L-type calcium channels", Front Endocrinol, 13, pp.896558~ (2022)[DOI]
- Torres AC, "Lysophosphatidic acid signaling in late cleavage and blastocyst stage bovine embryos", Mediat Inflamm, 2014, pp.678968~ (2014)[DOI]
- Lin T, "Procedure used for denuding pig oocytes influences oocyte damage, and development of in vitro and nuclear transfer embryos", Anim Reprod Sci, 152, pp.65~76 (2015)[DOI]
- Zhu X, "Early development of porcine parthenogenetic embryos and reduced expression of primed pluripotent marker genes under the effect of lysophosphatidic acid", Reprod Domest Anim, 53, pp.1191~1199 (2018)[DOI]
- Lin T, "Delayed blastocyst formation or an extra day culture increases apoptosis in pig blastocysts", Anim Reprod Sci, 185, pp.128~139 (2017)[DOI]
- Harada Y, "Selection of highquality and viable blastocysts based on timing of morula compaction and blastocyst formation", Reprod Med Biol, 19, pp.58~64 (2020)[DOI]
- Lechniak D, "Timing of the first zygotic cleavage as a marker of developmental potential of mammalian embryos", Reprod Biol, 8, pp.23~42 (2008)[DOI]
- Bohrer RC, "The incidence of DNA doublestrand breaks is higher in late-cleaving and less developmentally competent porcine embryos", Biol Reprod, 93, pp.59~ (2015)[DOI]
- Zhang L, "ROCK inhibitor Y-27632 suppresses dissociation-induced apoptosis of murine prostate stem", progenitor cells and increases their cloning efficiency, 2011, pp.6, e18271~
- DeChiara TM, "Parental imprinting of the mouse insulin-like growth factor II gene", Cell, 64, pp.849~859 (1991)[DOI]
- Berx G, "Involvement of members of the cadherin superfamily in cancer", Cold Spring Harb Perspect Biol, 1, pp.a003129~ (2009)[DOI]
- Markkula M, "Ratio of proliferating cell nuclear antigen-positive nuclei to total cell number is higher in day 7 than in day 8 vitrified in vitro-produced bovine embryos", Biol Reprod, 65, pp.52~59 (2001)[DOI]
- Gjørret JO, "Chronology of apoptosis in bovine embryos produced in vivo and in vitro", Biol Reprod, 69, pp.1193~1200 (2003)[DOI]
- Lin T, "α-Solanine impairs oocyte maturation and quality by inducing autophagy and apoptosis and changing histone modifications in a pig model", Reprod Toxicol, 75, pp.96~109 (2018)[DOI]
- Chen X, "Integration of external signaling pathways with the core transcriptional network in embryonic stem cells", Cell, 133, pp.1106~1117 (2008)[DOI]
- Liu S, "Sox2 is the faithful marker for pluripotency in pig : evidence from embryonic studies", Dev Dyn, 244, pp.619~627 (2015)[DOI]
- Loh YH, "The Oct4 and Nanog transcription network regulates pluripotency in mouse embryonic stem cells", Nat Genet, 38, pp.431~440 (2006)[DOI]
- Keramari M, "Sox2 is essential for formation of trophectoderm in the preimplantation embryo", PLOS ONE, 5, pp.e13952~ (2010)[DOI]
- Bui HT, "Epigenetic reprogramming in somatic cells induced by extract from germinal vesicle stage pig oocytes", Development, 139, pp.4330~4340 (2012)[DOI]
