家畜卵母细胞和胚胎去脂方法研究进展

doi:10.16431/j.cnki.1671-7236.2023.02.020

摘要/Abstract

摘要： 卵母细胞和胚胎冷冻技术可与超数排卵和胚胎移植等繁殖技术相结合,打破生理、地区和时间对母畜繁殖潜力的限制,对国际种源流通、优良家畜扩繁和濒危动物种质资源保存具有重要意义。然而,卵母细胞和胚胎中丰富的脂质在降温过程中容易引起膜结构损伤、内质网和线粒体损伤、脂质过氧化,降低其解冻后发育能力,这极大地限制了冷冻卵母细胞和胚胎的应用。众多研究表明,在冷冻前降低卵母细胞和胚胎的脂质含量有利于提高其解冻后的存活率、囊胚率和妊娠率。目前,常用的降脂方法包括3种,即离心极化脂滴后通过显微操作去除脂滴、添加化学物质促进脂质代谢以及通过调控脂质代谢相关基因的表达降低脂质含量。作者简要阐述了过高的脂质含量引起卵母细胞和胚胎冷冻损伤的机制,总结了离心去脂法与化学去脂法的原理、应用效果以及局限性,并探讨了通过调控脂质代谢相关基因的表达从而去脂的可能性,以期为开发更稳定的卵母细胞和胚胎冷冻技术提供一定的参考。

关键词: 卵母细胞; 胚胎; 去脂; 冷冻效率

Abstract: Cryopreservation of oocytes and embryos can be combined with reproduction techniques such as superovulation and embryo transfer to break the limitations of physiology,region and time on the reproductive potential of female animals,which is of great significance for international circulation of germplasm,propagation of excellent domestic animals and conservation of germplasm of endangered animals.However,oocytes or embryos are prone to membrane damage,endoplasmic reticulum and mitochondrial damage and lipid peroxidation during cryopreserved process due to the abundant lipid content,which reduces their developmental ability after thawing and greatly limits the application of cryopreserved oocytes and embryos.Many researches have shown that reducing the lipid content of oocytes or embryos before cryopreservation is beneficial for improving the survival rate,blastocyst rate,and pregnancy rate of them after thawing.At present,there are three popular methods to reduce lipid content such as removing lipid droplets by micromanipulation after polarization by centrifugation,promoting lipid metabolism by adding chemicals,and regulating the expression of genes related to lipid metabolism.In this review,the mechanisms of damages caused by high lipid content in oocyte and embryo were briefly expounded,the principles,effects and limitations of methods that reduce the content of lipids by centrifugation or chemicals were summarized,and the possibility of lipid removal by regulating the expression of genes related to lipid metabolism was discussed.This review could provide some reference for the development of more stable cryopreservation technology of oocyte and embryo.

Key words: oocyte; embryo; degreasing; refrigeration efficiency

中图分类号:

S814.8

徐茜, 杨宇泽, 郝海生, 杜卫华, 朱化彬, 杨柏高, 赵学明. 家畜卵母细胞和胚胎去脂方法研究进展[J]. 中国畜牧兽医, 2023, 50(2): 626-637.

XU Xi, YANG Yuze, HAO Haisheng, DU Weihua, ZHU Huabin, YANG Baigao, ZHAO Xueming. Advances in Oocyte and Embryo Defatting Methods for Domestic Animals[J]. China Animal Husbandry & Veterinary Medicine, 2023, 50(2): 626-637.

参考文献

[1] RIENZI L,GRACIA C,MAGGIULLI R,et al.Oocyte,embryo and blastocyst cryopreservation in ART:Systematic review and meta-analysis comparing slow-freezing versus vitrification to produce evidence for the development of global guidance[J]. Human Reproduction Update,2017,23(2):139-155.
[2] 周青顺,秦茂,秦世新,等.猪冻精推广应用的思考[J].湖北畜牧兽医,2021,42(3):20-22. ZHOU Q S,QIN M,QIN S X,et al.Reflections on the promotion and application of swine frozen semen[J]. Hubei Joumal of Animal and Veterinary Sciences,2021,42(3):20-22.(in Chinese)
[3] PRENTICE J R,ANZAR M.Cryopreservation of mammalian oocyte for conservation of animal genetics[J]. Veterinary Medicine International,2010,2011:146405.
[4] VINING L M,ZAK L J,HARVEY S C,et al.The role of apoptosis in cryopreserved animal oocytes and embryos[J]. Theriogenology,2021,173:93-101.
[5] ARGYLE C E,HARPER J C,DAVIES M C.Oocyte cryopreservation:Where are we now?[J]. Human Reproduction Update,2016,22(4):440-449.
[6] FRITZ R,JINDAL S.Reproductive aging and elective fertility preservation[J]. Journal of Ovarian Research,2018,11(1):66.
[7] ABIR R,BEN-AHARON I,GAROR R,et al.Cryopreservation of in vitro matured oocytes in addition to ovarian tissue freezing for fertility preservation in paediatric female cancer patients before and after cancer therapy[J]. Human Reproduction,2016,31(4):750-762.
[8] WHITTINGHAM D G.Fertilization in vitro and development to term of unfertilized mouse oocytes previously stored at -196 ℃[J]. Journal of Reproduction and Fertility,1977,49(1):89-94.
[9] PARKENING T A,CHANG M C.Effects of cooling rates and maturity of the animal on the recovery and fertilization of frozen-thawed rodent eggs[J]. Biology of Reproduction,1977,17(4):527-531.
[10] WHITTINGHAM D G,LEIBO S P,MAZUR P.Survival of mouse embryos frozen to -196 degrees and -269 degrees C[J]. Science,1972,178(4059):411-414.
[11] SOMFAI T,YOSHIOKA K,TANIHARA F,et al.Generation of live piglets from cryopreserved oocytes for the first time using a defined system for in vitro embryo production[J]. PLoS One,2014,9(5):e97731.
[12] SPRÍCIGO J F,MORATÓ R,ARCARONS N,et al.Assessment of the effect of adding L-carnitine and/or resveratrol to maturation medium before vitrification on in vitro-matured calf oocytes[J]. Theriogenology,2017,89:47-57.
[13] HOSSEINI S M,ASGARI V,OSTADHOSSEINI S,et al.Developmental competence of ovine oocytes after vitrification:Differential effects of vitrification steps,embryo production methods,and parental origin of pronuclei[J]. Theriogenology,2015,83(3):366-376.
[14] VICENTE J S,VIUDES-DE-CASTRO M P,CEDANO-CASTRO J I,et al.Cryosurvival of rabbit embryos obtained after superovulation with corifollitropin alfa with or without LH[J]. Animal Reproduction Science,2018,192:321-327.
[15] HAO T,ZHANG P,HAO H,et al.The combination treatment of cholesterol-loaded methyl-beta-cyclodextrin and methyl-beta-cyclodextrin significantly improves the fertilization capacity of vitrified bovine oocytes by protecting fertilization protein JUNO[J]. Reproduction in Domestic Animals,2021,56(3):519-530.
[16] CHINEN S,YAMANAKA T,HIRABAYASHI M,et al.Rescue of vitrified-warmed bovine mature oocytes by short-term recovery culture with resveratrol[J]. Cryobiology,2020,97:185-190.
[17] OLEXIKOVÁ L,DUJÍČKOVÁ L,KUBOVIČOVÁ E,et al.Development and ultrastructure of bovine matured oocytes vitrified using electron microscopy grids[J]. Theriogenology,2020,158:258-266.
[18] SANCHES B V,LUNARDELLI P A,TANNURA J H,et al.A new direct transfer protocol for cryopreserved IVF embryos[J]. Theriogenology,2016,85(6):1147-1151.
[19] SANTOS E,APPELTANT R,DANG-NGUYEN T Q,et al.The effect of resveratrol on the developmental competence of porcine oocytes vitrified at germinal vesicle stage[J]. Reproduction in Domestic Animals,2018,53(2):304-312.
[20] CASILLAS F,BETANCOURT M,CUELLO C,et al.An efficiency comparison of different in vitro fertilization methods:IVF,ICSI,and PICSI for embryo development to the blastocyst stage from vitrified porcine immature oocytes[J]. Porcine Health Management,2018,4(1):16.
[21] HWANG I S,PARK M R,KWAK T U,et al.Effect of cytochalasin B treatment on the improvement of survival rate in vitrified pig oocyte[J]. Development & Reproduction,2018,22(3):245-252.
[22] JIA B Y,XIANG D C,ZHANG B,et al.Quality of vitrified porcine immature oocytes is improved by coculture with fresh oocytes during in vitro maturation[J]. Molecular Reproduction and Development,2019,86(11):1615-1627.
[23] XIANG D C,JIA B Y,FU X W,et al.Role of astaxanthin as an efficient antioxidant on the in vitro maturation and vitrification of porcine oocytes[J]. Theriogenology,2021,167:13-23.
[24] CHEN Y N,DAI J J,WU C F,et al.Apoptosis and developmental capacity of vitrified parthenogenetic pig blastocysts[J]. Animal Reproduction Science,2018,198:137-144.
[25] TAJIMA S,UCHIKURA K,KURITA T,et al.The effect of artificial insemination prior to transfer of a limited number of vitrified and warmed porcine embryos by open pulled straw (OPS) method on their survival ability for farrowing[J]. Animal Science Journal,2020,91(1):e13325.
[26] TAJIMA S,UCHIKURA K,KURITA T,et al.Insemination of recipient sows improves the survival to term of vitrified and warmed porcine expanded blastocysts transferred non-surgically[J]. Animal Science Journal,2020,91(1):e13453.
[27] AHMADI E,SHIRAZI A,SHAMS ESFANDABADI N,et al.Antioxidants and glycine can improve the developmental competence of vitrified/warmed ovine immature oocytes[J]. Reproduction in Domestic Animals,2019,54(3):595-603.
[28] DAVOODIAN N,KADIVAR A,AHMADI E,et al.Quercetin effect on the efficiency of ovine oocyte vitrification at GV stage[J]. Theriogenology,2021,174:53-59.
[29] MOAWAD A R,CHOI I,ZHU J,et al.Caffeine and oocyte vitrification:Sheep as an animal model[J]. International Journal of Veterinary Science and Medicine,2018,6:S41-S48.
[30] SUDIMAN J,LEE A,ONG K L,et al.Tolerance of lamb and mouse oocytes to cryoprotectants during vitrification[J]. Zygote,2019,27(1):36-45.
[31] SANAEI B,MOVAGHAR B,REZAZADEH VALOJERDI M,et al.Developmental competence of in vitro matured ovine oocytes vitrified in solutions with different concentrations of trehalose[J]. Reproduction in Domestic Animals,2018,53(5):1159-1167.
[32] LEDDA S,KELLY J M,NIEDDU S,et al.High in vitro survival rate of sheep in vitro produced blastocysts vitrified with a new method and device[J]. Journal of Animal Science and Biotechnology,2019,10:90.
[33] PAPADOPOULOS S,RIZOS D,DUFFY P,et al.Embryo survival and recipient pregnancy rates after transfer of fresh or vitrified, in vivo or in vitro produced ovine blastocysts[J]. Animal Reproduction Science,2002,74(1-2):35-44.
[34] MARSICO T V,DE CAMARGO J,VALENTE R S,et al.Embryo competence and cryosurvival:Molecular and cellular features[J]. Animal Reproduction,2019,16(3):423-439.
[35] MEN H,AGCA Y,RILEY L K,et al.Improved survival of vitrified porcine embryos after partial delipation through chemically stimulated lipolysis and inhibition of apoptosis[J]. Theriogenology,2006,66(8):2008-2016.
[36] DIEZ C,HEYMAN Y,LE BOURHIS D,et al.Delipidating in vitro-produced bovine zygotes:Effect on further development and consequences for freezability[J]. Theriogenology,2001,55(4):923-936.
[37] GHANEM N,HA A N,FAKRUZZAMAN M,et al.Differential expression of selected candidate genes in bovine embryos produced in vitro and cultured with chemicals modulating lipid metabolism[J]. Theriogenology,2014,82(2):238-250.
[38] ZOLINI A M,CARRASCAL-TRIANA E,RUIZ D K A,et al.Effect of addition of L-carnitine to media for oocyte maturation and embryo culture on development and cryotolerance of bovine embryos produced in vitro[J]. Theriogenology,2019,133:135-143.
[39] BATISTA R I,RAPOSO N R,CAMPOS-JUNIOR P H,et al.Trans-10,cis-12 conjugated linoleic acid reduces neutral lipid content and may affect cryotolerance of in vitro-produced crossbred bovine embryos[J]. Journal of Animal Science and Biotechnology,2014,5(1):33.
[40] SANCHES B V,MARINHO L S,FILHO B D,et al.Cryosurvival and pregnancy rates after exposure of IVF-derived Bos indicus embryos to forskolin before vitrification[J]. Theriogenology,2013,80(4):372-377.
[41] WANG Y,CHANG H,HE Q,et al.Effect of oocyte vitrification on glucose transport in mouse metaphase Ⅱ oocytes[J]. Reproduction,2021,161(5):549-559.
[42] QIU J,HASEGAWA A,MOCHIDA K,et al.Equilibrium vitrification of mouse embryos using low concentrations of cryoprotectants[J]. Cryobiology,2021,98:127-133.
[43] SUTTIROJPATTANA T,JUANPANICH T,PARNPAI R,et al.Vitrification of mouse two-cell and blastocyst stage embryos in simplified closed system using either a hemi-straw or a hollow fiber device[J]. Animal Science Journal,2021,92(1):e13585.
[44] DUNNING K R,RUSSELL D L,ROBKER R L.Lipids and oocyte developmental competence:The role of fatty acids and beta-oxidation[J]. Reproduction,2014,148(1):R15-R27.
[45] STURMEY R G,REIS A,LEESE H J,et al.Role of fatty acids in energy provision during oocyte maturation and early embryo development[J]. Reproduction in Domestic Animals,2009,44 Suppl 3:50-58.
[46] ROMEK M,GAJDA B,KRZYSZTOFOWICZ E,et al.New technique to quantify the lipid composition of lipid droplets in porcine oocytes and pre-implantation embryos using nile red fluorescent probe[J]. Theriogenology,2011,75(1):42-54.
[47] MCEVOY T G,COULL G D,BROADBENT P J,et al.Fatty acid composition of lipids in immature cattle,pig and sheep oocytes with intact zona pellucida[J]. Journal of Reproduction and Fertility,2000,118(1):163-170.
[48] GENICOT G,LEROY J L,SOOM A V,et al.The use of a fluorescent dye,Nile red,to evaluate the lipid content of single mammalian oocytes[J]. Theriogenology,2005,63(4):1181-1194.
[49] WALTHER T C,FARESE R J.Lipid droplets and cellular lipid metabolism[J]. Annual Review of Biochemistry,2012,81:687-714.
[50] WELTE M A.Expanding roles for lipid droplets[J]. Current Biology,2015,25(11):R470-R481.
[51] ABE H,YAMASHITA S,SATOH T,et al.Accumulation of cytoplasmic lipid droplets in bovine embryos and cryotolerance of embryos developed in different culture systems using serum-free or serum-containing media[J]. Molecular Reproduction and Development,2002,61(1):57-66.
[52] AMSTISLAVSKY S,MOKROUSOVA V,BRUSENTSEV E,et al.Influence of cellular lipids on cryopreservation of mammalian oocytes and preimplantation embryos:A review[J]. Biopreservation and Biobanking,2019,17(1):76-83.
[53] OKOTRUB K A,AMSTISLAVSKY S Y,SUROVTSEV N V.Raman spectroscopy reveals the lipid phase transition in preimplantation mouse embryos during freezing[J]. Archives of Biochemistry and Biophysics,2017,635:37-43.
[54] LEE C H,LIN W C,WANG J.All-optical measurements of the bending rigidity of lipid-vesicle membranes across structural phase transitions[J]. Physical Review.E,Statistical,Nonlinear,and Soft Matter Physics,2001,64(2 Pt 1):20901.
[55] SYDYKOV B,OLDENHOF H,DE OLIVEIRA B L,et al.Membrane permeabilization of phosphatidylcholine liposomes induced by cryopreservation and vitrification solutions[J]. Biochimica et Biophysica Acta Biomembranes,2018,1860(2):467-474.
[56] DE KRUYFF B,VAN DIJCK P W,GODLBACH R W,et al.Influence of fatty acid and sterol composition on the lipid phase transition and activity of membrane-bound enzymes in acholeplasma laidlawii[J]. Biochimica et Biophysica Acta,1973,330(3):269-282.
[57] TONER M,CRAVALHO E G,KAREL M.Thermodynamics and kinetics of intracellular ice formation during freezing of biological cells[J]. Journal of Applied Physics,1990,67(3):1582-1593.
[58] ARAV A,ZERON Y,LESLIE S B,et al.Phase transition temperature and chilling sensitivity of bovine oocytes[J]. Cryobiology,1996,33(6):589-599.
[59] ZERON Y,SKLAN D,ARAV A.Effect of polyunsaturated fatty acid supplementation on biophysical parameters and chilling sensitivity of ewe oocytes[J]. Molecular Reproduction and Development,2002,61(2):271-278.
[60] WU L L,DUNNING K R,YANG X,et al.High-fat diet causes lipotoxicity responses in cumulus-oocyte complexes and decreased fertilization rates[J]. Endocrinology,2010,151(11):5438-5445.
[61] AARDEMA H,VOS P L,LOLICATO F,et al.Oleic acid prevents detrimental effects of saturated fatty acids on bovine oocyte developmental competence[J]. Biology of Reproduction,2011,85(1):62-69.
[62] JUNGHEIM E S,LOUDEN E D,CHI M M,et al.Preimplantation exposure of mouse embryos to palmitic acid results in fetal growth restriction followed by catch-up growth in the offspring[J]. Biology of Reproduction,2011,85(4):678-683.
[63] GUALTIERI R,KALTHUR G,BARBATO V,et al.Mitochondrial dysfunction and oxidative stress caused by cryopreservation in reproductive cells[J]. Antioxidants (Basel),2021,10(3):337.
[64] ZHAO X M,HAO H S,DU W H,et al.Melatonin inhibits apoptosis and improves the developmental potential of vitrified bovine oocytes[J]. Journal of Pineal Research,2016,60(2):132-141.
[65] ALI A A,BILODEAU J F,SIRARD M A.Antioxidant requirements for bovine oocytes varies during in vitro maturation,fertilization and development[J]. Theriogenology,2003,59(3-4):939-949.
[66] ALAM F,SYED H,AMJAD S,et al.Interplay between oxidative stress,SIRT1,reproductive and metabolic functions[J]. Current Research in Physiology,2021,4:119-124.
[67] MIHALAS B P,DE IULIIS G N,REDGROVE K A,et al.The lipid peroxidation product 4-hydroxynonenal contributes to oxidative stress-mediated deterioration of the ageing oocyte[J]. Scientific Reports,2017,7(1):6247.
[68] YANG H C,YU H,MA T H,et al.Tert-Butyl hydroperoxide (tBHP)-induced lipid peroxidation and embryonic defects resemble glucose-6-phosphate dehydrogenase (G6PD) deficiency in C.elegans[J]. International Journal of Molecular Sciences,2020,21(22).
[69] POLGE C.The freezing of mammalian embryos:Perspectives and possibilities[J]. Ciba Found Symp,1977(52):3-18.
[70] VALM A M,COHEN S,LEGANT W R,et al.Applying systems-level spectral imaging and analysis to reveal the organelle interactome[J]. Nature,2017,546(7656):162-167.
[71] ZHOU L,YU M,ARSHAD M,et al.Coordination among lipid droplets,peroxisomes,and mitochondria regulates energy expenditure through the CIDE-ATGL-PPARα pathway in adipocytes[J]. Diabetes,2018,67(10):1935-1948.
[72] 刘叶阳,许梦瑶,李蓬,等.脂滴与其他细胞器互作调控脂稳态的分子机制[J].中国科学:生命科学,2022,52(1):76-94. LIU Y Y,XU M Y,LI P,et al.The molecular mechanisms of contacts between lipid droplets and other organelles in lipid homeostasis regulation[J]. Scientia Sinica Vitae,2022,52(1):76-94.(in Chinese)
[73] 薛梦琦,周悦,刘可可,等.玻璃化冷冻对猪MⅡ期卵母细胞及其DNA的影响[J].中国畜牧兽医,2021,48(7):2475-2483. XUE M Q,ZHOU Y,LIU K K,et al.Effect of vitrification on MⅡ oocytes and their DNA of porcine[J]. China Animal Husbandry & Veterinary Medicine,2021,48(7):2475-2483.(in Chinese)
[74] HARA K,ABE Y,KUMADA N,et al.Extrusion and removal of lipid from the cytoplasm of porcine oocytes at the germinal vesicle stage:Centrifugation under hypertonic conditions influences vitrification[J]. Cryobiology,2005,50(2):216-222.
[75] NAGASHIMA H,HIRUMA K,SAITO H,et al.Production of live piglets following cryopreservation of embryos derived from in vitro-matured oocytes[J]. Biology of Reproduction,2007,76(5):900-905.
[76] NAGASHIMA H,KASHIWAZAKI N,ASHMAN R J,et al.Removal of cytoplasmic lipid enhances the tolerance of porcine embryos to chilling[J]. Biology of Reproduction,1994,51(4):618-622.
[77] USHIJIMA H,YOSHIOKA H,ESAKI R,et al.Improved survival of vitrified in vivo-derived porcine embryos[J]. The Journal of Reproduction and Development,2004,50(4):481-486.
[78] ESAKI R,UEDA H,KUROME M,et al.Cryopreservation of porcine embryos derived from in vitro-matured oocytes[J]. Biology of Reproduction,2004,71(2):432-437.
[79] REN L,FU B,MA H,et al.Effects of mechanical delipation in porcine oocytes on mitochondrial distribution,ROS activity and viability after vitrification[J]. Cryo-Letters,2015,36(1):30.
[80] NIU Y,WANG C,XIONG Q,et al.Distribution and content of lipid droplets and mitochondria in pig parthenogenetically activated embryos after delipation[J]. Theriogenology,2015,83(1):131-138.
[81] WANG C,NIU Y,CHI D,et al.Influence of delipation on the energy metabolism in pig parthenogenetically activated embryos[J]. Reproduction in Domestic Animals,2015,50(5):826-833.
[82] 刘凤娟,刘晋冀,李小兵,等.离心参数对猪卵母细胞早期去脂效果的影响[J].云南农业大学学报(自然科学),2018,33(2):240-248. LIU F J,LIU J J,LI X B,et al.The influence of centrifugal parameters on the performance of early-delipated pig oocytes[J]. Journal of Yunnan Agricultural University (Natural Science),2018,33(2):240-248.(in Chinese)
[83] SPATE L D,MURPHY C N,PRATHER R S.High-throughput cryopreservation of in vivo-derived swine embryos[J]. PLoS One,2013,8(6):e65545.
[84] TOMINAGA K,SHIMIZU M,OOYAMA S,et al.Effect of lipid polarization by centrifugation at different developmental stages on post-thaw survival of bovine in vitro produced 16-cell embryos[J]. Theriogenology,2000,53(8):1669-1680.
[85] BARCELÓ-FIMBRES M,BRINK Z,SEIDEL G J.Effects of phenazine ethosulfate during culture of bovine embryos on pregnancy rate,prenatal and postnatal development[J]. Theriogenology,2009,71(2):355-368.
[86] PEREIRA R M,BAPTISTA M C,VASQUES M I,et al.Cryosurvival of bovine blastocysts is enhanced by culture with trans-10 cis-12 conjugated linoleic acid (10t,12c CLA)[J]. Animal Reproduction Science,2007,98(3-4):293-301.
[87] PEREIRA R M,CARVALHAIS I,PIMENTA J,et al.Biopsied and vitrified bovine embryos viability is improved by trans10,cis12 conjugated linoleic acid supplementation during in vitro embryo culture[J]. Animal Reproduction Science,2008,106(3-4):322-332.
[88] 张紫薇,于博,戴佳格,等.小檗碱对玻璃化冷冻猪卵母细胞脂滴含量及胚胎发育的影响[J].北京农学院学报,2020,35(1):67-74. ZHANG Z W,YU B,DAI J G,et al.Effects of berberine on lipid droplets content and embryo development in vitrified frozen porcine oocytes[J]. Journal of Beijing University of Agriculture,2020,35(1):67-74.(in Chinese)
[89] RAZA S,ABD E A,ABDELNOUR S A,et al.The role of forskolin as a lipolytic stimulator during in vitro oocyte maturation and the in vitro embryo production of livestock[J]. Reproduction in Domestic Animals,2021,56(12):1486-1496.
[90] TAKAHASHI T,INABA Y,SOMFAI T,et al.Supplementation of culture medium with L-carnitine improves development and cryotolerance of bovine embryos produced in vitro[J]. Reproduction,Fertility,and Development,2013,25(4):589-599.
[91] CHANKITISAKUL V,SOMFAI T,INABA Y,et al.Supplementation of maturation medium with L-carnitine improves cryo-tolerance of bovine in vitro matured oocytes[J]. Theriogenology,2013,79(4):590-598.
[92] SPRICIGO J F,MORATO R,ARCARONS N,et al.Assessment of the effect of adding L-carnitine and/or resveratrol to maturation medium before vitrification on in vitro-matured calf oocytes[J]. Theriogenology,2017,89:47-57.
[93] VERMA M,PANDEY S,BHAT I A,et al.Impact of L-carnitine on lipid content and post thaw survivability of buffalo embryos produced in vitro[J]. Cryobiology,2018,82:99-105.
[94] DAI J,HUANG X,ZHANG C,et al.Berberine regulates lipid metabolism via miR-192 in porcine oocytes matured in vitro[J]. Veterinary Medicine Science,2021,7(3):950-959.
[95] SUDANO M J,PASCHOAL D M,RASCADO T S,et al.Lipid content and apoptosis of in vitro-produced bovine embryos as determinants of susceptibility to vitrification[J]. Theriogenology,2011,75(7):1211-1220.
[96] PANYABORIBAN S,THARASANIT T,CHANKITISAKUL V,et al.Treatment with chemical delipidation forskolin prior to cryopreservation improves the survival rates of swamp buffalo (Bubalus bubalis) and bovine (Bos indicus) in vitro produced embryos[J]. Cryobiology,2018,84:46-51.
[97] LOWE J L,BARTOLAC L K,BATHGATE R,et al.Cryotolerance of porcine blastocysts is improved by treating in vitro matured oocytes with L-carnitine prior to fertilization[J]. The Journal of Reproduction and Development,2017,63(3):263-270.
[98] SARAIVA H F R A,BATISTA R I T P,ALFRADIQUE V A P,et al.L-carnitine supplementation during vitrification or warming of in vivo-produced ovine embryos does not affect embryonic survival rates,but alters CrAT and PRDX1 expression[J]. Theriogenology,2018,105:150-157.
[99] DADASHPOUR DAVACHI N,FALLAHI R,DIRANDEH E,et al.Effects of co-incubation with conspecific ampulla oviductal epithelial cells and media composition on cryotolerance and developmental competence of in vitro matured sheep oocytes[J]. Theriogenology,2018,120:10-15.
[100] DE ANDRADE M F,POEHLAND R.Lipid metabolism in bovine oocytes and early embryos under in vivo, in vitro,and stress conditions[J]. International Journal of Molecular Sciences,2021,22(7):3421.
[101] WANG L F,WANG X N,HUANG C C,et al.Inhibition of NAMPT aggravates high fat diet-induced hepatic steatosis in mice through regulating Sirt1/AMPKα/SREBP1 signaling pathway[J]. Lipids in Health and Disease,2017,16(1):82.
[102] LI T,WEN L,CHENG B.Cordycepin alleviates hepatic lipid accumulation by inducing protective autophagy via PKA/mTOR pathway[J]. Biochemical and Biophysical Research Communications,2019,516(3):632-638.
[103] UDDIN G M,YOUNGSON N A,CHOWDHURY S S,et al.Administration of nicotinamide mononucleotide (NMN) reduces metabolic impairment in male mouse offspring from obese mothers[J]. Cells,2020,9(4):791.
[104] MIAO Y,CUI Z,GAO Q,et al.Nicotinamide mononucleotide supplementation reverses the declining quality of maternally aged oocytes[J]. Cell Reports,2020,32(5):107987.
[105] MIN H,LEE M,CHO K S,et al.Nicotinamide supplementation improves oocyte quality and offspring development by modulating mitochondrial function in an aged caenorhabditis elegans model[J]. Antioxidants (Basel),2021,10(4):519.
[106] WU C,GUO Y,SU Y,et al.Cordycepin activates AMP-activated protein kinase (AMPK) via interaction with the γ1 subunit[J]. Journal of Cellular and Molecular Medicine,2014,18(2):293-304.
[107] XU H,WU B,WANG X,et al.Cordycepin regulates body weight by inhibiting lipid droplet formation,promoting lipolysis and recruiting beige adipocytes[J]. Journal of Pharmacy and Pharmacology,2019,71(9):1429-1439.
[108] CAÑÓN-BELTRÁN K,GIRALDO-GIRALDO J,CAJAS Y N,et al.Inhibiting diacylglycerol acyltransferase-1 reduces lipid biosynthesis in bovine blastocysts produced in vitro[J]. Theriogenology,2020,158:267-276.
[109] HAEMMERLE G,LASS A,ZIMMERMANN R,et al.Defective lipolysis and altered energy metabolism in mice lacking adipose triglyceride lipase[J]. Science,2006,312(5774):734-737.
[110] HAEMMERLE G,ZIMMERMANN R,HAYN M,et al.Hormone-sensitive lipase deficiency in mice causes diglyceride accumulation in adipose tissue,muscle,and testis[J]. Journal of Biological Chemistry,2002,277(7):4806-4815.
[111] AHMADIAN M,DUNCAN R E,VARADY K A,et al.Adipose overexpression of desnutrin promotes fatty acid use and attenuates diet-induced obesity[J]. Diabetes,2009,58(4):855-866.
[112] 詹小立.牛 ATGL基因对牛脂肪细胞中甘油三酯积累的调控作用研究[D].杨凌:西北农林科技大学,2014. ZHAN X L.Role and mechanism of the adipose triglyceride lipase ATGL in adipocytes triacylglycerol accumulation[D].Yangling:Northwest A&F University,2014.(in Chinese)
[113] 周亭亭.广西巴马小型猪 HSL基因表达与2型糖尿病的相关性研究[D].南宁:广西大学,2016. ZHOU T T.Study on the relationship between the expression of HSL gene and type 2 diabetes mellitus in Guangxi Bama mini-pig[D].Nan Ning:Guangxi University,2016.(in Chinese)
[114] TIAN H,LUO J,ZHANG Z,et al.CRISPR/Cas9-mediated stearoyl-CoA desaturase 1 (SCD1) deficiency affects fatty acid metabolism in goat mammary epithelial cells[J]. Journal of Agricultural and Food Chemistry,2018,66(38):10041-10052.
[115] BECUWE M,BOND L M,PINTO A,et al.FIT2 is an acyl-coenzyme a diphosphatase crucial for endoplasmic reticulum homeostasis[J]. Journal of Cell Biology,2020,219(10):e202006111.
[116] MCINTOSH A L,SENTHIVINAYAGAM S,MOON K C,et al.Direct interaction of Plin2 with lipids on the surface of lipid droplets:A live cell FRET analysis[J]. American Journal of Physiology.Cell Physiology,2012,303(7):C728-C742.
[117] SCHWARZ K R L,DE CASTRO F C,SCHEFER L,et al.The role of cGMP as a mediator of lipolysis in bovine oocytes and its effects on embryo development and cryopreservation[J]. PLoS One,2018,13(1):e191023.
[118] MCMANAMAN J L,BALES E S,ORLICKY D J,et al.Perilipin-2-null mice are protected against diet-induced obesity,adipose inflammation,and fatty liver disease[J]. Journal of Lipid Research,2013,54(5):1346-1359.
[119] ZHANG R N,FU X W,JIA B Y,et al.Expression of perilipin 2 (PLIN2) in porcine oocytes during maturation[J]. Reproduction in Domestic Animals,2014,49(5):875-880.
[120] PAWLAK P,WARZYCH E,CIESLAK A,et al.The consequences of porcine IVM medium supplementation with follicular fluid become reflected in embryo quality,yield and gene expression patterns[J]. Scientific Reports,2018,8(1):15306.
[121] SASTRE D,COSTA N N D,SÁ A L A D,et al.Expression of PLIN2 and PLIN3 during oocyte maturation and early embryo development in cattle[J]. Theriogenology,2014,81(2):326-331.
[122] STAMATIADIS P,BOEL A,COSEMANS G,et al.Comparative analysis of mouse and human preimplantation development following POU5F1 CRISPR/Cas9 targeting reveals interspecies differences[J]. Human Reproduction,2021,36(5):1242-1252.
[123] GIM G M,KWON D H,EOM K H,et al.Production of MSTN-mutated cattle without exogenous gene integration using CRISPR-Cas9[J]. Biotechnology Journal,2022,17(7):2100198.
[124] WANG X,YU H,LEI A,et al.Generation of gene-modified goats targeting MSTN and FGF5 via zygote injection of CRISPR/Cas9 system[J]. Scientific Reports,2015,5:13878.