Verma Megha, Pandey Sriti, Bhat Irfan A, Mukesh B, Anand J, Chandra Vikash, Sharma G Taru
Division of Physiology & Climatology, ICAR-Indian Veterinary Research Institute, Izatnagar, 243 122, UP, India.
Division of Physiology & Climatology, ICAR-Indian Veterinary Research Institute, Izatnagar, 243 122, UP, India.
Cryobiology. 2018 Jun;82:99-105. doi: 10.1016/j.cryobiol.2018.04.001. Epub 2018 Apr 5.
The aim of the present study was to see the impact of L-Carnitine (LC) on lipid biosynthesis and metabolism of buffalo embryos, and post thaw blastocyst survivability. In vitro fertilized (IVF) embryos were derived from slaughterhouse derived COCs and cultured in different doses of LC i.e. 0, 1 mM, 1.5 mM, 2 mM starting at 48 h post IVF. Blastocyst rate was significantly (p < 0.05) higher in 1.5 mM group than control and 1.0 mM group. Lipid content was measured indirectly by fluorescent intensity of lipid droplets after Nile red staining, and it was lower (p < 0.05) in treated than control groups. CPT1B, DGAT2 and DGAT1 mRNA expression was up regulated (p < 0.05) while AMPKg1 expression was down regulated in 1.5 mM and 2 mM groups compared to other groups (p < 0.05). mRNA expression of GLUT1, OCT4 and IFN-tau was higher (P < 0.05) in 1.5 mM group than the control group. Expression of BAX was down regulated at 1.5 mM LC. Blastocyts were vitrified by a modified OPS method and post thaw survivability of blastocysts was higher (P < 0.05) in 1.5 mM LC than other groups. In post thaw blastocysts, mRNA expression of GLUT1, OCT4 and IFN-tau was higher (P < 0.05) in 1.5 mM than other groups. Thus, it can be concluded that supplementation of l-carnitine (1.5 mM) in embryo culture media improved the quality of buffalo embryo production and post thaw blastocysts survivability by reducing fatty acid synthesis, enhancing fatty acid metabolism, and reducing lipid droplet formation.
本研究的目的是观察左旋肉碱（LC）对水牛胚胎脂质生物合成与代谢以及解冻后囊胚存活率的影响。体外受精（IVF）胚胎来源于屠宰场采集的卵丘-卵母细胞复合体（COCs），在IVF后48小时开始在不同剂量的LC（即0、1 mM、1.5 mM、2 mM）中培养。1.5 mM组的囊胚率显著高于对照组和1.0 mM组（p < 0.05）。通过尼罗红染色后脂质滴的荧光强度间接测量脂质含量，处理组的脂质含量低于对照组（p < 0.05）。与其他组相比，1.5 mM和2 mM组中肉碱棕榈酰转移酶1B（CPT1B）、二酰甘油酰基转移酶2（DGAT2）和二酰甘油酰基转移酶1（DGAT1）的mRNA表达上调（p < 0.05），而腺苷酸活化蛋白激酶γ1（AMPKg1）的表达下调（p < 0.05）。1.5 mM组中葡萄糖转运蛋白1（GLUT1）、八聚体转录因子4（OCT4）和干扰素τ（IFN-tau）的mRNA表达高于对照组（P < 0.05）。在1.5 mM LC时，凋亡蛋白BAX的表达下调。采用改良的开放式拉长细管（OPS）法对囊胚进行玻璃化冷冻，1.5 mM LC组解冻后囊胚的存活率高于其他组（P < 0.05）。在解冻后的囊胚中，1.5 mM组中GLUT1、OCT4和IFN-tau的mRNA表达高于其他组（P < 0.05）。因此，可以得出结论，在胚胎培养基中添加左旋肉碱（1.5 mM）可通过减少脂肪酸合成、增强脂肪酸代谢和减少脂质滴形成来提高水牛胚胎生产质量和解冻后囊胚的存活率。
