Supplementing ±-Linolenic acid in the in vitro maturation media improves nuclear maturation rate of oocytes and early embryonic development in the Nili Ravi buffalo

Abstract

The present study was conducted to investigate the effect of omega-3 poly unsaturated fatty acid (PUFA), α-linolenic acid (ALA; 18:3 n-3) on the in vitro maturation (IVM) of buffalo oocytes and subsequent embryonic development. Buffalo cumulusoocyte complexes (COCs; n = 2282) were in vitro matured in TCM-199 (0.6% fatty acid free bovine serum albumin, 0.02 Units/ml FSH, 1 µg/ml 17-β-estradiol, 10 µg/ml epidermal growth factor, 50 µg/ml gentamicin) supplemented with 0 (control), 25, 50, 100, 150 or 300 µm ALA under an atmosphere of 5% CO 2 in air at 38.5ºC for 22-24 h. The matured oocytes were then fertilized in Tyrode's Albumin Lactate Pyruvate (TALP) medium and cultured in synthetic oviductal fluid (SOF) medium. Concentrations up to 100 μm ALA improves (P ≤ 0.05) the cumulus expansion compared to control. Higher percentage of oocytes reaching MII stage was observed at 50 μm and 100 μm of ALA compared to control (P ≤ 0.05) . Concentrations of 150 and 300 µm ALA were detrimental both for cumulus expansion and nuclear maturation rate of buffalo oocytes. Moreover, supplementation with 100 μm ALA improved (P ≤ 0.05) cleavage rate compared to control and treatment with 50 and 100 μm ALA yielded significantly higher morulae compared to control. The results of present study indicate that the supplementation with 100 μm ALA to the IVM medium improves nuclear maturation rate of buffalo oocytes and subsequent early embryonic development.

Synergistic impact of α-linolenic acid and α-tocopherol on in vitro maturation and culture of buffalo oocytes

“…Perhaps the concentration of ALA used in this study has not produced oxidative stress to a level that could have been compensated by the antioxidant. As, mentioned earlier (Azam et al, 2017), ALA at higher concentration (300 µM) reduced the number of COCs with fully expanded oocytes and the rate of oocytes reaching up to MII stage. This reduced oocyte maturation rate at higher concentration of ALA, might have been due to the production of ROS in the IVM medium.…”

“…Cumulus oocyte complexes (COCs) were collected by aspiration of 2-8 mm visible follicles present on the surface of ovaries as described earlier (Azam et al, 2017). The COCs were classified as grade A, B, C and D, based on cumulus mass investment and ooplasm homogeneity under stereomicroscope as described earlier (Azam et al, 2017). Only grade A and B oocytes were processed for IVM.…”

“…Likewise, the in vitro bovine, ovine and porcine oocyte maturation and high quality embryos are obtained when fatty acids supplemented media was used (Marei et al, 2009;Ghaffarilaleh et al, 2014;Jeon et al ., 2020). We have previously shown that α-linolenic acid (100 µM) enriched in vitro growth media enhanced the rates of oocyte maturation and pre-implantation embryo development in Nili Ravi buffalo (Azam et al, 2017). Apart from the useful role of α-linolenic acid, it is also susceptible to lipid peroxidation.…”

“…Brazilian Journal of Biology, 2024, vol. 84, e253514 3/7 Maturation of buffalo oocytes in supplemented media completely denuded, stained with 1% aceto-orcein and examined for metaphase II (MII) stage (Azam et al, 2017).…”

Stimulation of follicle growth and development during estrus in Ettawa Grade does fed a flushing supplement of different polyunsaturated fatty acids

“…Several studies have considered the utilization of different PUFA sources for improving reproductive functions in ruminants [2,[9][10][11][12]. Alpha (α)-linolenic acid supplementation at a dose of 100 µM during in vitro maturation was reported to increase the nuclear maturity rate of buffalo oocytes [13].…”