Effect of arachidonic acid supplementation and cyclooxygenase/lipoxygenase inhibition on the development of early bovine embryos

Abstract

-The effect of arachidonic acid (AA) cascade on bovine embryo development in a granulosa cell co-culture system was studied. Arachidonic acid (100 µM) was supplemented from 1-cell to 8-16 cell block stage (first three days of coculture) and from 1-cell to hatching. Specific cyclooxygenase (indomethacin, 28 µM) and lipoxygenase (nordihydroguaiaretic acid -NDGA, 28 µM) inhibitors were used from 1-cell to 8-16 cell block stage with AA. Embryo development was assessed by cleavage, day 7-day 8 and hatched embryo rates and by measuring growth rates through development stages found in days 7-10 of culture (day 0 = insemination day). Embryo quality was scored at day 8. A 6.5-10.4% increase on cleavage rate after AA supplementation was found. This AA supplementation from 1-cell to hatching delayed embryo growth rate beyond day 7 and a reduction on hatching rate was detected. When AA supplementation was restricted to the first three days of co-culture those negative effects were overcome. Also, indomethacin and NDGA prevented the positive effect of AA and induced a significant reduction on cleavage, respectively. NDGA further decreased day 7 embryo rate and quality. Results suggest that AA has a two-phase action on bovine embryos, promoting early development and impairing embryo growth from day 7 onwards and hatching rates. Both cyclooxygenase and lipoxygenase were found to be important pathways to promote cleavage.Key Words: arachidonic acid, bovine embryos, granulosa, in vitro Influência do ácido araquidónico e da inibição da ciclo-oxigenase ou lipooxigenase no desenvolvimento inicial de embriões bovinos RESUMO -Estudou-se a influência da cascata do ácido araquidónico (AA) no desenvolvimento de embriões bovinos produzidos in vitro em co-cultura com células da granulosa. Os embriões foram suplementados com AA (100 µM) desde o estádio de 1 célula até 8-16 células (primeiros três dias de co-cultura) ou até a eclosão. Introduziram-se inibidores específicos da ciclooxigenase (indometacina, 28 µM) e da lipo-oxigenase (ácido nordihidroguaiarético -NDGA, 28 µM), juntamente com o ácido araquidónico, desde o estádio de 1 célula até 8-16 células. O desenvolvimento embrionário foi avaliado pelas taxas de clivagem de embriões nos dias 7 e 8 (D0 = dia da inseminação) e eclodidos. Avaliou-se também a velocidade de crescimento dos embriões entre os dias 7 e 10 de cultura e a qualidade dos embriões no dia 8. A suplementação dos embriões com AA aumentou a taxa de clivagem (6,5 e 10,4%). Por outro lado, a presença deste percursor das prostaglandinas e dos leucotrienos com os embriões desde o estádio de 1 célula até a eclosão atrasou sua velocidade de crescimento a partir do dia 7 e reduziu a taxa de eclosão em 20.4%.Estes efeitos negativos desapareceram quando a suplementação do AA foi restringida apenas aos três primeiros dias de co-cultura.A indometacina evitou o efeito positivo do AA na clivagem, tendo o NDGA reduzido esta taxa, mesmo em presença do AA, assim como a taxa de embriões em D7 e a qualidade dos embriões D8. Os resu...

Anti-Aging Effect of Urolithin A on Bovine Oocytes In Vitro

“…After 48 h of the insemination, the cleavage rate (cleaved embryos per total inseminated oocytes) was assessed, and cleaved embryos were maintained in SOF supplemented with BSA and 10% of fetal bovine serum (FBS). Embryos were cultured for 12 days [41,43] to assess the blastocyst development rate (at days 7, 9, and 12; D7 embryos per cleaved embryos) and hatched embryo rate (hatched embryos per D7 embryos) and their quality [43]. Day 7 embryos were classified as grade 1 (good quality), 2 (fair quality), and grade 3 (bad quality) [40,44].…”

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Effect of feed with Pittosporum undulatum in vivo on bovine progesterone levels and embryos produced in vitro

“…This enzyme is essential for the metabolism of arachidonic acid to prostaglandins that are important for ovulation and function of the corpus luteum (Tsai and Wiltbank 1998;Calder et al 2001;Sales and Jabbour 2003). This enzyme is also important for development of embryos (Thatcher et al 1984;Pereira et al 2006); therefore, a suppression of COX-2 activity may explain the lower rate of cleavage observed in the experimental group.…”