Amino acid concentrations in fluids from the bovine oviduct and uterus and in ksom-based culture media

Abstract

AabstractOviductal (OF) and uterine (UF) fluids from slaughterhouse bovine reproductive tracts selected around the pre-implantation period of embryo development were analyzed for twenty-four amino acids. Amino acid concentrations in the OF and UF fluids were compared with those in a base culture medium (KSOM) supplemented with either FCS or Minimum Essential Medium amino acids (MEM-aa) in addition to PVA, FCS or BSA. Concentrations of 23 amino acids in UF were significantly (P<0.05) higher than those provided by KSOM supplemented with any of the supplements. Cystine was the only amino acid that was supplemented in comparable levels to those in UF. However, OF levels of CYS were higher than those in UF and supplemented KSOM. Compared to amino acid levels in OF, the supplementation of KSOM with either FCS or MEM-aa in addition to PVA, FCS or BSA resulted in significantly (P<0.05) lower levels of 15, 13, 13 and 10 amino acids, respectively. The results showed that non-essential (NE) amino acids are the most abundant in both OF and UF, and compared to essential (E) amino acids, they were present in both fluids at ratios of 5:1 and 2:1, respectively. Glycine (14.1 mM), GLU (5.5 mM) and ALA (3.7 mM) were present in high concentrations and comprised 72% of the free amino acid pool of OF. In the uterine fluid, concentrations of GLY (12.0 mM), GLU (4.2 mM), ALA (3.1 mM) and SER (2.7 mM) were the highest. Concentrations of all non-standard amino acids were significantly (P<0.05) higher in UF compared to KSOM supplemented with either FCS or FCS in addition to MEM-aa. In OF, only CIT and -ALA were significantly (P<0.05) higher in concentrations compared to KSOM under similar supplementation. In conclusion, concentrations of most of the amino acids in the bovine reproductive tract were substantially higher than those supplemented in culture media commonly used for in vitro development of bovine preimplantation embryos. Adjustment of amino acid concentrations in the culture medium according to those found in the bovine reproductive tract may improve embryo development in vitro. Further culture studies are currently investigating this hypothesis.

Effects of systemic progesterone during the early luteal phase on the availabilities of amino acids and glucose in the bovine uterine lumen

“…The identification of Gly as a highly abundant amino acid in female reproductive tract fluids is consistent with previous observations in cattle (Fahning et al 1967;Tiffin et al 1991;Moore and Bondioli 1993;Steeves and Gardner 1999;Elhassan et al 2001;Hugentobler et al 2007), sheep (Hill et al 1997;Moses et al 1997;Gao et al 2009b), horses (Engle et al 1984) and rabbits (Miller and Schultz 1987). The relatively high abundance of Gly in the uterine lumen across species suggests that it has an important role in embryo and/or conceptus development, and this is further supported by its abundance in the fetuses of swine throughout pregnancy (Wu et al 1999).…”

Concentration and composition of free amino acids and osmolalities of porcine oviductal and uterine fluid and their effects on development of porcine IVF embryos

“…The free amino acid concentration and composition between bovine oviductal fluid and Eagle's formulation are obviously different. The total amount of Eagle amino acids (3.17mmol/L) is almost 10 times lower than that in bovine oviductal fluid (32.2mmol/L, Elhassan et al 2001;31.6mmol/L, Li et al 2003), and the concentration of several amino acid, such as glycine, is about 140 times lower (Elhassan et al 2001;Li et al 2003). Some non-protein amino acids, such as taurine (Tau), exist in both bovine and murine reproduction tract fluid (Elhassan et al 2001;Li et al 2003;Harris et al 2005) but are not contained in the Eagle formulation (Eagle 1959).…”

“…The total amount of Eagle amino acids (3.17mmol/L) is almost 10 times lower than that in bovine oviductal fluid (32.2mmol/L, Elhassan et al 2001;31.6mmol/L, Li et al 2003), and the concentration of several amino acid, such as glycine, is about 140 times lower (Elhassan et al 2001;Li et al 2003). Some non-protein amino acids, such as taurine (Tau), exist in both bovine and murine reproduction tract fluid (Elhassan et al 2001;Li et al 2003;Harris et al 2005) but are not contained in the Eagle formulation (Eagle 1959). Two research groups (McKiernan et al 1995;Bavister & Arlotto 1990;Lane & Gardner 1997;Lane et al 2001) have optimized the composition and concentration of amino acids in the culture media for hamster and mouse embryos.…”

The organic osmolytes betaine and proline are transported by a shared system in early preimplantation mouse embryos

“…It has previously been found that mouse oviductal fluid contains about 140 mM proline (Guerin et al, 1995), equivalent to 1-3% of total amino acids (Baltz, 2001). Proline is also present at significant concentrations in tubal fluid of cow and rabbit, to a lesser extent in sheep, but little in pig (Guerin et al, 1995;Elhassan et al, 2001). There was no information available about betaine in oviducts.…”