Metabolism of arachidonic acid by porcine blastocysts and endometrium |
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Authors: | GS Lewis RA Waterman |
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Institution: | Reproduction Laboratory Animal Science Institute U.S. Department of Agriculture Beltsville, Maryland 20705, USA |
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Abstract: | Metabolism of radiolabeled arachidonic acid (1AA) by blastocysts and endometrial slices recovered from five gilts 16 days after detection of estrus was studies . Blastocysts from each gilt were divided into four 216 ± 18 mg, and each portion was placed into a separate petri dish containing 15 ml modified minimum essential medium (MEM)_. The incubates from each gilt received either 25, 50, 100 or 200 μg radioinert arachidonic acid (AA). Endometrium was dissected from each uterin horn, sliced and duplicate 509 ± 3 mg portions from each gilt were placed into petri dishes containing 15 ml MEM and 200 μm AA. All incubates received 5 νCi of 1AA (either 14C]-arichidonic acid or 3H]-arichidonic acid). The incubates were rocked at 37°C for 24 h in an atmosphere of 50% n2:45% O2:5% CO2. After incubation, tissues and MEM were separated by centrifugation. Metabolism of 1AA was assessed in extracts of MEM and tissue homogenates by separating 1AA and its metabolites on columns of Sephades LH-20. Blastocysts produced compounds that migrated with 3H]-13,14-dihydro-15-keto-PGF2α (1PGFM), 3H]-PGE2 (1PGE2) and 3H]-PGF2α (1PGF2α). The greatest (P<.05) proportion (35.7 ± 1.8%) of the radioactivity in blastocyst MEM was recovered as PGE2. In blastocyst homogenates, most (66.2 ± 3.3%; P<0.05) of the radioactivity was in a nonporal peak assumed to be arachidonate esters. The concentration of AA ni MEM did not alter metabolism of 1AA by blastocysts. Endometrial slices produced 1PGFM and 1PGE2 but only in small amounts, and they were capable of producing nonpolar, probably esterified, forms of 1AA. It was concluded that porcine blastocysts produced and metabolized prostaglandins and that they make a contribution to the uterine milieu during early pregnancy. |
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