Metabolic assessment of wallaby blastocysts during embryonic diapause and subsequent reactivation

Abstract

The metabolism of tammar wallaby (Macropus eugenii) blastocysts was analysed by means of quantitative fluorescence microscopy during embryonic diapause and 2, 3, 4, 5, 8 and 10 days after reactivation to determine nutrient preferences during metabolic reactivation of the blastocyst. The surface area of quiescent blastocysts was 0.16 +/- 0.02 mm2 (mean +/- s.e.m.), and increased to 0.44 +/- 0.04 mm2 (P < 0.05) by Day 8 after removal of the sucking stimulus of the pouch young (RPY). Day-10 blastocysts, analysed over two successive breeding seasons, were significantly different in size from each other (Group A, 1992: 4.44 +/- 1.47 mm2; Group B, 1993: 18.87 +/- 4.62 mm2; P < 0.01), and both groups were significantly different in size from diapausing blastocysts (P < 0.01). There was no significant difference in carbohydrate uptake or production by blastocysts during the first five days after RPY. Glucose uptake by blastocysts recovered 8 days after RPY (61.9 +/- 30.0 pmol embryo-1 h-1) was significantly greater than that by Day-0 blastocysts (17.9 +/- 5.5 pmol embryo-1 h-1) and glucose uptake by both groups of Day-10 blastocysts (Group A, 174.0 +/- 28.4 pmol embryo-1 h-1; Group B, 616.0 +/- 239.0 pmol embryo-1 h-1) was significantly different from that by Day-0 blastocysts (P < 0.01). Pyruvate uptake by Day-10 blastocysts (Group A, 46.0 +/- 32.2 pmol embryo-1 h-1; Group B, 250.0 +/- 136.0 pmol embryo-1 h-1; P < 0.01) increased significantly compared with that by Day-0 blastocysts (6.4 +/- 1.6 pmol embryo-1 h-1; P < 0.01). Lactate production by Day-10 blastocysts (Group A, 186.7 +/- 30.3 pmol embryo-1 h-1; Group B, 285 +/- 129 pmol embryo-1 h-1; P > 0.01) was also significantly different from that by quiescent blastocysts (41.20 +/- 9.6 pmol embryo-1 h-1). There was a linear relationship between surface area and glucose uptake and surface area and pyruvate uptake (r2 = 0.965 and r2 = 0.971 respectively). Despite increases in carbohydrate uptake, there was a proportional decrease in lactate production indicating an increase in oxidative metabolism during reactivation. This suggests that there may be a metabolic switch at, or around, Day 5 after RPY.