ABSTRACT
Post-diapause gemmules of the freshwater sponge Eunapius fragilis remained quiescent when maintained at 5°C. Germination occurred within 48 to 72 h following warming to 20°-23°C, culminating with the emergence of a new sponge from the collagenous capsule. Both heat dissipation and oxygen consumption climbed steadily during germination and eventually reached 600% of the starting values. By comparison, energy flow was much lower over the same period of time in diapausing gemmules, clearly demonstrating metabolic depression during diapause. The calorimetric:respirometric (CR) ratio increased significantly from -354 kJ/mol O2 to -541 kJ/mol O2 between hours 3.5 and 56.5 of germination, with an average value across this period of about -495 kJ/mol O2. The low CR ratio at hour 12.5 (-374 +/- 21; +/- 1 SE, n = 3) was statistically below the oxycaloric equivalent, which suggests that gemmules may have experienced hypoxia during the more than 3 months of storage at 5°C prior to experiments. The increase in metabolism during germination could be blocked by perfusing the gemmules with nitrogen-saturated medium (nominally oxygen free). Developing gemmules were able to survive oxygen limitation for several hours at least; during that time energy flow was depressed to 6% of normoxic values. During germination, the range of values was 3.5 to 4.0 nmol/mg protein for ATP, 0.2 to 0.4 nmol/mg protein for ADP, and 0.5 to 0.8 nmol/mg protein for AMP. Because ATP was high even before gemmules were warmed to room temperature, it is unlikely that levels were severely compromised during the diapause condition.
ABSTRACT
Post-diapausing gemmules of the freshwater sponge Eunapius fragilis were found to contain sorbitol and glycogen as their primary carbohydrates. The sorbitol probably acts to increase the tolerance of the gemmules to freezing and desiccation. During germination, average sorbitol levels--measured as micromoles of sorbitol per gram of fresh weight of gemmule tissue (µmol/gfw)--declined from a control value of 36 µmol/gfw to about 4 µmol/gfw. Concomitantly, average glycogen levels increased from a control value of 29 µmol/gfw to a steady-state level of 62 µmol/gfw. It is probable that glycogen is being synthesized at the expense of sorbitol. The breakdown of sorbitol was associated with an increase in the activity of sorbitol dehydrogenase from undetectable levels in dormant gemmules to a maximum of 0.2 µmol/ min · mg protein after 30 h of exposure to 20°C. Aldose reductase activity remained constant throughout germination. These data support the hypothesis that the decrease in sorbitol levels is the result of an increase in the rate of catabolism by sorbitol dehydrogenase. The total activity of glycogen synthase did not change during germination; however, the activity of glucose-6-phosphate-dependent glycogen synthase was about 18 times greater than the activity of glucose-6-phosphate-independent glycogen synthase. Total glycogen phosphorylase activity increased from about 1.6 nmol/min.mg protein to 3.6 nmol/min.mg protein during germination. At the same time, however, the percentage of glycogen phosphorylase a decreased from almost 100% to about 84%. This decrease would attenuate the apparent increase in activity. cAMP levels remained constant throughout germination. The observed changes in the level of glycogen in the gemmules are not simply due to changes in the activity of either glycogen phosphorylase or glycogen synthase.
ABSTRACT
Explanted oviducts from 13- and 16-day embryonic female chicks responded to estradiol (1 to 4 micrograms/ml) in the culture medium by forming a thickened epithelium folded into crypts and containing goblet cells and by development of the stroma. In medium containing 3 micrograms/ml estradiol and 0.125 micrograms/ml hydrocortisone, 13-day embryonic oviducts exhibited further development: The columnar epithelial cells formed cilia and tubular glands were formed.
