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Effect of microgravity on mammillary cones of Japanese Quail eggs

A study was conducted in cooperation with NASA, National Aeronautics and Space Administration, to determine if normal incubation of fertile eggs of Japanese quail could occur in microgravity. Our role was to examine the eggshells using scanning electron microscopy (SEM) to establish whether normal calcium loss had occurred. Based on a previous baseline study with Japanese quail, we predicted that the earliest calcium loss from mammillary cones of shells would be at 12 d of incubation. There were 5 experimental groups in the NASA study. The eggs of all groups were incubated at 37 to 37.5° C and turned hourly. Eggs in groups 1 and 2 were incubated in space in an Avian Development Facility (ADF) unit aboard the Orbiter Endeavour flight STS-108 for 12 days. Group 1 was in 0 g; group 2 was spun in a centrifuge at 1 g. Groups 3 and 4 were incubated in the ground control laboratory in an ADF unit. Group 3 was stationary at 1 g, while group 4 was spun vertically in a centrifuge, resulting in variable force from 0 to 2 g. Group 5 was incubated on Earth in a standard laboratory incubator. The incubation was interrupted at days 4, 7, and 12 in each of the test groups. The day 4 and 7 shells were injected with 4% paraformaldehyde in 50 Mm Cacodylate buffer, pH 7.4 to terminate incubation. Incubation of day 12 eggs was terminated by opening the shells; no preservative was used. Eggshell samples from each group were treated with 6% sodium hypochlorite for removal of shell membranes. The SEM examination revealed that calcium loss had occurred in some mammillary cones from each group, regardless of treatment or length of incubation. Fixative most likely played a role in the dissolution of Ca from the mammillary cones of shells from embryos incubated for 4 and 7 d. An ash analysis of the Ca content of the day 12 shells obtained from live embryos showed no significant difference among treatment groups. These results indicate that embryos incubated in microgravity for 12 d utilized Ca from the shell during development.