Effects of gravity perturbation on developing animal systems

Developing systems provide unique opportunities for analyzing the effects of microgravity on animals. Several unusual types of cells as well as various extraordinary cellular behavior patterns characterize the embryos of most animals. Those features have been exploited as test systems for space flight. The data from previous experiments are reviewed, and considerations for the design of future experiments are presented.     

The influence of gravity on the process of development of animal systems

The development of animal systems is described in terms of a series of overlapping phases: pattern specification; differentiation; growth; and aging. The extent to which altered (micro) gravity (g) affects those phases is briefly reviewed for several animal systems. As a model, amphibian egg/early embryo is described. Recent data derived from clinostat protocols indicates that microgravity simulation alters early pattern specification (dorsal/ventral polarity) but does not adversely influence subsequent morphogenesis. Possible explanations for the absence of catastrophic microgravity effects on amphibian embryogenesis are discussed.     

Early development of Xenopus embryos is affected by simulated gravity.

Early amphibian (Xenopus laevis) development under clinostat-simulated weightlessness and centrifuge-simulated hypergravity was studied. The results revealed significant effects on (i) "morphological patterning" such as the cleavage furrow pattern in the vegetal hemisphere at the eight-cell stage and the shape of the dorsal lip in early gastrulae and (ii) "the timing of embryonic events" such as the third cleavage furrow completion and the dorsal lip appearance. Substantial variations in sensitivity to simulated force fields were observed, which should be considered in interpreting spaceflight data.     

Subcellular components of the amphibian egg: insights provided by gravitational studies.

Most cytoplasmic regions of fertilized amphibian eggs move with respect to the gravity vector in experimentally gravity oriented eggs. The pattern and extent of this movement varies among different batches of eggs. This variation in apparent cytoplasmic viscosity (or, conversely, cytoplasmic mobility) can be correlated with variations in subsequent morphogenesis of experimental, gravitationally manipulated eggs. Therefore, the proper interpretation of gravity experiments with amphibian eggs requires that one understand the subcellular basis for this variation on cytoplasmic mobility. Variation in the packing of the major cytoplasmic organelle, the yolk platelets, or the organization and amount of cytoskeletal components may explain the variation in cytoplasmic mobility. Evidence is presented that the variation in yolk volume density (fraction of total cytoplasmic volume occupied by yolk platelets) does not account for the variation in cytoplasmic mobility in Xenopus laevis eggs. Experimental evidence from cold-shocked inverted eggs indicates that microtubules may be involved in determining cytoplasmic mobility. However, quantitative evidence that the microtubule levels and state of the microtubules (polymerized vs. non-polymerized) in the whole Xenopus laevis egg does not correlate directly with cytoplasmic mobility is presented. The apparent conflict these data represent regarding the role of the cytoskeleton in determining cytoplasmic mobility is discussed.     

The amphibian egg as a model system for analyzing gravity effects.

Amphibian eggs provide several advantageous features as a model system for analyzing the effects of gravity on single cells. Those features include large size, readily tracked intracellular inclusions, and ease of experimental manipulation. Employing novel gravity orientation as a tool, a substantial data base is being developed. That information is being used to construct a 3-D model of the frog (Xenopus laevis) egg. Internal cytoplasmic organization (rather than surface features) are being emphasized. Several cytoplasmic compartments (domains) have been elucidated, and their behavior in inverted eggs monitored. They have been incorporated into the model, and serve as a point of departure for further inquiry and speculation.     

Understanding the organization of the amphibian egg cytoplasm: gravitational force as a probe.

A combination of hypergravity (centrifugation) and hypogravity (clinostat) studies have been carried out on amphibian (frog, Xenopus) eggs. The results reveal that the twinning caused by centrifugation exhibits substantial spawning to spawning variation. That variation can be attributed to the apparent viscosity of the egg's internal cytoplasm. Simulated hypogravity results in a relocation of the egg's third (horizontal) cleavage furrow, towards the equator. Substantial egg-to-egg variation is also observed in this "cleavage effect". For interpreting spaceflight data and for using G-forces as probes for understanding the egg's architecture the egg variation documented herein should be considered.     

Amphibian egg cytoplasm response to altered g-forces and gravity orientation

Elucidation of dorsal/ventral polarity and primary embryonic axis development in amphibian embryos requires an understanding of cytoplasmic rearrangements in fertile eggs at the biophysical, physiological, and biochemical levels. Evidence is presented that amphibian egg cytoplasmic components are compartmentalized. The effects of altered orientation to the gravitational vector (i.e., egg inversion) and alterations in gravity force ranging from hypergravity (centrifugation) to simulated microgravity (i.e., horizontal clinostat rotation) on cytoplasmic compartment rearrangements are reviewed. The behavior of yolk compartments as well as a newly defined (with monoclonal antibody) non-yolk cytoplasmic compartment, in inverted eggs and in eggs rotated on horizontal clinostats at their buoyant density, is discussed.