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Neff, A. W.

Publications and source records attributed to Neff, A. W..

14 records
All sourcesNASA NTRSDOE OSTI
NASA NTRS1996

Optomotor behaviour in Xenopus laevis tadpoles as a measure of the effect of gravity on visual and vestibular neural integration

The ability of aquatic vertebrates to maintain their position requires integration of visual and vestibular sensory information. To understand better how aquatic animals integrate such information, we measured the optomotor behaviour of Xenopus laevis tadpoles raised in growth chambers in microgravity (< 10(-3)g), normal gravity (1 g), hypergravity (3 g) and on a slowly rotating clinostat (simulated microgravity). The goal of this research was to determine how development in an altered gravitational force field affects the visual- and vestibular-dependent behaviour of tadpoles. This research represents the first time that the optomotor behaviour of an organism raised from fertilization in microgravity has been tested. Significant differences were observed in the optomotor behaviour among the four gravity treatments. When first exposed to normal gravity, the microgravity-raised tadpoles exhibited the strongest (or most positive) optomotor behaviour, while the 3 g centrifuge tadpoles showed no optomotor response. Some abnormal behaviours (such as erratic swimming, lying motionless and abnormal swimming posture) were observed in the tadpoles raised in altered gravity on the initial day of testing. One day later, the tadpoles raised in hypergravity did not differ significantly in their optomotor behaviour from control tadpoles raised in normal gravity. However, tadpoles raised in microgravity still displayed an exaggerated optomotor response. One week after the tadpoles had been introduced to normal gravity, there was no longer a significant difference in optomotor behaviour among the different gravity treatments. This convergence of optomotor behaviour by tadpoles from the different treatment reflects the acclimation of their vestibular systems to normal gravity.

NASA Discipline Developmental Biology↗
NASA NTRS1989

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 three-dimensional 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.

Malacinski, G. M.↗
NASA NTRS1989

Subcellular components of the amphibian egg - Insights provided by gravitational studies

The variability in the response of Xenopus laevis eggs to a given force environment is studied. The roles of cytoplasmic organelle, the yolk platelets, and cytoskeletal components in varying in cytoplasmic mobility are examined. The data reveal that the packing of yolk platelets is not a major factor in causing cytoplasmic mobility differences and microtubules may affect cytoplasmic mobility.

Neff, A. W.↗
NASA NTRS1986

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) nonyolk cytoplasmic compartment, in inverted eggs and in eggs rotated on horizontal clinostats at their buoyant density, is discussed.

Neff, A. W.↗
NASA NTRS1986

Effects of gravity perturbation on developing animal systems

The use of developing animal systems to analyze the effects of microgravity on animals is discussed. Some of the key features of developing systems, especially embryos, are reviewed and relevant space data are summarized. Issues to be addressed in the design of future space experiments are discussed. It is noted that an embryo which exhibits ground based gravity effects should be selected for use as a model system and individual variation in gravity response among batches of embryos should be taken into account.

Malacinski, G. M.↗
NASA NTRS1985

Response of amphibian egg non-yolk cytoplasm to gravity orientation

In order to study amphibian egg cytoplasmic organization and egg symmetrization at the molecular level, a library of seventeen monoclonal antibodies (MoAbs) against Xenopus laevis non-yolk egg proteins was produced. Several of these MoAbs react with non-yolk cytoplasmic antigens which are unevenly distributed in the fertile Xenopus egg.

Smith, R. C.↗
NASA NTRS1985

Simulated Microgravity as a Probe for Understanding the Mechanisms of Early Pattern Specification

Early pattern specification (e.g., axial structure morphogenesis, doreal ventral polarity, etc.) is monitored in amphibian eggs which were subjected to microgravity simulation by constant rotation on a horizontal clinostat. In contrast to previous clinostat experiments, rotation is initiated either prior to fertilization or immediately thereafter. Large proportions of clinostated eggs developed normal axial structures. A model which employs a multiple set of signals for specifying early pattern is discussed. Effects of microgravity simulation on the earliest post fertilization pattern specification event dorsal/ventral polarization is analyzed in detail. Other models are developed and they are discussed. As a general mechanism for explaining the manner in which regional developmental patterns emerge from the initial, radially symmetrial egg, the density compartment model is described. The identification of the various zones or compartments of egg cytoplasm using inverted eggs is explained.

Neff, A. W.↗
NASA NTRS1984

Influence of clinostat rotation on fertilized amphibian egg pattern specification

Pattern specification in fertile Xenopus eggs rotated on horizontal clinostats was monitored with respect to primary embryonic axis formation, subsequent morphogenesis, and compartmentalization of the cytoplasm. At the speeds of 1 to 24 rpm (which are believed to simulate microgravity) a large percentage of eggs developed normal axial structures. Eggs clinostated at 12 rpm showed a randomization of dorsal/ventral polarity. The cytoplasmic compartments showed some clinostat effects but no abnormal mixing, disruption or dislocation of compartments. It is predicted that Xenopus eggs fertilized and allowed to develop in space will retain normal cytoplasmic density compartments, establish primary axes and undergo normal morphogenesis in space. Their dorsal/ventral polarity may not, however, be determined by the sperm entrance site (as is the case for 1 g eggs).

Neff, A. W.↗
NASA NTRS1984

Topology of the germ plasm and development of primordial germ cells in inverted amphibian eggs

Inverted Xenopus eggs have reduced numbers of primordial germ cells (PGCs). The extent of the reduction varies from spawning to spawning. Histologic examination revealed that PGC counts were lowest in inverted eggs which displayed the greatest amount of shift in the vegetal mass of large yolk platelets, although the germ plasm itself always remained localized in the egg's original vegetal hemisphere. Even at blastulation the germ plasm continued to be localized in the egg's original vegetal hemisphere. In many cases, however, it was confined to the periphery of the embryo, which probably accounts for the reduced PGC number in some tadpoles. In other cases it may have been dispersed and therefore not detectable in histologic analyses. Although the altered site of involution in inverted embryos did not influence PGC development, subsequent cell movement patterns apparently did. Those embryos which displayed the largest degree of pattern reversal at the tail-bud stage also exhibited the most extreme reduction in PGC numbers. A brief cold shock (4 degrees C, 10 min) prior to first cleavage leads to a further reduction in PGC numbers in inverted embryos, probably as a result of the displacement of the germ plasm away from its original vegetal pole location.

Cell Polarity/physiology↗
NASA NTRS1984

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.

Malacinski, G. M.↗
NASA NTRS1984

Delayed fertilization of anuran amphibian (Xenopus) eggs leads to reduced numbers of primordial germ cells

Several media were tested for the extent to which they promoted high fertilization efficiencies in ovulated, stripped Xenopus eggs. One medium was selected for maintaining eggs in a 'delayed fertilization' (DelF) condition. DelF eggs displayed several unusual characteristics, including shift of the center of gravity, prominent sperm entrance site, and occasional polyspermy. The frequency of normal pattern formation varied according to the length of time eggs were maintained in the DelF condition. Various developmental abnormalities were observed during gastrulation, neurulation, and organogenesis. Most abnormalities appeared, however, to be related to morphogenesis of the endoderm. Primordial germ cell (PGC) development was examined in DelF eggs which displayed normal external morphological features at the swimming tadpole stage. PGC counts were usually normal in short-duration (eg, 5 hr) DelF eggs, but frequently substantially reduced or completely diminished in longer-duration (eg, 25h) tadpoles. Six spawnings were compared and shown to exhibit considerable variability in fertility, morphogenesis, and PGC development. Yolk platelet shifts and developmental parameters were examined in two additional spawnings. The subcortical cytoplasm in which the germ plasm is normally localized appeared to be disrupted in longer duration DelF eggs. That observation may account for low PGC counts in DelF tadpoles.

NASA Discipline Number 40-20↗
NASA NTRS1983

Response of amphibian egg cytoplasm to novel gravity orientation and centrifugation

The effects of inversion and centrifugation on the compartmentalization of cytoplasm in Xenopus laevis eggs are investigated experimentally. The rearrangement of yolk-platelet compartments (YPC) characterized by morphology, density, and viscosity differences is studied in fertilized, unfertilized, and unfertilized electrically activated eggs in normal, and inverted positions and with and without centrifugation at 10-183 x g for 5 min. The eggs are fixed and embedded in plastic or paraffin prior to sagittal sectioning (0.5, 4, or 8 microns) and microscopic examination; the results are presented in a diagram and discussed. A density-compartment model combining both animal/vegetal and dorsal/ventral polarities is proposed: YPC determined without gravity orientation during oogenesis respond to both sperm entrance point and gravity after fertilization, and the response involves breaking of the radial symmetry of the egg. It is predicted that Xenopus eggs in a microgravity environment will encounter difficulties in establishing a primary embryonic axis.

Neff, A. W.↗
NASA NTRS1982

The intracellular responses of frog eggs to novel orientations to gravity

It is found that multiple short doses of ultraviolet light are as effective as a single large dose in producing neural defects. In addition, 180 deg rotation (inversion) of irradiated eggs reduces the ultraviolet effect. Since yolk platelets may be the gravity sensing mechanism, their size, density, and distribution in normal and inverted eggs are investigated. Large platelets are denser and for the most part are in a distinct zone in the vegetal hemisphere, whereas small platelets are less dense and occur in the animal hemisphere. When inverted, the large platelets flow into the animal hemisphere as a coherent mass and partially displace the small platelets. Inversion is thought to rearrange cytoplasmic components necessary for later neural development into an appropriate configuration.

Radice, G. P.↗