Search PubMed⌕ Search

Biomedical subjects

A W Neff

Publications and source records attributed to A W Neff.

At least 37 records · Page 2Linked to original sources
PubMed1989

Amphibian (urodele) myotomes display transitory anterior/posterior and medial/lateral differentiation patterns.

Myotome differentiation during Mexican axolotl (Ambystoma mexicanum) somitogenesis was analyzed by employing anti-actin and anti-myosin monoclonal antibodies as molecular probes. Myotome differentiation occurs after segmentation and proceeds in the cranial-to-caudal direction along the somite file. Within individual somites myotome differentiation displays distinct polarities. Examination of the somite file at the tailbud stage revealed that soon after segmentation, actin/myosin accumulate predominantly in the anterior and medial region of the myotome initially. Subsequently, cells within the myotome differentiate in an anterior-to-posterior and medial-to-lateral direction. Experimental analysis of presomitic paraxial mesoderm grafts before segmentation revealed that this transient myotome polarity is autonomous. Comparative analyses indicate that this myotome differentiation pattern is urodele specific. Cynops pyrrhogaster undergoes myotome differentiation like the axolotl, while two anurans, Xenopus laevis and Bombina orientalis, do not.

Actins↗
PubMed1989

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.

Animals↗
PubMed1989

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.

Animals↗
PubMed1986

Organisation of Xenopus egg cytoplasm: response to simulated microgravity.

The cytoplasm of Xenopus fertilised eggs appears to be organised into three major compartments based primarily on the uneven distribution of yolk platelets. There is a shift of these yolk compartments during the first cell cycle that is thought to be involved in the dorsal/ventral morphogenesis of the embryo. The involvement of gravity in Xenopus cytoplasmic organisation and in compartment shifts was addressed by examining, cytologically, the yolk compartments in embryos that developed under the simulated microgravity conditions of the horizontal clinostat. The cytoplasmic organisation into yolk compartments was found to be maintained, and the asymmetric movements of compartments still occurred in eggs that developed on the clinostat. It is suggested that the organisation of Xenopus egg cytoplasm into discrete compartments relies on forces other than those involving gravity (i.e., not density differences), and that the compartment shifts that take place during the first cell cycle are active movements. The variation in compartment size and composition observed from batch to batch of eggs, and to a lesser extent from egg to egg, during this study was addressed.

Animals↗
PubMed1986

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.

Animals↗
PubMed1986

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.

Animals↗
PubMed1985

Use of the fluorochrome propidium iodide for the identification of Xenopus germ plasm during immunofluorescence studies.

Addition of 0.5 microgram/ml of the red fluorescent dye propidium iodide to the penultimate wash of Xenopus early embryo serial sections during immunofluorescence studies causes germ plasm to fluoresce brightly. Germ plasm can be accurately and speedily identified in the serial sections. The use of this dye is a marked improvement over previous methods of germ plasm identification in unstained sections. Studies involving screening for germ-plasm-specific antibodies are greatly facilitated by staining germ plasm red with propidium iodide and searching for green colabelling by fluoresceinated antibodies.

Animals↗
PubMed1985

Microgravity simulation as a probe for understanding early Xenopus pattern specification.

Pattern specification in early amphibians (Xenopus) was monitored in embryos subjected to gravity compensation (microgravity simulation) by constant low-speed rotation on a horizontal axis (clinostat). The useful range of clinostat speeds was determined empirically. The results were interpreted in terms of a set of models which account for the reorganization of the egg cytoplasm that follows fertilization and that correlates with the establishment of dorsal/ventral polarity. Large percentages of clinostated eggs displayed a positive result (normal axial structure morphogenesis). Consequently, normal development of amphibian eggs in the microgravity environment of space should be possible. Models which depend upon gravity-driven rearrangements for cytoplasmic organization (e.g. dorsal/ventral polarization) of the early embryo should, therefore, not be favoured. At several clinostat speeds symmetrization of the egg in accordance with the site of sperm penetration, a natural phenomenon, was altered. The results at those clinostat speeds indicate that models which employ sperm entrance as an obligatory feature of the cytoplasmic rearrangements that generate egg polarity are not applicable.

Animals↗
PubMed1984

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.

Animals↗
PubMed1984

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.

Animals↗
PubMed1984

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.

Animals↗
PubMed1984

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 1g eggs).

Animals↗
PubMed1984

Experimental analyses of cytoplasmic rearrangements which follow fertilization and accompany symmetrization of inverted Xenopus eggs.

Cytoplasmic rearrangements which follow fertilization were monitored in inverted eggs. A set of yolk compartments was resolved by cytological analyses of both normally oriented and inverted eggs. Those compartments were characterized by their yolk platelet compositions and movement during egg inversion. In addition to the major yolk masses which contain either small, intermediate or large platelets, minor cytoplasmic compartments which line the egg cortex were also identified. During egg inversion the yolk compartments shift. Those yolk mass shifts occurred only after the inverted egg was activated (by sperm, electrical or cold shock). The direction of shift of the major yolk components, rather than the sperm entrance site (as in normal orientation eggs), determines the dorsal/ventral polarity of the inverted egg. Among different spawnings the rate of shift varied. Eggs that displayed the fastest rate of shift exhibited the highest frequency of developmental abnormalities during organogenesis. Isopycnic density gradient analysis of yolk platelets and blastula blastomeres showed that isolated yolk platelets and mid-blastula blastomeres are not of uniform buoyant density. Three major yolk platelet density bands were resolved. Large, intermediate, and small yolk platelets were found in all bands. The high density band had the largest proportion of the large yolk platelets and the low density fraction showed the largest proportion of the small yolk platelets. Interpretation of novel observations on cytoplasmic organization provided criticisms of some earlier models. A new 'Density Compartment Model' was developed and presented as a coherent way to view the organization of the egg cytoplasm and the development of bilateral symmetry.

Animals↗
PubMed1983

Pattern formation in amphibian embryos prevented from undergoing the classical "rotation response" to egg activation.

Fertile Xenopus laevis eggs were immobilized so that they were prevented from undergoing the "rotation response" to activation. Many of those unrotated eggs developed through organogenesis, indicating that egg rotation is not a prerequisite for normal early embryogenesis. Various aspects of the regulation of pattern formation were analyzed in unrotated eggs: It was discovered that a substantial rearrangement of yolk platelets occurred without affecting subsequent pattern formation. The germ plasm, however, remained localized in the vegetal hemisphere in inverted eggs. Cleavage furrows and the site of involution were both often observed in novel locations in inverted eggs which were prevented from rotating during activation.

Animals↗