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At least 55 records · Page 3

Amphibian fertilization and development in microgravity

During the year before launch, female frogs will be tested every 3 months for the quantity and quality of eggs produced. Two weeks or more prior to launch, male and female frogs will be transported to the John F. Kennedy Space Center (KSC). During the few weeks before launch, groups will be periodically tested for egg quality to assure that the frogs have adapted to the KSC laboratory environment. About 27 hours before launch, four females will be placed in a damp foam-lined box, called the Adult Frog Box (AFB), through which 100 cc/min of air wil be circulated. The AFB will be lowered into the Spacelab and loaded into the Frog Environmental Unit (FEU) during the final pre-launch preparations. A sperm suspension, for use in flight to fertilize the eggs, will also be prepared and loaded during the pre-launch period. The sperm suspension, together with a kit of syringes containing Human Chorionic Gonadotropin (HCG), will be stored in a refrigerator aboard the shuttle until needed in flight. On the first day of flight, the AFB will be transferred from the FEU to the General Purpose Work Station (GPWS), which is a type of glove box specially designed to allow the crew to use chemicals and biological materials during the flight without contaminating the shuttle/Spacelab environment. Inside the GPWS the four adult frogs will be injected with the HCG hormone and returned to the FEU. Approximately 16 hours after injection, ovulation should have taken place and 15 to 20 eggs from each frog will be placed on each of two egg baskets and covered with sperm for 10 minutes. The egg baskets are inserted into acrylic egg chambers and 50 ml of 'pond water' (20 percent strength Modified Ringers solution (is added. One of the chambers from each frog will be placed on a centrifuge within the FEU and rotated to simulate normal terrestrial gravity (1 g). The remaining chambers are incubated under microgravity conditions within the FEU. Forty minutes after fertilization, the four chambers exposed to 1 g will be removed and observed within the GPWS using a dissecting microscope and camera system. On the basis of egg rotation and egg appearance, the 'best' and 'second best' frogs will be selected to contribute additional eggs. Using the two best frogs, 22 egg chambers will be loaded with eggs and fertilized. Eleven chambers wil be incubated at microgravity and eleven will be incubated on the centrifuge. At various times during the flight, chambers will be removed from the FEU and transferred to the GPWS where a formaldehyde-based fixative will be injected in order to preserve important developmental stages for in depth study following the flight. Five of the 22 chambers plus the eight fertilization test chambers will be returned to Earth with live tadpoles. The swimming behavior of these free swimming tadpoles will be examined within several hours of landing and some will be fixed for a detailed analysis of their inner ear, the otolity, the animals 'balance system.' Lychakov and Vinikov reported an increase in otolith size in tadpoles that developed (but were not fertilized) in space. Live tadpoles from the SL-J flight will be raised through metamorphsis for studies of maturation including an analysis of the effects of space flight on their ability to produce normal progency. Following the flight, the fixed embryos will be serially sectioned and stained using procedures that were developed for this experiment. The embroys fixed at the 2-4 cell stage will be examined for the distribution of cytoplasmic contents, including the various classes of yolk platelets, and the location and shape of the cleavage furrows. The gastrulae will be assessed for the normality of the complex cellular rearrangements that constitute blastopore formation and involution, i.e., the differentiation of the embryo into its specialized body parts. Using a new procedure, the gastrulae will be bleached which will enable the initial sperm entry point (SEP) to be located. The correlation between the SEP and the dorsal lip of the blastopore will be determined. Under normal terrestrial conditions it was shown that the SEP typically is located on the side of the egg opposite the future dorsal side of the embryo. The neurulae will be examined for the normality and completeness of the neural plate and archenteron expansion. The tadpole stages will be used to study the allometry and morphology of the various organ systems.

Souza, Kenneth A.↗

Development of Gravity-Sensing Organs in Altered Gravity Conditions: Opposite Conclusions From an Amphibian and a Molluscan Preparation

Several components of the systems animals use to orient to gravity might develop differently in micrograms. If the growth of the "test masses" on which gravity acts (otoliths, in vertebrates, statoliths or statoconia in most invertebrates) is controlled on the basis of their weight, larger otoliths (or their analogs) would be expected to develop in micrograms. The vestibular systems in animals reared in altered gravity have been studied in several species, with varied results being reported. Early Russian reports of Xenopus larvae reared in space indicated no qualitative differences in the vestibular organs, compared to ground-reared controls. A similar lack of differences in Xenopus were reported. The ultricular otolith was 30% larger in space-reared Xenopus. No differences in saccular otolith volume between centrifuged and control adult rats were found. A delay in otoconial development in chick embryos reared at 2 grams on a centrifuge was reported but in a later report, no differences in otolith weight between 2 grams and control chicks were found. Increased optokinetic responses in flight-reared Xenopus tadpoles, suggesting that the animals reared in the absence of gravity made greater relative use of their visual system, rather than the vestibular system, in orienting to a moving stimulus was reported. To test early Japanese newt, CYnops pyrrhogaster, were maintained in orbit for 15 days on the IML-2 mission in 1994. All specimens reached orbit before any otoconia were formed and all major components of the inner ear were formed by the end of the flight. In ground-based studies of he Aplysia statocyst, the volume of the statolith in embryos and the number statoconia in post-metamorphic animals were compared between 1-gram controls and specimens reared at 2 to 5.7 grams.

Wiederhold, Michael L.↗

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↗

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↗

Winter Severity Affects Occupancy of Spring- and Summer-Breeding Anurans Across the Eastern United States

Climate change is an increasingly important driver of biodiversity loss. The ectothermic nature of amphibians may make them particularly sensitive to changes in temperature and precipitation regimes, adding to declines from other threats. While active season environmental conditions can influence growth and survival, effects of variation in winter conditions on population dynamics are less well-studied. Given that extreme winter temperatures can influence amphibian survival and fitness, we expected that increased winter severity—as measured by variability in winter temperatures and snow cover—would be associated with decreased occupancy, and that populations that experience more severe winters would have the largest sensitivities and show the greatest declines.

climate↗

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.↗

Calbindin and parvalbumin are early markers of non-mitotically regenerating hair cells in the bullfrog vestibular otolith organs

Earlier studies have demonstrated hair cell regeneration in the absence of cell proliferation, and suggested that supporting cells could phenotypically convert into hair cells following hair cell loss. Because calcium-binding proteins are involved in gene up-regulation, cell growth, and cell differentiation, we wished to determine if these proteins were up-regulated in scar formations and regenerating hair cells following gentamicin treatment. Calbindin and parvalbumin immunolabeling was examined in control or gentamicin-treated (GT) bullfrog saccular and utricular explants cultured for 3 days in amphibian culture medium or amphibian culture medium supplemented with aphidicolin, a blocker of nuclear DNA replication in eukaryotic cells. In control cultures, calbindin and parvalbumin immunolabeled the hair bundles and, less intensely, the cell bodies of mature hair cells. In GT or mitotically-blocked GT (MBGT) cultures, calbindin and parvalbumin immunolabeling was also seen in the hair bundles, cuticular plates, and cell bodies of hair cells with immature hair bundles. Thus, these antigens were useful markers for both normal and regenerating hair cells. Supporting cell immunolabeling was not seen in control cultures nor in the majority of supporting cells in GT cultures. In MBGT cultures, calbindin and parvalbumin immunolabeling was up-regulated in the cytosol of single supporting cells participating in scar formations and in supporting cells with hair cell-like characteristics. These data provide further evidence that non-mitotic hair cell regeneration in cultures can be accomplished by the conversion of supporting cells into hair cells.

Non-NASA Center↗

Orientation: Sensory basis; Proceedings of the Conference, New York, N.Y., February 8-10, 1971.

Topics related to photoreceptors are considered, giving attention to visual pattern recognition and directional orientation in insects, the sensory basis of orientation in amphibians, and the aerial and underwater visual acuity in the California sea lion as a function of luminance. Other subjects explored are in the fields of phonoreceptors, chemoreception, vestibular receptors, and electrical and magnetic sensitivity. Questions of the development and evolution of orientation are also investigated, taking into account field studies of mass emigration and orientation in the spiny lobster and investigations concerning the jumping behavior in the Gobiid fish. Individual items are announced in this issue.

Source record↗

Gravity and the cell: Intracellular structures and Stokes sedimentation

Plant and certain animal embryos appear to be responsive to the gravity vector during early stages of development. The convection of particle sedimentation as the basis for the sensing of gravity is investigated using the cells of wheat seedlings, amphibian embryos, and mammals. Exploration of the mammalian cell for sedimenting particles reveals that their existence is unlikely, especially in the presence of a network of microtubules and microfilaments considered to be responsible for intracellular organization. Destruction of these structures renders the cell susceptible to accelerations several times g. Large dense particles, such as chromosomes, nucleoli, and cytoplasmic organelles are acted upon by forces much larger than that due to gravity, and their positions in the cell appear to be insensitive to gravity.

Todd, P.↗

Until the sun dies

This book gives a popular account of the forces that have shaped human beings into their present form and created the power of human intelligence, and considers the prospects for intelligent life on other planets in the solar system and elsewhere in the universe. The chain of events leading from the big bang to the origin of life on earth is reviewed together with the observations that established the expansion of the universe. Philosophical difficulties with the concept of a universe that has both a beginning and an end are pondered, steady-state cosmology is briefly explained, and the discovery of the relict microwave background is discussed. The formation of the solar system is recounted along with the scientific view of the origin of terrestrial life. Attention is given to the origin of cells and the evolution of oxygen-breathing life, multicelled creatures, armored animals, fishes, amphibians, early reptiles, dinosaurs, and mammals. The development of mammalian intelligence is traced from the early tree dwellers through monkeys, apes, ape men, humanoid tool makers, and primitive members of the genus Homo, to Homo sapiens. Possible evidence for the existence of life on Mars is evaluated together with prospects for communication or other contact with extraterrestrial intelligence.

Jastrow, R.↗

The Development of the Vestibular Apparatus Under Conditions of Weightlessness

A series of experiments has been carried out on the effect of space flight conditions on morphogenesis and the structure of the vestibular apparatus in amphibian and fish larvae. Larval development proceeded in weightlessness without serious morphological defects. The vestibular apparatus developed; its organization in the experimental animals did not differ qualitatively from that in the controls. The specific external stimulus (gravitation) appears not to be a necessary condition for the development of a gravitation receptor in ontogenesis although the appearance of the vestibular apparatus in phylogenesis was apparently related to this stimulus.

Vinnikov, Y. A.↗

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 Workshop on Animal Gravity-Sensing Systems

The opportunity for space flight has brought about the need for well-planned research programs that recognize the significance of space flight as a scientific research tool for advancing knowledge of life on Earth, and that utilize each flight opportunity to its fullest. For the first time in history, gravity can be almost completely eliminated. Thus, studies can be undertaken that will help to elucidate the importance of gravity to the normal functioning of living organisms, and to determine the effects microgravity may have on an organism. This workshop was convened to organize a plan for space research on animal gravity-sensing systems and the role that gravity plays in the development and normal functioning of these systems. Scientists working in the field of animal gravity-sensing systems use a wide variety of organisms in their research. The workshop presentations dealt with topics which ranged from the indirect gravity receptor of the water flea, Daphnia (whose antennal setae apparently act as current-sensing receptors as the animal moves up and down in water), through specialized statocyst structures found in jellyfish and gastropods, to the more complex vestibular systems that are characteristic of amphibians, avians, and mammals.

Corcoran, M. L.↗

Limitations on K-T mass extinction theories based upon the vertebrate record

Theories of extinction are only as good as the patterns of extinction that they purport to explain. Often such patterns are ignored. For the terminal Cretaceous events, different groups of organisms in different environments show different patterns of extinction that to date cannot be explained by a single causal mechanism. Several patterns of extinction (and/or preservational bias) can be observed for the various groups of vertebrates from the uppermost Cretaceous Hell Creek Formation and lower Paleocene Tullock Formation in eastern Montana. The taxonomic level at which the percentage of survivals (or extinctions) is calculated will have an effect upon the perception of faunal turnover. In addition to the better known mammals and better publicized dinosaurs, there are almost 60 additional species of reptiles, birds, amphibians, and fish in the HELL Creek Formation. Simple arithmetic suggests only 33 percent survival of these vertebrates from the Hell Creek Fm. into the Tullock Fm. A more critical examination of the data shows that almost all Hell Creek species not found in the Tullock are represented in one of the following categories; extremely rare forms, elasmobranch fish that underwent rapid speciation taxa that although not known or rare in the Tullock, are found elsewhere. Each of the categories is largely the result of the following biases: taphonomy, ecological differences, taxonomic artifact paleogeography. The two most important factors appear to be the possible taphonomic biases and the taxonomic artifacts. The extinction patterns among the vertebrates do not appear to be attributable to any single cause, catastrophic or otherwise.

Archibald, J. David↗

Microgravity Experiments On Animals

Paper describes experiments on animal subjects planned for Spacelab Life Sciences 1 mission. Laboratory equipment evaluated, and physiological experiments performed. Represents first step in establishing technology for maintaining and manipulating rodents, nonhuman primates, amphibians, and plants during space flight without jeopardizing crew's environment. In addition, experiments focus on effects of microgravity on cardiopulmonary, cardiovascular, and musculoskeletal systems; on regulation of volume of blood and production of red blood cells; and on calcium metabolism and gravity receptors.

Dalton, B. P.↗

Gravity effects on reproduction, development, and aging

The effects of various levels of gravity force (obtained by rotation in clinostats or by centrifugation) and the near-weightlessness condition aboard orbiting spacecraft on the fertilization, embryonic development, maturation, and aging of animals are examined. Results obtained from the American and Soviet spaceborne biology experiments are presented including those on mammals, amphibians, fish, birds, invertebrates, and protozoa. Theoretical issues related to the effect of gravity on various physiological systems are discused together with the future research goals concerning human life in space. It is noted that life in space (after adaptation to near-weightlessness) might be significantly prolonged due to a reduction in metabolic rate and a concomitant decrease in oxygen radical reactions.

Miquel, Jaime↗

Studying the effects of microgravity on lower vertebrate development and behavior

Lower vertebrates have been used in space research for at least 3 decades, and have a number of advantages such as the ability to be maintained safely in space conditions, high development rates, easy observability, and small size. Several major investigations with lower vertebrates are in the flight queue in various countries, and some of this research with amphibians that has Canadian Space Agency sponsorship is reviewed. In connection with NASA's frog embryology experiment, Canadian scientists will conduct postflight experiments on live tadpoles brought back from space to determine whether larvae that develop from these embryos are behaviorally normal. Swimming kinematics in particular will be examined, since a distinctive looping behavior of Xenopus tadpoles under microgravity has been noted. A collaborative study with the Institute of Biomedical Problems in Moscow is designed to elucidate the relationship between buoyancy regulation and lung development in tadpoles, and is scheduled to fly on a Biocosmos satellite in 1992.

Wassersug, Richard J.↗