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At least 19 records

Membrane permeability and the loss of germination factor from Neurospora crassa at low water activities

Neurospora crassa conidia incubating in buffer at low water activities release a germination-essential component as well as 260-nm absorbing and ninhydrin-positive materials, regardless of whether an electrolyte or nonelectrolyte is used to reduce water activity. Chloroform and antibiotics known to increase cell-membrane permeability have a similar effect. This suggests that membrane damage occurs in media of low water activity and that an increase in permeability is responsible for the release of cellular components. The damage caused in media of low water activity is nonlethal in most cases, and the conidia recover when transferred to nutrient medium.

Charlang, G.↗

A circadian rhythm of conidiation in Neurospora crassa (L-12)

Two fungi growth chambers containing six growth tubes each are used in this experiment. One chamber is for the space experiment; the other is for the simultaneous ground control experiment. The hyphae of Neurospora crassa band A mutant are inoculated at one end of each tube. Both the chambers are kept at 3 C plus or minus 1.5 C to stop hyphae growth until the Spacelab is activated. After the activation, each chamber is transferred simultaneously to the Spacelab and a phytotron in KSC and kept in continuous light at the same temperature. After about 24 hours of light exposure, each chamber is inserted into a growth chamber bag to keep it in constant darkness. The circadian rhythm of conidiation is initiated by this light to dark transition. After the dark incubation for 5 days at room temperature, both the growth chambers are kept at 3 C plus or minus 1.5 C to stop growth of the hyphae. After the space shuttle lands, both conidiation patterns are compared and analyzed. It has been known that numerous physiological phenomena show circadian rhythms. They are characterized by the fact that the oscillation can persist under constant conditions of light and temperature. Therefore, it has been accepted by most investigators that the generation mechanism of the circadian rhythm is endogeneous. However, one cannot reject the possibility that these rhythms are caused by some geophysical exogeneous factor having a 24-hour period, such as atmospheric pressure, gravity, or electromagnetic radiation. We use Neurospora crassa band A mutual which shows an obvious circadian rhythm in its spore-forming (conidiation) on the ground, and we intend to attempt the conidation of this mutant in the Spacelab where 24-hour periodicity is severely attenuated and to elucidate the effect of the geophysical exogeneous factor in the generation mechanism of the circadian rhythm.

Miyoshi, Yashuhiro↗

Germination-defective mutant of Neurospora crassa that responds to siderophores

A conditionally germination-defective mutant of Neurospora crassa has been found to be partially curable by ferricrocin and other siderophores. The mutant conidia rapidly lose their membrane-bound siderophores when suspended in buffer or growth media. Germination is consequently delayed unless large numbers of conidia are present (positive population effect). This indicates that the mutant has a membrane defect involving the siderophore attachment site.

Charlang, G.↗

Isolation and identification of the conidial germination factor of Neurospora crassa

The germination-essential substance (germination factor /GF/) that is lost from conidia of Neurospora crassa on exposure to solutions of low water activity has been isolated and identified as a group of iron-transport compounds, or siderochromes. The principal siderochrome of conidia is ferricrocin, a cyclic hexapeptide. A closely related substance, ferrichrome C, is tentatively identified as a minor constituent. The same substances are also present in extracts of mycelium along with small amounts of a third siderochrome, which has not been identified. The GF activity of culture filtrates is due to coprogen, the only siderochrome previously identified with N. crassa.

Horowitz, N. H.↗

Radiation and zero-gravity effects on human leukocytes and Neurospora crassa

The two parts of this experiment have resulted in proof that neither orbital space flight nor any of the stresses connected with it produced significant, unpredicted genetic damage, at least insofar as chromosomal-aberration production is a valid measure of this general type of effect. Furthermore, the Gemini 11 results lead to the conclusion that no synergistic effect exists between radiation and factors that are associated with space flight. No significant difference was found between dose-effect curves for survival or mutation induction of the inflight and ground-based samples irradiated on filters. Thus, like the blood experiment, this part of the experiment failed to result in data that were appropriate for confirmation of the apparent synergism that was observed in the Gemini 3 blood experiment. Also, the Nuerospora crassa experiment provided conclusive data that there is no difference in the genetic effects of irradiation during space flight and the genetic effects obtained in ground-based experiments. Both the Gemini 11 blood experiment and the Neurospora crassa experiment thus have failed to result in data confirmatory of the apparent synergism observed on the Gemini 3 mission.

Bender, M. A.↗

Ionizing radiation-induced mutagenesis: radiation studies in Neurospora predictive for results in mammalian cells

Ionizing radiation was the first mutagen discovered and was used to develop the first mutagenicity assay. In the ensuing 70+ years, ionizing radiation became a fundamental tool in understanding mutagenesis and is still a subject of intensive research. Frederick de Serres et al. developed and used the Neurospora crassa ad-3 system initially to explore the mutagenic effects of ionizing radiation. Using this system, de Serres et al. demonstrated the dependence of the frequency and spectra of mutations induced by ionizing radiation on the dose, dose rate, radiation quality, repair capabilities of the cells, and the target gene employed. This work in Neurospora predicted the subsequent observations of the mutagenic effects of ionizing radiation in mammalian cells. Modeled originally on the mouse specific-locus system developed by William L. Russell, the N. crassa ad-3 system developed by de Serres has itself served as a model for interpreting the results in subsequent systems in mammalian cells. This review describes the primary findings on the nature of ionizing radiation-induced mutagenesis in the N. crassa ad-3 system and the parallel observations made years later in mammalian cells.

NASA Discipline Radiation Health↗

The biological clock of Neurospora in a microgravity environment

The effects of simulated microgravity (via the use of a slowly rotating clinostat), altered orientation (via alterations in the vector of a 1-g force), and hypergravity (via centrifugation) on the circadian rhythm of conidiation in Neurospora crassa were investigated. It was found that the clinostat, while capable of producing some of the microgravity effects seen in space (Sulzman, 1984), did not produce major repeatable results. Similarly, alterations of the vector of a 1-g gravity load were not adequate simulations of space flight on the conidiation rhythm. The results on the acute and chronic exposures to hypergravity demonstrated that chronic exposure of Neurospora to a 3-g force had no damping effect. On the other hand, an acute 10-min exposure to this hypergravity (the lift-off conditions) was found to cause significant damping on the circadian rhythm of conidiation. This effect was eliminated by a brief light pulse given 36 hrs after the exposure to 3 g.

Ferraro, James S.↗

Preliminary characterization of persisting circadian rhythms during space flight

In order to evaluate the function of the circadian timing system in space, the circadian rhythm of conidiation of the fungus Neurospora crassa was monitored in constant darkness on the STS 9 flight of the Space Shuttle Columbia. During the first 7 days of spaceflight many tubes showed a marked reduction in the apparent amplitude of the conidiation rhythm, and some cultures appeared arrhythmic. There was more variability in the growth rate and circadian rhythms of individual cultures in space than is usually seen on earth. The results of this experiment indicate that while the circadian rhythm of Neurospora conidiation can persist outside of the earth's environment, either the timekeeping process or its expression is altered in space.

Sultzman, F. M.↗

Life sciences experiments on Spacelab 1

The objectives and procedures regarding various biological experiments to be conducted on Spacelab 1 are reviewed. These include the mapping of the HZE cosmic ray particle flux within the Spacelab module, investigating the effects of nullgravity on circadian cycles in the slime mold, Neurospora crassa, and measuring nutations of the dwarf sunflower, Helianthus annus. Emphasis is placed on research regarding possible changes in vestibulocular reflexes, vestibulospinal pathways, cortical functions involving perception of motion and spatial susceptibility. Also discussed are experiments regarding erythrokinetics in man and the effects of prolonged weightlessness of the humoral immune response in humans.

Buderer, M. C.↗

Neurospora circadian rhythms in space - A reexamination of the endogenous-exogenous question

To test the functioning of circadian rhythms removed from periodicities of the earth's 24-hour rotation, the conidiation rhythm of the fungus Neurospora crassa was monitored in constant darkness during spaceflight. The free-running period of the rhythm was the same in space as on the earth, but there was a marked reduction in the clarity of the rhythm, and apparent arrhythmicity in some tubes. At the current stage of analysis of the results there is insufficient evidence to determine whether the effect seen in space was related to removal from 24-hour periodicities and whether the circadian timekeeping mechanism, or merely its expression, was affected.

Sulzman, F. M.↗

Spacelab 1 and the Life Sciences Flight Experiments Program

The Life Sciences Flight Experiments Program (LSFEP) was established by NASA in 1978 to plan and direct efforts necessary to conduct a continuing program of in-flight life science investigations throughout the Space Shuttle era. The Spacelab 1 (SL-1) mission, conducted from November 28 to December 8, 1983, was to verify Spacelab performance through a variety of scientific experiments including life science. A description is given of the seven NASA life sciences experiments, which consisted of four human experiments, a fungus experiment, a plant experiment, and radiation experiments. Ten life sciences experiments from the European Space Agency were also flown. The experiments include studies of the circadian rhythms in Neurospora crassa, the nutation of Helianthus annus, the vestibular function during weightlessness, the influence of space flight on erythrokinetics in man, and the adaptation of vestibulo-spinal reflex mechanisms during space flight.

Bush, W. H.↗

Characterization of neurospora circadian rhythms in space

To determine whether the circadian rhythm of conidiation in neurospora crassa is endogenously derived or is driven by some geophysical time cue, an experiment was conducted on space shuttle flight STS-9, where inoculated race tubes were exposed to the microgravity environment of space. The results demonstated that the rhythm can persist in space. However, there were several minor alterations noted; an increase in the period of the oscillation and the variability of the growth rate and a diminished rhythm amplitude, which eventually damped out in 25% of the flight tubes. On day seven of the flight, the tubes were exposed to light while their growth fronts were marked. It appears that some aspects of this marking process reinstated a robust rhythm in all the tubes which continued throughout the remainder of the flight. It was hypothesized that the damping found prior to the marking procedure on STS-9 may have been a result of the hypergravity pulse of launch and not due to the microgravity of the orbital lab; furthermore, that the marking procedure, by exposing the samples to light, had reinstated rhythmicity. To test this, an investigation was conducted into the effects of acute and chronic exposure to hypergravity.

Ferraro, James S.↗

The evolution of energy-transducing systems. Studies with an extremely halophilic archaebacterium

The halobacterial ATPase was labeled with C-14-dicyclohexylcarbodiimide and subunit 2 of the enzyme was prepared by electroelution. Subunit 2 was cleaved by several chemical and enzymatic procedures for further preparation of peptides. Immunoreactions (Western blotting) of halobacterial membranes were performed with an antiserum against subunit A of the vacuolar ATPase from Neurospora crassa. A 85 K band (subunit 1) from the membranes of H saccharovorum and from two halobacterial isolates, which were isolated from Permian salt sediments, reacted strongly with the antiserum. The ATPase from the latter isolates resembled the ATPase from H saccharovorum, but had a higher content of acidic amino acids. If it can be verified that the age of the bacterial isolates is in the same range as when deposition of salt occurred, an extremely interesting system for the study of evolutionary questions would be available, since the salt-embedded bacteria presumably did not undergo mutational and selectional events.

Stan-Lotter, Helga↗

Relationship of the membrane ATPase from Halobacterium saccharovorum to vacuolar ATPases

Polyclonal antiserum against subunit A (67 kDa) of the vacuolar ATPase from Neurospora crassa reacted with subunit I (87 kDa) from a membrane ATPase of the extremely halophilic archaebacterium Halobacterium saccharovorum. The halobacterial ATPase was inhibited by nitrate and N-ethylmaleimide; the extent of the latter inhibition was diminished in the presence of adenosine di- or triphosphates. 4-chloro-7-nitrobenzofurazan inhibited the halobacterial ATPase also in a nucleotide-protectable manner; the bulk of inhibitor was associated with subunit II (60 kDa). The data suggest that this halobacterial ATPase may have conserved structural features from both the vacuolar and the F-type ATPases.

Stan-Lotter, Helga↗

Relationship of the Membrane ATPase from Halobacterium saccharovorum to Vacuolar ATPases

Polyclonal antiserum against subunit A (67 kDa) of the vacuolar ATPase from Neurospora crassa reacted with subunit I (87 kDa) from a membrane ATPase of the extremely halophilic archaebacterium Halobacterium saccharovorum. The halobacterial ATPase was inhibited by nitrate and N-ethylmaleimide; the extent of the latter inhibition was diminished in the presence of adenosine di- or triphosphates. 4-Chloro-7-nitrobenzofurazan in- hibited the hatobacterial ATPase also in a nucleotide- protectable manner; the bulk of inhibitor was associated with subunit II (60 kDa). The data suggested that this halobacterial ATPase may have conserved structural features from both the vacuotar and the F-type ATPases.

Stan-Lotter, Helga↗