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Damage to neurons in the rat when the animals are subjected to a constant magnetic field is a function of their reactive capacity. The reactive stage is characterized by dystrophic changes without any definite signs of destruction of tissue.
In the investigation the established technique of direct microscopic viewing was combined with the use of a closed circuit television system and cinematography. The red cell flow patterns in all capillaries were found to be oscillatory with characteristic cycle frequencies and amplitudes for all concentrations of inspired oxygen greater than 8%. Generally, there was a transient decrease in mean flow rate with increasing severity of hypoxia, with a gradual return toward control values. Red cell flow patterns are discussed along with questions of red cell configuration.
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A myocardial transducer for simultaneously measuring force and displacement within a very small area of myocardium is disclosed. The transducer comprised of an elongated body forked at one end to form an inverted Y shaped beam with each branch of the beam constituting a low compliant tine for penetrating the myocardium to a predetermined depth. Bonded to one of the low compliance tines is a small piezoresistive element for converting a force acting on the beam into an electrical signal. A third high compliant tine of the transducer, which measures displacement of the myocardium in a direction in line with the two low compliant tines, is of a length that just pierces the surface membrane. A small piezoresistive element is bonded to the third tine at its upper end where its bending is greatest. Displacement of the myocardium causes a deformation in curvature of the third tine, and the second small piezoresistive element bonded to the surface of its curved end converts its deformation into an electrical signal.
Horizontally body-casted rhesus monkeys are used as an animal model in order to study the physiological changes known as cardiovascular deconditioning which occur during weightless conditions. No difference was found between the experimental and control animals in heart mitochondrial oxidative phosphorylation which indicates that no apparent changes occurred in the primary energy-producing system of the heart. A marked increase in cytochrome oxidase activity was observed in the casted primate heart mitochondria compared to controls, while a 25% decrease in respiratory substrate-supported calcium uptake was found in casted primate heart mitochondria compared to controls. Sacroplasmic reticulum isolated from the primate hearts revealed marked changes in calcium transport activities. It is concluded that the marked depression in cardiac sarcoplasmic reticulum functions indicates altered calcium homeostasis in the casted-primate heart which could be a factor in cardiovascular deconditioning.
The influence of rat suspension on soleus disuse and atrophy was investigated to measure changes in fiber area and number and to determine if suspension elicited changes in lysosomal protease activity and rate of calcium uptake by the sarcoplasmic reticulum. The infuence of rat suspension on myosin light chain phosphorylation and succinate dehydrogenase activity are determined.
Progress is reported in the following areas: (1) microgel electrophoresis identification of single fibers; (2) skinned fiber preparation; and (3) microbiochemical techniques for assaying important enzymes and substrates in single fibers.
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The effect of 28-day-long hind-limb suspension (HS) combined with 10 daily forceful lengthening contractions of the limb on the morphological and metabolic properties of individual fibers of the soleus was studied in rats, using quantitative histochemical techniques. Compared with nonsuspended controls (CON), soleus wet weights of HS rats were decreased by 49 percent; the fibers staining lightly for myosin ATPase ('light-ATPase' fibers) atrophied more than the 'dark-ATPase' fibers. Single-fiber alpha-glycerophosphate dehydrogenase (GPD) and succinate dehydrogenase (SDH) activities were higher in HS than in CON rats. Daily forceful lengthening contractions did not prevent the HS-induced changes. The results support the view that the soleus fibers can change from a slow-twitch oxidative to a fast-twitch oxidative-glycolytic profile, but rarely to a fast-twitch glycolytic one, and that the SDH and GPD activities per volume of tissue can be increased even when there are severe losses of contractile proteins.
Cocultured Xenopus neurons and myocytes were subjected to nonvectorial gravity by clinostat rotation to determine the effects of microgravity on cell development and communications. Observed effects included increases in the myocyte and its nuclear area, fragmentation of nucleoli, the appearance of neuritic aneurysms, decreased growth in the presence of trophic factors, and decreased yolk utilization. These effects were most notable at 1-10 rpm and depended on the onset and duration of rotation. It is found that, in microgravity, cell differentiation is altered by interference with cytoskeleton-related mechanisms. It is suggested that the alteration of the distribution of acetylcholine receptor aggregates on myocytes which occurs might indicate that microgravity affects brain development.
The effects of the insulin-like growth factor, multiplication-stimulating activity (MSA), on chick myotube cultures are studied. The results indicate that MSA is an effective anabolic agent regulating protein metabolism and amino acid uptake, but not sugar transport. Similar size effects on protein metabolism and amino acid uptake in serum-free media were observed in parallel studies with insulin, although insulin levels well in excess of the normal physiological range are required to produce significant effects. It is suggested that there is a generally low insulin sensitivity in cultured chick myotubes relative to adult tissues.
The effects of two weeks of hind-limb suspension (HS) on the functional properties of slow-twitch and fast-twitch single fibers isolated from the predominantly slow-twitch soleus and fast-twitch gastrocnemius of the suspended leg of rats were investigated. Single fibers were suspended between a motor arm and force transducer, and, after their functional properties were studied, the fiber type was established by the myosin heavy chain analysis. It was found that, after HS, the greatest decrease in diameter and a reduction in peak tension occurred in slow-twitch fibers from soleus, followed by slow-twitch fibers from gastrocnemius. Fast-twitch fibers from the red gastrocnemius showed a significant reduction in diameter but no change in peak tension. No effect of HS was observed on the diameter of the fast-twitch fibers from the white gastsrocnemius (which is known to contain 87 percent fast glycolytic fibers).
The left ventricle of hearts from rats flown on the Cosmos 1887 biosatellite for 12.5 days was compared to the same tissue of synchronous and vivarium control animals maintained in a ground based laboratory. The volume density of the mitochondria in the myocardium of the space-flown animals was statistically less (p equal less than 0.01) than that of the synchronous or vivarium control rats. Exposure to microgravity resulted in a certain degree of myocardial degeneration manifested in mitochondrial changes and accumulation of myeloid bodies. Generalized myofibrillar edema was also observed.
The cellular compartmentalization of the cyclic AMP-receptor proteins in heart ventricular tissue obtained from rats flown on the Cosmos 1887 is determined. Photoaffinity labeling of soluble and particular cell fractions with a (32P)-8-azido analog of cyclic AMP is followed by electrophoretic separation of the proteins and by autoradiographic identification of the labeled isoforms of cAPK R subunits. It is shown that RII in the particulate subcellular fraction was significantly decreased in heart cells from rats in the flight group when compared to controls. Protein banding patterns in both the cytoplasmic fraction and in a fraction enriched in chromatin-bound proteins exhibited some variability in tissues of individual animals, but showed no changes that could be directly attributed to flight conditions. No significant change was apparent in the distribution of RI or RII cyclic AMP binding in the soluble fractions. It is inferred that the cardiac cell integrity or its protein content is not compromised under flight conditions.
A complete description of an instrumented ergometer system, including the sensors, the data acquisition system, and the methodologies to calculate the kinematic parameters were initially developed at Tulane University. This work was continued by the PI at NASA Johnson Space Center, where a flight ergometer was instrumented and tested during a KC-135 Zero-Gravity flight. The sensors that form part of the system include EMG probes and accelerometers mounted on the subject using the ergometer, load cells to measure pedal forces, and encoders to measure position and orientation of the pedal (foot). Currently, data from the flight test is being analyzed and processed to calculate the kinematic parameters of the individual. The formulation developed during the initial months of the grant will be used for this purpose. The system's components are compact (all sensors are very small). A salient feature of the system and associated methodology to determine the kinematics is that although it uses accelerometers, position is not determined by integration. Position is determined by determining the angle of two frames of reference for which acceleration at one point is known in coordinates of both frames.
The effects of 1, 2, and 3 wk of hindlimb suspension (HS) on force-velocity and power characteristics of single rat soleus fibers were determined. After 1, 2, or 3 wk of HA, small fiber bundles were isolated, placed in skinning solution, and stored at -20 C until studied. Single fibers were isolated and placed between a motor arm and force transducer, functional properties were studied, and fiber protein content was subsequently analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Additional fibers were isolated from soleus of control after 1 and 3 wk of HS, and fiber type distribution and myosin light chain stoichiometry were determined from SDS-PAGE analysis. After 1 wk of HS, percent type I fibers declined from 82 to 74%, whereas hybrid fibers increased from 10 to 18%. Percent fast type II fibers increased from 8% in control and 1 wk of HS to 26% by 3 wk of HS. Most fibers showed an increased unloaded maximal shortening velocity (V sub O)), but myosin heavy chain remained entirely slow type I. The mechanism for increased V(sub O) is unknown. There was a progressive decrease in fiber diameter and peak force after 1, 2, and 3 wk of HS, respectively. One week of HS resulted in a shift of the force-velocity curve, and between 2 and 3 wk of HS the curve shifted further such that V(sub O) was higher than control at all relative loads less than 45% peak isometric force. Peak absolute power output of soleus fibers progressively decreased through 2 wk of HS but showed no further change at 3 wk. The results suggest that between 2 and 3 wk the HS-induced alterations in the force-velocity relationship act to maintain the power output of single soleus fibers despite a continued reduction in fiber force.
Quantitative autoradiographic analysis of receptors for GABA and acetylcholine in the forebrain of rats flown on COSMOS 2044 was undertaken as part of a joint US-Soviet study to determine the effects of microgravity on the central nervous system, and in particular on the sensory and motor portions of the forebrain. Changes in binding of these receptors in tissue from animals exposed to microgravity would provide evidence for possible changes in neural processing as a result of exposure to microgravity. Tritium-labelled diazepam and Quinuclidinyl-benzilate (QNB) were used to visualize GABA (benzodiazepine) and muscarinic (cholinergic) receptors, respectively. The density of tritium-labelled radioligands bound to various regions in the forebrain of both flight and control animals were measured from autoradiograms. Data from rats flown in space and from ground-based control animals that were not exposed to microgravity were compared.