The role of nervous systems in temperature adaptation of poikilotherms.
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Biomedical subjects
Publications and source records attributed to C L Prosser.
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Single-unit, extracellular recordings were made from spontaneously active, thermosensitive neurons in the preoptic region of green sunfish acclimated to 25 degrees C. Activity of single cells was monitored during increases and decreases in local brain temperature over approximately a 10 degrees C range. Deep-body and skin temperatures were maintained at 25 degrees C. Of 276 neurons, 81% were insensitive, 17% were warm sensitive, and 2% cold sensitive. Warm responses were grouped into three basic types: exponential, linear, and nonlinear. All cold-sensitive neurons responded in a similar nonlinear manner. Mean levels of firing rate of thermosensitive neurons at 25 degrees C brain temperature ranged from 6 +/- 1.1 impulses/s to 22.7 +/- 10.8 impulses/s. Thermosensitivities were as high as 5.2 +/- 0.9 impulses . -1 . degrees C-1. Anatomic location of these neurons within the preoptic region appears random with some trend for the exponentially responding, warm-sensitive neurons to be located more medial than the other thermosensitive cell types. A small number of neurons were located in the ventrolateral telencephalon. In general, the thermosensitive responses observed resemble those found in other ectotherms and mammals with some exceptions.
The liver mitochondrial and microsomal membranes of green sunfish and rat were examined by steady state polarisation and differential polarised phase fluorimetry to determine the effects of seasonal adaptation of membrane dynamic structure to temperature. Steady state polarisation studies indicated that the liver mitochondria of green sunfish acclimated to different temperatures showed a greater partial compensation of membrane fluidity for the fatty acid composition of both membrane preparations generally became more unsaturated at lower acclimation temperatures, though the differences between 5 degrees C and 25 degrees C acclimated fish were more pronounced in the mitochondrial fraction than in the microsomal fraction. Differential polarised phase fluorimetric studies indicated that the rotations of diphynylhexatriene in mitochondrial and microsomal membranes were highly hindered, though the hindrance offered by membranes of 25 degrees C acclimated green sunfish was far greater than that offered by the membranes of 5 degrees C acclimated fish, thus supporting the concept of homeoviscous adaptation. The absolute rotational rate was not consistently affected by acclimation treatment.
The surface-bound Ca of isolated circular smooth muscle of cat small intestine can be removed by substitution of LiCl for NaCl in Krebs solution. This substitution removed surface-bound Ca (45Ca) and allowed us to study transmembrane 45Ca efflux. Neither the resting membrane potential nor contractility changed when Li was substituted for Na. Li removed the same extracellular 45Ca store as did ethylene glycol-bis-(beta-aminoethylether)-N,N'-tetraacetic acid. The resting transmembrane 45Ca efflux was inhibited by La3+ and was unchanged in Li, tris(hydroxymethyl)aminomethane, arginine, and sucrose Krebs solution. The extra 45Ca efflux observed upon electrical stimulation was no greater in Na-Krebs than Li-Krebs, but during response to acetylcholine the extra 45Ca efflux was greater in Na-Krebs than Li-Krebs. We conclude that the surface-bound Ca is sensitive to external Na and that the transmembrane Ca efflux is not completely dependent on external Na.
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Temperature adaptation of biological membranes was examined by comparing the fragmented sarcoplasmic reticulum preparation of goldfish acclimated to different temperatures. Membrane fluidity was estimated using the fluorescence polarization technique. There was considerable variation between preparations, but no consistent differences in fluidity were observed between 5- and 25 degrees C-acclimated goldfish, fish species adapted over an evolutionary period to arctic or desert temperatures, and rat. The fatty acid composition of the sarcoplamic reticulum preparations of differently acclimated goldfish showed differences in the proportion of mono- and polyunsaturated fatty acids while the proportion of saturated fatty acids remained relatively constant. However, the fatty acid composition of sarcoplasmic reticulum phosphoglycerides became more unsaturated in the order rat, desert pupfish, arctic sculpin, which correlates with their respective environmental or body temperature. It is concluded that differences in membrane components other than fatty acids are important in determining membrane dynamic structure. The inability to demonstrate homeoviscous adaptation in sarcoplasmic reticulum is supported by other evidence suggesting that functions of the sarcoplasmic reticulum that are measured in vitro are not affected by such modifications of their phosphoglyceride fatty acid composition as occur during thermal acclimation.
The "fluidity" of brain synaptosomal membrane preparations of arctic and hot-springs fish species, two temperature water fish species acclimated to different seasonal temperatures, and two mammals was estimated using the fluorescence polarization technique. At all measurement temperatures, the fluidity decreased in the order: arctic sculpin, 5 degrees-acclimated goldfish, 25 degrees-acclimated goldfish, desert pupfish, and rat. This correlated with increasing adaptation or body (i.e., cellular) temperatures of 0 degrees, 5 degrees, 25 degrees, 34 degrees, and 37 degrees and suggested a partial compensation of membrane fluidity for environmental temperature that occurs over the evolutionary time period as well as during laboratory (seasonal) acclimation. Evolutionary adaptation of relatively stenothermal species to constant thermal environments resulted in a more complete compensation than laboratory (seasonal) acclimation. Each compensation is accompanied by differences in the saturation of membrane phosphoglycerides. At increased cellular temperatures the proportion of saturated fatty acids increased and the unsaturation index decreased; the correlation between these indices and the measured expression of membrane dynamic structure was highly significant. It is concluded that the homeoviscous compensation of synaptic membrane function is an important component of temperature adaptation.
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Electrotonic spread of applied potentials was observed between longitudinal and circular muscle layers of the small intestine with no rectification. Nexal junctions were demonstrated between muscle fibers of each layer. Connective tissue cells bridged between the two muscle layers. These showed structural characteristics of fibrocytes and of interstitial cells. Some nexuses were seen between connective tissue cells and between these cells and muscle fibers of each layer but in most junctions the membranes were 10-18 nm apart. Since connective tissue can serve for electrical conduction between cultured heart cells and since electrical properties of intestinal muscle permit transmission with low degrees of coupling, it is suggested that interstitial cells and fibrocytes may electrically couple longitudinal and circular muscle layers of cat intestine.
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1. Slow waves recorded from isolated longitudinal muscle averaged 13 mV and had slow rate of rise (0.04 V/sec) whereas when recorded from intact segments the amplitude averaged 27 mV and the rate of rise was more rapid (0.09 V/sec), often with a notch between the initial peak and the plateau. Membrane potentials of longitudinal muscle were similar in isolated and intact preparations (- 66 mV). Resting potentials of circular muscle averaged - 67 mV.2. Small bundles of circular muscle tested in the double sucrose gap produced activity, either spontaneously or in response to stimulation, which fell into three categories: fast spikes (50-200 msec duration), slow spikes (1-5 sec duration), and small graded responses. The duration of fast spikes could be increased severalfold by the addition of TEA; the graded responses were converted to full-sized spikes by TEA.3. Treatment of circular muscle with Ca-free Krebs solution eliminated spikes, and in intact preparations reduced the amplitude and rate of rise of slow waves and eliminated the notch on slow waves.4. Current-voltage curves of longitudinal muscle show delayed rectification in the depolarizing quadrant; similar curves of circular muscle show anomalous rectification, i.e. a region where a very small current causes a large voltage change.5. Non-polarized electrotonic coupling between longitudinal and circular layers indicates low-resistance pathways. Apparent space constants of longitudinal muscle are greater when attached to circular muscle than when isolated.6. It is concluded that small slow potentials originate rhythmically in longitudinal muscle, that these spread passively to circular muscle where a regenerative amplification occurs which depends on Ca conductance and the amplified slow waves spread back to the longitudinal layer. In the intact intestine pacemaking is, therefore, separate from propagation and the circular muscle provides the bulk of depolarizing current for propagation.
When stomach muscles of skate, toad, or frog or intestinal muscle of cat are treated with Ca-free physiological solutions containing 2-5 mM EGTA or EDTA, spontaneous spikes and slow waves disappear reversibly. With continued treatment, depolarization of 25-30 mV from resting potentials of -65 mV occurs and rhythmic prolonged potentials of several seconds duration appear. They show rapid depolarization to near zero and rapid repolarization and they may continue for several hours. The prolonged potentials disappear when Na is replaced by Li, Tris, or choline. They are insensitive to TTX. The EGTA-induced waves are abolished by Mn, Co, La, verapamil, and D 600. After 10-15 min in 5 mM EGTA, voltage-current and abolition of anomalous rectification. It is concluded that when bound Ca is removed by a chelator, nonspecific reduction in resistance occurs and Na ions may enter rhythmically through channels normally used by Ca.
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Intact muscle layers separated from the small intestine of the cat were mounted in a specially designed chamber to measure electrical slow waves and NADH fluorescence simultaneously. Cooling the muscle to 17 degrees eliminated slow waves and simultaneously increased the level of fluorescence. Likewise, superfusing the muscle with a N2-bubbled glucose-free Krebs solution decreased the amplitude of slow waves and concomitantly increased fluorescence emission. In both cases, return to normal conditions reversed the effects on both slow waves and fluorescence. When signals were averaged over 30-70 slow waves, a pattern emerged with the fluorescence oscillations in phase with the electric oscillations. The NADH:NAD+ ratio reached a maximum at the most depolarized point of the slow waves and a minimum at the most polarized point between slow waves. This indicates maximum ATP utilization during the repolarization process. The correlation between redox oscillations and electrical slow wave generation is associated with cell metabolism.
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