Nancy J. Berner

8-OH-DPAT-sensitive neurons in the nucleus raphe magnus modulate thermoregulatory output in rats.

The nucleus raphe magnus (NRM) is purported to be a relay through which peripheral thermoafferent information is transmitted to thermointegrative centers located in the preoptic/anterior hypothalamus (POAH). Therefore, suppression of neural activity in the NRM should reduce thermoregulatory responses to peripheral thermal challenges, but not affect responses elicited by manipulation of POAH temperature. At low ambient temperatures lidocaine injections into the NRM of nonanesthetized rats resulted in decreases in POAH temperature, oxygen consumption, and electromyographic activity. At a warm ambient temperature, lidocaine injections into the NRM decreased the elevations in oxygen consumption and electromyographic activity elicited by cooling the POAH. The effects of lidocaine injections were duplicated by injection of a 5-HT(1A) agonist 8-hydroxy-dipropylaminotetralin (8-OH-DPAT) into the NRM. The effect of 8-OH-DPAT was eliminated by pre-treatment with a selective autoreceptor antagonist. These results suggest that NRM 5-HT neurons are modulating the relationship between output of thermointegrative centers and thermoregulatory effector responses rather than processing thermoafferent information.

Does the preoptic anterior hypothalamus receive thermoafferent information

The preoptic anterior hypothalamus (POAH) is considered the thermointegrative center of the mammalian brain. Studies on anesthetized and unanesthetized animals have demonstrated neurons in the POAH that respond to changes in both POAH temperature (TPOAH) and skin temperature (Ts). In these studies, however, electroencephalographic (EEG) activity was not monitored. Recent work has revealed the potential for arousal state selectivity of neurons combined with thermal influences on arousal state to create the appearance that cells are thermosensitive or thermoresponsive when in fact they may not be responding directly to temperature or to thermoafferent input. It is therefore necessary to reexamine the influence of central and peripheral temperature on POAH cells. In the present study, 66 POAH cells were recorded from urethan-anesthetized rats while EEG, TPOAH, and Ts were monitored. Seventy-five percent (41 of 55) of the cells were EEG state responsive; 22% (6 of 27) were TPOAH sensitive; and 33% (19 of 58) appeared to be Ts responsive. However, when EEG state changes were taken into account, none of the cells that appeared to be Ts responsive were responding to Ts within any uniform EEG state. All changes in their firing rates were associated with EEG state changes. This study raises a question as to whether or not peripheral temperature information is integrated in the POAH. Consideration should be given to the possibility that Ts information is integrated lower in the neuroaxis. Monitoring EEG is essential in studies attempting to characterize the integrative properties of POAH neurons of anesthetized or unanesthetized animals. This caveat applies not just to thermoregulatory studies but to investigations of other integrative functions of the hypothalamus and many other brain regions as well.The preoptic anterior hypothalamus (POAH) is considered the thermointegrative center of the mammalian brain. Studies on anesthetized and unanesthetized animals have demonstrated neurons in the POAH that respond to changes in both POAH temperature (TPOAH) and skin temperature (Ts). In these studies, however, electroencephalographic (EEG) activity was not monitored. Recent work has revealed the potential for arousal state selectivity of neurons combined with thermal influences on arousal state to create the appearance that cells are thermosensitive or thermoresponsive when in fact they may not be responding directly to temperature or to thermoafferent input. It is therefore necessary to reexamine the influence of central and peripheral temperature on POAH cells. In the present study, 66 POAH cells were recorded from urethan-anesthetized rats while EEG, TPOAH, and Ts were monitored. Seventy-five percent (41 of 55) of the cells were EEG state responsive; 22% (6 of 27) were TPOAH sensitive; and 33% (19 of 58) appeared to be Tsresponsive. However, when EEG state changes were taken into account, none of the cells that appeared to be Ts responsive were responding to Ts within any uniform EEG state. All changes in their firing rates were associated with EEG state changes. This study raises a question as to whether or not peripheral temperature information is integrated in the POAH. Consideration should be given to the possibility that Ts information is integrated lower in the neuroaxis. Monitoring EEG is essential in studies attempting to characterize the integrative properties of POAH neurons of anesthetized or unanesthetized animals. This caveat applies not just to thermoregulatory studies but to investigations of other integrative functions of the hypothalamus and many other brain regions as well.

Modelling the anisotropic electrical properties of skeletal muscle

We present a numerical model used to analyse the anisotropic electrical properties of frog muscle, measured in vivo. The model represents the anisotropic, irregularly shaped muscle as a set of cubic elements. We develop a finite difference method to calculate the electrical resistance between two electrodes inserted longitudinally or transversely into the muscle in terms of longitudinal and transverse muscle conductivities. Comparison of the measured impedance values with the calculated resistances yields the separate variation with frequency of the two conductivity components. We also compare the results of the numerical, finite difference method with those of two simple, analytical models.

OXYGEN CONSUMPTION BY MITOCHONDRIA FROM AN ENDOTHERM AND AN ECTOTHERM

Comparisons of metabolic properties of mitochondria from an endothermic and an ectothermic vertebrate were performed. Oxygen (O2) consumption rates of liver mitochondria from laboratory mice and western fence lizard (Sceloporus occidentalis) were determined over a range of temperatures (10, 20, 30 and 37 degrees C) and in the presence of a variety of substrates. At 37 degrees C the O2 consumption rate of mouse mitochondria was 4-11 times higher than lizard mitochondria in the presence of five of eight substrates. This range of differences is similar to differences reported for O2 consumption of endothermic animals, tissues and cells over those of ectotherms. Thermal sensitivity of mitochondria was measured by calculation of Q10s for O2 consumption. Q10s were highest for mouse mitochondria overall. The range that showed the highest Q10s for the mouse mitochondria was 30-20 degrees C, whereas for the lizard mitochondria it was 20-10 degrees C. Thus, mitochondria from the ectotherm showed a lower degree of temperature sensitivity than did mitochondria from the endotherm. The preferred substrate for all mitochondria at all temperatures was succinate, but mouse mitochondria then showed some preference for alpha-ketoglutarate and citrate, whereas lizard mitochondria showed a preference for pyruvate and malate + pyruvate.

The low-frequency dielectric properties of octopus arm muscle measured in vivo

The conductance and capacitance of octopus arm are measured in vivo over the frequency range 5 Hz to 1 MHz. Measurement of these parameters for a number of electrode separations permits the determination of the variations in tissue conductivity and dielectric constant with frequency. In the range 1-100 kHz the conductivity is independent of the frequency f and the dielectric constant varies as f-1. These results, in conjunction with those reported previously for frog skeletal muscle, are consistent with the fractal model for the dielectric properties of animal tissue proposed by Dissado. Transformation of the results to complex impedance spectra indicates the presence of a dispersion above 100 kHz.

Phenotypic flexibility and thermoregulatory behavior in the eastern red‐spotted newt (Notophthalmus viridescens viridescens)

Eastern red-spotted newts are ectotherms, aquatic as adults, and active year-round, breeding even during winter under ice. Earlier research, with field-captured newts, showed a correlation between seasonal changes in the activity of some muscle metabolic enzymes and in the behavior of the newts in a thermal gradient. This study was undertaken to further characterize acclimatory responses in the newts in a more controlled laboratory environment. Newts were obtained during fall, and maintained at 15 degrees C with 12:12 LD for 4 weeks for SMR (at 8 and 26 degrees C) and temperature preference experiments. Subsequently, half the newts were exposed to summer conditions (26 degrees C, 14:10 LD) and half to winter conditions (8 degrees C, 10:14 LD). After 12 weeks, SMR and temperature preference experiments were repeated, and enzyme assays for cytochrome c oxidase (CCO), citrate synthase (CS), and lactate dehydrogenase (LDH) were performed on muscle tissue homogenates, also at 8 and 26 degrees C. Newts changed all three parameters in the laboratory. SMRs were highest in winter-acclimated newts and lowest in summer-acclimated newts, whereas temperature preference was lowest in winter-acclimated newts and highest in summer-acclimated newts. Finally, CCO activity was completely compensated in winter-acclimated newts, CS activity was partially compensated, and LDH activity was not seasonally sensitive. These results indicate a connection or relationship between changes in seasonal environmental conditions, and some aspects of the muscle biochemistry, SMR, and thermoregulatory behavior of these ectotherms.

Metabolic depression during aestivation does not involve remodelling of membrane fatty acids in two Australian frogs

Changes in membrane lipid composition (membrane remodelling) have been associated with metabolic depression in some aestivating snails but has not been studied in aestivating frogs. This study examined the membrane phospholipid composition of two Australian aestivating frog species Cyclorana alboguttata and Cyclorana australis. The results showed no major membrane remodelling of tissue in either frog species, or in mitochondria of C. alboguttata due to aestivation. Mitochondrial membrane remodelling was not investigated in C. australis. Where investigated in C. alboguttata, total protein and phospholipid content, and citrate synthase (CS) and cytochrome c oxidase (CCO) activities in tissues and mitochondria mostly did not change with aestivation in liver. In skeletal muscle, however, CS and CCO activities, mitochondrial and tissue phospholipids, and mitochondrial protein decreased with aestivation. These decreases in muscle indicate that skeletal muscle mitochondrial content may decrease during aestivation. Na+K+ATPase activity of both frog species showed no effect of aestivation. In C. alboguttata different fat diets had a major effect on both tissue and mitochondrial phospholipid composition indicating an ability to remodel membrane composition that is not utilised in aestivation. Therefore, changes in lipid composition associated with some aestivating snails do not occur during aestivation in these Australian frogs.

Thermal Acclimatization in Overwintering Tadpoles of the Green Frog, Lithobates clamitans (Latreille, 1801).

Seasonal acclimatization permits organisms to maintain function in the face of environmental change. Tadpoles of the green frog (Lithobates clamitans) overwinter as tadpoles in much of their range. Because they are active in winter, we hypothesized that green frog tadpoles would display acclimatization of metabolic and locomotor function. We collected tadpoles in Sewanee, Tennessee (35.2°N) in winter and summer. Tadpoles collected during each season were tested at both winter (8°C) and summer (26°C) temperatures. Winter tadpoles were able to maintain swimming performance at both temperatures, whereas swimming performance decreased at cold temperatures in summer tadpoles. There was no evidence for seasonal acclimatization of whole-animal metabolic rate. Although whole-animal metabolic acclimatization was not observed, the activities of cytochrome c oxidase, citrate synthase, and lactate dehydrogenase measured in skeletal muscle homogenates showed higher activity in winter-acclimatized tadpoles indicating compensation for temperature. Further, the composition of muscle membranes of winter tadpoles had less saturated and more monounsaturated fatty acids and a higher ω-3 balance, unsaturation index, and peroxidation index than summer tadpoles. These data indicate that reversible phenotypic plasticity of thermal physiology occurs in larval green frog tadpoles. They appear to compensate for colder temperatures to maintain burst-swimming velocity and the ability to escape predators without the cost of maintaining a constant, higher standard metabolic rate in the winter.

Impedance Spectroscopy of Newt Tails

We use impedance spectroscopy to characterize the electrical properties of newt tail. The newt model has attracted recent interest because the regeneration of newt tail and limbs may provide insights into tissue engineering for mammals. Impedance spectroscopy could provide a convenient means to monitor the progress of regeneration while various controlling stimuli are applied. In this initial phase of our research we compare the impedance spectra of healthy and necrotic tail tissue and identify a potentially confusing artifact in the spectra due to interneedle capacitance.