The posterior basal diencephalon of rats enhances expression of an activated state.

STUDY OBJECTIVES: Various ablation studies have implicated the posterior basal diencephalon in the promotion of wakefulness. Although many studies have examined the role of this structure in promotion of cortical arousal, few investigations have attempted to examine its importance in regulation of motor activation (behavioral arousal). In the current study, recordings of freely moving decerebrate rats with and without a posterior basal diencephalon were performed. These studies allowed determination of the behavioral states expressed by the preparations and whether removal of the posterior basal diencephalon completely eliminated expression of both activated state with and activated state without limb movements. DESIGN: Muscle activity was recorded from limb and neck muscles. Eye movements and heart rate were also monitored. The percentage of time spent in various behavioral states and the proportion of limb movements expressed in each of these states were determined. MEASUREMENTS AND RESULTS: Rats with an intact posterior basal diencephalon cycled between all behavioral states. However, they spent most of the recording time in an activated state. In contrast, removal of the posterior basal diencephalon produced rats that spent most of the recording period in a quiescent state. Limb movements were expressed mainly by animals with an intact posterior basal diencephalon, and only when these animals were in the activated state. CONCLUSIONS: The results of this study suggest that the posterior basal diencephalon is required for expression of an activated state and specifically provide evidence for a descending projection from this region required for expression of this state and associated motor activation.

Use of course-embedded projects for program assessment.

There is increasing demand on science faculty to develop authentic assessment measures for both individual courses and undergraduate programs. We report here on a quarter-long group project used in a neurophysiology course that can be used for either purpose. Small groups of four to five students critically analyze at least 10 articles from the primary scientific literature. The end result of this process is the equivalent of a scientific review article that is presented in two formats, a 10-min oral presentation and a scientific poster presentation. Students perform better on application tasks than on analysis, synthesis, or evaluation tasks associated with the project (P < 0.025) and generally respond positively to process questions (59-82%) but less positively to task questions (36-76%) about group dynamics. The cognitive skills and basic content knowledge required to complete this project are developed throughout the undergraduate program. Thus the project is a type of culminating program experience. However, the project also assesses basic course proficiency, because students cannot analyze primary neuroscience research without an understanding of neurophysiological principles.

Regulation of posterior lateral hypothalamic arousal related neuronal discharge by preoptic anterior hypothalamic warming.

The posterior lateral hypothalamus (PLH) is a brain region involved in cortical activation and maintenance of behavioral arousal. Spontaneous and evoked neuronal activities were recorded in the PLH of cats before and during local warming of the preoptic-anterior hypothalamus (POAH). Forty-six percent of PLH neurons recorded in awake cats decreased spontaneous discharge during POAH warming. In the absence of POAH warming, these same cells exhibited significantly lower discharge rates in non-rapid eye movement (NREM) sleep compared to waking. Responses of PLH neurons to POAH warming were also investigated in sleeping cats to determine whether hypothalamic thermosensitive neurons modulated sleep-wake regulation across arousal states. POAH warming had no influence on spontaneous activities of PLH neurons during NREM sleep, but suppressed activity of a small subset of neurons during rapid eye movement (REM) sleep. Thus, during wakefulness or REM sleep, hypothalamic thermosensitive neurons may modulate behavioral state via inhibition or disfacilitation of arousal systems.

Reticular formation activity during wakefulness and sleep in a hibernator (Spermophilus lateralis).

This study examined ground squirrel reticular formation (RF) cell activity as a function of: (1) arousal state, and, (2) phasic alterations in neck muscle activity. A total of 37 neurons were recorded from 11 behaving squirrels (Spermophilus lateralis). Five cells were located in the midbrain RF, 15 were in the lateral pontine site implicated in inhibition of motor output, i.e. the subcoeruleus area, 11 were in the medial pontine RF, and 6 were in the medullary RF. Regardless of location, the majority of the cells (81%; 30 of 37) exhibited greatest activity when the animals were awake and/or in rapid-eye-movement (REM) sleep. However, the relationship between neuronal activity and phasic alterations in neck muscle activity differed as a function of location. The activity of 53% of cells (8 of 15) located in the subcoeruleus area increased with phasic decreases in electromyographic (EMG) activity, whereas the activity of cells (22 of 22) in other RF sites decreased or did not change with phasic decreases in EMG activity. The increased activity of subcoeruleus area cells during phasic decreases in neck muscle activity is further evidence suggesting that the lateral pontine RF is involved in promotion of muscle atonia during REM sleep and possibly wakefulness.

Action potential duration increases as body temperature decreases during hibernation.

The effect of temperature on the duration of both population spikes and action potentials of single neurons has been investigated in a variety of in vitro preparations. A few studies have examined the influence of temperature on spike potentials from spinal motoneurons of intact, anesthetized mammals. In all cases, the duration of the action potential or population spike increased as temperature decreased. A similar increase in the duration of action potentials accompanied hibernation in the ground squirrel (Spermophilus lateralis). Oscilloscope traces of 2 brainstem reticular formations, and 8 posterior thalamic single units were photographed at a body temperature (Tb) of 34-36 degrees C during euthermia prior to entrance into hibernation and at Tb's ranging from 10 to 27 degrees C during hibernation. There was a significant increase in the duration of the second component of the diphasic action potential at the lower Tb (P less than 0.01). This temperature effect was reversible, i.e. action potential durations returned to preentrance euthermic values following arousal from hibernation (Tb = 34-36 degrees C). This study is the first to use behaving animals to demonstrate that changes in biophysical characteristics of central nervous system neurons occur at low Tb. These changes in membrane characteristics probably result in alterations in neuronal functioning and information processing during hibernation.

Neuronal activity during sleep and complete bouts of hibernation.

Changes in arousal state in a euthermic mammal exert powerful influences on major neural regulatory systems. Changes in behavioral state occur at body temperature (Tb) greater than 25 degrees C during hibernation. However, no information exists regarding alterations in arousal states during deep torpor. In this study we used a combination of electroencephalographic, electromyographic, and posterior thalamic neuronal activity in ground squirrels (Spermophilus lateralis) to evaluate arousal states during deep hibernation. No state homologous to rapid-eye-movement sleep was observed below Tb = 21 degrees C during hibernation. However, the animals did continue to cycle through states homologous to electrophysiologically defined wakefulness (AW) and non-rapid-eye-movement (NREM) sleep at all temperatures examined (Tb = 14-36 degrees C). These results extend previous observations that hibernation is not a homogeneous state. Instead, deep torpor consists primarily of a state similar to NREM sleep, interrupted periodically by short intervals of a form of AW. These periodic alterations in state should be accompanied by changes in the properties of many regulatory systems and must be accounted for in any theory of the neural control of hibernation.

Phosphagen and intracellular pH changes during contraction of creatine-depleted rat muscle.

To evaluate the functional role of phosphocreatine (PCr) and creatine in muscle metabolism, these compounds were depleted by feeding rats the creatine analogue, beta-guanidinopropionate (beta-GPA, 2% of diet). Changes in phosphate metabolites and intracellular pH were monitored in gastrocnemius muscle in situ by phosphorus nuclear magnetic resonance (31P-NMR) at 162 MHz using the surface coil technique. After 3 mo of feeding, 25 mumol/g of phosphorylated beta-GPA (beta-GPAP) had accumulated, and PCr, creatine, and ATP levels were reduced to 6, 17, and 56%, respectively, compared with muscles of control animals. In resting muscle, there was no measurable exchange of phosphate between beta-GPAP and ATP by the NMR saturation transfer method. During muscle stimulation at 1 and 5 Hz, the maximum net rate of beta-GPAP hydrolysis was 10% that of PCr in control muscles, so that after 150 s inorganic phosphate had increased to less than 50% of the level attained in control muscles. At both rates, peak twitch force declined toward a steady state more rapidly in beta-GPA-loaded muscles, but after 100 s force was either not different (1 Hz) or significantly greater (5 Hz) in the beta-GPA-fed animals. Intracellular pH initially decreased more rapidly during stimulation and recovered more rapidly afterward in the beta-GPA-loaded muscles compared with controls. This difference could be explained by the difference in expected proton consumption due to net PCr hydrolysis. However, despite buffering by PCr hydrolysis, pH ultimately decreased more in control muscle (6.1 vs. 6.3 for 5 Hz), indicating greater acid accumulation compared with beta-GPA-loaded muscles. In the superficial, predominantly fast-twitch glycolytic section of muscles clamp-frozen after 5-Hz stimulation for 150 s, lactate accumulation was twofold greater in controls. The results indicate that PCr is not essential for steady-state energy production but that the phosphate from PCr hydrolysis may be important for maximum activation of glycogenolysis and/or glycolysis.

Ketone body metabolism in a ground squirrel during hibernation and fasting.

Hibernating Belding's ground squirrels (Spermophilus beldingi) are ketotic relative to fed nonhibernators. Muscles from torpid individuals, when incubated in media containing physiological concentrations of glucose and ketone, show reduced uptake of glucose in the presence of ketone. The magnitude of the reduction is dependent on ketone concentration and reaches 60% in heart and 100% in pectoralis at 1.4 mM ketone. Fasted squirrels are also ketotic. However, ketone does not reduce glucose uptake in muscles from fed or fasted animals. Glucose utilization by muscles decreases during a long-term fast, but the reduction is independent of ketone. Thus both a long-term fast and hibernation lead to changes in muscle tissues that decrease their reliance on glucose as an energy source. Ketosis leads to glucose sparing during hibernation, whereas muscle glucose utilization is decreased independently of ketone during a fast. The glucose sparing achieved in both hibernation and fasting leads to conservation of body protein, the major source of gluconeogenic precursors in fasting mammals.

Plasma membrane expansion terminates in Saccharomyces cerevisiae secretion-defective mutants while phospholipid synthesis continues.

Phospholipid synthesis activity and plasma membrane growth have been studied in the Saccharomyces cerevisiae temperature-sensitive, secretion-defective mutants isolated by Novick and Schekman (Proc. Natl. Acad. Sci. U.S.A. 76:1858-1862, 1979; Novick et al., Cell 21:205-215, 1980). The mutants, sec1 through sec23, do not grow at 37 degrees C and exhibit lower rates of phospholipid synthesis than does the wild-type strain X2180. None of the mutants exhibits a decline in lipid synthesis rapid enough to explain secretion failure. Plasma membrane growth was assessed indirectly by examining the osmotic sensitivity of spheroplasts derived from cultures transferred from 24 to 37 degrees C. Spheroplasts from the normal-growing strain X2180 exhibited a small rapid increase in osmotic sensitivity and stabilized at a more sensitive state. Spheroplasts from the sec mutants exposed to the same temperature shift exhibited progressively increasing osmotic sensitivity. Cycloheximide treatment prevented progressive increases in osmotic fragility. These data are compatible with the hypothesis that plasma membrane expansion is restricted in the sec mutants. During incubation at 37 degrees C, the accumulation of intracellular materials within the no-longer expanding plasma membrane exerts osmotic stress on the membrane, increasing with time. The gene products defective in Novick and Schekman's sec mutants appear to be required for both extracellular protein secretion and plasma membrane growth in yeast cells.