Food and water restriction protocols: physiological and behavioral considerations.

Food and water restriction protocols are common in animal research, yet they often elicit discussion and controversy among institutional animal care and use committee members who review them. Determining a single standard by which all restriction protocols can be evaluated or performed may not be realistic. However, information about the physiologic and behavioral impact of food and water restriction can provide a basis for making rational judgments about these issues in general. This review will discuss the physiologic and behavioral consequences of food or water deprivation periods of 24 h or less and of chronic restriction schedules, with special reference to protocols that use food or water restriction as a motivational tool for behavioral training.

Modulation of sleep by cortisone in normal and bacterially infected rabbits.

Infectious disease is known to alter both sleep patterns and hydrocortisone (cortisol) concentrations in rabbits. Moreover, the sleep-altering effects of microbial infections are likely to be mediated via endogenous immune modulators whose actions are attenuated by glucocorticoids. To evaluate the relationships between sleep and glucocorticoids during infectious disease, the effects of cortisone administration (20 mg/kg, im) on sleep were examined before and after inoculation of rabbits with Staphylococcus aureus or Escherichia coli. When administered alone, cortisone did not alter the amount of time spent in slow-wave sleep (SWS) but did reduce electroencephalographic (EEG) slow-wave amplitudes during SWS, the number of bouts of SWS and the amount of time spent in rapid-eye-movement sleep. The duration of individual bouts of SWS was increased after cortisone treatment. Bacterially infected rabbits developed biphasic changes in sleep patterns that were characterized by an initial increase and a subsequent decrease both in SWS time and in EEG slow-wave amplitudes during sleep. Cortisone treatment attenuated these effects in S. aureus-inoculated rabbits. In contrast, cortisone treatment did not alter the initial phase of enhanced sleep in E. coli-inoculated rabbits but did attenuate the subsequent sleep suppression. These data indicate that glucocorticoid administration is associated with an attenuated sleep response in bacterially inoculated rabbits.

Visual and oculomotor functions of monkey subthalamic nucleus.

1. Single-unit recordings were obtained from the subthalamic nuclei of three monkeys trained to perform a series of visuooculomotor tasks. The monkeys were trained to fixate on a spot of light on the screen (fixation task). When the spot was turned off and a target spot came on, they were required to fixate on the target quickly by making a saccade. Visually guided saccades were elicited when the target came on without a time gap (saccade task). Memory-guided saccades were elicited by delivering a brief cue stimulus while the monkey was fixating; after a delay, the fixation spot was turned off and the monkey made a saccade to the remembered target (delayed saccade task). 2. Of 265 neurons tested, 95 showed spike activity that was related to some aspects of the visuooculomotor tasks, whereas 66 neurons responded to active or passive limb or body movements. The task-related activities were classified into the following categories: eye fixation-related, saccade-related, visual stimulus-related, target- and reward-related, and lever release-related. 3. Activity related to eye fixation (n = 22) consisted of a sustained spike discharge that occurred while the animal was fixating on a target light during the tasks. The activity increased after the animal started fixating on the target and abruptly ceased when the target went off. The activity was unrelated to eye position. It was not elicited during eye fixation outside the tasks. The activity decreased when the target spot was removed. 4. Activity related to saccades (n = 22) consisted of a phasic increase in spike frequency that was time locked with a saccade made during the tasks. The greatest increases occurred predominantly after saccade onset. This activity usually was unrelated to spontaneous saccades made outside the task. The changes in activity typically were optimal in one direction, generally toward the contralateral side. 5. Visual responses (n = 14) consisted of a phasic excitation in response to a visual probe stimulus or target. Response latencies usually were 70-120 ms. The receptive fields generally were centered in the contralateral hemifield, sometimes extending into the ipsilateral field. The receptive fields included the foveal region in seven neurons; most of these neurons responded best to parafoveal stimulation. Peripheral stimuli sometimes suppressed the activity of visually responsive neurons. 6. Activity related to target and reward (n = 29) consisted of sustained spike discharge that occurred only when the monkey could expect a reward by detecting the dimming of the light spot that he was fixating.(ABSTRACT TRUNCATED AT 400 WORDS)

Striatal neuronal activity during the initiation and execution of hand movements made in response to visual and vibratory cues.

Recordings were obtained from 146 neurons in the neostriatum of rhesus monkeys while they performed wrist movements in response to visual and vibratory cues. Of these, 75 putamena nd 29 caudate neurons exhibited changes in firing rate that were temporally related to the onset of the wrist movements and that began prior to movement onset. This premovement activity (PMA) usually was directionally specific, in that the magnitude or direction of change in firing rates was different during flexion trials as compared to trials involving wrist extension. PMA onset usually preceded movement onset by more than 100 ms and in most instances preceded the average onset of task-related changes in electromyographic (EMG) activity in muscles of the wrist and forelimb. For most neurons, the changes in neuronal activity that began prior to movement were maintained during movement execution. However, approximately one-third of the neurons that exhibited PMA changed their firing rate in the opposite direction, relative to their PMA and to their baseline rate of activity, once the movement began. Several other neurons either exhibited PMA only or they altered their discharge rates during movement execution but did not exhibit PMA. These observations suggest that, despite the close temporal relationship between the onset of PMA and the onset of wrist movement, the neuronal mechanisms mediating the PMA may differ from those that occur during movement execution. The PMA onset of neostriatal neurons occurred earlier in visually cued than in vibratory cued trials. These differences were statistically significant only for flexion trials, however, in which movements were made against a load and in the same direction as the palmar vibratory stimulus. For trials involving wrist extension, PMA onsets for visually cued as compared with vibratory cued trials were not statistically different. These findings contrast with data obtained previously from somatosensory cortical neurons during performance of the same behavioral task. On average, PMA in the putamen began earlier, relative to movement onset, than it did in the somatosensory cortex. Moreover, in the somatosensory cortex, PMA onset occurred earlier in vibratory cued than in visually cued trials, irrespective of movement direction (Nelson 1988; Nelson and Douglas 1989). For putamen neurons, but not for caudate or cortical neurons, the onset of PMA also occurred significantly earlier during extension trials than flexion trials, irrespective of the modality of the "go-cue".(ABSTRACT TRUNCATED AT 400 WORDS)

Single-unit activity in the globus pallidus and neostriatum of the rat during performance of a trained head movement.

Single-unit extracellular neuronal recordings were obtained from the globus pallidus (GP) and the neostriatum (NS) of rats while they performed a learned head movement in response to an auditory cue. In both GP and NS, units that altered their discharge rate in association with head movements and with the cues that triggered these head movements were prevalent. Frequently, the responses were directionally-specific (i.e., the magnitude or direction of change in firing rate of these neurons was substantially different for trials in which head movements were made to the left vs. the right). For some units, firing rates were altered only in response to the movement cue or only in association with head movements. However, the majority of neurons exhibited responses with both cue-related and movement-related components. Neuronal responses to the auditory cue usually were context-dependent, in that they did not occur if the same stimulus was presented when the animal was not performing the task. At least a small proportion of GP and NS neurons also appeared to exhibit context-dependent movement-related activity, in that responses occasionally were observed that were associated either with sensory-triggered head movements or with spontaneous head movements, but not with both. These data are consistent with previous suggestions that the activity of basal ganglia neurons during movement performance is highly dependent on the conditions associated with movement initiation. The data also indicate that the response characteristics of both GP and NS neurons in the rat are generally similar to those that have been described for basal ganglia neurons in primates and cats during sensory-triggered movement tasks. However, the proportion of task-related neurons that exhibited responses with both movement-related and cue-related components was greater than has generally been reported in studies of cats and primates, suggesting that neurons with these response properties may be more predominant in the rat basal ganglia.

Blockade of both D1- and D2-dopamine receptors inhibits amphetamine-induced ascorbate release in the neostriatum.

In vivo recordings with electrochemically modified microvoltammetric electrodes revealed that several neuroleptic drugs, including haloperidol, clozapine, and thioridazine, blocked the rise in extracellular ascorbate produced by amphetamine in the neostriatum of urethane-anesthetized rats. This effect was also observed in animals that received a combined injection of Sch-23390 and sulpiride, but not when either of these drugs were administered alone or in combination with the 5-HT2 blocker, ritanserin. These results indicate that a combined blockade of D1- and D2-dopamine receptors blocks amphetamine-induced ascorbate release.

Heterogeneous responses of neostriatal neurons to amphetamine in freely moving rats.

Single-unit activity was recorded from the neostriatum of unrestrained, behaving rats. Neuronal discharges were found to vary with specific motor responses, general changes in motor activity, or the presentation of orienting stimuli. In each case, however, 1.0 mg/kg D-amphetamine produced approximately equal numbers of excitations and inhibitions. A subsequent injection of a higher dose (5.0 mg/kg) either produced a greater change in firing rate in the same direction or reversed the direction of the low-dose response. Amphetamine, therefore, does not produce uniformly excitatory effects in the neostriatum of ambulant animals. In fact, the neuronal response to amphetamine appears to reflect a complex interaction of several factors, including ongoing behavior and drug dose.

Ascorbate antagonizes the behavioral effects of amphetamine by a central mechanism.

The behavioral response to amphetamine was monitored in rats that received simultaneous intraventricular infusions of saline or ascorbate. Both groups of animals displayed comparable responses, although ascorbate significantly delayed the onset of amphetamine-induced locomotion and rearing. In rats pretreated with a threshold dose of haloperidol (0.025 mg/kg), virtually all aspects of the amphetamine response were attenuated, and this effect was enhanced by ascorbate. In haloperidol-pretreated rats, ascorbate significantly lowered sniffing and forepaw shuffling throughout the amphetamine response. These results suggest that ascorbate antagonizes dopaminergic transmission by a central mechanism.

Enhanced sodium appetite in rats with lesions centered on nucleus medianus.

Recent experiments have demonstrated that rats with lesions of the ventral portion of nucleus medianus (vNM) frequently exhibit a chronic and robust hyperdipsia, which occurs only at night. This study indicates that the same brain damage may produce a nocturnal appetite for sodium that is similarly pronounced and persistent. Of 68 rats with vNM lesions, 33 were observed to drink at least 15 ml of 0.51 M NaCl solution per day, and 11 of them consumed more than 30 ml daily. The basis for this impressive consumption of saline is uncertain; the brain-damaged rats had normal sodium concentrations, renin activities, and aldosterone levels in plasma during basal maintenance conditions, and they conserved sodium in urine when maintained on a sodium-deficient diet. Nevertheless, the present results indicate that vNM and/or local fibers of passage may play an important role in the control of sodium appetite, as it does in the control of thirst.

Oxytocin and vasopressin secretion in response to stimuli producing learned taste aversions in rats.

Administration of lithium chloride, copper sulfate, and apomorphine to rats each stimulated the secretion of oxytocin (OT) and, to a much lesser degree, arginine vasopressin. These agents are assumed to cause visceral illness in rats because of their effectiveness in promoting the acquisition of learned taste aversions. CuSO4 had a greater effect on plasma OT levels when administered ip rather than iv, whereas LiCl did not, results suggesting that LiCl probably stimulates OT secretion by central chemoreceptor activation whereas CuSO4 acts predominantly by local peritoneal irritation. A causal role for circulating OT in the acquisition of learned taste aversions was not found. These and other findings suggest that peripheral levels of OT may represent a quantifiable marker of visceral illness in rats and therefore might be useful in the interpretation of behavioral studies in which learned taste aversions are produced, provided that other stimuli of neurohypophyseal secretion are absent.

Short-term effects of dopamine-depleting brain lesions on spontaneous activity of striatal neurons: relation to local dopamine concentration and behavior.

The spontaneous activity of striatal neurons was measured after dopamine (DA)-depleting brain lesions were produced in rats by the neurotoxin 6-hydroxydopamine. The extent of DA depletion was determined using tissue punches from the same regions of striatum in which cell activity was recorded. Results showed that the spontaneous activity of Type II neurons in either the medial or lateral striatum increased only when local DA depletions exceeded 90%; when local depletions were less than 90%, spontaneous firing rates of Type II neurons were equivalent to control values. This finding was consistent with additional observations that ingestive and motor behaviors of the same animals were not disrupted until striatal DA depletions exceeded 90%. It also was found that spontaneous firing rates of neurons in the lateral but not the medial striatum could be at control levels in animals clearly exhibiting aphagia, adipsia and akinesia.

Impaired secretion of vasopressin and oxytocin in rats after lesions of nucleus medianus.

Regulation of posterior pituitary secretion of vasopressin (AVP) and oxytocin (OT) was studied in rats given electrolytic lesions of ventral nucleus medianus (vNM). As described previously, rats with such lesions were chronically hypernatremic and showed impaired drinking responses to an osmotic challenge. AVP secretion in response to osmotic stimuli also was significantly blunted, although sufficient increases in plasma AVP levels did occur, in association with an abnormally high range of plasma sodium concentrations, to allow urinary concentration comparable to control animals. These findings suggest that vNM lesions cause an upward resetting of the osmotic threshold for AVP secretion. In contrast, hypovolemia, produced by subcutaneous polyethylene glycol treatment, and hypotension, produced by phentolamine treatment, both evoked AVP responses in rats with vNM lesions that were equivalent to those seen in control animals. Plasma OT responses to osmotic and hemodynamic stimuli were analogous to the AVP responses. These findings reproduce the major clinical features observed in humans with the disorder of essential hypernatremia and by doing so support proposals that this disorder is caused by lesions in the vicinity of the anterior hypothalamus that result in selective destruction of afferent osmosensitive inputs to the neurohypophysis.

Modulation of neostriatal activity by iontophoresis of ascorbic acid.

Iontophoresis (20-80 nA) of ascorbic acid (AA) accelerated the firing rate of approximately one-third of the neurons tested in the anteromedial neostriatum of anesthetized rats. When administered to neostriatal neurons that were activated by the simultaneous ejection of glutamic acid (GLU), AA excited more than two-thirds of the cells examined, including many that were not excited by AA alone. At ejection currents above 80 nA, AA further increased the activity of some GLU-activated neurons, but suppressed the firing rate of others. Electrochemical quantification of AA ejection during iontophoresis indicated that the concentration of AA at the tip of the recording electrode was within reasonable physiological limits. It is concluded that endogenous AA may modulate neuronal activity in the neostriatum.

Impaired drinking responses of rats with lesions of nucleus medianus: circadian dependence.

The drinking behavior of rats with electrolytic lesions of ventral nucleus medianus (vNM) was examined during acute hyperosmolality and hypovolemia. The brain-damaged animals were impaired in their drinking responses to systemic treatment with hypertonic saline or polyethylene glycol solution when they were tested during the day. However, apparently normal drinking responses to both dipsogenic challenges were observed when the same animals were pretreated with the stimulant drug, caffeine, or when they were tested at night. These results suggest that lesions of vNM may produce complex alterations in the control of drinking behavior rather than the destruction of sensory receptors. The lesions appear to disrupt both circadian influences on drinking and activational components of drinking that normally serve to facilitate the behavioral response. The present results, together with similar findings for rats given lesions of the subfornical organ, support recent proposals that periventricular tissue bordering the rostral wall of the third cerebral ventricle plays an important role in the central control of drinking.

Hyperdipsia in rats after electrolytic lesions of nucleus medianus.

Ablation of the ventral portion of nucleus medianus (vNM) in rats produced a temporary adipsia or hypodipsia that was accompanied by pronounced urinary fluid losses. When ad libitum drinking resumed, about half of the brain-damaged animals became hyperdipsic, exhibiting chronic two- to threefold elevations in their daily water intakes during the nocturnal hours of the day-night cycle. Rats that remained normodipsic after vNM ablation usually exhibited hyperdipsia if they were food-deprived overnight. The basis for the hyperdipsia produced by vNM ablation was not clear. The elevated water intakes appeared not to result from chronic urinary fluid losses, because hyperdipsic rats were able to concentrate their urine during the day, when they drank little. Moreover, the animals did not seem to be volume depleted; their plasma renin activities were not elevated, and they drank normally in association with meals. These and other findings suggest that vNM lesions damage neural substrates that control drinking behavior, and the hyperdipsia results from this rather than from physiological changes produced by the lesion.