Stereotaxic Apparatus Research Articles

Acetylcholine release in the rostral ventrolateral medulla of spontaneously hypertensive rats.

1. Central acetylcholine (ACh) has been implicated in the pathogenesis of experimental hypertension and the rostral ventrolateral medulla (RVLM) is an important area for cardiovascular regulation. The purpose of this study was to determine whether cholinergic neurotransmission in the RVLM is altered in spontaneously hypertensive rats (SHR). 2. Experiments were performed on male SHR (12-16 weeks) and age-matched Wistar-Kyoto rats (WKY). The rats anaesthetized with pentobarbital were placed in a stereotaxic apparatus. For determining the release of ACh in the RVLM, a dialysis probe was introduced into the RVLM. 3. An RVLM microinjection of cholinergic agents elicited an increase in blood pressure. The pressor response to physostigmine was greater in SHR than that of WKY whereas there was no difference in the pressor response to ACh or carbachol between SHR and WKY. 4. The release of ACh in the RVLM was greater in SHR than that of WKY. Physostigmine (0.5 mg/kg, i.p.) produced increases in medulla ACh contents. The increase in ACh content of the rostroventral medulla including the RVLM was greater in SHR than that of WKY whereas there was no difference in ACh contents of the other three parts of the medulla oblongata between SHR and WKY. 5. Depressor responses to scopolamine injected bilaterally into the RVLM were greater in SHR than those of WKY. 6. These results suggest that ACh release is enhanced specifically in the RVLM of SHR. It appears that this enhanced release of ACh in the RVLM of SHR contributes to the maintenance of hypertension in SHR.

Baclofen-induced block of the Hering–Breuer expiratory-promoting reflex in rats

Baclofen, a gamma-aminobutyric acid (GABA) agonist acting on GABAB receptors, has profound effects on respiratory function. We tested a hypothesis that GABAB receptors are involved in modulation of the respiratory reflexes mediated by pulmonary slowly adapting stretch receptors. Urethane-anesthetized (1.2-1.6 g/kg; i.p.) Wistar rats with body weight 230-320 g were breathing room air supplemented with oxygen. The gas mixture was applied to the T-shaped tracheal cannula at a flow rate of 1.5 L/min. The head of the animals was mounted in a stereotaxic apparatus, and the medulla was exposed. We recorded integrated diaphragmatic EMG (Di) and tracheal and arterial blood pressures. Injections of 100 nL of saline into the nucleus tractus solitarii (NTS) preceded injections of 60 or 110 pmol baclofen in group 1 (n = 14), and 2.8 nmol CGP 35348, a GABAB receptor antagonist, in group 2 (n = 5). Saline had no effects on baseline Di. Occlusion of the tracheal cannula caused an increase in tracheal pressure to 8 cmH2O (1 cmH2O = 98.1 Pa) and provoked a prolongation of expiration (the Hering-Breuer expiratory-promoting reflex) in all drug-free rats. Baclofen abolished the Hering-Breuer reflex in all rats. CGP 35348 reversed the block of the reflex. In group 2, CGP 35348 prevented the effects of baclofen, and alone did not affect respiration. These results support our hypothesis and suggest that GABAB receptors are present on the medullary pathway of slowly adapting mechanoreceptor input. However, GABAB receptors in the rat NTS may not be involved in modulation of the fundamental respiratory function.

Priming of locomotor initiation by electrical stimulation in the hypothalamus and preoptic region in the anesthetized rat

electrical stimulation at a locomotor site can prime (i.e., shorten the latency to initiate) stepping elicited by subsequent stimulation of the same or a different site. We tested for the priming effect in representative sites along the medial forebrain bundle, and determined if its magnitude showed regional differences. Rats ( n = 20) were anesthetized with Nembutal and held in a stereotaxic apparatus over a wheel. Stepping was detected by accelerometers attached to the hindlimbs. Priming and test trains of stimulation (0.5-ms cathodal pulses, 50 Hz, 25–75 MA, 7–9-s train duration) separated by 20 s were delivered every 90 s. When the priming and test stimulations were applied to the same site, the priming effects were similar along the entire extent of the medial forebrain bundle. When the priming and test sites were different, the priming effect depended on their relative positions. Anterior stimulation primed posterior sites at magnitudes comparable to those produced by stimulating the same posterior site. Posterior stimulation primed anterior sites at a level half of that produced by stimulation of the same anterior site. This pattern was found for priming and test sites that were ipsilateral and contralateral. Priming is a general and robust phenomenon with properties that may be useful for studying locomotor initiation pathways.

Expression of developmentally specific isoforms on non-NMDA mRNAs in the hippocampus of schizophrenics

In order to investigate the process of gene expression of N-methyl-d-aspartate glutamate receptor (NMDAR) subunits in the rat neostriatum and how this relates to motor behaviors, a single dose of antisense phosphodiester oligodeoxynucleotide specific for NMDAR1 was unilaterally applied in the neostriatum in a stereotaxic apparatus. After one day of antisense treatment, ipsilateral rotation behaviors that were induced by apomorphine were found in the treated animals. Reductions in the levels of expression of NMDAR1 and NMDAR2A messenger RNAs (NMDAR1: 20.6%; NMDAR2A: 19.7%) were found in the antisense-treated striatal tissues by reverse transcriptase–polymerase chain reaction. There was no change in the levels of NMDAR2B, NMDAR2C and NMDAR2D messenger RNAs. After two days, western blotting experiments showed that there were decreases in the levels of expression of NMDAR1 (decreased 27.6%) and NMDAR2A (decreased 19.2%) proteins in the NMDAR1 antisense-treated striatal tissues. In addition, NMDAR1 immunoreactivity was found to decrease in intensity in the NMDAR1 antisense-treated neostriatum. At the cellular level, the intensity of NMDAR1 immunoreactivity in perikarya of presumed medium spiny neurons was found to decrease.These results indicate that a single dose of NMDAR1 antisense modifies the expression of NMDAR1 messenger RNA and protein in neurons in the neostriatum. The modification in the expression of NMDAR1 has differential effects in the expression of NMDAR2 subunits. Gene expression of the native NMDAR subunits is likely to be a dynamic process. The change in gene expression of the NMDAR subunits in the neostriatum may have a profound effect on the motor behaviors of rats.

Activity of neurons in the lateral preoptic area during drinking behavior by the rat

Rats were trained to drink water, sucrose solution, and saline while their heads were painlessly held in a stereotaxic apparatus with an attachment fixed to the skull. A total of 286 neurons in the lateral preoptic area (LPO) were recorded during the drinking behavior. During water intake, 40% of the neurons tested were either excited or inhibited. Sucrose elicited responses in 25% of the neurons, and saline in 34%. Of the 52 neurons recorded while rats, drank all test solutions, 8 specifically responded to water, 3 were sucrose specific, 4 were saline specific, 12 responded to all solutions, and the remaining 25 did not respond to any solutions. Artificial cerebrospinal fluids (ACSFs) with physiologically hypertonic (+30 mOsm/kg, by NaCl or mannitol) and hypotonic (−30 mOsm/kg) osmotic pressure were applied to 88 neurons through a multibarrel micropipette. Of these neurons, 15 (17%) were hypotonic-sensitive; they were excited by hypotonic ACSF and/or inhibited by hypertonic ACSFs. There were 4 (5%) that were hypertonic-sensitive; they were excited by hypertonic ACSFs and/or inhibited by hypotonic ACSF. Of six hypotonic-sensitive neurons examined, three were water specific, and none were sucrose nor saline specific. Of four hypertonic-sensitive neurons, one was water specific, and another was saline specific. These results showed activity of some LPO neurons specifically related to water intake. Osmosensitive neurons in the LPG may regulate water intake.

Minimally Invasive Neurosurgery Using CRW-3 Stereotaxy

The Cosman-Roberts-Wells (CRW-3) scanner independent stereotactic system is a recent arc-radius design developed from the Brown-Roberts-Wells system. The results of 74 supratentorial non-basal tumor suspect cases treated with this second generation computed tomography (CT) guided stereotactic apparatus are presented. Using a simple stereotactic target classification coupled with biopsy, trephine or mini-craniotomy, it has been possible to biopsy, excise and develop new strategies using microneurosurgical techniques with extremely low morbidity.

Stimulation of prolactin secretion in the pig: central effects of relaxin and the antiprogesterone RU 486

Our previous study has shown that oral administration of a potent progesterone antagonist, RU 486, caused a marked elevation of plasma concentrations of both PRL and progesterone in hysterectomized pigs bearing aging corpora lutea. Hysterectomized pigs (hysterectomy performed on day 8; estrus = day 0) were subjected to cranial surgery for chronic placement of a head-mounted stereotaxic apparatus for intracerebroventricular (icv) administration of relaxin (300 U once daily on days 111 and 113; n = 6) and RU 486 (4 mg once daily on days 111, 113, and 115; n = 5) to test whether relaxin and RU 486 exert their actions within the central nervous system and/or pituitary gland to affect PRL and GH secretion. Control pigs (n = 3) received icv injection of vehicle. Intensive blood sampling revealed that icv injection of relaxin on day 111 markedly increased the plasma PRL concentration from 8 to 38 ng/ml within 10 min (P < 0.01). An identical icv injection of relaxin on day 113 caused only a modest increase in PRL, but the overall mean concentration of PRL after relaxin treatment was greater than that before treatment (14 vs. 8 ng/ml; P < 0.05). Intracerebroventricular injection of RU 486 on day 111 greatly elevated plasma PRL. The increase in PRL lasted more than 2 h, with several peak increases of 18-29 ng/ml (P < 0.01). The PRL response to subsequent icv infusion of RU 486 on days 113 and 115 was blunted, but the overall mean concentration of PRL (14 ng/ml) after icv injection of RU 486 remained greater (P < 0.01) than that before treatment (9 ng/ml). In contrast, PRL concentrations in the control group remained unchanged after injection. Plasma concentrations of GH, relaxin, and progesterone were significantly altered in neither hormone- nor vehicle-treated groups during this brief period of sequential blood sampling. This study provides strong evidence that relaxin has a central role in modulating PRL secretion in the pig. In addition, the antagonistic effects on progesterone receptor by RU 486 in the central nervous system and/or pituitary gland caused an abrupt increase in PRL secretion in these hysterectomized gilts.

Stimulation of prolactin secretion in the pig: central effects of relaxin and the antiprogesterone RU 486.

Our previous study has shown that oral administration of a potent progesterone antagonist, RU 486, caused a marked elevation of plasma concentrations of both PRL and progesterone in hysterectomized pigs bearing aging corpora lutea. Hysterectomized pigs (hysterectomy performed on day 8; estrus = day 0) were subjected to cranial surgery for chronic placement of a head-mounted stereotaxic apparatus for intracerebroventricular (icv) administration of relaxin (300 U once daily on days 111 and 113; n = 6) and RU 486 (4 mg once daily on days 111, 113, and 115; n = 5) to test whether relaxin and RU 486 exert their actions within the central nervous system and/or pituitary gland to affect PRL and GH secretion. Control pigs (n = 3) received icv injection of vehicle. Intensive blood sampling revealed that icv injection of relaxin on day 111 markedly increased the plasma PRL concentration from 8 to 38 ng/ml within 10 min (P < 0.01). An identical icv injection of relaxin on day 113 caused only a modest increase in PRL, but the overall mean concentration of PRL after relaxin treatment was greater than that before treatment (14 vs. 8 ng/ml; P < 0.05). Intracerebroventricular injection of RU 486 on day 111 greatly elevated plasma PRL. The increase in PRL lasted more than 2 h, with several peak increases of 18-29 ng/ml (P < 0.01). The PRL response to subsequent icv infusion of RU 486 on days 113 and 115 was blunted, but the overall mean concentration of PRL (14 ng/ml) after icv injection of RU 486 remained greater (P < 0.01) than that before treatment (9 ng/ml). In contrast, PRL concentrations in the control group remained unchanged after injection. Plasma concentrations of GH, relaxin, and progesterone were significantly altered in neither hormone- nor vehicle-treated groups during this brief period of sequential blood sampling. This study provides strong evidence that relaxin has a central role in modulating PRL secretion in the pig. In addition, the antagonistic effects on progesterone receptor by RU 486 in the central nervous system and/or pituitary gland caused an abrupt increase in PRL secretion in these hysterectomized gilts.

Microstimulation mapping of the basal forebrain in the anesthetized rat: The “preoptic locomotor region”

Previous studies have indicated that the basal forebrain at the level of the preoptic area contains neurons which participate in the initiation of locomotion. This study attempted to localize those neurons by mapping sites at which 25- and 50-μA stimulation (50 Hz, 0.5 ms cathodal pulses, 10-s trains) initiated hindlimb stepping. Anesthetized rats were held in a stereotaxic apparatus supported by a sling so that stepping movements rotated a wheel. Anesthesia was maintained by periodic injections of Nembutal (7 mg/kg) supplemented by lidocaine injections. Stimulation was applied through 50–70-μm diameter pipettes filled with 2 M NaCl at approximately 1600 sites in the basal forebrain, adjacent thalamus, and striatum. A circumscribed grouping of 25-μA locomotor sites, centered in the lateral preoptic area, defined the preoptic locomotor region. It extended into the ventral bed nucleus of the stria terminalis, the lateral part of the medial preoptic area, the anterior hypothalamic area, the medial and rostral parts of the ventral pallidum, medial substantia innominata, and the horizontal limb of the diagonal band. This general region is known to project to the midbrain locomotor region and the ventral tegmental area; it is proposed to initiate locomotion in service of primary motivational systems. Among the structures generally negative for locomotor sites were the dorsal and ventral striata, septal complex, bed nucleus of stria terminalis, and lateral ventral pallidum and substantia innominata. These findings indicate that low current stimulation applied to a circumscribed area centered in the lateral preoptic area produces locomotor stepping in the anesthetized rat. Whether the activated elements in this preoptic locomotor region are cells or fibers is not yet known. The degree of localization afforded by these findings indicates that the areas that are most likely to contain the mediating elements are quite limited in extent.

Arousal-dependent properties of medial septal neurons in the unanesthetized rat

We have performed a qualitative and quantitative analysis of the electrophysiological properties of medial septal neurons in the unanesthetized rat. The rat's head was held in a stereotaxic apparatus by a painless head-restrained system that was implanted seven days prior to the recording sessions. Extracellular recordings were made in a mixed population of antidromically identified septohippocampal neurons and unidentified medial septal neurons in different states of arousal and in response to peripheral and reticular stimulations. The spontaneous activity as well as the percentage of rhythmically bursting septal neurons varied significantly according to the state of arousal. Higher values were noted in paradoxical sleep (28 imp/s and 94% of bursting neurons) as compared with wakefulness with hippocampal theta rhythm (17.4 imp/s and 64.2% of bursting neurons) and slow wave sleep (12.3 imp/s and 8% of bursting neurons). The frequency of the bursts was significantly higher during paradoxical sleep. In individual medial septal neurons, arousing stimuli and paradoxical sleep could induce rhythmic bursting activity in previously non-bursting neurons provided that they were fast-firing neurons. No differences were noted in the functional characteristics of neurons in the medial septal nucleus as compared with the diagonal band of Broca. When the unanesthetized rats were compared with a group of urethane-anesthetized rats, the spontaneous activity was higher and more irregular in the absence of anesthesia. The percentage of the bursting neurons was significantly lower in the unanesthetized rats (32.3% vs 43.3%). However, the frequency of the bursts was higher (5.9 ± 0.1 Hz vs3.5 ± 0.1 Hz). Since the patterns of activity of medial septal neurons fluctuate in different physiologically relevant states, previous classifications of these neurons made by ourselves and other authors, in urethane-anesthetized rats, may not be appropriate

Decreased cardiorespiratory effects of neuropeptide Y in the nucleus tractus solitarius in diabetes.

It has been observed that diabetes results in increased neuropeptide Y (NPY) in various brain regions, especially the paraventricular nucleus, which projects to the nucleus of the solitary tract (NTS). Because previous studies indicated a pathophysiological relationship between diabetes and NPY, we investigated the effect of diabetes on the sensitivity of NTS-mediated responses to NPY administration. Rats were made diabetic using streptozocin (55 mg/kg iv) and maintained for 48 to 50 days. Normal and streptozocin-diabetic rats were anesthetized with urethan and alpha-chloralose, instrumented for cardiovascular and respiratory monitoring, and positioned in a stereotaxic apparatus. The brain stem was exposed surgically. NPY (0.15 nmol/kg) was microinjected into the NTS and the cardiovascular and respiratory parameters were monitored for 60 min. Diabetes increased systolic (SAP), diastolic (DAP), and mean (Pa) blood pressure but not pulse pressure (PP) and heart rate (HR). Respiratory parameters were not altered. NPY significantly decreased SAP, DAP, Pa, PP, HR, respiratory rate, and minute volume in normal animals. In diabetic animals, NPY also decreased SAP, DAP, and Pa but pronouncedly increased PP. Although NPY decreased the SAP and Pa in diabetic animals, the response was attenuated compared with normal animals. The respiratory parameters and HR of diabetic animals, unlike normal animals, did not respond to NPY administration. We conclude that chronic diabetes results in a decreased sensitivity to NTS-mediated responses and that the hyporesponsiveness of the NTS to NPY modulation may be important in the tendency toward elevated blood pressure and hypertension in diabetes.

Locomotor sites mapped with low current stimulation in intact and kainic acid damaged hypothalamus of anesthetized rats

To determine whether local neurons mediated the locomotor effects of electrical stimulation of the lateral hypothalamus, kainic acid injections (0.5-1.25 micrograms), intended to destroy neural somata as opposed to fibers of passage, were made unilaterally in the tuberal-posterior hypothalamus of 22 rats. The area of lesion and its contralateral homolog were mapped for locomotor stepping sites in Nembutal-anesthetized rats mounted in a stereotaxic apparatus such that locomotor stepping rotated a wheel. Stimulation (25 and 50 microA, 50 Hz, 0.5-ms cathodal pulses, 10-s trains) was delivered through 50-80 microns glass pipettes filled with 2 M saline. Contralateral to the lesion, locomotor stepping sites were common in the perifornical lateral and medial hypothalamus and less dense in the zona incerta. On the side of the kainic-acid lesion, locomotor sites were generally absent in the central part of the damaged area. If they did appear within the area of lesion, they tended to be near the border with intact tissue. In a few cases, locomotor stepping sites were found centrally located in the lesion amidst widespread loss of somata. In four rats, additional maps of anterior locomotor regions in the preoptic area ipsilateral to the lesion suggested that their descending fibers were largely spared by the kainic lesions. Local neurons appear to be major contributors to the locomotion elicited by electrical stimulation of the lateral hypothalamus, but fibers of passage may also participate.

A stereotaxic atlas and implantation technique for nuclei of the diencephalon of Atlantic salmon (Salmo salar) parr

A stereotaxic apparatus and technique for its implantation in diencephalic nuclei of Atlantic salmon parr of 20 to 30 g body weight is described. An atlas of nuclei in the diencephalon is also presented.

Stimulation of the brainstem reticular formation evokes locomotor activity in embryonic chicken (in ovo)

This study was designed to examine the period of embryonic chick development during which descending brainstem-spinal projections, originating from defined avian brainstem locomotor regions, become functionally active. Locomotor activity was examined using a new in ovo preparation for the focal electrical stimulation of embryonic brainstem locomotor regions. Embryos or hatchlings were anesthetized and mounted in a stereotaxic apparatus. Leg and wing muscle electromyographic (EMG) recordings were used to monitor any brainstem-stimulated motor activity. At present, we have been successful in demonstrating coordinated brainstem-evoked locomotion in embryos as early as embryonic day 15. The patterns of evoked locomotor activity were similar to locomotion evoked in hatchling chicks and were of 4 types: (1) alternating hindlimb movements (‘stepping’), (2) synchronous (in-phase) hindlimb movements (‘hatching’), (3) synchronous wing movements (‘flapping’), and (4) simultaneous ‘stepping’ and ‘flapping’. The cycle durations of evoked embryonic hindlimb movements are shorter than those observed for hatchling chicks. The present results are the first direct demonstration of functional connections between descending supraspinal neurons and spinal locomotor circuits at such an early stage of embryonic development. With modifications in technique, it may be possible to demonstrate functional connections at even earlier stages of embryonic development.

Direct Cerebrospinal Fluid Delivery of an Antiretroviral Agent Using Multivesicular Liposomes

The use of multivesicular liposomes for administration of antiviral agents into cerebrospinal fluid was explored in a Sprague-Dawley rat model. DDC (2',3'-dideoxycytidine) was encapsulated into multivesicular liposomes made from dioleoyl lecithin, dipalmitoyl phosphatidylglycerol, cholesterol, and triolein. The half-lives of drug leakage in human plasma and in 0.9% NaCl were 15 and 47 h, respectively. After intraventricular injection with a stereotaxic apparatus, DDC levels within the central nervous system decreased exponentially, with a half-life of 1.1 h for the unencapsulated DDC and 23 h for the liposome-encapsulated DDC. There were no abnormalities observed in the behavior of the rats. Encapsulation of a more hydrophilic antiviral agent is expected to increase the half-life even further. The results of this study offer the possibility of a practical intrathecal drug delivery for drugs that do not cross the blood-brain barrier.

Latency to initiate locomotion elicited by stimulation of the diencephalon positively correlates in awake and anesthetized rats

Locomotor stepping can be elicited by brain stimulation at various diencephalic sites under moderate levels of Nembutal. This study determined if locomotor initiation measured under anesthesia provides a valid measure of the intersite factors which determine initiation in the awake condition. We compared the latencies to inititate locomotor stepping elicited by electrical stimulation (50 μA, 0.5-msec pulses, 10 to 160 Hz) by rats tested while awake and unrestrained in a rotary runway or anesthetized and held in a stereotaxic apparatus. In the latter tests, initial anesthesia was provided by Nembutal (25 mg/kg) and 2% halothane and maintenace anesthesia was provided by 7 mg/kg as needed and local injections of lidocaine. For 30 sites in 16 rats, average locomotor initiation latency in the awake condition and the shortest latencies in the anesthetized condition were positively correlated (r=.78). Locomotion at sites with long latencies in the awake condition was frequently blocked in the anesthetized condition, but sites with short latencies were rarely blocked. The results indicate that the shortest locomotor latencies in the anesthetized condition approximate the latencies measured in the awake condition. It is concluded that the anesthetized condition can provide valid initiation measures, but sites with long latencies in the awake condition are prone to depression under anesthesia.

Locomotor stepping elicited by electrical stimulation of the hypothalamus persists after lesion of descending fibers of passage

Locomotion initiated by electrical stimulation of the lateral hypothalamus could be due to activation of local neurons or of fibers of passage descending from locomotor regions in the basal forebrain. This study mapped hypothalamic sites for electrically elicited locomotion in six rats with electrolytic lesions of the ipsilateral basal forebrain sources of descending fibers of passage. For mapping, anesthetized rats were held in a stereotaxic apparatus supported by a sling so that stepping movements rotated a wheel. Anesthesia was maintained by periodic injections of Nembutal (7 mg/kg) supplemented by lidocaine injections. Stimulation (25 and 50 μA, 50 Hz, 0.5 msec cathodal pulses, 10 sec trains) was applied through 50–80 μm diameter pipettes filled with 2 M saline. In all cases, locomotor stepping could be elicited by stimulation in sites ipsilateral to the lesion at currents of 50 μA or less. In the one case in which 25 μA sites were not found in the lateral hypothalamus, the lesion extended caudally to within 1 mm of the stimulation sites. These findings do not exclude a locomotor role for fibers of passage but they suggest that activation of lateral hypothalamic neurons is sufficient to initiate locomotion.

Effects of iontophoretically applied monoamines on somatosensory cortical neurons of unanesthetized rats

The response of somatosensory cortical neurons to iontophoretic applications of monoamines was studied in unanesthetized rats. The animal's head was held in a stereotaxic apparatus by means of a painless head-restraining system implanted 8 days prior to the recording sessions. The electrodes consisted of a recording micropipette attached to a multibarreled iontophoresis micropipette. The electrode penetrations were reconstructed on camera lucida drawings of frontal brain sections. The percentage of cortical neurons responding to application of monoamines were 76% after noradrenaline, 58% after dopamine and 66% after serotonin. The differences observed among percentages of responses seemingly correlate with the relative abundance of terminal axons and receptors for each of the three monoamines in the somatosensory cortex. The vast majority of the responding neurons were inhibited by monoamines and this inhibitory effect was independent of the level of spontaneous activity. The depressant effect of the monoamines on glutamate and acetylcholine-evoked responses supports a modulatory role for these substances. Serotonin was the most potent, followed by noradrenaline and dopamine. The present study shows that when the influence of anesthesia is eliminated, the predominant effect of monoamines on cortical first somatosensory neurons is one of inhibition. These findings contrast with results obtained under some anesthetic conditions, as well as under in vitro conditions.

Effects of acetylcholine on single cortical somatosensory neurons in the unanesthetized rat

Experiments have been performed on unanesthetized rats using a chronic restraint system. The animal's head was held in a stereotaxic apparatus by means of two metallic tubes fixed on the skull with dental cement. Electrodes consisted of a recording micropipette (filled with 1 M NaCl and 2% Pontamine Blue) attached to a multibarreled micropipette for iontophoresis. Electrode penetrations were reconstructed on camera lucida drawings of frontal brain sections. The overall percentage of spontaneously active somatosensory neurons was 77% with a mean spontaneous activity of 5.9 impulse/s ( n = 405). Yet differences were observed in the proportions of active cells as well as in the mean spontaneous activity between cortical layers (both parameters being significantly higher in layers V and VI). Comparison with results obtained under urethane anesthesia [Dykes R.W. and Lamour Y. (1988) J. Neurophysiol. 60, 703–724] shows that the percentage of the spontaneously active neurons and the mean spontaneous activity were both significantly higher in unanesthetized rats (77 vs 36%; 5.9 vs 2.6 impulse/s). Nevertheless, the laminar distribution of the most active cells was similar under both conditions. In the present study, 52.3% of the neurons ( n = 380) were excited by acetylcholine and 46% ( n = 198) by carbachol. Significantly larger percentages of neurons excited by acetylcholine were found in layers Vb and VIb. These effects of cholinergic agonists—observed for the first time in unanesthetized rats—differed significantly from those previously obtained under anesthesia (33 and 34% of neurons excited by acetylcholine and carbachol, respectively) [Lamour Y. et al. (1982) Neuroscience 7, 1483–1494]. In the absence of anesthesia, cholinoceptive neurons were significantly more numerous in layers II–III. However, under both conditions, the highest percentages of excitatory responses were observed in layers Vb and VIb. A strong relation between the level of spontaneous unitary activity and the probability of being excited by acetylcholine was observed.

Locomotor stepping initiated by glutamate injections into the hypothalamus of the anesthetized rat.

Glutamate (50 mM, 50 nl) injected into the tuberal and posterior hypothalamus was tested for capacity to elicit locomotor stepping. Rats (n = 23) were anesthetized with Nembutal and suspended by a sling in a stereotaxic apparatus such that locomotor stepping rotated a wheel. In 61 of 275 sites tested, stepping was initiated by glutamate injections within 60 s. Positive sites were widespread and contained in the lateral hypothalamus, the perifornical area, the dorsomedial nucleus, and the zona incerta. The perifornical and lateral hypothalamic sites were most likely to have locomotor responses and the shortest latencies. These findings indicate that selective activation of hypothalamic neurons as opposed to fibers of passage can initiate locomotion.