Suppression of locomotor activity produced by acute injections of kainic acid into the median raphe nucleus

Several lines of evidence suggest that cells within the immediate vicinity of the median raphe nucleus may exert an inhibitory influence on locomotor activity. If this theory were correct, one would expect that excitation of neurons within the median raphe would have a suppressive effect on behavior. This possibility was examined in the present study where low doses of the glutamate analogue kainic acid were injected into the median raphe nucleus of methylphenidate-pretreated rats. Our results indicate that these injections were able to antagonize, in a dose-dependent manner, both the horizontal locomotor activity and the nose poking induced by methylphenidate. These results provide further support for the existence of inhibitory median raphe influences on locomotion and suggest the possibility that endogenous excitatory amino acids may play a role in raphe functioning.     

Hippocampal theta activity and behavioral sequences in a reward-directed approach locomotor task

Hippocampal rhythmic slow wave activity (theta) has been implicated in the processing of stimuli associated with movement. This study determined whether the theta rhythm showed phase relationships or changes in amplitude and frequency with the onset of stimuli and behavioral sequences in a skilled locomotor approach task. Rats with bipolar electrodes spanning CA1 approached a stall, turned to enter it, approached and depressed a treadle, waited 1.35 s, and approached a milk reward located forward either to the right or to the left. Auditory cues indicated the location of the reward during the waiting period and at the reward onset. A video capture system (20-ms sampling) was synchronized to the hippocampal recording system (10-ms sampling). Behavioral events identified by motion analysis were used to generate averages of hippocampal slow wave activity, theta peak amplitudes, and intervals between peaks. Theta activity at 8-10 Hz was almost continuous during the behavioral sequences. Phase relations with stimuli or movement onsets occurred infrequently and were not consistent across the four subjects. Theta peak amplitude and frequency decreased as the rat slowed locomotion in the stall and reached the treadle. Onset of locomotion directed to a reward location occurred on a positive peak of averaged theta activity. When locomotion had short latencies, increases in theta frequency appeared after the onset but, when it had longer latencies, frequency increases appeared 200 ms before onset. The results indicate that the execution of instrumental movement modulates both theta amplitude and frequency, and that the preparation for locomotion modulates theta frequency.     

Hippocampal innervation by serotonin neurons of the midbrain raphe in the rat.

The organization of the brainstem serotonin neuron projection to the hippocampal formation was analyzed in the rat. This projection arises in the raphe nuclei of the midbrain. Following destruction of the midbrain raphe nuclei, chiefly nucleus centralis superior, there is a 72% decrease in hippocampal serotonin content. Injection of tritiated amino acid into the midbrain raphe nuclei results in transport of tritiated protein to the hippocampal formation and this transport is blocked in animals pretreated by intraventricular administration of 5,6-dihydroxytryptamine (5,6-DHT). Autoradiographic analysis indicates that the transport reaches the hippocampal formation primarily via two major pathways, the cingulum and the fornix. Cingulum fibers terminate predominantly in the dorsal hippocampus whereas the fornix distributes throughout the entire hippocampal formation. Some fibers reach the ventral hippocampus from the entorhinal area. Within the hippocampus there is dense labeling in a restricted lamina of the CA1 stratum lacunosum-moleculare with moderate labeling in stratum radiatum. Stratum oriens is sparsely labeled in CA1 and moderately so in CA2 and CA3. Stratum radiatum and stratum lacunosum-moleculare are moderately densely labeled in CA2 and Ca3. The area dentata is sparsely to moderately labeled in the molecular layer and heavily labeled in a thin lamina of the hilar zone immediately beneath the granule cell layer. The remaining hilar zone is moderately labeled. All of the discrete labeling of the hippocampus and area dentata described above is absent in animals pretreated with 5,6-DHT. These observations indicate that serotonin neurons of the midbrain raphe provide a highly organized innervation of the hippocampal formation in the rat.     

Hippocampal theta activity in monkeys

The hippocampal theta rhythm has been extensively studied in many subprimate mammals. Considering the technical difficulties involved in recording from freely moving animals during voluntary motion and REM sleep, it was thought that urethane anesthesia might be appropriate for initial studies of the primate hippocampal EEG. Three of three macaques and one of two squirrel monkeys showed clear rhythmic hippocampal EEG activity. One very old squirrel monkey (a 16-year-old female) showed no theta activity in the hippocampal EEG. Similarities of the monkey theta activity with theta rhythm of urethane-anesthetized rats included: (1) a high coherence between recordings from electrodes separated by several millimeters within the hippocampal formation; (2) sensitivity of the theta activity to muscarinic drugs; and (3) its correlation with spontaneous movements during light anesthesia. Important differences were: (1) the frequency of the monkey theta activity was 7-9 Hz compared to the 4-5 Hz found in rats; (2) theta activity was not detected in the distal apical dendritic regions of CA1 or dentate in the monkey; (3) considerable amounts of low-frequency EEG co-existed with the monkey theta activity; and (4) the durations of bouts of theta activity in monkeys were much shorter than in rats. We conclude that primates generate hippocampal theta activity homologous, but not identical, to that of rats.     

Association of the mesencephalic locomotor region with locomotor activity induced by injections of amphetamine into the nucleus accumbens

Injections of amphetamine into the nucleus accumbens increased locomotor activity of rats. Subsequent injections of procaine into the midbrain, in the region of the pedunculopontine nucleus, significantly reduced the amphetamine-induced locomotor activity. Control experiments showed that procaine injections into the contralateral pedunculopontine nucleus had little or no effect, as well as ipsilateral injections dorsal and ventral to the pedunculopontine nucleus. These findings suggest that release of dopamine from amphetamine injections into the accumbens gives rise to ipsilateral descending influences on the region of the pedunculopontine nucleus, a major component of the mesencephalic locomotor region. Descending influences from the nucleus accumbens to mesencephalic locomotor region may serve as a link for limbic-motor integration in behavioral response initiation.     

Deficits in locomotor behaviour and thermoregulation produced by intrahypothalamic dopamine injections

When nigrostriatal dopamine neurones degenerate, a loss of functional dopamine in the striatum occurs and is accompanied by increased dopamine in the degenerating axons which traverse the hypothalamus. While the behavioural deficits which occur after nigrostriatal degeneration have been attributed to the loss of functional dopamine neurotransmission, evidence produced by us suggests that the increased levels of amines in the degenerating axons may be neuroactive and participate in the production of these behavioural deficits. To test this hypothesis further, albino rats were injected bilaterally with 200 nmol of dopamine in a location just rostral to the diencephalon/mesencephalon border, where amine accumulation is commonly observed following lateral hypothalamic damage. The effect of these injections upon open field performance, thermoregulation and motor reflex control was determined 40 min after dopamine injection. In a second study, pargyline (15 mg/kg. i.p.) was administered 30 min before intracerebral dopamine to determine whether this treatment would increase the severity of motor and thermoregulatory deficits which occurred after dopamine injections alone. Deficits in locomotion, rearing and the ability to regulate body temperature were seen after the dopamine injections while motor reflex control in these animals was similar to that seen in vehicle-injected controls. The behavioural deficits displayed by pargyline pretreated, dopamine injected animals were slightly but not significantly more severe than those displayed by animals receiving dopamine injections alone. Fluorescent histochemical assessment of injection sites revealed that dopamine injection produced an increase in fluorescence or “amine accumulation” at the site of injection but this was considerably less than that seen after catecholamine degeneration. These results add further support to the hypothesis that amines which accumulate in degenerating neurones may be neuroactive and may thereby participate in the production of behavioural deficits attributed previously to the loss of functional dopamine neurotransmission.     

Hippocampal theta activity in the acute precollicular rat

In eleven acute precollicular rats, cortical and hippocampal EEG activity was recorded. Hippocampal theta activity of low frequency (mean 4.5 c/sec) was found in nine rats and in four occupied above 50% of the recording time. In contrast, in the cortex the synchronized EEG activity dominated. The electrical stimulation of the posterior hypothalamus induced the theta rhythm, or increased its frequency. The large amounts of theta activity in some rats suggests that the forebrain theta pacemakers, partly modulated by the posterior hypothalamus, are released from brain stem inhibitory influences.     

Circadian activity rhythms in rats with midbrain raphe lesions

The dorsal and median mesencephalic raphe nuclei provide a robust projection to the hypothalamic suprachiasmatic nucleus, the site of a putative neuronal circadian pacemaker. Although it has been suggested that the raphe may play a role in the circadian timing system, this role has not yet been specified. In the present report, we examined the circadian activity patterns of rats with large midbrain lesions aimed at the median and dorsal raphe nuclei under conditions of light-dark entrainment, and while free-running in constant light and constant darkness. The results indicate that midbrain raphe lesions may interface with the expression of free-running circadian activity rhythms.     

Muscle tone suppression and stepping produced by stimulation of midbrain and rostral pontine reticular formation

Stimulation of the midbrain retrorubral (RRN), ventral paralemniscal tegmental field (vFTP), reticular tegmental (TRN), and pedunculopontine tegmental (PPN) nuclei was found to produce bilateral suppression of muscle tone in the unanesthetized, decerebrate animal. The RRN is the most rostral area shown to produce such suppression. This muscle tone suppression was frequency- and intensity-dependent. At low stimulus intensities, bilateral suppression was produced at these sites. At higher current and frequency levels, 2 types of muscle responses were found, excitation in PPN and RRN and initial suppression followed by excitation in TRN and vFTP. The mean latency to muscle tone suppression was not significantly different in TRN (36.8 msec) and RRN (36.5 msec). However, muscle tone suppression latency was significantly shorter in vFTP (31 msec) and PPN (27.1 msec). In addition to muscle tone suppression, stepping-like activity could be elicited at the same points by consecutive train stimulations in PPN and single train stimulation in TRN and vFTP. Thus, systems producing atonia are colocalized with those producing locomotion. We hypothesize that the midbrain atonia regions control more caudal regions producing muscle tone suppression in REM sleep, and that the locomotor and atonia eliciting regions are normally coactivated during REM sleep.     

Effects of midbrain raphe stimulation and lesion on EEG activity in rats

In order to clarify whether the dorsal raphe (d-R) and medial raphe nuclei (m-R) of the midbrain play a different role in EEG activity, the effects of either electrical stimulation or lesion of each raphe nucleus were examined in rats. After m-R stimulation, EEG activity changed to a marked drowsy pattern. D-R stimulation, however, failed to produce such effects. M-R lesion elicited a significant arousal in EEG. The change in hippocampal EEG was the most characteristic, i.e., synchronized waves with an extreme increase in voltage. D-R lesion did not produce such a change. The hippocampal EEG arousal was easily abolished by L-5-hydroxytryptophan without any change in the cortical EEG or behavior. These results suggest that the m-R plays a more important role than the d-R in regulating the hippocampal EEG activity, especially in inhibiting the appearance of high voltage synchronized waves in the hippocampus.     

GABA injected into the anterior dorsal tegmentum (ADT) of the midbrain blocks stepping initiated by stimulation of the hypothalamus

Previous work showed that the activity rates of certain neurons in the anterior dorsal tegmentum (ADT) of the midbrain correlated with the onset of stepping elicited by hypothalamic stimulation. This study determined if reversible inactivation of the ADT would block locomotion elicited by hypothalamic stimulation of anesthetized rats (urethane, 800 mg/kg). GABA (concentrations 0.25–1.0 mg/μl in saline) were injected in 52 sites in 21 rats. GABA at volumes of 0.1 or 0.2 μl blocked hindlimb stepping in 18 cases. Locomotor blocks occurred within 5 min of the injection, and typically recovered within 10–20 min. The effective blocking sites were clustered around the interstitial nucleus of the medial longitudinal fasciculus. Sites more dorsal and more anterior were not as effective as sites in and ventral to this nucleus. The data are consistent with a role for the ADT of the midbrain in locomotor initiation.     

Hippocampal theta activity related to elicitation and inhibition of approach locomotion

This study determined if the hippocampal theta rhythm showed phase relationships or changes in amplitude and frequency with the onset of stimuli and locomotion in a task in which auditory cues initiated and suppressed approach locomotion. Rats with electrodes in the dorsal hippocampus lapped at a milk dipper and were presented a tone which predicted the delivery of a food pellet. In some trials the pellet cue tone was negated by 60-Hz clicks beginning 0.3 s after onset, and no pellet was delivered. A video capture system (20-ms sampling) synchronized to the hippocampal recording system (10-ms sampling) was used to determine the onset of locomotor approach to the pellet area. The findings failed to support proposals that phase-related mechanisms play a role in encoding and retrieval of movement-related information. Neither the pellet cue nor the negating cue reset the theta rhythm, and they did not produce differential evoked potentials. During milk lapping, theta amplitude increased in the 1/2 s prior to all pellet cues regardless of their locomotor effect. Frequency also rose but only when a non-negated pellet elicited short-latency locomotion. During locomotor execution, theta peak amplitude peaked earlier than theta frequency by approximately one period. In general during performance of this task, increasing theta amplitude reflected a general preparation to process the cue and increasing theta frequency reflected the readiness to respond to the cue with locomotion.     

Identification of the midbrain locomotor region and its relation to descending locomotor pathways in the Atlantic stingray, Dasyatis sabina.

The midbrain locomotor region (MLR) in the Atlantic stingray, Dasyatis sabina, was identified and characterized. Stimulation (50-100 microA, 60 Hz) of the midbrain in decerebrated, paralyzed animals was used to elicit locomotion monitored as alternating activity in nerves innervating an antagonist pair of elevator and depressor muscles. Effective sites for evoking locomotion in the midbrain included parts of several nuclei: the caudal portion of the interstitial nucleus of the medial longitudinal fasciculus and the caudomedial parts of the cuneiform and subcuneiform nuclei. This region did not include the red nucleus, any parts of the optic tectum or the medial or lateral mesencephalic nuclei. Electrical stimulation in the MLR evokes locomotion in either the ipsi- or contralateral pectoral fin, whereas stimulation in the medullary reticular formation evokes locomotion only in the contralateral fin. Lesion experiments were performed to identify the location of descending pathways from the midbrain to the medullary reticular formation. To abolish locomotion evoked by electrical stimulation in the MLR, the medial reticular formation in the rostral medulla had to be lesioned bilaterally, or the ipsilateral medial medullary reticular formation and fibers projecting from the MLR to the contralateral midbrain had to be disrupted. Injections of HRP into the magnocellular/gigantocellular reticular formation confirmed that this area received bilateral projections from the MLR. The MLR of the Atlantic stingray appears to be similar to the lateral component of the mammalian MLR and to the MLR in other non-mammalian vertebrates.     

Interactions between naloxone and GABA in the control of locomotor activity in the rat

Summary It was found that naloxone causes a small but significant reduction of motility. The GABA_B agonist baclofen and the GABA transaminase inhibitor-acetylenic GABA (GAG) also reduced locomotor activity. When a subeffective dose of baclofen was combined with naloxone 0.8 or 3.2 mg/kg, baclofen significantly inhibited motility beyond the inhibition caused by naloxone + saline. GAG, in a dose of 12.5 mg/kg, was also potentiated by naloxone, 3.2 mg/kg. The locomotion reducing effects of naloxone could be blocked by either picrotoxin or bicuculline. It is concluded that GABAergic mechanisms participate in the inhibition of locomotor activity provoked by naloxone. The possibility that this drug disinhibits GABAergic neurons is discussed.     

Identification of the midbrain locomotor region and its relation to descending locomotor pathways in the Atlantic stingray,Dasyatis sabina

The midbrain locomotor region (MLR) in the Atlantic stingray,Dasyatis sabina, was identified and characterized. Stimulation (50–100 μA, 60 Hz) of the midbrain in decerebrated, paralyzed animals was used to elicit locomotion monitored as alternating activity in nerves innervating an antagonist pair of elevator and depressor muscles. Effective sites for evoking locomotion in the midbrain included parts of several nuclei: the caudal portion of the interstitial nucleus of the medial longitudinal fasciculus and the caudomedial parts of the cuneiform and subcuneiform nuclei. This region did not include the red nucleus, any parts of the optic tectum or the medial or lateral mesencephalic nuclei. Electrical stimulation in the MLR evokes locomotion in either the ipsi- or contralateral pectoral fin, wheres stimulation in the medullary reticular formation evokes locomotion only in the contralateral fin. Lesion experiments were performed to identify the location of descending pathways from the midbrain to the medullary reticular formation. To abolish locomotion evoked by electrical stimulation in the MLR, the medial reticular formation in the rostral medulla had to be lesioned bilaterally, or the ipsilateral medial medullary reticular formation and fibers projecting from the MLR to the contralateral midbrain had to be disrupted. Injections of HRP into the magnocellular/gigantocellular reticular formation confirmed that this area received bilateral projections from the MLR. The MLR of the Atlantic stingray appears to be similar to the lateral component of the mammalian MLR and to the MLR in other non-mammalian vertebrates.     

GABA antagonists reverse the somatostatin dependent attenuation of rat locomotor activity

Somatostatin infusion in rat ventral pallidum (VP) led to the attenuation of locomotor activity (Marazioti, A., Kastellakis, A., Antoniou, K., Papasava, D., Thermos, K., 2005. Somatostatin receptors in the ventral pallidum/substantia innominata modulate rat locomotor activity. Psychopharmacology 181, 319–326). In the present study, we investigated the putative circuitry involved in somatostatin’s actions by examining the involvement of GABAergic neurotransmission in locomotor activity subsequent to somatostatin’s infusion into the VP. Male Sprague–Dawley rats, 300–350 g, were used for all experiments. Saline or somatostatin (240 ng/0.5 μl/side) in the absence or presence of bicuculline (GABA-A antagonist; 5 mg/kg/ml, i.p.; 120 ng/side nucleus accumbens (NAc)) or phaclofen (GABA-B antagonist; 10 mg/kg/ml, i.p.; 120 ng/side NAc) were infused bilaterally, and the locomotor activity measured for 60 min using a rectangular activity cage. Somatostatin infused in the VP decreased the locomotor activity of the rat in a statistically significant manner. Bicuculline (i.p., and in the NAc) and phaclofen (only i.p.) reversed SRIF’s actions, when administered prior to somatostatin’s infusion in the VP. The present study provides further information on somatostatin’s involvement in the VP-NAc circuitry, and implicates the GABAergic system in somatostatin’s actions in the VP.     

Metabotropic glutamate receptors and the generation of locomotor activity: interactions with midbrain dopamine

Interactions between excitatory amino acid (EAA) and dopamine (DA) pathways in the basal ganglia have been known for some time to contribute importantly to the generation of motor behaviors. In particular, the role played by ionotropic glutamate receptors (iGluRs) in such interactions and in the production of locomotion has received considerable attention, particularly in brain areas such as the ventral tegmental area (VTA) where EAA afferants are known to modulate the activity of DA neurons and the nucleus accumbens (NAcc) where descending EAA projections and ascending DA mesencephalic projections come in close apposition to each other and co-innervate intrinsic neurons projecting to motor output regions. Recently, the growing importance of the metabotropic glutamate receptor (mGluR) in the generation of motor behaviors and various forms of plasticity has begun to emerge. The known coupling of the mGluR to second messenger systems and its demonstrated role in the long-term modulation of synaptic transmission make it a logical candidate not only for the generation of locomotion involving EAA-DA interactions, but also for the induction and expression of locomotor plasticity involving these neurotransmitters. In this review, we examine the evidence supporting a role for mGluRs in the generation of DA-dependent locomotion as well as in one form of locomotor plasticity: the sensitization of locomotor activity by psychomotor stimulant drugs.