Hyposialorrhea as an early manifestation of Parkinson disease.

We sought to determine whether hyposialorrhea is an early manifestation of Parkinson disease (PD). We measured basal and citric acid stimulated secretion of whole saliva in 20 patients with early stage (Hoehn-Yahr I-II) PD who had motor symptoms for less than 1 year and were on no medication and 11 age matched controls. Compared to controls, PD patients had significant reduction of both basal (0.0964+/-0.08 vs 0.293+/-0.112 ml/min, p<0.001) and reflex (0.263+/-0.213 vs 0.537+/-0.313 ml/min, p<0.001) salivary secretion. Our findings confirm that hyposialorrhea is an early autonomic manifestation of PD.

Turning behaviour induced by stimulation of the 5-HT receptors in the subthalamic nucleus.

The basal ganglia, which receive a rich serotonergic innervation, have been implicated in hyperkinetic and hypokinetic disorders. Moreover, a decrease in subthalamic nucleus (STN) activity has been associated with motor hyperactivity. To address the role of subthalamic serotonergic innervation in its motor function, turning behaviour was studied in rats with stimulation of the subthalamic serotonin (5-HT) receptors by intracerebral microinjections. The intrasubthalamic administration of 5-HT induced dose-dependent contralateral turning behaviour, with a maximal effect at a dose of 2.5 microg in 0.2 microL. Similar results were observed with microinjections of other 5-HT receptor agonists: quipazine (a 5-HT2B/C/3 agonist), MK-212 (a 5-HT2B/C agonist) and m-chlorophenylbiguanidine (a 5-HT3 agonist), while microinjections of 5-HT into the zona incerta or in the previously lesioned STN were ineffective. The effect of 5-HT was blocked by coadministration of the antagonist mianserin. Stimulation of subthalamic 5-HT receptors in animals bearing a lesion of the nigrostriatal pathway did not modify the motor response, which indicates that the dopamine innervation of the nucleus is not involved in this effect. Kainic acid lesion of the substantia nigra pars reticulata (SNr) suppressed the contralateral rotations elicited by stimulation of 5-HT2B/C/3 subthalamic receptors. This suggests a role of the subthalamic-nigral pathway in the turning activity. Furthermore, the partial blockade of glutamatergic receptors in the SNr by the antagonist DNQX increased the contralateral circling elicited by stimulation of 5-HT receptors in the STN. We concluded that the activation of the 5-HT2B/C and 5-HT3 subthalamic receptors elicited contralateral turning behaviour, probably via the subthalamic-nigral pathway.

High-frequency stimulation of the subthalamic nucleus silences subthalamic neurons: a possible cellular mechanism in Parkinson's disease.

The subthalamic nucleus participates in the control of movement and is considered a surgical target in the treatment of parkinsonian symptoms. Using the rat brain in vitro slice technique we show that sustained high-frequency (>100 Hz) electrical stimulation (i.e., 'tetanic stimulation') of the nucleus, as used in humans to treat Parkinson's disease, silenced subthalamic neurons. Two main cell types were identified. 'Tonic cells' (68%) showed delayed inward rectification, fired continuously, switched to bursting and stopped firing when strongly depolarized with injected current. Tetanic stimulation of the nucleus induced a steady depolarization (approximately 18 mV) that triggered action potentials at a high rate followed by bursts and finally (approximately 25 s) totally silenced tonic cells. The control tonic activity was recovered rapidly (<10 s) after ending stimulation. 'Phasic cells' (25%) discharged a single initial brief burst of action potentials both when depolarized by prolonged current injection and tetanic stimulation and did not show inward rectification. An infrequent cell type called 'phasic-tonic' (7%) showed a mixed discharge. We suggest that the silencing effect of tetanic stimulation is not a frequency-dependent presynaptic depression and could result from the gradual inactivation of Na+-mediated action potentials. These findings suggest that the remission of parkinsonian symptoms by treatment with high-frequency electrical stimulation of the subthalamic nucleus in humans may primarily reside on its capacity to suppress the action potential activity of subthalamic neurons.

Stimulation of dental pulp with simultaneous recording of the jaw opening reflex in the rat.

The processing of nociceptive information in the central nervous system has been analysed in most studies by activation of peripheral nerves. However, the limitation of this method is the simultaneous activation of noxious and inocuous fibers. Nevertheless, the stimulation of the tooth pulp is believed to activate mainly nociceptive fibers which could be the method of choice. On the other hand, the response to nociceptive activation of the dental pulp is easily quantified by the amplitude of the jaw opening reflex, a nocifensive flexion withdrawal reflex. In this protocol we describe a technique to manufacture and implant electrodes in lower incisors of the rat and a method to prepare and insert stainless steel twisted bipolar electrodes to record the electromyographic activity of both digastric muscles, in response to nociceptive dental pulp stimulation. This approach was applied in the study of the analgesic effects of the rat's striatal stimulation.

Subthalamic nucleus lesions reduce low frequency oscillatory firing of substantia nigra pars reticulata neurons in a rat model of Parkinson's disease.

Single unit recordings performed in animal models of Parkinson's disease revealed that output nuclei neurons display modifications in firing pattern and firing rate, which are supposed to give rise to the clinical manifestations of the illness. We examined the activity pattern of single units from the substantia nigra pars reticulata, the main output nuclei of the rodent basal ganglia, in urethane-anesthetized control and 6-hydroxydopamine-lesioned rats (a widespread model of Parkinson's disease). We further studied the effect of a subthalamic nucleus lesion in both experimental groups. Subthalamic nucleus lesion produces behavioral improvement in animal models of Parkinson's disease, and was expected to reverse the changes induced by 6-hydroxydopamine lesions. A meticulous statistical investigation, which included a non-biased classification of the recorded units by means of cluster analysis, allowed us to identify a low frequency oscillation of firing rate ( approximately 0.9 Hz) occurring in approximately 35% of the units recorded from 6-hydroxydopamine-lesioned rats, as the main feature differentiating 6-hydroxydopamine-lesioned and control rats. Subthalamic nucleus lesions significantly reduced the proportion of oscillatory units in 6-hydroxydopamine-lesioned rats. However, the population of nigral units recorded from rats bearing both lesions still differed significantly from control units. These results suggest that oscillatory activity in the basal ganglia output nuclei may be related to some clinical features of parkinsonism, and suggest a putative mechanism through which therapeutic interventions aimed at modifying subthalamic nucleus function produce clinical benefit in Parkinson's disease.

Pharmacological involvement of the calcium channel blocker flunarizine in dopamine transmission at the striatum.

Single intrastriatal microinjections of 25, 50 and 100nmol/microl of flunarizine in normal rats produced a dose-dependent turning behavior toward the injected side when they were challenged with apomorphine (1mg/kg, s.c). This effect was seen at 1, 3 and 7 days following administration of the high dose of flunarizine, but had subsided by 24h after administration of the intermediate dose; the low dose was ineffective. However, intrastriatal injection of the high dose of flunarizine resulted in a local lesion and thereafter this dose was not used. A similar dose-response relationship was determined for nifedipine, an L-type calcium channel antagonist. Injection of this antagonist did not result in apomorphine-elicited rotational behavior, reflecting its lack of antidopaminergic action. Intrastriatal injections of haloperidol (5microg/microl), an antagonist of dopamine D(2) receptors, or the sodium channel blocker lidocaine (40microg/microl), were given in order to compare their effects to those observed with flunarizine. Intracerebral injection of haloperidol produced ipsilateral turning in response to systemic administration of apomorphine given 60min after. The same response was obtained with the injection of apomorphine 10min after the injection of intracerebral lidocaine. This effect was no longer apparent 24h after the microinjection of haloperidol and 60min after the injection of lidocaine. In rats rendered hemiparkinsionian by lesioning the nigrostriatal pathway with 6OHDA, intrastriatal microinjection of flunarizine (50nmol/microl) significantly reduced apomorphine (0.2mg/kg, s.c.)-elicited turning behavior towards the non-lesioned side. These results suggest an antidopaminergic effect of flunarizine mediated by antagonistic action of post-synaptic striatal dopamine receptors. However, an action of the drug on sodium channels may not be ruled out. These studies offer additional supporting evidence for the induction or aggravation of extrapyramidal side-effects in patients receiving flunarizine.

Striatal modulation of the jaw opening reflex.

The effect of striatal electrical and chemical conditioning stimulation (L-glutamate 80-160 nmoles/0.5 microl) on the jaw opening reflex (JOR) was studied in Sprague-Dawley male rats anesthetized with urethane. The JOR was evoked by stimulation of the tooth pulp of lower incisors. This response was suppressed by transection of the dental root, which indicates according with the bibliography, a specific activation of the pulp nerves. Three type of responses were obtained on the evoked JOR by conditioning stimulation of the striatum; being the main one the suppression of the reflex elicited by tooth pulp activation. A second type of response was an increase of the tooth-JOR amplitude. This effect was observed more frequently with glutamate stimulation rather than with electrical activation of the striatum. A third response was observed with chemical stimulation but not by electrical stimulation of the striatum. This was a triphasic response which consisted in an increase followed by an inhibition and a late increase of the tooth-JOR amplitude. A biphasic effect, an increase prior to a decrease of the JOR amplitude, was also recorded with a minor frequency. The distribution of effective sites for electrical and chemical stimulation within the striatum are mainly similar located in the rostral aspect of the nucleus, with the inhibitory sites in the middle of the nucleus and intermingled with the excitatory ones. The complex responses (tri/biphasic) were observed ventrally and caudally in the nucleus. On the basis of the results mentioned above, one could assume that the striatum is related to the modulation of the JOR evoked probably by nociceptive stimulation. However, activation of other type of fibers could not be ruled out.

Substantia nigra pars reticulata units in 6-hydroxydopamine-lesioned rats: responses to striatal D2 dopamine receptor stimulation and subthalamic lesions.

In order to increase our understanding of Parkinson's disease pathophysiology, we studied the effects of intrastriatally administered selective dopamine receptor agonists on single units from the substantia nigra pars reticulata of 6-hydroxydopamine (6-OHDA)-lesioned rats with or without an additional subthalamic nucleus lesion. Nigral pars reticulata units of 6-OHDA-lesioned rats were classified into two types, showing regular and bursting discharge patterns, respectively ('non-burst' and 'burst' units). Non-burst and burst units showed distinct responses to intrastriatal quinpirole (the former were excited and burst units inhibited). Furthermore, subthalamic nucleus lesions significantly decreased the number of nigral units showing a spontaneous bursting pattern, and reduced the proportion of units that responded to quinpirole. In contrast, subthalamic lesions did not alter the proportion of nigral units that responded to SKF38393, although the lesions changed some response features, e.g. response type and magnitude. Burst analysis showed that quinpirole did not modify the discharge pattern of burst units, whereas SKF38393 produced a shift to regular firing in 62% of the burst units tested. In conjunction, our results support that: (i) the subthalamic nucleus has an important influence on output nuclei firing pattern; (ii) striatal D2 receptors have a strong influence on nigral firing rate, and a less relevant role in controlling firing pattern; (iii) burst and non-burst units differ in their response to selective stimulation of striatal dopamine receptors; (iv) the effects of striatal D2 receptors on nigral units are mainly, though not exclusively, mediated by the subthalamic nucleus; and (v) nigral responses to SKF38393 involve the subthalamic nucleus.

D1-D2 dopamine receptor interaction: an in vivo single unit electrophysiological study.

After intrastriatal administration of selective dopamine receptor agonists only a small percentage of substantia nigra pars reticulata single units showed changes in firing rate (23% after SKF38393 and 17% after quinpirole). After their intrastriatal co-administration, however, or after the application of the non-selective dopamine receptor agonist apomorphine, 72% and 69% of units responded, respectively. This result confirms the participation of the striatum in the phenomenon of D1-D2 receptor interaction, and show that co-activation of both receptor subtypes produced a maximal effect on basal ganglia output nuclei.

Substantia nigra pars reticulata single unit activity in normal and 60HDA-lesioned rats: effects of intrastriatal apomorphine and subthalamic lesions.

The spontaneous activity and the response to intrastriatal application of apomorphine of substantia nigra pars reticulata (SNpr) single units was studied in four experimental groups of rats: (1) normal rats; (2) subthalamic nucleus (STN) lesioned rats; (3) rats bearing a 6-hydroxydopamine (60HDA) lesion; and (4) 60HDA-lesioned animals with an additional STN lesion. Thirty-eight percent of units from 60HDA-lesioned rats showed a bursting pattern of spontaneous activity, which was never found in normal rats. STN lesions had no effect on the spontaneous activity of SNpr units from normal rats, but reduced the percentage of burst units in 60HDA-lesioned animals. Intrastriatal apomorphine produced responses in 62% of SNpr units from normal rats and 85% of units from 60HDA-lesioned animals (P < 0.05). In addition, the modifications in the firing rate and in the coefficient of variation of the interspike intervals induced by intrastriatal apomorphine were significantly greater for the units isolated from 60HDA-lesioned rats. In particular, it was noted that all the burst units responded to apomorphine, showing the highest changes in firing rate and coefficient of variation. However, intrastriatal apomorphine did not always turn the activity of burst units into a more physiological pattern. STN lesions reduced the percentage of units responding to intrastriatal apomorphine in normal rats. In 60HDA-lesioned rats, STN lesions reduced the number of responsive units, and their change in mean firing rate and coefficient of variation. Our results show that the STN participates in the genesis of the bursting pattern of activity of SNpr units in 60HDA-lesioned rats, and that STN lesions can partially revert the abnormal spontaneous and apomorphine-induced responses of SNpr units in these animals.

Turning behavior induced by injections of glutamate receptor antagonists into the substantia nigra of the rat.

We have found recently that muscimol microinjections into the subthalamic nucleus produce contralateral turning activity [Murer and Pazo (1993) NeuroReport, 4:1219-1222]. To test the hypothesis that a reduced glutamate action on substantia nigra pars reticulata neurons mediates this turning response, we examined the effect of unilateral intranigral microinjections of the AMPA/kainate receptor antagonist 6,7-dinitro-quinoxaline-2,3-dione (DNQX) and the competitive N-methyl-D-aspartate (NMDA) receptor antagonist DL-2-amino-5-phosphonovaleric acid (AP-5). DNQX and AP-5 both produced a dose-dependent contralateral turning response, while vehicle administration did not induce turning activity. Application of glutamate receptor antagonists at adjacent regions of the mesencephalic tegmentum were also ineffective. Coadministration of NMDA or AMPA significantly reduced the turning response induced by AP-5 or DNQX, respectively. Lesions of the nigrostriatal pathway by 6-hydroxydopamine did not modify the response to DNQX or AP-5 administration into the nigra. However, their behavioral effects were significantly reduced by a lesion of the ipsilateral subthalamic nucleus. Our results show that the blockade of a tonic input acting on AMPA/kainate and NMDA receptors located at the substantia nigra produces contralateral turning behavior. The effect seems to involve pars reticulata cells since this area remains unchanged after destruction of dopaminergic neurons. The subthalamic nucleus seems to be the endogenous source of the agonist acting on the nigral glutamate receptors related to turning behavior.

Defensive behavior induced by stimulation of the ventral striatum in the cat.

The behavioral responses induced by electrical stimulation of the ventral striatum were studied in 15 freely moving cats, chronically implanted with multiwire electrodes. Two types of defensive responses could be evoked from the ventral striatum, an arrest response and an escape or flight reaction. Stimulation of the dorsal striatum and the internal capsule induced contralateral head turning and movements of the face or limbs. The number of ventral striatal sites eliciting escape responses increased along the sessions, while their threshold decreased. Lesioning the ventral pallidal region with kainic acid increased significantly the threshold for defensive responses, whereas head turning responses showed no change. The results of this study indicate that the ventral striatum could be part of the neural substrate for defensive behavior, and that the ventral pallidal region plays a role in its mediation.

Turning behavior in rats with unilateral lesions of the subthalamic nucleus: synergism between D1 and D2 receptors.

Rats with unilateral kainic acid lesion of the subthalamic nucleus showed a dose dependent rotational response to the lesioned side (ipsilateral) after systemic administration of the non-selective dopaminergic agonist apomorphine. Both D2 and D1 selective antagonists ((-)sulpiride and SCH23390) inhibited the response to apomorphine in these rats. Selective D2 and D1 agonists (quinpirole and SKF38393) were unable to induce turning behavior. However, an ipsilateral circling response was obtained after the simultaneous application of both agonists. The interaction mechanism between dopaminergic receptor subtypes seems to be similar to that of other normosensitive models of turning previously studied (Barone et al., 1986; Robertson and Robertson, 1986; Arnt and Perregard, 1987; Asim et al., 1990; Pazo et al., 1993). It is proposed that the ipsilateral turning response to dopaminergic agonists in rats with subthalamic nucleus lesion results from an impaired behavioral expression of the action of dopaminergic agonists on one side, leading the rats to turn away from the intact hemisphere.

Behavioral responses induced by electrical stimulation of the caudate nucleus in freely moving cats.

The caudate nucleus and adjacent structures of 26 freely moving cats were stimulated through multiwire electrodes chronically implanted. Two main effects here observed with trains of pulses of high frequency (100 Hz) and short duration (1 s): (1) contralateral head turning and (2) arrest reaction, which was associated with crouching and escape behavior. The responses follow a certain topographic distribution. Head turning was elicited with the lowest mean threshold in sites located in the internal two-thirds and caudal region of the caudate nucleus, while the arrest reaction was elicited from the ventromedial region of the caudate and adjacent nucleus accumbens. Stimulation of the corpus callosum and internal capsule produces postural instability, ventral flexion of the head and flexion of the contralateral limb. The extra-caudate responses were accompanied by contralateral head turning when the stimulated points were near of the caudate border. Experimental evidence suggested that striatal responses were not due to current spread to adjacent structures or to activation of corticofugal fibers. The head rotation was suppressed following interruption of the ipsilateral striatal outflow by electrolytic lesion of the globus pallidus and adjacent internal capsule. The chemical lesion of the substantia nigra and the ventral pallidum produced a significant increase in the stimulation threshold for head turning and arrest reaction, respectively. These results suggest a topographic arrangement of the responses evoked by electrical stimulation of the caudate nucleus in the cat, which are mediated by the substantia nigra pars reticulata and the ventral pallidum.

Circling behaviour induced by activation of GABAA receptors in the subthalamic nucleus.

The behavioural effect of drugs acting on GABAA receptors in the subthalamic region and the entopeduncular nucleus was studied in freely moving cats. Microinjections of muscimol into the zona incerta-Forel's fields produced tilt of the head to the injected site, while injections of bicuculline or picrotoxin produced the same effect but to the opposite side. Microinjections of muscimol into the subthalamic nucleus elicited circling behaviour to the contralateral side, while bicuculline and picrotoxin produced ipsilateral turning. However, when high doses of the antagonists were injected, contralateral circling was observed. Activation or blockade of GABAA receptors into the entopeduncular nucleus was ineffective. These results imply that GABAergic mechanisms in the subthalamic nucleus and the zona incerta-fields of Forel are involved in circling behaviour and vertical head movements, respectively.

D1 and D2 receptors and circling behavior in rats with unilateral lesion of the entopeduncular nucleus.

The role of D1 and D2 striatal dopamine receptors on circling behavior was studied in a normosensitive model obtained by unilateral kainic acid lesion of the entopeduncular nucleus. In this model, the sensitivity of striatal dopamine receptors was preserved, because kainic acid destroyed the neurons of the entopeduncular nucleus and left undamage the fibers of passage and axon terminals. Systemic administration of SKF 38393 to these animals fails to induce circling activity. In contrast, administration of quinpirole elicited rotation toward the lesioned side, which was increased by concurrent injection of SKF 38393. This behavior was inhibited by pretreatment with either a specific D1 (SCH 23390) or D2 (-sulpiride) antagonist. The apomorphine also induced ipsilateral circling that was abolished by pretreatment with D1 or D2 antagonists. The above results suggest that coactivation of both D1 and D2 striatal dopamine receptors are necessary to induce rotation in this normosensitive model.

Spontaneous and evoked activity of the caudate neurons to central and peripheral stimuli after brain lesions.

The involvement of the cerebral cortex, commissural fibers and thalamus on caudate-caudate relations was studied in locally anesthetized, paralysed and artificially ventilated cats. This type of experimental preparation was necessary since a complete suppression of spontaneous and evoked activity is produced by subanesthetic doses of general anesthesia. Two types of caudate action potentials were encountered on the basis of their waveform characteristics: biphasic and triphasic spikes, the former being the largest population (80%). These waveforms were independent of the microelectrode resistance and the distance to recorded neurons. However, their responses were very similar to both central and peripheral stimuli. Caudate stimulation depressed the spontaneous discharges of the majority of the responsive units recorded within the opposite nucleus, while striatal neurons were activated by stimulation of the contralateral cortex. Decortication, thalamic lesion (motor nuclei and massa intermedia) and section of the corpus callosum decrease the firing rates of caudate neurons with biphasic spikes, while the discharges of the neurons with triphasic action potentials remained unchanged. Bilateral ablation of the cerebral cortex decreased the responsiveness of striatal neurons to contralateral nucleus and sciatic nerve and reduced the number of spontaneously active cells per recording tract. Section of the commissural fibers also depressed the caudate responses to the contralateral nucleus, and to the opposite precruciate cortex, although thalamic lesion did not affect the responsiveness of caudate cells to both central and peripheral stimuli.(ABSTRACT TRUNCATED AT 250 WORDS)

Role of the mesopontine area in the circling behavior induced by apomorphine in rats bearing unilateral lesion of the entopeduncular nucleus.

The role of the mesopontine area and superior colliculus in turning behavior induced by systemic administration of apomorphine was studied in rats bearing a unilateral entopeduncular lesion. Bilateral electrolytic damage of the superior colliculus resulted in an enhancement of the ipsilateral circling response to apomorphine, perhaps as a consequence of an increased locomotor drive in such animals. Bilateral electrolytic lesions of the mesopontine area decreased apomorphine-induced turning in entopeduncular rats, while a bilateral kainic acid lesion of the same region was ineffective. It was concluded that the pedunculopontine nucleus and adjacent reticular formation are not an essential link for the striopallidal complex output mediating circling in this model. Fibers running through this region could be implicated in the expression of the behavior under study. Since a unilateral electrolytic lesion of the mesopontine area contralateral to the damaged entopeduncular nucleus reduced drug-induced turning, we propose that an uncrossed pathway from the intact striopallidal complex mediates circling in our rats.

Effects of central and peripheral inputs on single pineal cell activity in the rat.

The influences exerted by central and peripheral afferents to the pineal gland have been studied in rats anesthetized with urethane (1.2 g/kg, i.p.). Spontaneous action potentials arising from the pinealocytes were recorded by means of glass micropipettes filled with 3 M NaCl containing a dye. The electrical stimulation of suprachiasmatic nucleus, superior cervical ganglia, sciatic nerve and retina evoked discharge changes in a significant number of pineal cells. However, a relatively higher proportion of pinealocytes failed to respond to these afferents. Three types of responses could be observed. Inhibitions were the predominant response patterns to suprachiasmatic nucleus, superior cervical ganglia and sciatic nerve, while excitations were mainly elicited following photic stimulation, whereas the remaining evoked activity was biphasic responses, which were observed in a small number of cells after stimulation of suprachiasmatic nucleus, superior cervical ganglia and retina. These data confirm some previous neural inputs to the pineal and demonstrate the existence of a modulatory effect of the suprachiasmatic nucleus on pinealocyte discharges as well as somatosensory afferents to the gland by way of the sciatic nerve.

Responses elicited by species-specific models in the cichlid Crenicichla lepidota (Heckel).

Species recognition by the cichlid fish C. lepidota was studied by measuring species-specific aggressive behavior toward either live conspecifics (control) or toward different two-dimensional models. A plain fish-shaped model elicited aggressive behavior, but the responses were mostly absent when the model was a rectangle. Adding heavy stripes to the fish-shaped form increased the responses, which were further enhanced by adding a spot. Comparable aggressive responses were also induced by adding spots and eye to dummies with thin stripes. We conclude that in our experimental conditions, fish-like shape and contrast seem to be of great significance for recognition of conspecifics in the cichlid C. lepidota (Heckel).