Dopamine agonist-mediated rotation in rats with unilateral nigrostriatal lesions is not dependent on net inhibitions of rate in basal ganglia output nuclei.

Current models of basal ganglia function predict that dopamine agonist-induced motor activation is mediated by decreases in basal ganglia output. This study examines the relationship between dopamine agonist effects on firing rate in basal ganglia output nuclei and rotational behavior in rats with nigrostriatal lesions. Extracellular single-unit activity ipsilateral to the lesion was recorded in awake, locally-anesthetized rats. Separate rats were used for behavioral experiments. Low i.v. doses of D1 agonists (SKF 38393, SKF 81297, SKF 82958) were effective in producing rotation, yet did not change average firing rate in the substantia nigra pars reticulata or entopeduncular nucleus. At these doses, firing rate effects differed from neuron to neuron, and included increases, decreases, and no change. Higher i.v. doses of D1 agonists were effective in causing both rotation and a net decrease in rate of substantia nigra pars reticulata neurons. A low s.c. dose of the D1/D2 agonist apomorphine (0.05 mg/kg) produced both rotation and a robust average decrease in firing rate in the substantia nigra pars reticulata, yet the onset of the net firing rate decrease (at 13-16 min) was greatly delayed compared to the onset of rotation (at 3 min). Immunostaining for the immediate-early gene Fos indicated that a low i.v. dose of SKF 38393 (that produced rotation but not a net decrease in firing rate in basal ganglia output nuclei) induced Fos-like immunoreactivity in the striatum and subthalamic nucleus, suggesting an activation of both inhibitory and excitatory afferents to the substantia nigra and entopeduncular nucleus. In addition, D1 agonist-induced Fos expression in the striatum and subthalamic nucleus was equivalent in freely-moving and awake, locally-anesthetized rats. The results show that decreases in firing rate in basal ganglia output nuclei are not necessary for dopamine agonist-induced motor activation. Motor-activating actions of dopamine agonists may be mediated by firing rate decreases in a small subpopulation of output nucleus neurons, or may be mediated by other features of firing activity besides rate in these nuclei such as oscillatory firing pattern or interneuronal firing synchrony. Also, the results suggest that dopamine receptors in both the striatum and at extrastriatal sites (especially the subthalamic nucleus) are likely to be involved in dopamine agonist influences on firing rates in the substantia nigra pars reticulata and entopeduncular nucleus.

Multisecond oscillations in firing rate in the basal ganglia: robust modulation by dopamine receptor activation and anesthesia.

Multisecond oscillations in firing rate in the basal ganglia: robust modulation by dopamine receptor activation and anesthesia. Studies of CNS electrophysiology have suggested an important role for oscillatory neuronal activity in sensory perception, sensorimotor integration, and movement timing. In extracellular single-unit recording studies in awake, immobilized rats, we have found that many tonically active neurons in the entopeduncular nucleus (n = 15), globus pallidus (n = 31), and substantia nigra pars reticulata (n = 31) have slow oscillations in firing rate in the seconds-to-minutes range. Basal oscillation amplitude ranged up to +/-50% of the mean firing rate. Spectral analysis was performed on spike trains to determine whether these multisecond oscillations were significantly periodic. Significant activity in power spectra (in the 2- to 60-s range of periods) from basal spike trains was found for 56% of neurons in these three nuclei. Spectral peaks corresponded to oscillations with mean periods of approximately 30 s in each nucleus. Multisecond baseline oscillations were also found in 21% of substantia nigra dopaminergic neurons. The dopamine agonist apomorphine (0.32 mg/kg iv, n = 10-15) profoundly affected multisecond oscillations, increasing oscillatory frequency (means of spectral peak periods were reduced to approximately 15 s) and increasing the regularity of the oscillations. Apomorphine effects on oscillations in firing rate were more consistent from unit to unit than were its effects on mean firing rates in the entopeduncular nucleus and substantia nigra. Apomorphine modulation of multisecond periodic oscillations was reversed by either D1 or D2 antagonists and was mimicked by the combination of selective D1 (SKF 81297) and D2 (quinpirole) agonists. Seventeen percent of neurons had additional baseline periodic activity in a faster range (0.4-2.0 s) related to ventilation. Multisecond periodicities were rarely found in neurons in anesthetized rats (n = 29), suggesting that this phenomenon is sensitive to overall reductions in central activity. The data demonstrate significant structure in basal ganglia neuron spiking activity at unexpectedly long time scales, as well as a novel effect of dopamine on firing pattern in this slow temporal domain. The modulation of multisecond periodicities in firing rate by dopaminergic agonists suggests the involvement of these patterns in behaviors and cognitive processes that are affected by dopamine. Periodic firing rate oscillations in basal ganglia output nuclei should strongly affect the firing patterns of target neurons and are likely involved in coordinating neural activity responsible for motor sequences. Modulation of slow, periodic oscillations in firing rate may be an important mechanism by which dopamine influences motor and cognitive processes in normal and dysfunctional states.

Intracellularly recorded response of rat striatal neurons in vitro to fenoldopam and SKF 38393 following lesions of midbrain dopamine cells.

The effect of long-term (6-19 weeks) 6-hydroxydopamine-induced (6-OHDA) lesions of midbrain dopamine cells on dopamine D1-like agonist-induced changes in the excitability of rat striatal neurons was investigated in vitro using tissue slices and intracellular recording techniques. Fenoldopam and (+/-)-SKF 38393 predominantly decreased excitability in control preparations including striatal neurons located contralateral to 6-OHDA injection sites and neurons obtained from rats receiving sham injections or no treatment. Fenoldopam also inhibited neurons ipsilateral to lesions of midbrain dopamine cells. (+/-)-SKF 38393, unlike fenoldopam, produced predominantly increases in the excitability of ipsilateral striatal neurons. Superfusion of the D1 receptor antagonist, SCH 23390, blocked fenoldopam-induced decreases in excitability but not the (+/-)-SKF 38393-induced excitation of neurons ipsilateral to the lesion. Sequential application of fenoldopam and quinpirole, a D2/D3 receptor agonist, produced responses to both drugs in a majority of neurons. The results demonstrate that inhibitory responses to fenoldopam are mediated by D1 receptors, while excitatory effects of (+/-)-SKF 38393 in the striatum ipsilateral to the lesion are apparently not dependent on D1 receptor activation. These findings also suggest that dopamine D1 and D2/D3 receptors are able to concurrently influence the excitability of striatal neurons in the dopamine deafferentated striatum. Similar regulation of striatal neurons in vivo may contribute to dopaminergic regulation of basal ganglia output and the ability of dopaminomimetic agents to ameliorate symptoms of dopaminergic deficiency in Parkinson's disease.

Electrophysiological characterization of rat striatal neurons in vitro following a unilateral lesion of dopamine cells.

The effects of a unilateral 6 to 19-week lesion of dopamine cells on the excitability of rat striatal neurons were investigated in vitro using the intracellularly recorded membrane properties of neurons obtained ipsilateral and contralateral to 6-hydroxydopamine (6-OHDA) injection sites. Neurons ipsilateral to the lesion site and in striatal tissue depleted of dopamine exhibited resting membrane potentials and membrane resistances similar to those recorded in contralateral striatal neurons. Denervation appeared to have no appreciable effect on the proportion of neurons exhibiting various patterns of neuronal spiking (repetitive, bursting, or single spike) evoked by depolarizing current pulses. Current-voltage determinations revealed nominal rectification in the majority of neurons and marked nonlinearity consistent with inward rectification at potentials hyperpolarized and depolarized to rest in a large proportion of the remaining neurons. Neurons ipsilateral to 6-OHDA lesion sites exhibited these relationships in the same proportion as contralateral control cells. However, ipsilateral neurons with nominal rectification exhibited an average rate constant for the early onset of small hyperpolarizing membrane transients which was significantly smaller than that of controls. This finding suggests that intrinsic membrane parameters regulating the excitability of certain striatal neurons may be under the influence of dopamine or other factors closely associated with nigrostriatal nerve terminals.

Morphological and electrophysiological evidence for electrotonic coupling of rat dorsolateral septal nucleus neurons in vitro.

Intracellular injections of Lucifer Yellow were utilized to evaluate the incidence of dye-coupling among dorsolateral septal nucleus (DLSN) neurons recorded from slice preparations of adult rat septal nuclei. Twenty percent of single injections of Lucifer Yellow resulted in pairs of labeled neurons. These dye-coupled cells were morphologically heterogeneous and did not exhibit any morphological characteristics that could be used to distinguish them from non dye-coupled neurons. The spatial separation of cell bodies and close apposition of dendrites within each pair indicated that the dye transfer site(s) were situated at dendrodendritic and/or dendrosomatic rather than somatosomatic junctions. The main axon of some dye-coupled neurons gave rise to intrinsic axon collaterals prior to exiting the nucleus indicating that these coupled neurons function as projection neurons as well as local circuit interneurons. Electrophysiological recordings of the passive membrane properties and spontaneous activity of individual dye-coupled neurons revealed no significant difference from non dye-coupled cells in the DLSN. Some neurons exhibited spontaneously occurring fast potentials which presumably represent electrotonic potentials. These fast potentials were often tightly coupled with action potentials but could be distinguished from synaptic potentials by their shape and their lack of voltage-dependent changes in amplitude. These morphological and supportive electrophysiological data provide the first indirect evidence for electrotonic coupling of dorsolateral septal neurons. The functional significance of this coupling may lie in the potential for synchronization of the output of the DLSN which could play an important role in the septal maintenance and modulation of hippocampal Theta rhythm.

Spontaneous bursting and non-bursting activity in morphologically identified neurons of the rat dorsolateral septal nucleus, in vitro.

Membrane potential-dependent changes in the repetitive firing properties of morphologically identified rat dorsolateral septal nucleus neurons were investigated in a submerged slice preparation using intracellular recording techniques and lithium acetate-Lucifer Yellow-filled microelectrodes. The results indicate that the majority of dorsolateral septal nucleus neurons are capable of burst firing and suggest, moreover, the existence of neuronal subtypes with distinct differences in spike waveform and the pattern of spontaneous activity. In the largest proportion of neurons, single spike activity predominated at membrane potentials near rest while burst-like discharges prevailed at more hyperpolarized membrane potentials. Less frequently observed were neurons exhibiting different burst waveforms at various membrane potentials. In a few neurons, hyperpolarization slowed neuronal firing but did not elicit burst-like discharges. Characteristics such as the presence of burst or single spike discharges, spike afterpotentials, and the membrane potential dependence of repetitive firing patterns did not appear to be closely associated with membrane time constant, membrane resistance, or resting membrane potential. A detailed examination of the somatodendritic and axonal morphology of the Lucifer Yellow-filled cells revealed that these electrophysiologically identified neurons in the dorsolateral septal nucleus are morphologically heterogeneous. However, there did not appear to be any correlation between a particular somatodendritic morphology and the expression of a distinct spontaneous firing pattern. The present findings demonstrate that neurons in the rat dorsolateral septal nucleus are morphologically diverse and capable of intrinsically generating rhythmic neuronal activity. Similar patterns of rhythmic neuronal firing in vivo may provide a substrate for the integration of afferent neuronal activity and have a central role in intraseptal circuitry necessary for generation of hippocampal theta rhythm.

Somatostatin induced hyperpolarization of septal neurons is not blocked by pertussis toxin.

The coupling of postsynaptic somatostatin receptors to pertussis toxin (PTX) sensitive guanine nucleotide regulatory proteins (G proteins) was investigated in dorsolateral septal nucleus (DLSN) neurons using a submerged brain slice preparation and intracellular recording techniques. Rats were pretreated with PTX i.c.v. and neuronal responsivity to somatostatin and baclofen, a selective GABAB receptor agonist, tested using a submerged brain slice preparation and intracellular recording techniques. In tissue obtained from rats pretreated with PTX (2.5 micrograms) for 2-5 days, somatostatin applied by superfusion (0.1 microM) produced membrane hyperpolarization and decreased the membrane resistance of DLSN neurons. Hyperpolarizing effects of somatostatin persisted in the presence of tetrodotoxin (0.3 microM) blocking synaptic transmission. Current-voltage relations of the somatostatin-induced, PTX-resistant hyperpolarization indicated a reversal potential close to the equilibrium potential for potassium ions. Membrane hyperpolarizations in PTX treated tissue were similar to those recorded in tissue from vehicle control or untreated rats. Hyperpolarizing responses to the selective GABAB receptor agonist baclofen, however, were blocked by the PTX treatment used in the present study. Our findings suggest that the postsynaptic inhibitory effects of somatostatin in the DLSN is not mediated by a somatostatin receptor coupled to PTX-sensitive G proteins. These G proteins, however, appear to be an essential link in the postsynaptic GABAB receptor-mediated response of DLSN neurons.

A calcium-dependent slow afterdepolarization recorded in rat dorsolateral septal nucleus neurons in vitro.

1. Conventional intracellular and single-electrode voltage-clamp recordings were obtained from rat brain slices containing dorsolateral septal nucleus (DLSN) neurons in vitro. 2. We observed a slow afterdepolarizing potential (slow-ADP) that lasted up to several seconds (half-decay time was in the range of 0.7-1.4 s) in almost 15% of DLSN neurons; these same neurons could exhibit burst firing activity. The amplitude of this slow-ADP was not affected by hyperpolarization of the membrane potential. 3. The slow-ADP was associated with an increased membrane conductance. Hybrid voltage clamping of the slow-ADP revealed a transient slow inward current (slow-ADC). The current-voltage relationship of the slow-ADC was linear between -40 and -100 mV and generated an extrapolated reversal potential of -30 mV. 4. We investigated the ionic mechanism of the slow-ADP in the rat DLSN. Slow-ADPs were not blocked by 1 microM tetrodotoxin (TTX) but were markedly depressed by 200 microM Cd2+, Ca2(+)-free, low-Na+ solutions, and the intracellular injection of ethylene glycol-bis(B-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA). Neither diltiazam (10 microM), an L-type Ca2+ channel blocker nor omega-conatoxin (0.2-2.5 microM), an N-type Ca2+ channel blocker affected the slow-ADP. Similarly, the slow-ADP was not affected in a low-Cl- solution. On the other hand, the slow-ADP was enhanced in a K(+)-free solution. In addition, the slow-ADP was not affected by 1 mM kynurenic acid, a broad-spectrum excitatory amino acid antagonist. 5. We conclude that the slow-ADP in the rat DLSN is mediated by a novel Ca2(+)-dependent, Na(+)-dependent, and nonsynaptic inward current that may be similar to the Ca2(+)-activated nonspecific cation channel currents (i.e., CAN-currents) described in various tissues. This current appears to underlie some forms of spontaneous bursting activity recorded from rat DLSN neurons. It may also be responsible for some types of bursting activity recorded in other CNS neurons.

Somatostatin depresses GABA receptor-mediated inhibition in the rat dorsolateral septal nucleus.

The effect of somatostatin-14 (SS-14) on gamma-aminobutyric acid (GABA)-mediated inhibitory neurotransmission in the dorsolateral septal nucleus (DLSN) was investigated using a submerged slice preparation and intracellular recording techniques. Somatostatin-14 applied by superfusion or by pressure ejection from micropipettes predominantly inhibited the intracellularly recorded fast inhibitory postsynaptic potential (fIPSP) and late hyperpolarizing potential (LHP) elicited by focal electrical stimulation of the DLSN. The decreases in LHP and fIPSP amplitude occurred at low concentrations of peptide, in the absence of appreciable changes in the passive-membrane properties of postsynaptic neurons, and outlasted the membrane hyperpolarizing effect produced by SS-14 at higher concentrations. The ability of SS-14 to modulate postsynaptic GABA receptor responses underlying the fIPSP and LHP were investigated by applying baclofen, a selective GABAB receptor agonist, and isoguvacine, a selective GABAA receptor agonist, by pressure ejection. Hyperpolarizing responses to GABAA and GABAB receptor stimulation were significantly decreased during superfusion of SS-14. Tetrodotoxin applied by superfusion blocked electrically evoked synaptic potentials but not the depressant effect of SS-14 on baclofen- or isoguvacine-induced hyperpolarization. Facilitation of the fIPSP or LHP by SS-14 also occurred but less frequently and consistently than the depressant action. Excitatory postsynaptic potentials and membrane response to NMDA or quisqualate appeared unaltered by bath-applied SS-14. These findings suggest a novel postsynaptic action of SS-14 leading to depression of synaptic responses mediated by GABAA and GABAB receptors. Synaptically released SS-14 in the DLSN may participate in modulation of feedforward and/or feedback inhibitory mechanisms coordinating DLSN function in the septo-hippocampal system.

Somatostatin hyperpolarizes neurons and inhibits spontaneous activity in the rat dorsolateral septal nucleus.

Intracellular recordings were made from rat brain neurons in a submerged slice preparation containing the dorsolateral septal nucleus (DLSN). Somatostatin-14 (SS-14) was applied to these neurons by superfusing solutions containing known concentrations of the peptide or by pressure ejection from micropipettes. With either method of treatment, SS-14 produced membrane hyperpolarization and decreased membrane resistance in a concentration-dependent manner. The hyperpolarizing response to SS-14 occurred in virtually all neurons tested and appeared to result from a direct action on DLSN neurons mediated by an increased permeability to potassium ions. The SS-14-induced membrane hyperpolarization was not blocked by naloxone, bicuculline, tetrodotoxin, or calcium-free, high-magnesium superfusion media. In a small number of neurons, SS-14 application produced a membrane depolarization which did not exhibit clear concentration-dependence and was blocked by superfusion of calcium-free, high-magnesium media indicating an indirect action. These findings reveal that SS-14 is a potent inhibitor of DLSN neurons in vitro and provide the first evidence that receptors for this putative neurotransmitter are located on postsynaptic neurons in this nucleus. Synaptically released SS-14 may play an important role in the modulation of septohippocampal function.

Projection neurons in the rat dorsolateral septal nucleus possess recurrent axon collaterals.

Projection neurons in the rat dorsolateral septal nucleus (DLSN) were labeled intracellularly with horseradish peroxidase (HRP) in an in vitro slice preparation. The labeled neurons exhibited widespread 'isodendritic' type dendritic fields. Each of the neurons was identified as a projection neuron by the tracing of its main axon out of DLSN. The axons of these neurons gave rise to intrinsic collaterals which branched to form an extensive axon plexus which was confined to DLSN. These axon collaterals exhibited numerous en passant swellings suggestive of boutons. It is proposed that the recurrent axon collaterals of DLSN projection neurons may form an anatomical substrate for local inhibition within DLSN.

Analogue separates biological effects of salmon calcitonin on brain and renal cortical membranes.

The conformation-activity relationship of salmon calcitonin in kidney and brain was investigated with regard to effects on membrane binding and adenylate cyclase activity. Since an amphipathic alpha-helical conformation on the calcitonin molecule is associated with high potency in lowering serum calcium, the activity of the parent peptide was compared to that of [Gly8, D-Arg24]des-Leu16-salmon calcitonin, a calcitonin analogue (CTA) with less helix forming potential. The results indicate that while salmon calcitonin possesses similar potency in brain and kidney, CTA is effective only in brain. Furthermore, CTA did not inhibit the binding of 125I-labeled human calcitonin gene-related peptide (HCGRP) to brain membranes. Our findings suggest that the specific binding and effects of salmon calcitonin on adenylate cyclase activity in brain do not depend on conformational features in the middle region of the molecule, although the alpha-helical structure in this region does appear to be an important property for salmon calcitonin binding to renal cortical membranes.

Neuronal membrane sensitivity to a salmon calcitonin analogue with negligible ability to lower serum calcium.

The effects of [Gly8-des-Leu16-D-Arg24]-salmon calcitonin (CTA) on spontaneous extracellular activity and the passive membrane properties of rat forebrain neurons were studied in vivo and in vitro. This analogue had negligible ability to lower serum calcium relative to salmon calcitonin (SCT), however, the pattern of neuronal membrane sensitivity to CTA was similar to that produced by SCT and calcitonin generelated peptide. Depression of extracellularly recorded spontaneous action potential firing rate was the predominant response of neurons to microiontophoretic application of each peptide in vivo. Intracellular recordings from septal neurons suggest that the extracellularly observed inhibitory effects may be due to changes in membrane potential. We conclude that different structure-activity relationships may apply to the pharmacological effects of calcitonin in brain and its serum calcium-lowering action.

Effects of human pancreatic growth hormone-releasing hormone and fragments of rat hypothalamic growth hormone-releasing hormone on the activity of rat brain neurons.

The effect of human pancreatic growth hormone-releasing factor (hpGRF), rat hypothalamic GRF1-29 (rhGRF1-29) and rhGRF3-40 on spontaneous extracellular activity was studied in urethane anesthetized, male rats. The results show that these peptides similarly depress the firing rate of responsive neurons in the amygdala, caudate-putamen and globus pallidus. A single neuron was excited by hpGRF and it was localized to the medial amygdala. These findings are in line with the hypothesis that GRF has effects on the central nervous system in addition to regulating pituitary growth hormone secretion.

Antagonistic behavioral effects of calcitonin and amphetamine in the rat.

Using an automated testing apparatus, the hypermotility induced by amphetamine had previously been found to be inhibited by intracerebroventricular (ICV) administration of salmon calcitonin (CT). The present study used a computer-supported direct observational method to characterize further the interactions of CT and amphetamine. After treatment with amphetamine (1.5 mg/kg, IP), the incidence of rearing, nose poking, and locomotion was reduced in rats that were pretreated with 85 pmol salmon CT ICV; the incidence of sniffing and grooming remained unchanged. CT-induced dyskinesia, a unique consequence of central CT treatment, was attenuated but not abolished by administration of amphetamine. These results support the premise that a compound with receptor recognition characteristics similar to those of salmon CT may act as a neurotransmitter-modulator in the central nervous system.

Motor effects of calcitonin administered intracerebroventricularly in the rat.

In rats treated with salmon calcitonin (CT) administered intracerebroventricularly (i.c.v., 85 or 8.5 pmol), spasmodic body movements, hopping and tail jerks, collectively termed dyskinesia, appeared within 1 h of administration and persisted for at least 24 h. In addition, spontaneous grooming, rearing and locomotion occurred less often in CT-treated rats than in vehicle-injected animals, while the incidence of both sniffing and nose poking remained essentially unchanged. The CT failed to displace either [3H]dopamine or [3H]spiperone from striatal membranes, and the behavioral effects were not blocked by haloperidol or SCH 23390, suggesting that the peptide did not directly affect dopamine receptors. The dyskinesia was not blocked by scopolamine, atropine, muscimol, diazepam or ketanserin. These data are consistent with the hypothesis that a compound with recognition characteristics similar to those of salmon CT may function as a neurotransmitter-modulator in the central nervous system.

Sensitivity of rat forebrain neurons to growth hormone-releasing hormone.

The effects of iontophoretically applied human pancreatic growth hormone-releasing factor (hpGRF), peptide histidine isoleucine (PHI-27), and somatostatin (SS) on the extracellular activity of single cells in the hypothalamus, thalamus, and cortex of the rat brain were studied in urethane-anesthetized, male rats. Neurons with membrane sensitivity to hpGRF, PHI-27, and SS were present in each brain region. Although neurons excited by these peptides were encountered in thalamus and hypothalamus, depression of neuronal firing was the predominant response observed. Overall, the neurons responding to hpGRF also possessed membrane sensitivity to PHI-27, whereas, the hpGRF sensitive neurons appeared to be more divided as to their ability to respond to SS. The results clearly demonstrate that hpGRF and PHI-27 are capable of affecting the membrane excitability of neurons in several brain regions. The distribution of neurons sensitive to hpGRF suggests that hypothalamic GRF, in addition to its well documented role in the regulation of pituitary growth hormone secretion, may subserve other physiological events in the rat central nervous system as a neurotransmitter and/or neuromodulator.

Calcitonin and calcitonin gene-related peptide alter the excitability of neurons in rat forebrain.

Individual neurons in the hypothalamus, thalamus, cortex, and other forebrain areas of urethane-anesthetized, male rats were iontophoretically tested for their membrane sensitivity to salmon calcitonin (CT), human CT, and CT gene-related peptide (CGRP). Extracellular recording of unit activity revealed that depression of neuronal firing was the predominant effect of iontophoretically applied salmon CT (35 of 74 cells tested). Few neurons responded to salmon CT with an increase in firing rate (N = 3). When CGRP was iontophoretically applied a pattern of response resembling that of salmon CT was observed. CGRP was predominantly inhibitory and excited those neurons whose firing rate was increased by salmon CT. Inhibition was also the predominant effect of human CT. However, no neurons were excited by human CT. The results clearly demonstrate that a subpopulation of neurons with membrane sensitivity to salmon CT, human CT, and CGRP are present in the rat forebrain. This finding suggests that modulation of neuronal activity may underlie the behavioral and biochemical effects of these peptides when administered centrally. Endogenous CGRP and CT-like peptides in rat brain may be capable of regulating these events as neurotransmitters or neuromodulators.