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Functional organization within the medullary reticular formation of the intact unanesthetized cat. III. Microstimulation during locomotion.

1. This article presents the results from stimulation in 21 loci within the medullary reticular formation (MRF; between 0.5 and 2.5 mm from the midline) and in 5 loci in the medial longitudinal fasciculus (MLF) of four intact, unanesthetized cats during locomotion. Stimulus trains (11 pulses, 0.2-ms duration, 330 Hz, stimulus strength 35 microA) were applied at those loci in each track at which the most widespread effects in each of the four limbs were obtained with the cat at rest. Electromyograms were recorded from flexor and extensor muscles of each limb. 2. As previously reported, stimulation with the cat at rest generally evoked brief, short-latency, twitch responses in both flexor and extensor muscles of more than one limb. In contrast, stimulation during locomotion evoked a more complex pattern of activity in which responses were normally evoked in one or other of the muscle pairs and incorporated into the locomotor pattern. 3. In the majority of sites, the stimulation evoked excitatory responses in the flexor muscles of each of the four limbs during that period of the step cycle in which each respective muscle was naturally active; stimulation in the stance phase of locomotion, although less effective, was also capable of producing responses in these muscles. All three ipsilateral extensor muscles studied [long and lateral heads of triceps and vastus lateralis (Tri, TriL, and VL, respectively)] were normally inhibited during their phase of muscle activity, although excitatory responses were occasionally seen. Responses in the contralateral (co) Tri were invariably excitatory and were largest during the period of muscle activity, whereas responses during the period of activity of the coVL were mixed, with both excitatory and inhibitory responses being seen from any one locus. 4. Excitatory responses were normally largest when stimulation was applied during the time that the muscle was active during the locomotor cycle. Responses evoked at times when the muscle was inactive were sometimes larger than those evoked with the animal at rest; such responses were most commonly seen in the hindlimb flexors and in the coVL. 5. In both flexors and extensors of each of the four limbs, the latency of the responses was greatest when the cat was at rest and least for stimuli given during the period of activity of the respective muscle. Average latencies during the period of muscle activity ranged from a minimum of 9.0 +/- 2.6 (SD) ms for inhibitory responses in the ipsilateral Tri and TriL to a maximum of 17.1 +/- 3.0 ms for the responses evoked in the ipsilateral semitendinosus.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Role of dorsal mesencephalic reticular formation and deep layers of superior colliculus in turning behaviour elicited from the striatum.

Kainate or electrolytic lesions were placed unilaterally in the dorsal mesencephalic reticular formation (MRF) or in the deep layers of the superior colliculus (DLSC) on the same side of a unilateral lesion of the medial forebrain bundle with 6-OHDA. Before the lesions the rats turned contralaterally when challenged with 0.25 mg/kg of apomorphine. After lesions of the MRF most rats turned ipsilaterally in response to the same dose of apomorphine. After lesions of the DLSC apomorphine-induced contralateral turning was significantly reduced but not abolished. The results indicate that the MRF and DLSC play a primary role in the expression of turning originated from the striatum.

Animals↗

[Role of dorsal regions of the hippocampus and midbrain reticular formation in escape reaction development in rabbits].

Experiments on unrestrained rabbits were carried out to study the role of the dorsal hippocampus (DH) and midbrain reticular formation (MRF) in escape reaction elicited by electrical stimulation of the ventromedial hypothalamus (VMH). DH stimulation made ipsi- or contralaterally to the stimulated hypothalamic center of "affective reactions" increased while MRF stimulation reduced the latency of escape. Coagulation of the structures inhibited escape reaction in rabbits. The inhibition of the latter reaction in hippocampectomized animals was characterized by the increased general motor activities, whereas rabbits with lesions in the MRF were adynamic. That the DH and MRF play a different role in the mechanisms by which excitations spread from the VMH to the neocortical areas was revealed after computer analysis of the power of the main EEG rhythms.

Animals↗

Blockade of reticular formation activity, due to carisoprodol maternal administration, and its effects on rat skeleton development.

The purpose of this experiment was to study the influence of reticular formation blockade, due to carisoprodol maternal administration, on rat skeleton development. The drug was administered three times a day orally by stomach tube at doses: T1--20 mg/kg/24 h, T2--200 mg/kg/24 h, T3--400 mg/kg/24 h. The fetuses obtained on 21st day of gestation were counted and macroscopically examined. Placental and fetal weight, fetal and tail length were checked. After fixation in 95% ethanol the fetuses were stained under single alizarin red S Dawson method and examined under a stereo-dissection microscope. Morphological examination revealed no major malformations. Insignificant number of subcutaneous ecchymose and various skeleton anomalies were observed. The experiment revealed that carisoprodol has no influence on rat skeleton development.

Abnormalities, Drug-Induced↗

Actions of 3 alpha,5 alpha-tetrahydrodeoxycorticosterone on single neurones of the mesencephalic reticular formation in the rat.

The effects of microinjections of 3 alpha,5 alpha-tetrahydrodeoxycorticosterone (3 alpha-THDOC) on single neurones of the mesencephalic reticular formation (MRF) were investigated in rats anaesthetized with urethan. Microinjections of approximately 100 nl of 0.5-2 microM 3 alpha-THDOC inhibited firing of 105 of 112 neurones (94%). Microinjections of approximately 100 nl of 100-250 nM of 3 alpha-THDOC did not alter neuronal activity, but in 52 of 68 cases (76%) it potentiated the inhibitory action of microiontophoretically applied gamma-aminobutyric acid (GABA). The 3 beta-isomer of tetrahydrodeoxycorticosterone did not elicit any changes in neuronal firing. The effects of 3 alpha-THDOC were reversibly antagonized by microiontophoretically applied bicuculline. This 'in vivo' study supports the hypothesis that 3 alpha-THDOC may function as endogenous modulator of GABAA-mediated inhibition in various physiopathological conditions.

Action Potentials↗

Actions of neurotransmitters on pontine medical reticular formation neurons of the cat.

The actions of several neurotransmitters were determined on 43 antidromically identified reticulospinal neurons and 72 unidentified neurons in the paraabducens reticular formation of the anesthetized cat. All neurons were excited by glutamate and aspartate, both of which caused brief, high-frequency responses. In 80% of the reticulospinal neurons glutamate was more potent than aspartate, whereas in 61% of the unidentified neurons aspartate was more potent. Glutamate responses were reversibly antagonized by curare applied by pressure injection. Fast inhibitory responses were obtained on all neurons tested to gamma-aminobutyric acid, glycine, and norepinephrine. Some neurons showed similarly fast inhibitory responses to acetylcholine and serotonin. Acetylcholine and serotonin both acted on most neurons, but approximately equal numbers of neurons showed a relatively fast inhibition, a relatively slow and long-lasting excitation, and a biphasic combination of inhibition and slow-excitation responses. The pattern of responses to acetylcholine and serotonin is consistent with a spatial separation of excitatory and inhibitory receptors on different portions of the cell, possibly reflecting different inputs that use the same transmitter but have effects of opposite electrical and functional polarity. Although complicated by the phenomenon of excitatory and inhibitory responses to the same transmitter, these results are compatible with the Hobson-McCarley model of generation of desynchronized sleep.

Acetylcholine↗

Convulsive seizures induced by N-methyl-D-aspartate microinjection into the mesencephalic reticular formation in rats.

Effects of microinjections of a single 2 or 10 nmol dose of N-methyl-D-aspartate (NMDA) into the unilateral mesencephalic reticular formation (MRF) on behavior and electroencephalogram were examined in rats (n=18) during a 15 min period (Exp. 1), and subsequent effects of sound stimulation with key jingling applied at 15, 30, and 45 min after the injections were observed (Exp. 2). The microinjections of 2 nmol dose of NMDA (n=10) induced hyperactivity (9 of 10 rats) and running/circling (8 of 10 rats) in Exp. 1, and hyperactivity (3 of 10 rats) in Exp. 2. Moreover, the microinjections of 10 nmol dose of NMDA (n=8) induced not only hyperactivity (8 of 8 rats) and running/circling (7 of 8 rats) but also generalized tonic-clonic seizures (GTCS) (5 of 8 rats) in Exp. 1; these seizure patterns were also elicited by sound stimulation in Exp. 2. The seizure patterns were accompanied by electroencephalographic seizure discharges in the MRF and the motor cortex. In contrast, the control group rats (n=10) which received a single dose of saline microinjection into the unilateral MRF showed no behavioral or electroencephalographic changes in both Exp. 1 and 2. These findings suggest that the MRF has an important role in the development of GTCS, which follows hyperactivity and running/circling, and that potentiation of excitatory neurotransmission in the MRF participates in the development of audiogenic seizures as well as GTCS.

Acoustic Stimulation↗

Global increase in cerebral metabolism and blood flow produced by focal electrical stimulation of dorsal medullary reticular formation in rat.

We sought to determine whether the increases in local cerebral blood flow (LCBF) elicited by focal electrical stimulation within the dorsal medullary reticular formation (DMRF), are secondary to or independent of, increased local cerebral glucose utilization (LCGU). Rats were anesthetized (chloralose), paralyzed, artificially ventilated and arterial pressure and blood gases controlled. LCBF and LCGU were determined in two separate groups of animals, using the autoradiographic [14C]iodoantipyrine and [14C]2-deoxyglucose methods, respectively. In unstimulated controls, LCBF (n = 5) and LCGU (n = 5) were linearly related (r = 0.780; P less than 0.001) in the 27 brain regions studied. During DMRF stimulation LCGU increased significantly in 21 of the 27 regions, including cerebral cortex (up to 168% of control), thalamic nuclei (up to 161%) and selected ponto-medullary regions (e.g. parabrachial complex: 212%; vestibular complex: 147%). Along with LCGU, LCBF rose significantly in 25 regions (sensory motor cortex: 163%; anterior thalamus: 161%; parabrachial complex: 186%). Correlation analysis demonstrated that, during DMRF stimulation, the close relationship between LCBF and LCGU is preserved (r = 0.845; P less than 0.001) and that, in addition, the increase in LCBF (delta LCBF) is proportional to the increase in LCGU (delta LCGU) (delta LCBF = 2.18 delta LCGU + 6.92; r = 0.7729; P less than 0.001). Excitation of neurons or fibers within DMRF increases brain metabolism globally and blood flow secondarily. The DMRF appears to modulate cerebral metabolism globally, by as yet undefined pathways.

Animals↗

Contribution of the superior colliculus and the mesencephalic reticular formation to gaze control.

Converging lines of evidence support a role for the intermediate and deep layers of the superior colliculus (SC) and the mesencephalic reticular formation (MRF) in the control of combined head and eye movements (i.e., gaze). Recent microstimulation, single-cell recording, and lesion experiments are reviewed in which monkeys are free to move their heads. Cells in the SC discharge in advance of combined head and eye movements and most likely provide a gaze error signal to downstream structures. In contrast, the neurons in the MRF are of at least two types. Eye cells have features that are similar to neurons in the rostral portion of the SC, but fire before the onset of horizontal eye movments. A second group of MRF neurons begin to fire after the onset of the gaze shift and are most closely associated with movements of the head. The peak discharge of these late-onset MRF neurons occurs near the peak head velocity. Stimulation in the rostral SC generates eye movements with fixed amplitude and direction. A similar response is noted after stimulation of the more dorsal portion of the caudal MRF. Stimulation in the caudal portion of the SC produces combined head and eye movements of fixed amplitude. Electrical activation of the more ventral portions of the caudal MRF generates goal-directed and centering eye movements. Temporary inactivation of the SC with the GABA agonist muscimol generated hypometria and curved trajectories of contralateral eye movements. Inactivation of the caudal MRF produced contralateral hypermetria and ipsilateral hypometria of saccades. Release of the monkey's head demonstrated a profound contralateral head tilt. Taken together, these data suggest that the gaze signal generated in the SC is filtered by neurons in the MRF to generate a feedback signal of eye motor error. The head signal found in the MRF could cancel a portion of the gaze signal coming from the SC in the form of head velocity feedback.

Animals↗

Neurotransmitter phenotypes of intermediate zone reticular formation projections to the motor trigeminal and hypoglossal nuclei in the rat.

Numerous studies suggest an essential role for the intermediate (IRt) and parvocellular (PCRt) reticular formation (RF) in consummatory ingestive responses. Although the IRt and PCRt contain a large proportion of neurons with projections to the oromotor nuclei, these areas of the RF are heterogeneous with respect to neurotransmitter phenotypes. Glutamatergic, GABAergic, cholinergic, and nitrergic neurons are all found in the PCRt and IRt, but the projections of neurons with these phenotypes to the motor trigeminal (mV) and hypoglossal nucleus (mXII) has not been fully evaluated. In the present study, after small injections of Fluorogold (FG) into mV and mXII, sections were processed immunohistochemically to detect retrogradely labeled FG neurons in combination with the synthetic enzymes for nitric oxide (nitric oxide synthase) or acetylcholine (choline acetyltransferase) or in situ hybridization for the synthetic enzyme for GABA (GAD65/67) or the brainstem vesicular transporter for glutamate (VGLUT2). In three additional cases, FG injections were made into one motor nucleus and cholera toxin (subunit b) injected in the other to determine the presence of dual projection neurons. Premotor neurons to mXII (pre-mXII) were highly concentrated in the IRt. In contrast, there were nearly equal proportions of premotor-trigeminal neurons (pre-mV) in the IRt and PCRt. A high proportion of pre-oromotor neurons were positive for VGLUT2 (pre-mXII: 68%; pre-mV: 53%) but GABAergic projections were differentially distributed with a greater projection to mV (25%) compared to mXII (8%). Significant populations of cholinergic and nitrergic neurons overlapped pre-oromotor neurons, but there was sparse double-labeling (<10%). The IRt also contained a high proportion of neurons that projected to both mV and MXII. These different classes of premotor neurons in the IRt and PCRt provide a substrate for the rhythmic activation of lingual and masticatory muscles.

Acetylcholine↗

Involvement of cortical, thalamic and midbrain reticular formation neurons in spike and wave discharges: extracellular study in feline generalized penicillin epilepsy.

Extracellular activity of single units, simultaneously recorded in cortex, thalamus, and midbrain reticular formation was investigated during feline generalized penicillin epilepsy. The firing activity of neurons recorded in the cortex was invariably and consistently enhanced in coincidence with the positive peak and the positive-negative transient of the "spike" of the spike and wave complex, and it was greatly decreased during the wave. In the nonspecific thalamic nuclei three classes of neurons were identified according to their patterns of activity during the spike and wave complex: (i) neurons behaving like cortical units, (ii) neurons with enhanced firing activity during the wave and a decreased activity during the "spike," and (iii) unmodified neurons. In the nucleus lateralis posterior neurons of the third class were not found. Most midbrain reticular neurons could be classified in the same three classes of the nonspecific thalamic nuclei; however, 11% of those units increased their activity 20 to 30 ms earlier than did the cortical units (class IV). Investigation of the activities of all these neuronal populations immediately prior to a spike and wave discharge showed that the rhythmic cycle of excitation-inhibition commenced earlier in the cortical neurons than in any other subcortical neuron. Moreover, there were some nonspecific thalamic neurons of class II with an inhibitory phase exactly coincident with the activation of class IV midbrain reticular neurons. These data suggest (i) a leading role of cortical neurons in initiating and maintaining a spike and wave burst; (ii) the involvement of a corticothalamocortical circuit in timing the bursts, and (iii) an accessory reticulothalamic loop also involved in regulating the intraburst frequency of the spike and wave complex.

Action Potentials↗

Different effects of electrical stimulation of the mesencephalic and pontine reticular formation on the release of dopamine and acetylcholine in the cat caudate nucleus.

The effects of unilateral electrical stimulation of the pontine (PRF) and mesencephalic (MRF) reticular formation on the release of acetylcholine (ACh) and of [3H]dopamine continuously synthesised from [3H]tyrosine were examined in both caudate nuclei of halothane-anaesthetised cats implanted with push-pull cannulae. Stimulation of PRF led to a prolonged bilateral increase in the release of [3H]dopamine, whereas a significant reduction in [3H]amine release was observed in the ipsilateral caudate nucleus following stimulation of the MRF. Changes in ACh release were also seen, but they seemed to be independent from those in dopamine release: the release of ACh was enhanced markedly in both caudate nuclei following stimulation of the MRF, whereas a more moderate increase in the release of ACh occurred ipsilaterally following stimulation of the PRF. These data indicate that both the MRF and the PRF are involved in the control of dopaminergic and cholinergic transmission in the basal ganglia.

Acetylcholine↗

Enhancement of rapid eye movement sleep in the rat by cholinergic and adenosinergic agonists infused into the pontine reticular formation.

The cholinergic agonist carbachol (1.1 mM) and the adenosinergic agonist cyclohexyladenosine (0.1 mM) were microinjected (60 nl) into the region of the caudal, oral pontine reticular formation of the rat. Local intracerebral infusion of each receptor agonist resulted in significant, long-lasting (at least 8 h) elevations in rapid eye movement sleep without reduction in latency to onset. The effects of carbachol were reduced by the muscarinic receptor antagonist atropine, while those of cyclohexyladenosine were reduced by the adenosinergic receptor antagonist 8-cyclopentyltheophylline. Atropine failed to antagonize the long-term induction of rapid eye movement sleep following cyclohexyladenosine, but did appear to suppress increases in the first 2 h. Similarity of effects on sleep parameters and the lack of additivity when injected consecutively are consistent with these agonist ligands targeting the same cellular mechanisms through their respective receptors. These findings suggest that transitory increases in the pons of either acetylcholine or adenosine may underlie long-lasting elevations in the amount of rapid eye movement sleep. Adenosine may play a role in the increased rapid eye movement sleep following prolonged wakefulness, as well as following conditions of stress and learning.

Adenosine↗

Studies of the role of the paramedian pontine reticular formation in the control of head-restrained and head-unrestrained gaze shifts.

Results of three experiments related to the role of the paramedian pontine reticular formation (PPRF) in the control of gaze are described. (1) Chronic unit recording methods, used to study the on-directions of short-lead burst neurons in head-restrained monkeys, and (2) reversible inactivation techniques confirmed the traditional view of the importance of PPRF in the control of horizontal eye movements. Reversible inactivation of neurons in the vicinity of identified short-lead burst neurons produced dramatic reductions in the speed of saccades to horizontal target displacements. The reductions in velocity were largely compensated for by an increase in saccade duration. Only minor, if any, effects were observed upon the velocity, duration, and amplitude of saccades to upward target displacements. (3) Microstimulation was applied to omnipause neurons to gate activity of excitatory burst neurons that discharge during coordinated eye-head movements. The microstimulation failed to noticeably slow (prevent) head movements when stimulation was applied during (prior to onset of) gaze shifts, suggesting that signals relayed to motoneurons innervating the neck muscles are not inhibited by the omnipause neurons. In other words, the desired gaze signal is parsed into eye and head pathways upstream of the excitatory burst neurons.

Animals↗

Enhanced GABA function in the angular complex (lateral periaqueductal grey matter and adjacent reticular formation) alters the postural component of striatal- or nigral-derived circling.

Unilateral injection of muscimol into the angular complex (lateral periaqueductal grey matter and adjacent reticular formation) caused ipsiversive rotation. Focal injection of picrotoxin into the same site produced contraversive rotation. Administration of apomorphine to animals with a unilateral 6-OHDA lesion of the left medial forebrain bundle caused contraversive rotation. Focal injection of muscimol into the angular complex reversed the direction of rotation such that apomorphine administration now produced ipsiversive circling. Unilateral injection of muscimol into substantia nigra zona reticulata caused contraversive rotation. Focal injection of picrotoxin into the same site produced ipsiversive rotation. The prior injection of muscimol into the ipsilateral angular complex prevented the contraversive rotation induced by intranigral administration of muscimol such that animals now showed ipsiversive circling. In both 6-OHDA-lesioned animals and animals receiving intranigral muscimol, focal injection of muscimol into the angular complex caused a reversal in the direction of circling through loss of the postural component with no obvious change in locomotor activity. Bilateral electrolytic lesions of the angular complex overall had no effect on amphetamine-induced locomotion. Manipulation of GABA function in the angular complex alters circling behaviour initiated from the striatum or substantia nigra by altering the postural component without affecting the locomotor response of the animals. The data suggest a critical role for the angular complex as an outflow station from basal ganglia.

Animals↗

Multi-segmental and generalized suppression of postural muscle tone evoked by stimulating the dorsal tegmental field and the medial pontine reticular formation in acute decerebrate cats.

In acute decerebrate cat, stimulation of the dorsal tegmental field (DTF) of the caudal pons along its midline and the medial pontine reticular formation (mPRF) evokes generalized suppression of postural muscle tone. In this study, the effects of stimulation of the DTF area were compared with those evoked by stimulating the mPRF unilaterally or bilaterally. By stimulating the mPRF bilaterally, simultaneous suppression of tonic activities in the neck, lumbar back, forelimb and hindlimb muscles, bilaterally, was evoked as that evoked by stimulating the DTF area alone.

Animals↗

Extracellular levels of serotonin in the medial pontine reticular formation in acute decerebrate cats with a microdialysis technique.

Extracellular levels of serotonin (5-HT) and 5-hydroxyindolacetic acid (5-HIAA) were measured in the medial pontine reticular formation of acute decerebrate cats. The mean basal levels of 5-HT and 5-HIAA were 26 fmol/20 microliters and 15 pmol/20 microliters. Perfusion of the dialysis probe with high K+ and Ca(2+)-free Ringer solution for 60 min resulted in 4.8-8.5 x increase and 25-48% decrease in the extracellular levels of 5-HT, respectively, in comparison to the basal 5-HT levels. Perfusion with TTX-added Ringer solution for 60 min resulted in a consistent decrease in the extracellular levels of 5-HT.

Animals↗