Blink reflex elicited by auditory stimulation in the rabbit

The pathway of the blink reflex, elicited by auditory stimulation, was investigated electrophysiologically. The reflex was recorded as microvibrations of the eyelid and was named the auditory-evoked eyelid microvibration (AMV). Pharmacophysiological studies suggest that AMV is closely related to the midbrain reticular formation and studies of electrical lesions in the midbrain reticular formation support this. Lesions in several parts of the central nervous system provide evidence that the inferior colliculus has an important role in AMV, and the cerebral cortex may have an inhibitory influence. Studies of brainstem transections indicate that the reflex pathway of AMV exists between the lower midbrain and the upper medulla. Because of its ease and simplicity, AMV is believed to be a useful test for evaluation of the function of the brainstem.     

Potentiation of fentanyl suppression of the jaw-opening reflex by transcranial electrical stimulation

Stinus et al. [L. Stinus, M. Auriacombe, J. Tignol, A. Limoge, M. Le Moal, Transcranial electrical stimulation with high frequency intermittent current (Limoge's) potentiates opiate-induced analgesia: blind studies, Pain, 42 (1990) 351–363.] observed that transcranial electrical stimulation (TCES) with high-frequency intermittent current potentiated opiate-induced analgesia using the tail-flick test. In unanesthetized, chronic preparations, electrical stimulation (0.5 Hz) of the lower incisor pulp of rats elicits a short-(6 ms) and a long-latency (12–18 ms) jaw-opening reflex (JOR) without any evidence of aversive behavior [J. Azérad, F. Fuentes, I. Lendais, A. Limoge, B. Pollin, Methods for selective tooth pulp stimulation in acute and chronic preparations in rats, J. Physiol., 406 (1988) 3P.]. Fentanyl increases thresholds of both reflexes and transiently suppresses the long-latency JOR. We then decided to look at the influence of TCES on both drug-induced mean of maximal threshold variation (MMTV) and duration of JOR suppression period. These parameters have been investigated in 43 Wistar rats with or without TCES administered for 3 h before the drug injection and throughout the testing period. TCES alone has no effect. In contrast, it significantly increases the duration of the reflex suppression period (149 ± 5% vs. control, P < 0.001) while fentanyl-increased reflex thresholds remain unchanged. The fentanyl-induced JOR suppression period returns to the control values 2 days later. When a second 3-h TCES session is delivered 2 or 4 days after the first TCES session, a similar increase of this suppression period is observed. Moreover, 2 days after a second TCES session, an increase of the duration of the fentanyl-induced JOR suppression period is systematically observed. In contrast, a 6-h TCES session never induces such effects. These results confirm a potentiating effect of TCES on opioid action and demonstrate the value of repeated TCES sessions.     

Modulation of a viscerosomatic reflex by electrical and chemical stimulation of hypothalamic structures in the rat

Ventromedial forebrain structures were stimulated electrically with short (10-ms) trains of pulses to test for effects on a viscerosomatic reflex. Stimulation at many hypothalamic sites led to an attenuation or even a complete inhibition of reflex activity. The most sensitive sites, however (i.e. those requiring currents of 50 μA or less to inhibit the reflex), were located in the anterior hypothalamus/preoptic area (AH/POA) and rostrally in the diagonal band of Broca (DBB). At certain sites the effects of electrical stimulation were compared with those of microinjection of an excitatory amino acid (DL-homocysteic acid) which is known to excite neuronal cell bodies and not axons. The results of this part of the study indicated that activation of cell bodies located in the ventromedial AH/POA (from the level of the optic chiasma caudally to the level of DBB rostrally) mediate, at least in part, the inhibitory effects on visceral afferent processing. These data are discussed in relation to a possible role of AH/POA in the spinal processing of nociceptive information of visceral origin.     

Potentiation by vestibular stimulation of the immobility reflex elicited by clamping in developing and adult rats

We have previously described (Exp. Neurol., 97: 315-326, 1987) that clamping of the neck elicits a profound immobility with flexion of limbs and ventroflexion of head and neck ('carrying posture'). We also observed that when a clamped rat pup is carried by the experimenter's hand, such carrying posture was potentiated. In the present paper we observed that in adult rats vestibular stimulation by circular acceleration potentiates the duration of immobility reflex by clamping and intensifies the limb flexion. In rat pups from 10 to 20 days, vestibular stimulation potentiated only the carrying posture but not the duration of the immobility reflex by clamping.     

Activation by high intensity peripheral nerve stimulation of adrenergic and opioidergic inhibition of a spinal reflex in the decerebrated rabbit

The short-latency sural to gastrocnemius reflex in the decerebrated rabbit was depressed for 20-30 min following high intensity conditioning stimulation of the common peroneal nerve. This effect was observed in animals with or without spinal section, but was greater in non-spinalized preparations. Graded conditioning stimuli showed that it was necessary to activate fine myelinated common peroneal axons to inhibit the reflex. In spinalized rabbits, maximal inhibition was achieved with conditioning stimulation of fine myelinated axons and was completely reversed by the opioid antagonist naloxone. In non-spinalized rabbits, maximal inhibition was only obtained with conditioning stimuli which activated non-myelinated axons. In these preparations the effects of common peroneal nerve stimuli were only blocked by co-administration of naloxone with the alpha 2-adrenoceptor antagonist idazoxan. Thus high intensity peripheral nerve stimuli activated a segmental opioidergic and a supraspinal adrenergic suppression of the sural-gastrocnemius withdrawal reflex. Such long-lasting suppression of reflex excitability may contribute to recovery from intensely noxious stimuli.     

Medial hypothalamic stimulation suppresses nociceptive spinal dorsal horn neurons but not the tail-flick reflex in the rat

This study investigated the potential analgesic effects of medial hypothalamic stimulation (HS) on a measure of nocifensive behavior (tail-flick test (TF] in awake rats, and potential inhibitory effects of identical HS on spinal dorsal horn neuronal responses to noxious skin heating in the same animals anesthetized with sodium pentobarbital. Sixty-five male Sprague-Dawley rats implanted with a bipolar stimulation electrode in histologically verified medial hypothalamic sites were tested behaviorally for TF suppression during HS (100 ms trains at 100 Hz, 3/s, 100-1100 microA) in 2-4 consecutive weekly test sessions. Thirty-three of these rats were then used in electrophysiological experiments to record responses of 36 dorsal horn units to noxious skin heating (48-54 degrees C, 10 s/2 min) of the hindfoot pad in the absence of and during HS. Behaviorally, 31/65 rats had no TF suppression at the highest HS intensity tested (1100 microA), 24/65 rats exhibited aversive behavior or motor activity which disallowed reliable TF testing, and only 10/65 rats showed TF suppression in at least one test session. In electrophysiological experiments, the heat-evoked responses of 25/36 dorsal horn units were inhibited to at least 50% of control during HS. The responses of 11 units remained at 65-100% of the control responses during HS of up to 1100 microA. In rats demonstrating TF suppression, 4/7 units were inhibited. In rats with no TF suppression, 10/15 units were inhibited, and in rats showing aversive behavior, 11/14 units were inhibited by HS. These data indicate that although HS suppresses spinal nociceptive neurons, it does not cause reliable TF suppression in unanesthetized rats and bring into question the often-held assumption that stimulation-evoked descending inhibition of spinal nociceptive neurons implies behavioral analgesia.     

Noxious stimulation of the toes evokes long-lasting, naloxone-reversible suppression of the sural-gastrocnemius reflex in the rabbit

Repetitive stimulation of the small myelinated and non-myelinated afferents of the common peroneal (c.p.) nerve evokes a long-lasting (20-25 min), naloxone-reversible inhibition of the sural-gastrocnemius reflex in the decerebrated and spinalized rabbit. Altering the number and frequency of stimuli applied to the c.p. nerve showed that this inhibition was dependent on temporal summation of afferent input from that nerve, and that the optimum frequency for producing the effect was between 2 and 10 Hz. Application of natural conditioning stimuli in and around the receptive field of the c.p. nerve showed that noxious, but not innocuous, mechanical and thermal stimuli could evoke long-lasting inhibition of the sural-gastrocnemius reflex. Thermal stimuli produced a biphasic change in the excitability of the reflex with facilitation followed by inhibition. The opioid antagonist naloxone (250 micrograms.kg-1) blocked all suppression resulting from these natural noxious stimuli. Chemical stimulation of the skin with mustard oil did not evoke naloxone-reversible inhibition of the reflex. These results indicate that intensely noxious stimuli can promote the release of opioid peptides in the spinal cord, and that one of the functions of these peptides may be to regulate the level of excitability in withdrawal reflex pathways.     

Lateral hypothalamic modulation of the gustatory-salivary reflex in rats

It is well recognized that the basic mechanism for the gustatory-salivary reflex is located in the lower brainstem and that suprabulbar structures possibly influence this mechanism. This study is designed to evaluate the neurophysiological mechanism underlying the effect of the lateral hypothalamic area (LHA) on the bulbar gustatory-salivary reflex. Submandibular salivary secretion and the electrical activity of the preganglionic parasympathetic fibers innervating the submandibular and sublingual glands were recorded in anesthetized rats. Stimulation of the tongue with high concentrations of chemical solutions (1 M and 2 M NaCl and 0.01 M and 0.05 M HCl) and/or pinching the tongue with a small clamp induced a profuse salivary secretion (3 to 28.5 microliter/5 min) recorded from a unilateral submandibular gland. The preganglionic fibers consisted of three types: taste-related fibers, which increased their firing rate by taste stimuli; pinch-related fibers, which increased their firing rate by pinching; and unidentified fibers, which did not respond to taste or pinching stimulations of the tongue. Electrical stimulation of the ipsilateral LHA caused the secretion of a small amount of saliva (1.5 microliter/5 min), and it appeared that taste-related fibers more often received polysynaptic connections from the LHA than other types of fibers. Conditioning stimulation of the LHA increased the magnitude of impulse discharges by the test stimulation of the tongue in the taste-related fibers. Therefore, the results of this study demonstrate that the LHA enhances the activity of the gustatory-salivary reflex in the lower brainstem.     

Modulation of the jaw-opening reflex by peripheral electrical stimulation

Modulation of the jaw-opening reflex (JOR) by peripheral electrical stimulation was studied in the rat. The JOR was evoked by electrical stimuli delivered to the tongue, infraorbital nerve, or tooth pulp chamber, and single-pulse conditioning stimuli were delivered to the forelimb, hind limb, or tail. Threshold current for eliciting the JOR was modulated in a biphasic manner with facilitation when the delay between conditioning and test stimuli was short (peaking at 10 to 15 ms) and inhibition at longer intervals (peaking at 40 to 60 ms). Modulation was similar for all peripheral conditioning sites and was not affected by Fentanyl, naloxone, or picrotoxin. Thus, the modulation of the JOR by single-pulse peripheral electrical stimulation is a widespread, nonsegmental phenomenon, and is probably not associated with the endogenous opiate system. Data collected during the course of this study call into question the usefulness of the JOR elicited by electrical stimulation in the rat incisor tooth pulp chamber as a pain model.     

Conditioning and reflex modification of the rabbit nictitating membrane response using electrical stimulation in auditory nuclei

Electrical brain stimulation (EBS) was applied to four nuclei in the auditory system, namely, the cochlear nucleus (CN), superior olive (SO), inferior colliculus, and medial geniculate. EBS was also applied to the pontine nuclei, which are the main relays for transmitting auditory conditioned stimuli (CSs) into the cerebellar pathways for conditioning of the nictitating membrane response (NMR). EBS of the CN, but no other site, yielded reflex modification, which was an increase in the unconditioned NMR to an airpuff unconditioned stimulus (US) when preceded by EBS. Throughout the experiment, EBS of the SO produced a distinctive distribution of NMRs, in which a high proportion had latencies less than 50 ms. When EBS was repeatedly paired with the airpuff US, conditioned responses (CRs) were acquired to comparable levels across all sites. At each site, response likelihood was an increasing function of the EBS parameters of pulse amplitude, pulse frequency, and pulse width. Combined with anatomical findings, these results indicate that multiple encodings of an auditory CS are sent to the pathways for the NMR.     

Potentiation of GABA inhibitory action in cerebellum by locus coeruleus stimulation

In cerebellum, excitatory and inhibitory responses of Purkinje cells, produced both synaptically and by microiontophoresis of putative amino acid neurotransmitters, have been shown previously to be enhanced during NE iontophoresis. The influence of locus coeruleus conditioning stimulation on Purkinje cell responses to GABA iontophoresis was examined to determine whether endogenous NE, released from synaptic terminals, could exert similar modulatory effects. Locus coeruleus stimulation at current intensities which alone elicited no direct depression of Purkinje cell spontaneous discharge potentiated the inhibition produced by GABA. Iontophoretic application of sotalol, a specific β-adrenergic blocker, antagonized this enhancement of GABA inhibition. Repetitive activation of the classic non-adrenergic cerebellar afferents did not enhance the GABA response, despite causing a direct depression in spontaneous rate. A neuromodulatory role is suggested for tonic adrenergic input in the mammalian central nervous system.     

Changes in the masseteric reflex during hypothalamic aphagia in cats

Experiments were conducted on 13 adult cats, each bearing a bipolar stimulating electrode in the mesencephalic nucleus of the trigeminal nerve, a bipolar recording electrode in the ipsilateral masseter muscle, bipolar cortical recording electrodes, and two monopolar electrodes implanted bilaterally in hypothalamic sites. In each animal, the masseteric reflex (MR), evoked by stimulation of the mesencephalic nucleus, was recorded during five daily sessions, which included periods of eating and sleeping. Subsequently, unilateral or bilateral electrolytic lesions were made in the hypothalamus through the implanted electrodes. Recording was continued for 24 h immediately after the lesions and then daily for 3 h. It was found that bilateral lesions in the lateral or far-lateral hypothalamus resulted both in aphagia and a decrease in the MR in nine of 13 cats. In two cats a strong aphagia was accompanied by a large decrease in the MR, and in three cats a mild aphagia or hypophagia was observed with a small decrease in the MR. The changes in feeding behavior in the remaining cats were not proportional to the changes in the MR. In three of these cats, the lesions resulted in a strong decrease in the MR but only in a mild decrease in feeding. In another animal a strong aphagia was accompanied only by a mild decrease in the MR. In two other cats an increase in the MR was observed while the animals were hypophagic. Unilateral lesions in the far-lateral hypothalamus in two cats resulted in hypophagia, but produced no change in the MR. From these data we conclude that alterations in the masseteric reflex, which is known to be directly related to food intake, may be dependent on brain mechanisms different from those responsible for the control of food intake, and that changes in oral motor activity related to mastication are not an indispensable component of hypothalamic aphagia.     

Phase changes induced by ketamine in the vertical vestibulo-ocular reflex in the rabbit

The effect of ketamine has been tested on the phase of the vertical vestibulo-ocular reflex of rabbits sinusoidally oscillated at various frequencies. A significant phase lag, predominantly affecting the macular component of the reflex, was observed. This action resembles that induced by Nembutal in the same preparation. A specific action of ketamine on synaptic transmission is suggested. Erroneous phase relationship between natural stimuli and responses can be obtained in experiments employing ketamine.     

Attenuation by spinal cord stimulation of a nociceptive reflex generated by colorectal distention in a rat model

The mechanisms underlying the cause and treatment of visceral pain of gastrointestinal origin are poorly understood. Previous clinical studies have shown that spinal cord stimulation (SCS) attenuates neuropathic and ischemic pain, and animal experiments have provided knowledge about probable physiological mechanisms. The goal of the present study was to investigate whether SCS influences colonic sensitivity in a conscious rat. A visceromotor behavioral response (VMR), induced by colorectal distention, was used to quantify the level of colonic sensitivity. Under anesthesia, an electrode (cathode) was placed on the dorsal surface of the spinal cord at L1. One week after implantation of the SCS electrode, the effects of stimulation delivered with different intensities (50 Hz, 0.2 ms for 30 min) on colonic sensitivity were determined. Nociceptive levels of colorectal distention (60 mm Hg for 10 min) induced an enhanced VMR quantified as an increased number of abdominal muscle contractions compared to controls in which the balloon catheter was inserted into the colorectal region but not distended. Colonic sensitization with acetic acid increased the VMR to innocuous levels of colorectal distention (30 mm Hg for 10 min). We found that SCS induced a significant depression of the VMR produced by colorectal distention in both normal rats and those with sensitized colons. The suppressive effect of SCS on colonic sensitivity suggests that SCS may have therapeutic potential for the treatment of visceral pain of gastrointestinal origin associated with abdominal cramping and painful abdominal spasms.     

Potentiation of rabbit trigeminal responses to capsaicin in a low pH environment

The sensitivity of 35 adult rabbit trigeminal ganglion neurons to low pH (pH 6.0), 10 μM capsaicin (CAP) and 10 μM capsaicin at low pH (CAP@pH6.0) was studied using voltage-clamp whole-cell recording techniques. Neurons responded to pH 6.0 with a transient inward current, followed by a more slowlt activating (sustained) net inward current. Responses to capsaicin showed only a sustained current. Capsaicin caused an increase in membrane conductance, whereas responses to low pH were associated with either a net increase or decrease in conductance. A subset of neurons (n = 14) responded to CAP@pH6.0 with a sustained current which exceeded the sum of the peak sustained currents evoked by CAP and pH 6.0 applied singularly by approximately a factor of 4. The current was associated with a substantial increase in membrane conductance. The present results indicate that, in addition to a direct conductance activating effect, protons have the ability to enhance the current evoked by capsaicin.     

Excitation of hypothalamic paraventricular neurons by stimulation of the raphe nuclei

Extracellular recordings were made from 467 anti-dromically identified neurosecretory neurons and 148 non-neurosecretory neurons in the paraventricular nucleus of the hypothalamus of hemispherectomized cats under pentobarbital anesthesia. Stimulation of the dorsal, median, and pontine raphe nuclei excited 31%, 26%, and 12% of neurosecretory neurons tested, respectively, and inhibited 9%, 7%, and 8%. The excitatory responses in 13 of 14 neurons tested were blocked by either of two intravenously administered 5-HT2 antagonists, cyproheptadine or methysergide. The 5-HT1A antagonist, (-)pindolol, partially blocked the excitatory responses elicited by raphe stimulation in three of five neurons tested. The inhibitory responses to raphe stimulation were not affected by application of these antagonists. More non-neurosecretory neurons than neurosecretory neurons were excited in response to raphe stimulation and these excitatory responses were also blocked by these antagonists. We conclude that most electrically stimulated synaptic inputs from the midbrain raphe nuclei to the hypothalamic paraventricular nucleus are excitatory and are mainly mediated by 5-HT2 receptors.     

Facilitation of baroreflex-induced bradycardia by stimulation of specific hypothalamic sites in the rat

Hypothalamic stimulation generally inhibits baroreflex-induced bradycardia. However, we have noted discrete areas of the rat hypothalamus which facilitate reflex bradycardia. The effects of hypothalamic stimulation on baroreflex-induced changes in heart rate were investigated in urethane-anesthetized rats (1.2 g/kg, i.p.; n = 6) instrumented with femoral arterial and venous catheters. Bipolar electrodes (250 micron diameter) were implanted stereotaxically in the hypothalamus. Baroreflex-induced bradycardia was elicited by phenylephrine (PE) injection (8-20 micrograms/kg). Responses to stimulation (STIM) (50-150 microA, 80 Hz, 0.5 ms), PE, and Stim + PE were studied for 1 min. In the ventral medial and anterior hypothalamus, STIM caused transient increases in blood pressure and no changes in heart rate. Peak blood pressure was lower during STIM + PE than during PE (144 +/- 5 vs 164 +/- 3 mm Hg; P less than 0.05). However, STIM + PE resulted in a lower heart rate compared to PE (194 +/- 22 22 vs 270 +/- 17 bpm; P less than 0.05). At 1 min, the heart rate in STIM + PE rats remained lower than in PE rats (205 +/- 37 vs 319 +/- 16 bpm; P less than 0.05). Atropine administration indicated that the facilitation was primarily parasympathetic in nature. These results identify specific hypothalamic regions which facilitate baroreflex-induced bradycardia by parasympathetic mechanisms.