Intracellular recording and HRP-staining of cortical neurons in feline ganglioside storage disease

To test whether there were any functional differences between the central gray substance (PVG) and other neural structures for the production of ultrasonic vocalization in the rat, an electrical stimulation experiment was systematically undertaken in diencephalic and mesencephalic tegmental regions. The sound production sensitivity to electrical stimulation was the highest in PVG. This suggested that the periventricular fiber system of Schu¨tz's bundle might be a possible neural structure underlying sound emission in the rat.     

Nociceptive projection from tooth pulp to the lateral hypothalamus in rats

The response of the rat lateral hypothalamic (LHA) neurons to tooth pulp electrical stimulation and the sensory projection pathway from the incisor pulp to the LHA were studied by electrophysiology and histology. 1) LHA neurons that responded to contralateral lower incisor pulp stimulation were found in the lateral part of the LHA. These neurons also responded to intensive tail pinch, but not to innocuous stimuli nor to applied glucose. 2) Histological study after injection of WGA-HRP into the lateral part of the LHA revealed many retrogradely labeled neurons in the ventral part of the periventricular gray (PVG) in the mesencephalon. 3) The PVG neurons responded antidromically to LHA stimulation and to contralateral lower incisor pulp stimulation. 4) After injection of WGA-HRP into the ventral portion of the PVG, many labeled cells were found in the contralateral subnucleus caudalis in the spinal tract of the trigeminal nucleus (NTST) where termination of the pulpal afferent was previously reported. It is thus suggested that the PVG is the most likely site of transmission relay of nociceptive inputs from incisors to the LHA.     

Modulatory effects of somatosensory electrical stimulation on neural activity of the dorsal cochlear nucleus of hamsters

The effects of somatosensory electrical stimulation on the dorsal cochlear nucleus (DCN) activity of control and tone-exposed hamsters were investigated. One to three weeks after sound exposure and control treatment, multiunit activity was recorded at the surface of the left DCN before, during, and after electrical stimulation of the basal part of the left pinna. The results demonstrated that sound exposure induced hyperactivity in the DCN. In response to electrical stimulation, neural activity in the DCN of both control and exposed animals manifested four response types: S-S, suppression occurring during and after stimulation; E-S, excitation occurring during stimulation and suppression after; S-E, suppression occurring during stimulation and excitation after; and E-E, excitation occurring during and after stimulation. The results showed that there was a higher incidence of suppressive (up to 70%) than of excitatory responses during and after stimulation in both groups. In addition, there was a significantly higher degree of suppression after, rather than during stimulation. At high levels of electrical current, the degree of the induced suppression was generally higher during and after stimulation in exposed animals than in controls. The similarity of our results to those of previous clinical studies further supports the view that DCN hyperactivity is a direct neural correlate of tinnitus and that somatosensory electrical stimulation can be used to modulate DCN hyperactivity. Optimization of stimulation strategy through activating only certain neural pathways and applying appropriate stimulation parameters may allow somatosensory electrical stimulation to be used as an effective tool for tinnitus suppression.     

The effects of septal stimulation on spontaneous and tail-shock evoked neuronal activity in the brainstem of the rat

The effects of stimulation of the lateral septum (LS) and the medial septum (MS) with single pulses of current were assessed on the neural activity evoked by a noxious tail shock as well as on spontaneous activity in 72 cells located in the periventricular gray (PVG), periaqueductal gray (PAG), dorsal raphe (DR) and nucleus raphe magnus (NRM) of the anesthetized rat. Pronounced inhibitory effects of evoked activity, occurring with a 10–15 ms latency and lasting 20–100 ms, were found primarily in the DR and ventrolateral PAG. Inhibition was not confined to nociceptive cells, and could also be observed in some units unresponsive to noxious stimulation. Short-lived excitatory effects were encountered mostly in the PVG and dorsolateral PAG units. The results were interpreted as support for the notion of septal participation in pain inhibition, and the hypothesis was advanced that its mode of action may involve suppression of aspects of nociception at the supraspinal level.     

Somatotopic organization of the human periventricular gray matter.

The periventricular gray (PVG) matter is an established anatomical target for chronic deep brain stimulation (DBS) in the treatment of certain intractable pain syndromes. Data relating to the representation of pain and other somatosensory modalities within the PVG in humans are negligible. We examined the character and location of somatosensory responses elicited by electrical stimulation along the length of the PVG in a patient who underwent unilateral DBS for intractable nociceptive head pain. Consistent responses were obtained and indicated the presence of a somatotopic representation in this region. The contralateral lower limb was represented cranially, followed by the upper limb and trunk, with the face area located caudally, near the level of the superior colliculi. Bilateral representation was only observed in the forehead and scalp.     

Serotonin involvement in descending inhibition of spinal nociceptive transmission produced by stimulation of medial diencephalon and basal forebrain

The responses of single lumbar dorsal horn neurons to noxious radiant heat stimuli (50 degrees C, 10 sec) applied to glabrous footpad skin were recorded in cats anesthetized with sodium pentobarbital and 70% N2O. Responses were markedly reduced during electrical stimulation (100-msec trains at 100 Hz, 3/sec, up to 400 microA) at sites in the medial diencephalic periventricular gray (PVG), preoptic area, and basal forebrain. A role for serotonin (5-hydroxytryptamine, 5-HT) was investigated by determining whether descending inhibition from these areas could be affected by (1) acute systemic administration of the 5-HT antagonist methysergide, or (2) depletion of central 5-HT levels by pretreatment with the 5-HT synthesis inhibitor p-chlorophenylalanine (PCPA; 500 mg/kg, i.p.). Inhibition produced by stimulation at these sites was reduced or abolished in 22 cases following administration of methysergide (0.2 to 1 mg/kg) to non-pretreated cats. In the PCPA-pretreated cats, stimulation in preoptic or basal forebrain areas inhibited the responses of 26 units to noxious skin heating to varying degrees; PVG stimulation inhibited the responses of 14 of 26 units, while the remainder were unaffected. The mean current threshold for inhibition produced by PVG or preoptic/basal forebrain stimulation was significantly higher, while mean inhibition at 200 microA was significantly lower, in units from PCPA-pretreated cats compared to those from non-pretreated cats. The results indicate that 5-HT may be involved in the mediation of spinal inhibition produced by medial diencephalic and basal forebrain stimulation.     

Integration of neural responses originating from different regions of the cortical somatosensory map

The neural pathways responsible for detecting peripheral tactile stimuli are well known; however, the interactions between different somatosensory regions have been less well investigated. This study demonstrates how the contralateral sensory response of rat barrel cortex to whisker stimulation is affected by stimulation of contralateral forepaw and ipsilateral whisker and forepaw. The barrel cortex in the right hemisphere was located using optical imaging. A 16-channel multielectrode was used to measure field potentials evoked by contralateral electrical stimulation of the whisker pad. A standard response in the right barrel cortex to single pulse electrical stimulation of the contralateral whisker pad was modulated by applying conditioning stimulation to one of three other regions of the body (the ipsilateral whisker pad, the ipsilateral or contralateral forepaws). In conditions where the standard contralateral whisker stimulus preceded the conditioning pulse, the size of response was identical to when it was stimulated alone. However, when the ipsilateral whisker and contralateral forepaw conditioning stimuli preceded the contralateral whisker pad stimulation, up to a 35% reduction in the contralateral whisker response was observed. These results confirm and extend previous studies [Proc. Natl. Acad. Sci. U. S. A. 97 (2000) 11026-11031; J. Neurosci. 21 (2001) 5251-5261], which show bilateral integration of neural activity within the rat somatosensory system. Furthermore, the longer latency of the inhibition following stimulation of the contralateral forepaw suggests the possible involvement of extracortical circuitry.     

Cardiovascular autonomic effects of transcutaneous auricular nerve stimulation via the tragus in the rat involve spinal cervical sensory afferent pathways

BackgroundElectrical stimulation on select areas of the external auricular dermatome influences the autonomic nervous system. It has been postulated that activation of the Auricular Branch of the Vagus Nerve (ABVN) mediates such autonomic changes. However, the underlying neural pathways mediating these effects are unknown and, further, our understanding of the anatomical distribution of the ABVN in the auricle has now been questioned.ObjectiveTo investigate the effects of electrical stimulation of the tragus on autonomic outputs in the rat and probe the underlying neural pathways.MethodsCentral neuronal projections from nerves innervating the external auricle were investigated by injections of the transganglionic tracer cholera toxin B chain (CTB) into the right tragus of Wistar rats. Physiological recordings of heart rate, perfusion pressure, respiratory rate and sympathetic nerve activity were made in an anaesthetic free Working Heart Brainstem Preparation (WHBP) of the rat and changes in response to electrical stimulation of the tragus analysed.ResultsNeuronal tracing from the tragus revealed that the densest CTB labelling was within laminae III-IV of the dorsal horn of the upper cervical spinal cord, ipsilateral to the injection sites. In the medulla oblongata, CTB labelled afferents were observed in the paratrigeminal nucleus, spinal trigeminal tract and cuneate nucleus. Surprisingly, only sparse labelling was observed in the vagal afferent termination site, the nucleus tractus solitarius. Recordings made from rats at night time revealed more robust sympathetic activity in comparison to day time rats, thus subsequent experiments were conducted in rats at night time. Electrical stimulation was delivered across the tragus for 5 min. Direct recording from the sympathetic chain revealed a central sympathoinhibition by up to 36% following tragus stimulation. Sympathoinhibition remained following sectioning of the cervical vagus nerve ipsilateral to the stimulation site, but was attenuated by sectioning of the upper cervical afferent nerve roots.ConclusionsInhibition of the sympathetic nervous system activity upon electrical stimulation of the tragus in the rat is mediated at least in part through sensory afferent projections to the upper cervical spinal cord. This challenges the notion that tragal stimulation is mediated by the auricular branch of the vagus nerve and suggests that alternative mechanisms may be involved.     

Electrical stimulation of the septal area in the rat: Prolonged suppression of water intake and correlation with self-stimulation

The role of the septal region in the control of drinking and self-stimulation behavior in the rat was investigated under conditions of forced and self-delivered brain stimulation. Sixty minutes of intermittent septal stimulation significantly reduced post-stimulation water consumption in water deprived rats without affecting the consumption of solid food or a liquid diet following food deprivation. In addition, those animals demonstrating the greatest suppression of water intake after forced stimulation also emitted the highest response rates to obtain rewarding brain stimulation from the same electrodes. These results support the hypothesis that the septal region plays an inhibitory role in the control of drinking behavior and suggests a relationship between the reward obtained by electrical brain stimulation and the central neural systems controlling thirst in the rat.     

Identification of ultrasonic vocalization substrates determined by electrical stimulation applied to the medulla oblongata in the rat

The aim of the present investigation was to identify the neural structures, within the rat medulla, that are responsible for rodent ultrasound production. Sound producing substrates were found to be located in the reticular formation and some cranial nerve nuclei as well as several other nuclei in the lateral and dorsomedial portions of the medulla. To estimate the degree of involvement in the generation of ultrasound signals, the sound response latencies were measured for each structure. The lateral reticular nucleus and the facial nucleus showed latencies that were significantly shorter than those for other nuclei, and they were assumed to have a primary part in rodent ultrasound production. Audible sounds of considerably longer latencies were produced exclusively by stimulation of the trigeminal spinal tract nucleus. No ultrasounds could be obtained in this region. These results were discussed in terms of innervations of the facial and laryngeal musculature by the specific neural structures. Present results were also discussed with reference to the roles of the bulbar monoaminergic neurons projecting to the spinal cord and the role of ascending nociceptive pathways.     

Reduced Oxytocin Release from the Neural Lobe of Lactating Rats is Associated with Reduced Pituitary Content and does not Reflect Reduced Excitability of Oxytocin Neurons

Lactating rats show reduced oxytocin release compared with virgin female rats in response to a variety of stimuli, including stress and osmotic stimulation. We sought to establish whether this is a consequence of a reduced response in the oxytocin cells, or of a change in stimulus-secretion coupling at the level of the neurosecretory terminals in the neural lobe. Blood sampling experiments in anaesthetized rats showed that systemic administration of cholecystokinin resulted in significantly less oxytocin release in lactating rats than in virgin female rats. Electrophysiological recordings of single cells in the supraoptic nucleus, however, showed no difference in the responsiveness of oxytocin cells to this stimulus. Oxytocin release evoked by electrical stimulation or by depolarization with high potassium solutions was lower in isolated neural lobes from lactating rats than in glands from non-lactating rats, whereas evoked vasopressin release was similar in the two groups. The lactating rat neural lobes had a reduced oxytocin content: to study the consequences of depletion we compared hormone release evoked by electrical stimulation in vitro in neural lobes from normal male rats, and from male rats given 2% NaCI to drink for 2 or 4 days. Saline drinking resulted in a reduction in gland content of both oxytocin and vasopressin, and the evoked release of both hormones was also significantly reduced when expressed as a percentage of the gland content, as was also seen for oxytocin release for glands from lactating rats. Finally, measurement of the extracellular potassium response to stimulation of the isolated neural lobe as an index of the excitability of neural lobe neurosecretory axons was unchanged in lactating rats compared with virgin female rats. Together, the data indicate that reduced oxytocin release observed in lactating rats is a simple consequence of reduced oxytocin content in the neural lobe rather than of a reduced excitability of the oxytocin neurons.     

Behavioral thresholds in the cat to frequency modulated sound and electrical stimulation of the auditory nerve

This behavioral study has helped confirm that cats can perceive low rates of electrical stimulation of the basal or high frequency end of the cochlea. The upper limit on the rate of stimulation that could be perceived was 600–800 pulse/sec. It was also shown that the behavioral threshold for low rates of change of a frequency modulated electrical stimulus was similar to that of sound. In the case of an electrical stimulus it was 85 pulses/sec/sec, and in the case of sound it was 97 cycles/sec/sec.     

Functional parallels between the neural and environmental antecedents of aggression

A series of experiments studied the elicitation of biting to electrical brain stimulation in acute squirrel monkey and rat preparations. Stimulation of several neural areas produced biting during stimulation while stimulation at other loci produced biting upon stimulation offset. A second study demonstrated that brain stimulation that produced biting at its offset also could reinforce biting, and here biting could be brought under discriminative environmental stimulus control. Further studies found that partially restrained but intact monkeys and rats would approach and bite inanimate targets during or following brain stimulation. Subsequent experiments demonstrated that unrestrained rats, depending again upon stimulation electrode placement, attacked mice following the offset of reinforcing brain stimulation, and this stimulation offset also produced differential electromyographic activity in masseter versus other muscle groups. Based upon both a literature survey and these findings, it was concluded that the neural mediation of aggressive behavior produced by brain stimulation was congruent with the causal relationships between the onset of aversive and offset of reinforcing antecedent environmental events and aggression.     

Repetitive vocalizations evoked by electrical stimulation of avian brains. IV. Evoked and spontaneous activity in expiratory and inspiratory nerves and muscles of the chicken (Gallus gallus).

The activity in respiratory nerves and muscles in response to electrical stimulation of vocal substrates in the brain and to CO2 stimulation of the respiratory centers was studied in 28 adult chickens. It was found that the same nerves and muscles were active during both vocalization and respiration. Stimulation of vocal substrates resulted in short latency bursting in the expiratory nerves and muscles. As stimulation intensity increased, progressively longer duration bursts composed of numerous subbursts were produced. By relating muscle activity with sound production , such bursting was shown to underlie evoked vocalizations. Background activity in inspiratory nerves and muscles continued uninterruptedly past stimulus onset only stopping when expiratory activity began. Thereafter inspiratory bursting reciprocated with expiratory bursting and was shown to underlie the intervals between vocalizations. The pattern of activity which was evoked by stimulating vocal substrates was found to strongly interact with the pattern of activity evoked by CO2 stimulation of the respiratory system. Simultaneous records of respiratory and tracheal muscles demonstrated that the same information was sent to both groups of muscles during evoked vocalization. Activity in the respiratory muscles was recorded during spontaneous vocalization of a free-moving bird and was found to resemble that recorded from anesthetized birds. Finally the activity of single units in the obex region of the medulla was recorded during electrical stimulation of vocal substrates and during CO2 stimulation of the respiratory system. Rhythmically active units were found only in the medulla. Unit activity paralleled that found in the nerves and muscles. On the basis on the data accumulated, two models of the chicken vocal system are presented. The first is a model of the sound-producing structures of the chicken. The second is a model of the neural machinery which controls the sound-producing structures. The two models are used as a basis for an explanation of the production of voclizations by the chick of the same species.     

Cortical microstimulation in auditory cortex of rat elicits best-frequency dependent behaviors

Electrical activation of the auditory cortex has been shown to elicit an auditory sensation; however, the perceptual effects of auditory cortical microstimulation delivered through penetrating microelectrodes have not been clearly elucidated. This study examines the relationship between electrical microstimulus location within the adult rat auditory cortex and the subsequent behavior induced. Four rats were trained on an auditory frequency discrimination task and their lever-pressing behavior in response to stimuli of intermediate auditory frequencies was quantified. Each trained rat was then implanted with a microwire array in the auditory cortex of the left hemisphere. Best frequencies (BFs) of each electrode in the array were determined by both local field potential and multi-unit spike-rate activity evoked by pure tone stimuli. A cross-dimensional psychophysical generalization paradigm was used to evaluate cortical microstimulation-induced behavior. Using the BFs of each electrode, the microstimulation-induced behavior was evaluated relative to the auditory-induced behavior. Microstimulation resulted in behavior that was dependent on the BFs of the electrodes used for stimulation. These results are consistent with recent reports indicating that electrophysiological recordings of neural responses to sensory stimuli may provide insight into the sensation generated by electrical stimulation of the same sensory neural tissue.     

A preparation for studying electrical stimulation of the retina in vivo in rat

A remaining challenge to the development of electronic prostheses for vision is improving the effectiveness of retinal stimulation. Electrode design and stimulus parameters need to be optimized such that the neural output from the retina conveys information to the mind's eye that aids the patient in interpreting his or her environment. This optimization will require a detailed understanding of the response of the retina to electrical stimulation. The identity and response characteristics of the cellular targets of stimulation need to be defined and evaluated. Described here is an in vivo preparation for studying electrical stimulation of the retina in rat at the cellular level. The use of rat makes available a number of well-described models of retinal disease that motivate prosthesis development. Artificial stimulation can be investigated by adapting techniques traditionally employed to study the response of the retina to photic stimuli, such as recording at the cornea, single-cell recording, and pharmacological dissection of the response. Pilot studies include amplitude-intensity response data for subretinal and transretinal stimulation paradigms recorded in wild-type rats and a transgenic rat model of autosomal dominant retinitis pigmentosa. The ability to record single-unit ganglion cell activity in vivo is also demonstrated.     

Vocalization after electrostimulation of the brain of pigeons in relation to heart- and breathing-rates

Vocalizations elicited by electrical stimulation in the midbrain of pigeons were studied in connection with heart- and breathing-rates. Most of the sites where sound production could be elicited were found in the nucleus intercollicularis (ICo), lying rostromedial to the nucleus mesencephalicus lateralis, pars dorsalis (Mld). All vocalizations were accompanied by an increase of heart- and breathing-rates. Electrical stimulation elicits predominantly changes in breathing-rate which can be accompanied by passive vocalization.     

Vagal stimulation-induced gastric acid secretion in the anesthetized rat

The dynamics of gastric acid secretion induced by electrical stimulation of the cervical vagus in the anesthetized rat were investigated using a continuous collection-titration system permitting high temporal resolution. Stimulation with pulse rates of 2 or 4 impulses/s (pps) produced maximal gastric acid responses with small cardiovascular effects. With continuous stimulation, secretion was sustained for at least 1 h. Frequency-response profiles suggested that the parietal cells are innervated predominantly by fine-caliber C-fibers. Continuous stimulation was 3 times as effective as stimulation in bursts of higher frequencies. The minimal latency for the onset of secretion was 2.6 min at 4 pps, however, one- and two-min stimulations still produced proportionate but delayed secretory responses. It is concluded that, with low frequency cervical vagus stimulation, the rat stomach preparation described and employed in the present experiment is a useful model for further studies on the interaction of neural and humoral factors on gastric acid secretion.     

体外低频电刺激促进背根神经元突起的生长：上调钙离子介导的脑源性神经营养因子表达可为其途径

摘要 背景：短时低频电刺激已被证明可显著促进周围神经系统损伤后轴突的再生,目前对电刺激是如何促进其突起生长还有待证实。 目的：体外培养背根神经元,观察短时低频电刺激对神经元突起生长的影响,探讨电刺激发挥作用可能的细胞信号分子。 设计、时间及地点：体外培养背根神经元及离体电刺激处理,于2007-05/2008-10在上海交通大学医学院完成。 材料：新生48h Sprague-Dawley大鼠20只(中科院上海生命科学研究所动科所)。 方法：体外培养背根神经元,随机分为两组,正常对照组(n = 6)及电刺激组(n = 8)。电刺激组施予离体电刺激(20Hz, 100μs, 3V),持续作用1h。为探讨电刺激发挥作用经由的细胞信号分子,在施予电刺激前预先加入钙离子通道阻滞剂Nifedipine孵育4小时,再给予电刺激,再次检测各组神经元突起的生长情况。 主要观察指标：β-tubulin染神经元,测量各组神经元突起的长度。RT-PCR、 western blot和ELISA分别检测神经元BDNF的表达和分泌。 结果：短时低频电刺激促进神经元突起的生长,增强其表达和分泌BDNF (P < 0.05)。Nifedipine的使用削弱了电刺激对神经元突起生长及BDNF合成的促进作用 (P < 0.05)。 结论：短时低频电刺激促进体外培养的背根神经元突起的生长及BDNF的合成,初步认为电刺激对神经元突起生长的促进作用,至少通过促发钙内流所致BDNF表达和分泌增多所致。 关键词：电刺激;背根神经元;突起生长;BDNF;Ca2+