Fictive respiratory rhythm in the isolated brainstem of frogs

Spontaneous rhythmically bursting activity was recorded from the trigeminal, vagal and hypoglossal nerve roots of the isolated brainstem from the frogsRana catesbeiana andRana pipiens superfused with a bicarbonate-free HEPES-buffer solution. Burst frequency, burst duration and the activity profile of the spontaneous neural discharges in vitro resembled those of a less radical preparation, the decerebrate, fictively breathing frog. After complete midsagittal section, each half of the isolated brainstem generated its own rhythmic neural activity which resembled that of the intact isolated brainstem. The spontaneous activity generated within each half of the brainstem is probably coordinated by decussating axons or by groups of neurons located along the midline of the brainstem. Our results suggest that these coordinating entities extend the length of the brainstem (in a rostro-caudal dimension) and the degree of contact rather than the location of the contact between the two halves of the brainstem determines the synchronization of the right and left halves. Burst frequency of both the intact and hemisected brainstem preparation was decreased by alkaline challenge and increased by acid challenge. We conclude that this endogeneous rhythmic activity represents the efferent motor output underlying lung ventilation in these animals.Abbreviations EMG electromyogram - ENG electroneurogram - V trigeminal nerve - Vmd mandibular branch of trigeminal nerve - X vagal nerve - X1 laryngeal branch of vagal nerve - H hypoglossal nerve - Hsh sternohyoid branch of hypoglossal nerve - Hm main branch of hypoglossal nerve     

Cerebellar afferents in the frogs,Rana esculenta and Rana temporaria

Summary Afferents to the cerebellum in frogs (Rana esculenta, Rana temporaria) were studied by use of retrograde transport of horseradish peroxidase. Following injections restricted to the molecular layer of the cerebellum cell labelling was found in the contralateral inferior olive and the ventral portion of the caudal medullary raphe. Injections involving the granular layer resulted in labelling in the ventral horn of the cervical spinal cord, the caudal spinal trigeminal nucleus, the nucleus caudalis and the medial portion of the nucleus ventralis of the vestibular nerve, the inferior reticular nucleus and the nucleus of the fasciculus longitudinalis medialis. Following larger injections, which may have spread significantly into the cerebellar, secondary gustatory, trigeminal or vestibular nuclei, labelled cell bodies were also found in the nucleus ruber, nucleus solitarius, the rostral spinal trigeminal nucleus and the rostral rhombencephalic reticular formation. It is unclear whether the fibers from these latter areas innervate the cerebellum of the frog, as they do in mammals, or only reach the underlying areas. This situation emphasizes a limitation of the HRP technique when applied to small structures as is often the case in lower vertebrates.Supported by Grant Gr 276 to U. G.-C. from the Deutsche Forschungsgemeinschaft.     

Waveform generation in Rhamphichthys rostratus (L.) (Teleostei,Gymnotiformes)

Rhamphichthys rostratus (L.) emits brief pulses (2 ms) repeated very regularly at 50 Hz. The electric organ shows a heterogeneous distribution of the electrocyte tubes and the occurrence of three electrocyte types (caudally innervated, rostrally innervated and marginallycaudally innervated). In the sub-opercular region the electric organ consists of a pair of tubes containing only caudally innervated electrocytes. At the abdominal region the EO consists of three pairs of tubes. Each pair contains one of the described electrocyte types. The number of electrocyte tubes increases toward the tail to reach nine or ten pairs in the most caudal segments. In the intermediate region most tubes contain doubly innervated electrocytes except the ventral pair that contains caudally innervated electrocytes. The caudal 25% contains exclusively caudally innervated electrocytes. The electric organ discharge consists of five wave components (V1 to V5). Electrophysiological data are consistent with the hypothesis that V1 results from the activity of the rostral faces of rostrally innervated electrocytes. V2 results from the activities of rostral faces of marginally-caudally innervated electrocytes while V3 results from the activities of caudal faces of most electrocytes. Curarization experiments demonstrated that V4 and V5 result from action potential invasion and are not directly elicited by neural activity.Abbreviations AEN1 anterior electromotor nerve 1 - AEN2 anterior electromotor nerve 2 - BMB boraxic methylene blue - CIE caudally innervated electrocytes - EMF electromotive force - EO electric organ - EOD electric organ discharge - I current amplitude - MCIE marginally-caudally innervated electrocytes - MT medial tubes - PEN posterior electromotor nerve - R _n internal impedance - RIE rostrally innervated electrocytes - Rl load resistor - SAT short abdominal tubes - V voltage amplitude     

Ventrolateral medullary respiratory network participation in the expiration reflex in the cat.

The expiration reflex is a distinct airway defensive response characterized by a brief, intense expiratory effort and coordinated adduction and abduction of the laryngeal folds. This study addressed the hypothesis that the ventrolateral medullary respiratory network participates in the reflex. Extracellular neuron activity was recorded with microelectrode arrays in decerebrated, neuromuscular-blocked, ventilated cats. In 32 recordings (17 cats), 232 neurons were monitored in the rostral (including B?tzinger and pre-B?tzinger complexes) and caudal ventral respiratory group. Neurons were classified by firing pattern, evaluated for spinal projections, functional associations with recurrent laryngeal and lumbar nerves, and firing rate changes during brief, large increases in lumbar motor nerve discharge (fictive expiration reflex, FER) elicited during mechanical stimulation of the vocal folds. Two hundred eight neurons were respiratory modulated, and 24 were nonrespiratory; 104 of the respiratory and 6 of the nonrespiratory-modulated neurons had altered peak firing rates during the FER. Increased firing rates of bulbospinal neurons and expiratory laryngeal premotor and motoneurons during the expiratory burst of FER were accompanied by changes in the firing patterns of putative propriobulbar neurons proposed to participate in the eupneic respiratory network. The results support the hypothesis that elements of the rostral and caudal ventral respiratory groups participate in generating and shaping the motor output of the FER. A model is proposed for the participation of the respiratory network in the expiration reflex.     

Enhanced baroreflex-mediated inhibition of respiration after muscimol dialysis in the rostroventral medulla.

The rostral ventral medulla (RVM) may be important in the control of cardiorespiratory interactions. We hypothesized that inhibition of the RVM would enhance inhibition of breathing associated with transient blood pressure elevations. In 25 piglets 3-16 days of age, we studied the effect of acutely increasing blood pressure, by systemic infusion of phenylephrine, on respiratory activity before and after inhibition of neural activity in the RVM by dialysis of 10 mM muscimol, a GABA(A)-receptor agonist. Muscimol dialysis through probes that were placed along the ventral medullary surface from approximately 1 mm rostral to the facial nucleus to approximately 0.5 mm caudal to the facial nucleus augmented the respiratory inhibition associated with acute increases in blood pressure. No similar enhancement of respiratory inhibition after phenylephrine treatment was seen in six control animals that did not receive muscimol dialysis. We conclude that the piglet RVM participates in cardiorespiratory interactions and that dysfunction of homologous regions in the human infant could lead to cardiorespiratory instability and may be involved in the pathogenesis of sudden infant death syndrome.     

Characterization of respiratory-related neurons in the isolated brainstem of the frog

Using intra- and extracellular recording techniques we examined the spontaneous discharge and membrane properties of respiratory-related neurons in isolated brainstem preparations of the frogs Rana catesbeiana and Rana pipiens that display spontaneous respiratory related activity in vitro. We observed neurons that depolarize during the fictive lung ventilation cycle as well as neurons that depolarize during the non-lung ventilation phase. Respiratory-related neurons demonstrated significant decreases in membrane input resistance during the fictive lung ventilation cycle but showed no evidence of voltage-dependent membrane conductances activated near resting membrane potential. Furthermore, respiratory neurons showed little spike frequency adaptation, their oscillatory activity was not dissociated from the global respiratory motor output following imposed changes in membrane potential, and spontaneous fluctuations in membrane potential were not observed following reversible interruption of respiratory burst activity by application of solutions low in calcium and high in magnesium. Taken together these results suggest that bulbar respiratory neurons in the isolated frog brainstem sampled in our study do not display endogenous bursting characteristics. Rather, they are strongly influenced by synaptic input. Accepted: 20 March 1997     

Nitric oxide changes its role as a modulator of respiratory motor activity during development in the bullfrog (Rana catesbeiana)

Nitric oxide (NO) is a unique chemical messenger that has been shown to play a role in the modulation of breathing in amphibians and other vertebrates. In the post-metamorphic tadpole and adult amphibian brainstem, NO, acting via the neuronal isoform of nitric oxide synthase (nNOS), is excitatory to the generation of lung burst activity. In this study, we examine the modulation of breathing by NO during development of the amphibian brainstem. Isolated brainstem preparations from pre-metamorphic and late-stage post-metamorphic tadpoles (Rana catesbeiana) were used to determine the role of NO in modulating central respiratory neural activity. Respiratory neural activity was monitored with suction electrodes recording extracellular activity of cranial nerve rootlets that innervate respiratory musculature. Brainstems were superfused with an artificial cerebrospinal fluid (aCSF) at 20-22 degrees C containing l-nitroarginine (l-NA; 1-10 mM), a non-selective NOS inhibitor. In pre-metamorphic tadpoles, l-NA increased fictive gill ventilation frequency and amplitude, and increased lung burst frequency. By contrast, l-NA applied to the post-metamorphic tadpole brainstem had little effect on fictive buccal activity, but significantly decreased lung burst frequency and the frequency of lung burst episodes. These data indicate that early in development, NO provides a tonic inhibitory input to gill and lung burst activity, but as development progresses, NO provides an excitatory input to lung ventilation. This changing role for NO coincides with the shift in importance in the different respiratory modes during development in amphibians; that is, pre-metamorphic tadpoles rely predominantly on gill ventilation whereas post-metamorphic tadpoles have lost the gills and are obligate air-breathers primarily using lungs for gas exchange. We hypothesize that NO provides a tonic input to the respiratory CPG during development and this changing role reflects the modulatory influence of NO on inhibitory or excitatory modulators or neurotransmitters involved in the generation of respiratory rhythm.     

Are pacemaker properties required for respiratory rhythm generation in adult turtle brain stems in vitro?

The role of pacemaker properties in vertebrate respiratory rhythm generation is not well understood. To address this question from a comparative perspective, brain stems from adult turtles were isolated in vitro, and respiratory motor bursts were recorded on hypoglossal (XII) nerve rootlets. The goal was to test whether burst frequency could be altered by conditions known to alter respiratory pacemaker neuron activity in mammals (e.g., increased bath KCl or blockade of specific inward currents). While bathed in artificial cerebrospinal fluid (aCSF), respiratory burst frequency was not correlated with changes in bath KCl (0.5-10.0 mM). Riluzole (50 microM; persistent Na(+) channel blocker) increased burst frequency by 31 +/- 5% (P < 0.05) and decreased burst amplitude by 42 +/- 4% (P < 0.05). In contrast, flufenamic acid (FFA, 20-500 microM; Ca(2+)-activated cation channel blocker) reduced and abolished burst frequency in a dose- and time-dependent manner (P < 0.05). During synaptic inhibition blockade with bicuculline (50 microM; GABA(A) channel blocker) and strychnine (50 muM; glycine receptor blocker), rhythmic motor activity persisted, and burst frequency was directly correlated with extracellular KCl (0.5-10.0 mM; P = 0.005). During synaptic inhibition blockade, riluzole (50 microM) did not alter burst frequency, whereas FFA (100 microM) abolished burst frequency (P < 0.05). These data are most consistent with the hypothesis that turtle respiratory rhythm generation requires Ca(2+)-activated cation channels but not pacemaker neurons, which thereby favors the group-pacemaker model. During synaptic inhibition blockade, however, the rhythm generator appears to be transformed into a pacemaker-driven network that requires Ca(2+)-activated cation channels.     

Control of phrenic nerve activity and blood pressure by the medullary raphe nuclei in cats

Electrical and chemical stimulation methods were used to determine the topographic organization of the medullary raphe nuclei (MRN) in controlling the systemic arterial blood pressure (BP) and phrenic nerve activities (PNA). Decerebrated, unanesthetized and bilateral vagotomized cats were used. Effective points in the MRN were systematically explored with constant current stimulation. We found stimulation of the rostral MRN produced a decrease in PNA amplitude and increase in BP and PNA frequency. Stimulation of the caudal MRN produced increases in BP and the amplitude and frequency of PNA. Microinjection of glutamate solution into the caudal or the rostral MRN points produced qualitatively similar results. Thus, we concluded that the caudal MRN neurons had excitatory connections whereas the rostral MRN neurons had excitatory and inhibitory connections to the cardiovascular preganglionic neurons and the phrenic nerve motoneurons.     

The whole-body shortening reflex of the medicinal leech: motor pattern,sensory basis,and interneuronal pathways

The leech whole-body shortening reflex consists of a rapid contraction of the body elicited by a mechanical stimulus to the anterior of the animal. We used a variety of reduced preparations — semi-intact, body wall, and isolated nerve cord — to begin to elucidate the neural basis of this reflex in the medicinal leech Hirudo medicinalis. The motor pattern of the reflex involved an activation of excitatory motor neurons innervating dorsal and ventral longitudinal muscles (dorsal excitors and ventral excitors respectively), as well as the L cell, a motor neuron innervating both dorsal and ventral longitudinal muscles. The sensory input for the reflex was provided primarily by the T (touch) and P (pressure) types of identified mechanosensory neuron. The S cell network, a set of electrically-coupled interneurons which makes up a fast conducting pathway in the leech nerve cord, was active during shortening and accounted for the shortest-latency excitation of the L cells. Other, parallel, interneuronal pathways contributed to shortening as well. The whole-body shortening reflex was shown to be distinct from the previously described local shortening behavior of the leech in its sensory threshold, motor pattern, and (at least partially) in its interneuronal basis.Abbreviations conn connective - DE dorsal excitor motor neuron - DI dorsal inhibitor motor neuron - DP dorsal posterior nerve - DP:B1 dorsal posterior nerve branch 1 - DP:B2 dorsal posterior nerve branch 2 - MG midbody ganglion - VE ventral excitor motor neuron - VI ventral inhibitor motor neuron     

Sound production evoked by electrical stimulation of the forebrain in the oyster toadfish

In mammals, birds and amphibians the neural pathways controlling sound production descend from higher centers in the forebrain, whereas in fishes only brainstem and spinal centers have been explicitly implicated in sound production. We now report that electrical stimulation of the forebrain of the oyster toadfish (Opsanus tau) readily evokes both the agonistic grunt and the courtship boatwhistle. Boatwhistles are more realistic than ones previously evoked from lower centers. Positive stimulation sites are localized in the preoptic area (nucleus preopticus parvocellularis anterior) and the supracommissural nucleus of the ventral telencephalon, a likely homologue of the amygdala. Both sites contain gonadal steroid-concentrating neurons and play a central role in fish courtship behavior. Evoked sounds form a continuum from knock grunts, burst grunts, transition boatwhistles to complete boatwhistles; sound pressure level (SPL), fundamental frequency and duration increase consistently within the continuum. For all sound types, SPLs exhibit the smallest variation (coefficients of variation of 2.7 to 5.7%), fundamental frequency is intermediate (5 to 13%) and durations vary most widely (18 to 60%). Boatwhistles, with the smallest variation and greatest amplitude, are likely generated by a maximal output of the CNS and sonic muscles. Grunt SPLs however, vary over a range of 26 dB for all fish and by as much as 18 dB in an individual, suggesting recruitment of variable numbers of motor units despite electrical coupling within the sonic motor nucleus.Abbreviations AC anterior commissure - CNS central nervous system - DHT dihydrotestosterone - Dm medial nucleus of dorsal telenecphalon - DTAM dorsal tegmental area of medulla - E estrogen - HRP horseradish peroxidase - PM nucleus praeopticus magnocellularis - POA preoptic area - PPa nucleus praeopticus parvocellularis anterior - SMA sonic motor area - SMN sonic motor nucleus - SPL sound pressure level - T testosterone - VS supracommissural nucleus of ventral telencephalon - Vv ventral nucleus of ventral telencephalon     

The distribution of several amino acids in specific ganglia and nerve bundles of the lobster

Using the technique of measuring DNP-amino acid methyl esters by gas-liquid chromatography, the distribution of alanine, proline, glycine, GABA, glutamate and aspartate was determined in individual ganglia and the associated nerve bundles between these ganglia after isolation from the nervous system of the lobster, Homarus americanus. The brain or supraesophageal ganglion (27.2 mg) and the next 5 thoracic ganglia (varying from 24 to 10 mg in a rostral–caudal direction) as well as the nerve bundles connecting these ganglia were used. GABA and aspartate values varied the most among the individual ganglia; highest values were found in the second and third thoracic ganglia. The levels of alanine, proline, glycine and glutamate varied very little from ganglion to ganglion; however, the values for these amino acids did exhibit some variability among the individual connectives. The highest value for each was in the nerve bundle between the first and second thoracic ganglion. Glycine was present at the highest level of any of the amino acids whereas GABA was at the lowest level in the individual structures assayed.     

Blocking of glutamate ionotropic receptors: Influence on the respiratory activity generated by medullo-spinal preparations of rats in hypoxia

We studied the influences of a non-competitive blocker of glutamate NMDA-receptors ketamune and of a competitive blocker of AMPA-kainate non-NMDA receptors, CNQX, on the respiratory activity generelated by superfusedin situ semi-isolated medullo-spinal preparations (SIMSP) of 3- to 4-day-old rats. We compared the ampes recorded under conditions of superfusion, a standard solution and the solution saturated with an anoxic isocapine gas mixture were compared; pO₂ in these solutions were 440±22 and 41±8 mm Hg, respectively. The experments were carried out with the ventrolateral medullary region (VLMR) left intact or after separation of its rostral part, which propertchonally corresponded to the chemosensitiveM zone. A 3-min-long hypoxic test initially evoked an increase in the frequency of inspiratory discharges (IR) in the phrenic nerve followed by a frequency drop within the final half of the test. After the rostral VLMR had been separated, the hypoxic test did not elicit a significant decrease in the IR frequency. After preliminary application of 1.0 or 10.0 μM ketamine or CNQX on intact preparations, the IR frequency under hypoxic conditions dropped within the first half of the test and increased in the second half, while the amplitude and integral intensity of these discharges were depressed more intensively than in hypoxia with no applications. Using ketamme and CNQX in the same concentrations resulted in significant drops in the amplitude, frequency, and integral intensity of IR recorde din the hypoxic test. Our experiments showed that in the early postnatal period glutamate ionotropic receptors of rostral VLMR neurons are involved in the control of IR frequency under hypoxic conditions. The possible role of glutamatergic control of the respiratory rhythm and mechanisms of the influences resulting from blocking of NMDA and non-NMDA receptors on the parameters of respiratory activity are discussed.     

Modulation of swimming rhythmicity by 5-hydroxytryptamine during post-embryonic development in Xenopus laevis.

During the first 24 h of post-embryonic development in Xenopus laevis, a rapid change in the neural activity underlying swimming occurs in which the duration of ventral root discharge on each cycle increases from a single compound impulse to discrete bursts of activity. Moreover, this change in motor output progresses rostrocaudally, suggesting that it could result from the influence of a descending neural pathway upon the spinal rhythm-generating circuitry during early post-embryonic development. To begin to examine whether serotonergic neurons of brainstem raphe nuclei might have a role in this swimming development, we have studied the effects of 5-hydroxytryptamine (5HT) on fictive swimming in embryonic and larval animals. As previously demonstrated for other vertebrate locomotor rhythms, we find that bath-applied 5HT enhances the duration of motor activity on each cycle of larval fictive swimming. In addition, our results show that the sensitivity of the swimming rhythm to exogenous 5HT follows a strict rostrocaudal gradient. In young embryos (stages 32-36) 5HT does not affect the duration of ventral root impulses per cycle; by the time of hatching (stage 37/38), rostral but not caudal discharge is enhanced, and by stage 42 (24 h post-hatching) 5HT can increase motor burst durations along most of the length of the animal. These reversible changes induced by bath-applied 5HT closely resemble the normal rostrocaudal development of burst discharge during swimming in animals some 12 h older.(ABSTRACT TRUNCATED AT 250 WORDS)     

Waveform generation of the electric organ discharge inGymnotus carapo

Summary The electric organ (EO) ofGymnotus carapo was studied using different neurohistological techniques including conventional electron microscopy. The electric tissue extends along the fish body from the pectoral girdle to the tip of the tail, constituting a single, undivided organ. However, taking into account the number, arrangement, and innervation of the electrocytes, it is possible to divide the EO into three different portions. The more rostral portion is included within the ventral wall of the abdominal cavity. It consists of singly and doubly innervated electrocytes arranged in two rows at each side of the midline. Innervation of this zone is supplied by the first 5–7 segmental nerves and by the anterior electromotor nerves. Segmental nerves terminate on the rostral faces of doubly innervated electrocytes; axons stemming from the anterior electromotor nerves end on the caudal faces of both doubly and singly innervated electrocytes. There is an intermediate body-tail region in which the electrocytes are arranged in four dorsoventral tubes (tubes 1 to 4) on each side of the midline. In this zone, doubly innervated electrocytes (confined within tube 1) coexist together with singly innervated ones, receiving nerve terminals on their caudal faces (tubes 2, 3, and 4). The innervation characteristics appear modified at more distal portions of the tail where the doubly innervated electrocytes of tube 1 are replaced by singly innervated units. The most distal portion of the EO (approximately its terminal 30%) consists of numerous, homogeneously innervated electrocytes with nerve endings distributed exclusively on their caudal faces. Nerve supply to the intermediate and distal regions derives from the posterior electromotor nerves (PENs) which appear as well-defined anatomical entities beyond the level of metamere XXVII. At the bodytail and more distal regions the innervation pattern of the EO is particularly complex. Thin nerve trunks arise from the PENs and project ventrally toward the electrocyte tubes. Before reaching the electric tissue the electromotor axons branch frequently. Our anatomical studies indicate that the EO is heterogeneous, a feature consistent with most recent electrophysiological and biophysical experiments.Abbreviations AEN anterior electromotor nerve - EMN electromotoneurons - EO electric organ - EOD electric organ discharge - LLN lateral line nerve - PEN posterior electromotor nerve     

Developmental nicotine exposure alters cholinergic control of respiratory frequency in neonatal rats

Prenatal nicotine exposure with continued exposure through breast milk over the first week of life (developmental nicotine exposure, DNE) alters the development of brainstem circuits that control breathing. Here, we test the hypothesis that DNE alters the respiratory motor response to endogenous and exogenous acetylcholine (ACh) in neonatal rats. We used the brainstem‐spinal cord preparation in the split‐bath configuration, and applied drugs to the brainstem compartment while measuring the burst frequency and amplitude of the fourth cervical ventral nerve roots (C4VR), which contain the axons of phrenic motoneurons. We applied ACh alone; the nicotinic acetylcholine receptor (nAChR) antagonist curare, either alone or in the presence of ACh; and the muscarinic acetylcholine receptor (mAChR) antagonist atropine, either alone or in the presence of ACh. The main findings include: (1) atropine reduced frequency similarly in controls and DNE animals, while curare caused modest slowing in controls but no consistent change in DNE animals; (2) DNE greatly attenuated the increase in C4VR frequency mediated by exogenous ACh; (3) stimulation of nAChRs with ACh in the presence of atropine increased frequency markedly in controls, but not DNE animals; (4) stimulation of mAChRs with ACh in the presence of curare caused a modest increase in frequency, with no treatment group differences. DNE blunts the response of the respiratory central pattern generator to exogenous ACh, consistent with reduced availability of functionally competent nAChRs; DNE did not alter the muscarinic control of respiratory motor output. © 2016 Wiley Periodicals, Inc. Develop Neurobiol 76: 1138–1149, 2016     

Fluorescence location of RVLM kainate microinjections that alter the control of breathing

Kainic acid (4.7 mM) applied to the rostral ventrolateral medulla (RVLM) surface decreases phrenic output, CO2 sensitivity, and blood pressure in chloralose-urethan-anesthetized, vagotomized, paralyzed, glomectomized, servoventilated cats. In this study using the same preparation, bilateral 50- to 100-nl kainate injections just below the RVLM surface better localized these responses topographically. The physiological responses to unilateral 10-nl kainate injections were then correlated with anatomic location determined by fluorescent microbeads (0.5 micron diam). Many sites were associated with no effect, a few rostral and caudal sites with increased phrenic activity, and cluster of sites with decreased phrenic activity often to apnea, decreased CO2 sensitivity, and decreased responses to carotid sinus nerve stimulation. Blood pressure was unaffected. These sites, within 400 microns of the surface, were ventral to the facial nucleus, ventrolateral to the nucleus paragigantocellularis lateralis, caudal to the superior olive, and rostral to the retrofacial nucleus. They appeared to be within the recently described retrotrapezoid nucleus, which contains cells with respiratory-related activity and projections to the dorsal and ventral respiratory groups. Cells within this site appear able to provide tonic input to respiration and to affect peripheral and central chemoreception.     

Developmental nicotine exposure enhances inhibitory synaptic transmission in motor neurons and interneurons critical for normal breathing

Nicotine exposure in utero negatively affects neuronal growth, differentiation, and synaptogenesis. We used rhythmic brainstems slices and immunohistochemistry to determine how developmental nicotine exposure (DNE) alters inhibitory neurotransmission in two regions essential to normal breathing, the hypoglossal motor nucleus (XIIn), and preBötzinger complex (preBötC). We microinjected glycine or muscimol (GABA_A agonist) into the XIIn or preBötC of rhythmic brainstem slices from neonatal rats while recording from XII nerve roots to obtain XII motoneuron population activity. Injection of glycine or muscimol into the XIIn reduced XII nerve burst amplitude, while injection into the preBötC altered nerve burst frequency. These responses were exaggerated in preparations from DNE animals. Quantitative immunohistochemistry revealed a significantly higher GABA_A receptor density on XII motoneurons from DNE pups. There were no differences in GABA_A receptor density in the preBötC, and there were no differences in glycine receptor expression in either region. Nicotine, in the absence of other chemicals in tobacco smoke, alters normal development of brainstem circuits that are critical for normal breathing. © 2015 Wiley Periodicals, Inc. Develop Neurobiol 76: 337–354, 2016     

Investigation on Acetylcholine,Aspartate, Glutamate and GABA Extracellular Levels from Ventral Hippocampus During Repeated Exploratory Activity in the Rat

The extracellular levels of aspartate, glutamate, -aminobutyric acid (GABA), and acetylcholine (ACh) were investigated by microdialysis, coupled with HPLC, in the ventral hippocampus of rats during two 30-min exploration periods. Motor activity was monitored. During exploration I, an increase in motor activity associated with a 315% increase in aspartate, 181% in glutamate, and 264% in ACh levels, occurred during the first 10 min. The increase in GABA level reached a maximum of 257% during the second 10 min. The neurotransmitter levels returned to basal values within 40 min. During exploration II, 1 h later, a smaller increase in neurotransmitter levels and motor activity was observed. In both explorations, the increase in neurotransmitter levels was completely abolished by 1 and 3 M TTX. A statistically significant relationship was found between neurotransmitter extracellular levels and motor activity, for aspartate and glutamate in exploration I, and for ACh in exploration I and II. In conclusion, exploratory activity is associated with or depends on the activation of neuronal systems in the ventral hippocampus releasing aspartate, glutamate, GABA, and ACh. The activation is dampened by habituation.