Neurotensin modulates synaptic transmission in the nucleus of the solitary tract of the rat

Whole-cell patch clamp recordings were made from neurons of the rat subpostremal region of the nucleus tractus solitarius (NTS) in transverse brainstem slices. Neurotensin (NT) enhanced the firing rate of action potentials from 0.8 +/- 0.4 Hz in control to 1.9 +/- 1.3 Hz (n = 9) and increased their decay time. The peak amplitude of the after-hyperpolarization was decreased by 34+/-5% (n = 9). These effects were associated with a depolarization of 4 +/- 1 mV (n = 10) in the resting membrane potential and an increase in the input resistance (from 768 +/- 220 MOmega to 986+/-220 MOmega; n = 5) and were compensated by manually hyperpolarizing the cell to control values. In voltage clamp experiments NT decreased an outward current (from 488 +/- 161 to 340 +/- 96 pA at +40 mV; n = 5) which reversed near the potassium equilibrium potential. In addition, NT increased the frequency of both excitatory and inhibitory spontaneous synaptic currents, an effect blocked by tetrodotoxin, and did not change the evoked excitatory or inhibitory postsynaptic currents. The selective NTR1 receptor antagonist SR48692 reversibly blocked the effects of NT on both action potentials and spontaneous synaptic currents. Our results suggest that NTR1 receptors can modulate post-synaptic responses in neurons of the subpostremal NTS by increasing cell excitability as a result of blockade of a potassium conductance.     

Dopamine modulates synaptic transmission in the nucleus of the solitary tract

10.1152/jn.00224.2002. Dopamine (DA) modulates the cardiorespiratory reflex by peripheral and central mechanisms. The aim of this study was to examine the role of DA in synaptic transmission of the nucleus tractus solitarius (NTS), the major integration site for cardiopulmonary reflexes. To examine DA's role, we used whole cell, voltage-clamp recordings in a rat horizontal brain stem slice. Solitary tract stimulation evoked excitatory postsynaptic currents (EPSCs) that were reduced to 70 +/- 5% of control by DA (100 microM). The reduction in EPSCs by DA was accompanied by a decrease in the paired pulse depression ratio with little or no change in input resistance or EPSC decay, suggesting a presynaptic mechanism. The D1-like agonist SKF 38393 Br (30 microM) did not alter EPSC amplitude, whereas the D2-like agonist, quinpirole HCl (30 microM), depressed EPSCs to 73 +/- 4% of control. The D2-like receptor antagonist, sulpiride (20 microM), abolished DA modulation of EPSCs. Most importantly, sulpiride alone increased EPSCs to 131 +/- 10% of control, suggesting a tonic D2-like modulation of synaptic transmission in the NTS. Examination of spontaneous EPSCs revealed DA reversibly decreased the frequency of events from 9.4 +/- 2.2 to 6.2 +/- 1.4 Hz. Sulpiride, however, did not alter spontaneous events. Immunohistochemistry of NTS slices demonstrated that D2 receptors colocalized with synaptophysin and substance P, confirming a presynaptic distribution. D2 receptors also localized to cultured petrosal neurons, the soma of presynaptic afferent fibers. In the petrosal neurons, D2 was found in cells that were TH-immunopositive, suggesting they were chemoreceptor afferent fibers. These results demonstrate that DA tonically modulates synaptic activity between afferent sensory fibers and secondary relay neurons in the NTS via a presynaptic D2-like mechanism.     

Kv1.3 channels regulate synaptic transmission in the nucleus of solitary tract

The voltage-gated K(+) channel Kv1.3 has been reported to regulate transmitter release in select central and peripheral neurons. In this study, we evaluated its role at the synapse between visceral sensory afferents and secondary neurons in the nucleus of the solitary tract (NTS). We identified mRNA and protein for Kv1.3 in rat nodose ganglia using RT-PCR and Western blot analysis. In immunohistochemical experiments, anti-Kv1.3 immunoreactivity was very strong in internal organelles in the soma of nodose neurons with a weaker distribution near the plasma membrane. Anti-Kv1.3 was also identified in the axonal branches that project centrally, including their presynaptic terminals in the medial and commissural NTS. In current-clamp experiments, margatoxin (MgTx), a high-affinity blocker of Kv1.3, produced an increase in action potential duration in C-type but not A- or Ah-type neurons. To evaluate the role of Kv1.3 at the presynaptic terminal, we examined the effect of MgTx on tract evoked monosynaptic excitatory postsynaptic currents (EPSCs) in brain slices of the NTS. MgTx increased the amplitude of evoked EPSCs in a subset of neurons, with the major increase occurring during the first stimuli in a 20-Hz train. These data, together with the results from somal recordings, support the hypothesis that Kv1.3 regulates the duration of the action potential in the presynaptic terminal of C fibers, limiting transmitter release to the postsynaptic cell.     

Potentiation of GABAergic synaptic transmission in the rostral nucleus of the solitary tract

Whole-cell recordings were made from neurons in the rostral nucleus of the solitary tract in horizontal brainstem slices. Monosynaptic GABAA receptor-mediated inhibitory postsynaptic potentials were evoked by single stimulus shocks or by high-frequency tetanic stimulation in the presence of glutamate receptor blockers. While single stimulus-evoked inhibitory postsynaptic potentials had variable amplitudes, tetanic stimulation-induced, hyperpolarizing postsynaptic potentials were of a more constant amplitude. Furthermore, tetanic stimulation resulted in potentiation of the amplitude of single stimulus shock-evoked inhibitory postsynaptic potentials. Of 55 neurons that were tested, potentiation lasted over 30 min for 11, 10-30 min for 13, less than 10 min for 23 and no potentiation occurred in eight. Tetanic stimulation did not result in potentiation of the tetanic stimulus-evoked hyperpolarizing postsynaptic potentials. Both the single stimulus shock- and tetanic stimulus-evoked potentials had similar inhibition concentration-response curves to the GABAA antagonist, bicuculline methiodide (EC50 = 0.75 and 0.83, respectively), indicating that they were mediated by the same postsynaptic receptors. By comparing the effect of bicuculline methiodide on the amplitude of the single stimulus shock-evoked inhibitory postsynaptic potentials and the tetanic stimulus-evoked hyperpolarizing potentials, we concluded that a single stimulus shock does not activate all postsynaptic GABAA receptors. However, tetanic stimulation results in activation of all postsynaptic GABAA receptors and induces long-lasting changes in the presynaptic GABAergic neuron. These long-lasting changes of the presynaptic neuron facilitate the release of GABA during single stimulus shock and, as a consequence, more postsynaptic receptors are activated during single stimulus shock-evoked synaptic transmission. This conclusion is supported by the results of experiments in which the extracellular Ca2+ concentration was manipulated to change the amount of neurotransmitter released from the presynaptic GABAergic terminals. The single stimulus shock-evoked inhibitory postsynaptic potentials were sensitive to the extracellular Ca2+ concentration, whereas tetanic stimulus-evoked inhibitory post-synaptic potentials were essentially insensitive to extracellular Ca2+ concentration. The relationship between the single stimulus shock-evoked inhibitory postsynaptic potential amplitude and extracellular Ca2+ concentration indicates that, in control physiological saline containing 2.5 mM Ca2+, a single stimulus shock activates less than half the postsynaptic GABA receptors. The phenomenon of long-lasting potentiation of inhibitory transmission within the rostral nucleus of the solitary tract may be important in the processing of gustatory information and play a role in taste-guided behaviors.     

Frequency dependence of synaptic transmission in nucleus of the solitary tract in vitro

Afferent fibers from visceral sensory receptors enter the medulla oblongata, form the solitary tract, and synapse with neurons in the nucleus of the solitary tract. In the present study longitudinal slices were prepared from guinea pig medulla in order to examine the properties of transmission at these synapses in vitro. Synaptic responses to selective stimulation of solitary tract fibers were recorded intracellularly from neurons in an area, close to the obex and immediately medial and lateral to the tract, where arterial baroreceptor fibers are known to terminate. The amplitude of maximally evoked postsynaptic potentials (PSPs) in solitary tract neurons was strongly dependent on stimulus frequency. On increasing frequency from 0.5 to 20 Hz, a PSP depression of 80% was reached in 4-8 s. The mean depression was 35% at 5 Hz and 60% at 10 Hz. Sufficient local connections were retained in vitro that solitary tract stimulation evoked disynaptic inhibitory potentials and long latency, possibly polysynaptic, excitatory potentials in some neurons. The possibility that frequency-dependent changes in the efficacy of these local synaptic circuits contributed to PSP depression was examined. The role of postsynaptic inhibition in synaptic depression was tested by examining the frequency dependence of PSPs at membrane potentials close to the reversal of their excitatory component. The resulting hyperpolarizing PSPs were also depressed suggesting that a facilitation of postsynaptic inhibition at high frequency does not underlie the depression. The contribution of depression in multisynaptic excitatory pathways to PSP depression was assessed by exclusion. At low stimulus intensities, excitatory synaptic events with no long latency components were evoked. These events exhibited a similar frequency dependence to that of maximal PSPs. These results suggest that mechanisms operating at synapses made by solitary tract fibers are responsible for the frequency dependence of PSPs recorded in solitary tract neurons. Such mechanisms might contribute to the adaptation of some cardiovascular reflexes initiated by baroreceptors.     

Postnatal development of inhibitory synaptic transmission in the rostral nucleus of the solitary tract

To explore the postnatal development of inhibitory synaptic activity in the rostral (gustatory) nucleus of the solitary tract (rNST), whole cell and gramicidin perforated patch-clamp recordings were made in five age groups of rats [postnatal day 0-7 (P0-7), P8-14, P15-21, P22-30, and P >55]. The passive membrane properties of the developing rNST neurons as well as the electrophysiological and pharmacological characteristics of single and tetanic stimulus-evoked inhibitory postsynaptic potentials (IPSPs) were studied in brain slices under glutamate receptor blockade. During the first postnatal weeks, significant changes in resting membrane potential, spontaneous activity, input resistance, and neuron membrane time constant of the rNST neurons occurred. Although all the IPSPs recorded were hyperpolarizing, the rise and decay time constants of the single stimulus shock-evoked IPSPs decreased, and the inhibition response-concentration function to the gamma-aminobutyric acid (GABA) receptor antagonist bicuculline methiodide (BMI) shifted to the left during development. In P0-7 and P8-14, but not in older animals, the IPSPs had a BMI-insensitive component that was sensitive to block by picrotoxin, suggesting a transient expression of GABA(C) receptors. Tetanic stimulation resulted in both short- and long-term changes of inhibitory synaptic transmission in the rNST. For P0-7 and P8-14 animals tetanic stimulation resulted in a sustained hyperpolarization that was maintained for some time after termination of the tetanic stimulation. In contrast, tetanic stimulation of neurons in P15-21 and older animals resulted in hyperpolarization that was not sustained but decayed back to a more positive level with an exponential time course. Tetanic stimulation resulted in potentiation of single stimulus shock-evoked IPSPs in ~50% of neurons in all age groups. These developmental changes in inhibitory synaptic transmission in the rNST may play an important role in shaping synaptic activity in early development of the rat gustatory system during a time of maturation of taste preferences and aversions.     

Hydrogen sulfide augments synaptic neurotransmission in the nucleus of the solitary tract

Within the brain stem, the nucleus tractus solitarii (NTS) serves as a principal central site for sensory afferent integration from the cardiovascular and respiratory reflexes. Neuronal activity and synaptic transmission in the NTS are highly pliable and subject to neuromodulation. In the central nervous system, hydrogen sulfide (H?S) is a gasotransmitter generated primarily by the enzyme cystathionine-β-synthase (CBS). We sought to determine the role of H?S, and its generation by CBS, in NTS excitability. Real-time RT-PCR, immunoblot, and immunohistochemistry analysis identified the presence of CBS in the NTS. Patch-clamp electrophysiology in brain stem slices examined excitatory postsynaptic currents (EPSCs) and membrane properties in monosynaptically driven NTS neurons. Confocal imaging of labeled afferent synaptic terminals in NTS slices monitored intracellular calcium. Exogenous H?S significantly increased the amplitude of evoked solitary tract (TS)-EPSCs, frequency of miniature (m)EPSCs, and presynaptic terminal calcium fluorescence in the NTS. H?S did not alter action potential discharge or postsynaptic properties. On the other hand, the CBS inhibitor aminooxyacetate (AOA) significantly reduced the amplitude of TS-EPSCs and presynaptic terminal calcium fluorescence in the NTS without altering postsynaptic properties. Taken together, these data support a presynaptic role for endogenous H?S in modulation of excitatory neurotransmission in the NTS.     

Inhibition by endomorphin-1 and endomorphin-2 of excitatory transmission in adult rat substantia gelatinosa neurons

Intrathecally-administered endomorphin-1 and endomorphin-2 produce antinociceptive effects which are different from each other. In order to elucidate a cellular basis for this result, we examined the effects of endomorphin-1 and endomorphin-2 on holding currents and spontaneous glutamatergic excitatory transmission in substantia gelatinosa neurons of adult rat spinal cord slices by use of the whole-cell patch-clamp technique. In about half of the neurons examined, endomorphin-1 and endomorphin-2 produced an outward current having a similar amplitude (25-27 pA at 1 microM) at -70 mV with almost the same value of effective concentration producing half-maximal response (0.19-0.21 microM). Both of them reversed at a potential close to the equilibrium potential for K+, and had the slope conductance that was larger at negative (-120 to -140 mV) than positive potentials (-60 to -90 mV). The endomorphin-1 and endomorphin-2 currents were reduced in amplitude by K+-channel inhibitors, Ba2+ (100 microM) and 4-aminopyridine (1 mM), and also by mu-opioid receptor antagonist D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH2 (1 microM) to a similar extent. The endomorphin-2 but not endomorphin-1 current amplitude was increased by dipeptidyl peptidase IV inhibitor diprotin A (30 microM). One micromolar endomorphin-1 and endomorphin-2 reduced the frequency of spontaneous excitatory postsynaptic current with a similar time course and extent without altering its amplitude; these actions were not in the presence of D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH2 (1 microM). We conclude that endomorphin-1 and endomorphin-2 hyperpolarize membranes by opening inwardly-rectifying K+ channels and attenuate the spontaneous release of L-glutamate from nerve terminals in the substantia gelatinosa, both of which are mediated by mu-opioid receptors, in a manner quantitatively similar to each other. The difference in antinociceptive effects between endomorphin-1 and endomorphin-2 could not be attributed to a distinction in their effects on excitatory transmission in substantia gelatinosa neurons, and may be explained by a difference in their enzymatic degradation.     

Projections from the nucleus of the solitary tract in the rat

The axonal projections of neurons in and near the nucleus of the solitary tract have been visualized using titrated amino acid autoradiography. Axons of neurons of this nucleus ramify extensively within the nucleus itself, but much less so in the nucleus commissuralis. They also enter cranial motor nuclei within the medulla. Axons originating in the anterior part of the nucleus of the solitary tract extend to the hypoglossal, facial and probably trigeminal motor nuclei, but not to the dorsal motor nucleus of the vagus or the nucleus ambiguus. The posterior part of the nucleus of the solitary tract projects to all these motor nuclei. In the spinal cord solitary nucleus axons remain in the medial gray directly caudal to the solitary nucleus itself. The distribution becomes very weak by C₃ after some fibers spread laterally into the caudal trigeminal nucleus. Fibers are labeled in the contralateral ventral columns, but they could not be unequivocably attributed to solitary neurons. Axons ascending from the nucleus of the solitary tract extend no further rostrally than the pons, where they terminate in the caudal end of the parabrachial nuclei.Although often treated as entirely separate systems, the present results indicate that secondary gustatory neurons in the anterior solitary nucleus and secondary visceral afferent neurons in the posterior solitary nucleus have very similar rostral and caudal projections. The pontine parabrachial nuclei, the rostral termination of solitary nucleus neurons, have extensive direct connections to the thalamus, the hypothalamus and the limbic forebrain. Assuming similar connections occur in other mammals, these findings establish the existence of di-synaptic visceral afferent access to the highest autonomic integrative centers in the brain.     

Prostaglandin E2 depresses solitary tract-mediated synaptic transmission in the nucleus tractus solitarius

Prostaglandin E(2) (PGE(2)) is a prototypical inflammatory mediator that excites and sensitizes cell bodies [Kwong K, Lee LY (2002) PGE(2) sensitizes cultured pulmonary vagal sensory neurons to chemical and electrical stimuli. J Appl Physiol 93:1419-1428; Kwong K, Lee LY (2005) Prostaglandin E(2) potentiates a tetrodotoxin (TTX)-resistant sodium current in rat capsaicin-sensitive vagal pulmonary sensory neurons. J Physiol 56:437-450] and peripheral nerve terminals [Ho CY, Gu Q, Hong JL, Lee LY (2000) Prostaglandin E (2) enhances chemical and mechanical sensitivities of pulmonary C fibers in the rat. Am J Respir Crit Care Med 162:528-533] of primary vagal sensory neurons. Nearly all central nerve terminals of vagal afferents are in the nucleus tractus solitarius (NTS), where they operate with a high probability of release [Doyle MW, Andresen MC (2001) Reliability of monosynaptic sensory transmission in brain stem neurons in vitro. J Neurophysiol 85:2213-2223]. We studied the effect of PGE(2) on synaptic transmission between tractus solitarius afferent nerve terminals and the second-order NTS neurons in brain stem slices of Sprague-Dawley rats. Whole-cell patch recording in voltage clamp mode was used to study evoked excitatory postsynaptic glutamatergic currents (evEPSCs) from NTS neurons elicited by electrical stimulation of the solitary tract (ST). In 34 neurons, bath-applied PGE(2) (200 nM) decreased the evEPSC amplitude by 49+/-5%. In 22 neurons, however, PGE(2) had no effect. We also tested 15 NTS neurons for capsaicin sensitivity. Seven neurons generated evEPSCs that were equally unaffected by PGE(2) and capsaicin. Conversely, evEPSCs of the other eight neurons, which were PGE(2)-responsive, were abolished by 200 nM capsaicin. Furthermore, the PGE(2-)induced depression of evEPSCs was associated with an increase in the paired pulse ratio and a decrease in both the frequency and amplitude of the spontaneous excitatory postsynaptic currents (sEPSCs) and TTX-independent spontaneous miniature excitatory postsynaptic currents (mEPSCs). These results suggest that PGE(2) acts both presynaptically on nerve terminals and postsynaptically on NTS neurons to reduce glutamatergic responses.     

Development of taste responses in rat nucleus of solitary tract

Extracellular responses from neurons in the nucleus of the solitary tract (NST) were studied in rats aged 5 days to adulthood during chemical stimulation of the tongue with monochloride salts, citric and hydrochloric acids, sucrose, sodium saccharin, and quinine hydrochloride. Multiunit taste responses were recorded in rats at 5-7 days of age and single-unit responses were recorded from 111 neurons in four other age groups of 14-20 days, 25-35 days, 50-60 days, and adult. NST neurons in rats aged 5-7 days consistently responded to relatively high concentrations (0.5 M) of NH4Cl and KCl and to citric and hydrochloric acid. However, they often did not respond to 0.5 M NaCl or to 0.1 M NH4Cl. Single NST neurons in rats aged 14 days and older characteristically responded to all 0.1 and 0.5 M salts and to both acids. At least 75% of neurons also responded to sucrose and sodium saccharin, and 46% responded to all of these stimuli and quinine hydrochloride. After 14 days, no developmental changes occurred in the number of stimuli to which neurons responded. There were substantial developmental alterations in the response magnitudes to some chemical stimuli. Average response frequencies increased after 35 days of age for 0.1 and 0.5 M NaCl, LiCl, KCl, and for sucrose and sodium saccharin. Response frequencies for NH4Cl, citric and hydrochloric acid, and quinine hydrochloride, however, did not change throughout development. The proportion of single NST neurons that responded maximally to specific monochloride salts did not change during development. Most single neurons in all age groups responded equally well to NH4Cl, NaCl, and LiCl. No NST neuron responded maximally to KCl. There were also no developmental differences in response latencies in rats aged 14 days and older. Response frequencies of second-order NST neurons generally reflect changes in responses from the primary afferent, chorda tympani fibers, throughout development; however, the increases in salt response frequencies from NST neurons occur comparatively later in development. Furthermore, at all ages, the taste responses to monochloride salts include higher response frequencies and a general loss in response specificity in NST compared to chorda tympani neurons.(ABSTRACT TRUNCATED AT 400 WORDS)     

Presynaptic inhibition of synaptic transmission by adenosine in rat subthalamic nucleus in vitro

Whole-cell patch clamp recordings were made from the subthalamic nucleus in rat brain slice preparations to examine the effect of adenosine on inhibitory and excitatory synaptic transmission. Adenosine reversibly inhibited both GABA-mediated inhibitory and glutamate-mediated excitatory postsynaptic currents. Adenosine at 100 microM reduced the amplitude of inhibitory and excitatory postsynaptic currents by 42+/-5% and 34+/-6%, respectively. Reductions in the amplitude of both inhibitory and excitatory postsynaptic currents were accompanied by increases in paired-pulse ratios. In addition, adenosine decreased the frequency of spontaneous miniature excitatory postsynaptic currents but had no effect on their amplitude. These results are consistent with a presynaptic site of action. The adenosine A(1) receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine completely reversed the adenosine-induced attenuation of inhibitory and excitatory postsynaptic currents, but 8-cyclopentyl-1,3-dipropylxanthine alone had no effect on synaptic currents evoked at 0.1 Hz. However, 8-cyclopentyl-1,3-dipropylxanthine inhibited a time-dependent depression of excitatory postsynaptic currents that was normally observed in response to a 5 Hz train of stimuli, suggesting that endogenous adenosine could be released during higher frequencies of stimulation. These results suggest that adenosine inhibits synaptic release of GABA and glutamate by stimulation of presynaptic A(1) receptors in the subthalamic nucleus.     

Proteolysis of endomorphin-1 by brain synaptic membrane-associated peptidases

The biotransformation of endomorphin-1 (EM-1) by brain synaptic membranes has been studied. Peptide fragments of EM-1 that were formed during incubation with membrane preparations in vitro were isolated by High-performance liquid chromatography (HPLC) and characterized using determination of amino acid composition. At pH 7.4, two degradation products, EM-1(1-3) and EM-1(1-2), were identified. EM-1 was degraded 77.5% at 30 min incubation with synaptic membranes. The time course of the experiments and the effect of carboxypeptidase inhibitor (CPI) demonstrated that the proteolysis reaction involves the participation of carboxypeptidase activity.     

Cross-correlation analysis of taste neuron pairs in rat solitary tract nucleus

1. Experiments were conducted to examine the possibility that the taste-sensitive neurons with similar taste-selectivity are preferentially innervated by common driving neurons whose taste-selectivity is also similar. 2. Multiple microelectrodes, in most cases a pair of glued electrodes, were inserted into the unilateral solitary tract nucleus (NTS) of the rat, and simultaneous recordings were made in neuron pairs responding to the four basic taste stimuli. The spike response density (RD) of each neuron during tastant stimulation was determined. Correlation coefficients of spike occurrence were calculated for each neuron pair during application of tastants and distilled water and also during spontaneous background activity. The frequency of correlated discharge (FC) of a neuron pair was measured as the area of the peak appearing on the cross-correlogram (CC). The FC value was divided by the RD value to calculate the weight of the correlated discharges in the output of each neuron (WC value). 3. Eleven pairs showed peaks in the CC constructed during tastant stimulation, whereas in other 11 pairs no peaks were found. The cross-correlation-positive group with peaks was composed of 18 NaCl-best (responding most vigorously to NaCl) and 4 HCl-best neurons, whereas the negative group without peaks included 9 NaCl-best, 9 HCl-best, and 4 sucrose-best neurons. 4. In the cross-correlation-positive pairs the taste quality most effective for one of the component neurons was often (13 NaCl-best and 2 HCl-best, 15/22 = 0.681) identical to the taste quality giving the highest probability of correlated discharge, i.e., highest FC value, in the neuron pair. 5. There were five cross-correlation-positive pairs (5/11 = 0.455) in which both of the component neurons were NaCl-best and the FC value was highest during NaCl stimulation. 6. The CCs constructed during water application exhibited peaks for all the pairs which gave positive cross-correlation in response to stimulation with tastants, whereas all pairs with negative cross-correlation during tastant stimulation never gave a detectable peak during water application. 7. In three pairs of the cross-correlation-positive group, the CCs constructed during spontaneous background activity without application of any liquid showed a small peak. 8. During NaCl stimulation some neurons exhibited relatively high FC values, but the WC values were always low. In contrast, during sucrose stimulation, the FC value was always low, but the WC value was quite high in some neurons.(ABSTRACT TRUNCATED AT 400 WORDS)     

Dopamine depresses glutamatergic synaptic transmission in the rat parabrachial nucleus in vitro

Nystatin-perforated patch recordings were made from rat parabrachial neurons in an in vitro slice preparation to examine the effect of dopamine on parabrachial cells and on excitatory synaptic transmission in this nucleus. In current clamp mode, dopamine reduced the amplitude of the evoked excitatory postsynaptic potential without significant change in membrane potential. In cells voltage-clamped at -65 mV, dopamine dose dependently and reversibly decreased evoked, pharmacologically isolated, excitatory postsynaptic currents with an EC50 of 31 microM. The reduction in excitatory postsynaptic current was accompanied by an increase in paired pulse ratio (a protocol used to detect presynaptic site of action) with no change in the holding current or in the decay of the evoked excitatory postsynaptic currents. In addition, dopamine altered neither postsynaptic (+/-)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate-induced currents, nor steady-state current voltage curves. Miniature excitatory postsynaptic current analysis revealed that dopamine caused a rightward shift of the frequency-distribution curve with no change in the amplitude-distribution curve, which is consistent with a presynaptic mechanism. The dopamine-induced attenuation of the excitatory postsynaptic current was almost completely blocked by the D1-like receptor antagonist SCH23390 (10 microM), although the D2-like antagonist sulpiride (10 microM) also partially blocked it. Combined application of both antagonists blocked all dopamine-induced synaptic effects. The synaptic effect of dopamine was mimicked by the D1-like agonist SKF38393 (50 microM), but the D2-1ike agonist quinpirole (50 microM) also had a small effect. Combined application of both agonists did not produce potentiated responses. Dopamine's effect on the excitatory postsynaptic current was independent of serotonin, GABA and adenosine receptors, but may have some interactions with adrenergic receptors. These results suggest that dopamine directly modulates excitatory synaptic events in the parabrachial nucleus predominantly via presynaptic D1-like receptors.     

Effects of corticosteroids on synaptic transmission in rat dorsolateral septal nucleus

The effects of corticosteroids on synaptic transmission in the rat dorsolateral septal nucleus (DLSN) were examined, in vitro, by using intracellular and voltage-clamp recording methods. Prednisolone (100 microM) increased the amplitude of excitatory postsynaptic potential (EPSP) and depressed both fast and slow inhibitory postsynaptic potentials (IPSP). Under voltage-clamp conditions, prednisolone (100 microM) increased the amplitude of excitatory postsynaptic current (EPSC) and depressed the fast and slow inhibitory postsynaptic currents (IPSCs). Corticosterone (100 microM) mimicked the effects of prednisolone on the postsynaptic currents (PSCs). To examine the direct effects of prednisolone on the EPSC and slow IPSC, the fast IPSC was blocked by bicuculline (20 microM). Under these experimental conditions, prednisolone (100 microM) did not alter the isolated EPSC but depressed slow IPSC by 22 +/- 3% (n = 10). The fast IPSC was isolated by pretreatment with kynurenic acid and CGP55845A, where the EPSC and slow IPSC were blocked. Prednisolone (100 microM) depressed the isolated fast IPSC in DLSN neurons. Prednisolone (100 microM) did not change either the inward current produced by glutamate or the outward current produced by gamma-aminobutyric acid (GABA). The results suggest that corticosteroids facilitate excitatory synaptic transmission in the DLSN by reducing the release of GABA from the presynaptic nerve terminals of interneurons.     

Modulation of peripheral inflammation by locally administered endomorphin-1

OBJECTIVES: Neurogenic inflammation is mediated via sensory peptides released from the peripheral terminals of sensory nerves and can be modulated by locally released opioid peptides at the site of injury. Endomorphins are recently discovered endogenous opioid peptides with high selectivity and affinity for the mu-opioid receptor. The aim of this study was to examine the ability of endomorphin-1 (EM-1) to modulate the inflammatory response under different injury conditions. METHODS: A vacuum-induced blister model in anaesthetised rats (nembutal 60 mg/kg i.p.) was used to examine the effect of EM-1 on the acute inflammatory response induced by; (1) electrical stimulation (ES) of the distal portion of the exposed/cut sciatic nerve at 20 V, 5 Hz, 2 ms for 1 min or; (2) superfusion of substance P (SP) over the blister base. In addition, the effect of EM-1 on the inflammatory response to SP was examined under acute, recurrent (repeated blister induction) and chronic (chronic sciatic nerve lesion) injury conditions. RESULTS: Prior and concomitant perfusion of EM-1 (100 microM) significantly inhibited the vascular response to ES by 58% compared to controls. EM-1 also inhibited the inflammatory response to SP (both vasodilatation and plasma extravasation) in a dose-dependent manner. Significant inhibition was achieved at 100 microM and 1 mM concentrations of EM-1. Naloxone (1 mg/kg i.v.) reversed the inhibitory effect of EM-1 on the inflammatory response to SP. EM-1 (100 microM) was equally potent in inhibiting the inflammatory response to SP under acute (34% inhibition) recurrent (39%) and chronic (42%) injury conditions. CONCLUSIONS: The current results demonstrate a greater inhibitory effect of EM-1 on the inflammatory response to electrical nerve stimulation (58% inhibition) compared to SP (34% inhibition) suggesting the involvement of both pre- and post-terminal mechanisms in the inhibitory modulatory actions of EM-1. Evidence is provided for the involvement of opioid receptors in this inhibitory effect. The results also suggest that EM-1 is equipotent in inhibiting the inflammatory response under different injury conditions.     

Chemosensory control by commissural nucleus of the solitary tract in rats

The commissural nucleus of the solitary tract (commNTS) is a main area that receives afferent signals involved in the cardiovascular and respiratory control like those related to chemoreceptor activation, however, the importance of the commNTS for the cardiorespiratory responses to chemoreceptor activation is still controversial. In the present study, we investigated the cardiorespiratory responses to hypoxia or hypercapnia in anesthetized and conscious rats treated with injections of the GABA-A agonist muscimol into the caudal portion of the commNTS. Male Holtzman rats (280-300 g) were used. In conscious rats that had a stainless steel cannula previously implanted into the commNTS, the injection of muscimol (2 mM) into the commNTS reduced the pressor response (16±2 mmHg, vs. saline: 36±3 mmHg) and the increase in ventilation (250±17 ml/min/kg, vs. saline: 641±28 ml/min/kg) produced by hypoxia (8-10% O(2)). In urethane anesthetized rats, the injection of muscimol into the commNTS eliminated the pressor response (5±2 mmHg, vs. saline: 26±5 mmHg) and the increase in phrenic nerve discharge (PND) (20±6%, vs. saline: 149±15%) and reduced the increase in splanchnic sympathetic nerve discharge (sSND) (93±15%, vs. saline: 283±19% of baseline) produced by hypoxia. However, muscimol injected into the commNTS did not change hypercapnia (8-10% CO(2)) induced pressor response or the increase in the sSND or PND in urethane anesthetized rats or the increase in ventilation in conscious rats. The present results suggest that the cardiorespiratory responses to hypoxia are strongly dependent on the caudal portion of the commNTS, however, this area is not involved in the responses to hypercapnia.