Differential sensitivity of spinal neurones to amino acids: an intracellular study on the frog spinal cord

Intracellular recordings from in vitro neurones of the frog spinal cord slice preparation were performed in order to examine the mechanism of action of gamma-aminobutyrate and glutamate on two distinct neuronal populations in the same region of the central nervous system. Amino acids were superfused at fast rate and low temperature (7 degrees C) to reduce their uptake process. On interneurones, the inhibitory action of gamma-aminobutyrate was characterized by a large input conductance increase while on motoneurones the conductance change was much smaller. Glutamate excited interneurones which greatly increased their input conductance and showed burst firing; motoneurones were also excited by glutamate but usually did not fire repeatedly nor showed large conductance changes. In spite of these differences the amplitude of depolarization in the presence of the same concentration of glutamate was similar for motoneurones and interneurones. It is suggested that amino acids (particularly glutamate) may act through different membrane mechanisms on two neuronal populations in the same region of the spinal cord.     

Analysis of synaptic transmission at single identified boutons on rat spinal neurons in culture

The spatial organization of receptor channels has a major influence on the speed and possible plasticity of synaptic signal transmission. We have studied glutamatergic synapses on neurons in organotypic cultures of rat spinal cord. In order to avoid the problems related to the analysis of currents of unknown origin within a neuron, we chose to examine the functional properties of single identified synapses. Iontophoretic mapping of the cell surface revealed hot spots of high glutamate sensitivity coincident with presynaptic boutons stained with the dye FM 1–43. Local application of KCl to these sites caused bursts of synaptic release. Hot spots typically consisted of 330 receptors with an average single-channel conductance of 8.3 pS. Evoked synaptic currents involved only about 40–50 receptors and nevertheless showed characteristics of saturation. This suggests that glutamate receptor clusters at sites of presynaptic terminals are organized into well separated subclusters opposite release sites.This award-winning article is published as received and has not been subjected to the normal peer review process     

Inhibition of nitric oxide synthase desensitizes retinal ganglion cells to light by diminishing their excitatory synaptic currents under light adaptation

The effect of inhibiting nitric oxide synthase (NOS) on the visual responses of mouse retinal ganglion cells (RGCs) was studied under light adaptation by using patch-clamp recordings. The results demonstrated that NOS inhibitor, l-NAME, reduced the sensitivity of RGCs to light under light adaptation at different ambient light conditions. These observations were seen in all cells that recordings were made from. l-NAME diminished the excitatory synaptic currents (EPSCs), rather than increasing the inhibitory synaptic currents, of RGCs to reduce the sensitivity of RGCs to light. Cones may be the sites that l-NAME acted to diminish the EPSCs of RGCs.     

γ-氨基丁酸受体和甘氨酸受体在大鼠丙泊酚麻醉中的作用

目的研究丙泊酚对大鼠海马CA1区电刺激诱发兴奋性突触后电流(EPSC)的影响,分析γ-氨基丁酸(GABA)受体和甘氨酸受体在丙泊酚麻醉中的作用。方法断头法分离wistar大鼠(13～19d)海马半脑,切出400μm厚度的海马脑片,全细胞膜片钳技术记录CA1区锥体神经元EPSC。80张脑片分为八组:脂肪乳剂组,50μmol/L丙泊酚组,100μmol/L丙泊酚组,200μmol/L丙泊酚组,SR95531组,士的宁组,SR95531 100μmol/L丙泊酚组,士的宁 100μmol/L丙泊酚组,每组10张。SR95531 100μmol/L丙泊酚组和士的宁 100μmol/L丙泊酚组先在循环液中加入10μmol/LSR95531或4μmol/L士的宁预孵脑片30min。八组均记录基础EPSC10min,然后加入不同药物,继续记录EPSC40min。膜钳制电压为-70mV。结果脂肪乳剂、SR95531和士的宁对EP-SC幅值无影响;丙泊酚呈剂量依赖性的抑制EPSC幅值,50、100、200μmol/L丙泊酚最大抑制EPSC幅值为14.4%、52.3%、67.8%;SR95531 100μmol/L丙泊酚组加入丙泊酚后,EPSC幅值基本无改变;士的宁 100μmol/L丙泊酚组加入丙泊酚后,EPSC幅值仍然下降,最大抑制程度为34.7%。结论丙泊酚主要通过增强GABAA受体功能使兴奋性突触活动降低,甘氨酸受体在其中起到协同和调节作用。     

Differential expression of NMDA receptors in serotonergic and/or GABAergic neurons in the midbrain periaqueductal gray of the mouse

N-methyl-d-aspartate (NMDA) receptors expressed in the midbrain periaqueductal gray (PAG) exert various physiological functions. The PAG contains various neurotransmitter phenotypes, which include GABAergic neurons and serotonergic neurons. In the present experiments, we made tight-seal whole-cell recordings from GABAergic and/or serotonergic neurons in mouse PAG slices and analyzed NMDA and non-NMDA receptor-mediated excitatory postsynaptic currents (EPSCs) evoked by electrical stimulation. The NMDA/non-NMDA ratio of EPSC amplitude was high and the decay time course of NMDA-EPSC was slow in non-serotonergic/GABAergic neurons. In contrast, serotonergic neurons exhibited a low NMDA/non-NMDA ratio and a fast decay time course of NMDA-EPSC. Peripheral nerve ligation-induced chronic pain was associated with an increased NMDA/non-NMDA ratio in serotonergic neurons. Additionally, single-cell real-time RT-PCR analysis showed that peripheral nerve ligation up-regulated NR2B subunit expression in non-serotonergic/non-GABAergic neurons. Such changes in NMDA receptor expression in the PAG result in an alteration of the descending modulation of nociception, which might be an underlying mechanism for peripheral nerve injury-evoked persistent pain. Finally, the expression of NMDA receptors seems differentially regulated among neurons of different neurotransmitter phenotypes in the PAG.     

Codeine presynaptically inhibits the glutamatergic synaptic transmission in the nucleus tractus solitarius of the guinea pig

Although codeine is the most prominent and centrally acting antitussive agent, the precise sites and mode of its action have not been fully understood yet. In the present study, we examined the effects of codeine on synaptic transmission in second-order neurons of the nucleus tractus solitarius (NTS), which is the first central relay site receiving tussigenic afferent fibers, by using whole-cell patch-clamp recordings in guinea-pig brainstem slices. Codeine (0.3-3 mM) significantly decreased the amplitude of excitatory postsynaptic currents (EPSCs) evoked by electrical stimulation of the tractus solitarius in a naloxone-reversible and concentration-dependent manner, but it had no effect on the decay time of evoked EPSCs (eEPSCs). The inhibition of eEPSCs was accompanied by an increased paired-pulse ratio of two consecutive eEPSCs. The inward current induced by application of AMPA remained unchanged after codeine application. A voltage-sensitive K+ channel blocker, 4-aminopyridine (4-AP) attenuated the inhibitory effect of codeine on eEPSCs. These results suggest that codeine inhibits excitatory transmission from the primary afferent fibers to the second-order NTS neurons through the opioid receptors that activate the 4-AP sensitive K+ channels located at presynaptic terminals.     

Subunit composition and kinetics of the Renshaw cell heteromeric nicotinic receptors

In Renshaw cells (RCs) of newborn mice, activation of motoneurons elicits a four-component synaptic current (EPSC) mediated by two glutamate receptors and two nicotinic receptors (nAChRs). We have analyzed the nicotinic component of the EPSC which is blocked by dihydro-beta-erythroidine (DHβE) with the dual objective of identifying the nAChR subunits involved and of understanding the kinetics of the response.     

Complex EPSCs evoked in substantia nigra reticulata neurons are disrupted by repetitive stimulation of the subthalamic nucleus

Although substantia nigra reticulata (SNR) neurons fire bursts of action potentials during normal movement, excessive burst firing correlates with symptoms of Parkinson's disease. A major excitatory output from the subthalamic nucleus (STN) to the SNR is thought to provide the synaptic impetus for burst firing in SNR neurons. Using patch pipettes to record from SNR neurons in rat brain slices, we found that a single electrical stimulus delivered to the STN evokes a burst of action potentials. Under voltage-clamp conditions, STN stimulation evokes a complex EPSC that is comprised of an initial monosynaptic EPSC followed by a series of late EPSCs superimposed on a long-lasting inward current. Using varied stimulation frequencies, we found that the initial EPSC was significantly reduced or abolished after 2 s of 50-100 Hz STN stimulation. However, only 4 s of 1 Hz stimulation was required to abolish the late component of the complex EPSC. We suggest that differential effects of repetitive STN stimulation on early and late components of complex EPSCs may help explain the frequency-dependent effects of deep brain stimulation of the STN that is used in the treatment of Parkinson's disease.     

Adaptations of glutamatergic synapses in the striatum contribute to recovery from cerebellar damage

Recent findings proposed that the cerebellum and the striatum, key structures in motor control, are more interconnected than commonly believed, and that the cerebellum may influence striatal activity. In the present study, the possible changes of synaptic transmission in the striatum of hemicerebellectomized rats have been investigated. Neurophysiological recordings showed a significant facilitation of glutamate transmission in the contralateral striatum occurring early following hemicerebellectomy. This process of synaptic adaptation appears to be relevant for the compensation of cerebellar deficits. Accordingly, pharmacological blockade of glutamate N-methyl-d-aspartate (NMDA) receptors with MK-801 prevented the rearrangement of excitatory synapses in the striatum and interfered with the recovery from motor disturbances in rats with cerebellar lesions. Hemicerebellectomy also perturbed gamma-aminobutyric acid (GABA) transmission in contralateral but not ipsilateral striatum. The present findings advance the role of striatal excitatory transmission in the compensation of cerebellar deficits, providing support to the notion that adaptations of striatal function exert a role in the recovery of cerebellar symptoms.