Actions of D-Ala2-D-Leu5-enkephalin and dynorphin A (1–17) on neocortical neurons in vitro

Intracellular recordings were made from neocortical neurons in vitro. Application of D-Ala2-D-Leu5-enkephalin (DADL) by different methods produced a decrease in EPSP amplitude and in the amplitude of L-glutamate-induced depolarizations without changes in membrane potential or membrane input resistance. The DADL effects were blocked by naloxone and persisted when synaptic transmission was depressed, suggesting DADL acts on postsynaptically located opiate receptors. With dynorphin A (1–17), depolarizations, hyperpolarizations, decreases and increases in EPSP were observed, but never an anti-glutamate effect

Responses of substantia gelatinosa neurons to putative neurotransmitters in an in vitro preparation of the adult rat spinal cord

Extracellular recordings were performed from neurons of the substantia gelatinosa (SG) in an in vitro preparation obtained from the spinal cord of adult rats. About 40% of neurons were spontaneously active. They could be synaptically influenced by low and high threshold fiber input entering the spinal cord through dorsal and ventral and ventral roots. Repetitive low threshold stimulation led to a transient increase in activity of a number of these neurons, whereas high intensity stimulation predominantly reduced excitability. The majority of non-spontaneously active neurons responded to an increase of stimulus intensity covariantly with an increase in firing rate. The excitatory effect of phoretically administeredl-glutamate as well as synaptically induced and spontaneous activity was reduced or abolished by phoretically administered GABA, glycine or the enkephalin-analogued-Ala2-d-Leu5-enkephalin. The actions of the enkephalin analogue were blocked by phoretically applied naloxone. The findings are consistent with the notion from in vivo investigations of a structurally and functionally heterogeneous population of neurons which display a responsiveness to microtopically applied putative neurotransmitters resembling dorsal horn neurons in deeper layers

Disinhibition of hippocampal CA3 neurons induced by suppression of an adenosine A1 receptor-mediated inhibitory tonus: Pre- and postsynaptic components

Intracellular recordings were performed on hippocampal CA3 neuronsin vitro to investigate the inhibitory tonus generated by endogenously produced adenosine in this brain region. Bath application of the highly selective adenosine A1 receptor antagonist 1,3-dipropyl-8-cyclopentylxanthine at concentrations up to 100 nM induced both spontaneous and stimulus-evoked epileptiform burst discharges. Once induced, the 1,3-dipropyl-8-cyclopentylxanthine-evoked epileptiform activity was apparently irreversible even after prolonged superfusion with drug-free solution. The blockade of glutamatergic excitatory synaptic transmission by preincubation of the slices with the amino-3-hydroxy-5-methyl-4-isoxazolpropionic acid receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (10 μM), but not with theN-methyl-d-aspartate receptor antagonistd-2-amino-5-phosphonovaleric acid (50/μM), prevented the induction of epileptiform activity by 1,3-dipropyl-8-cyclopentylxanthine. The generation of the burst discharges was independent of the membrane potential, and the amplitude of the slow component of the paroxysmal depolarization shift increased with hyperpolarization, indicating that the 1,3-dipropyl-8-cyclopentylxanthine-induced bursts were synaptically mediated events. Recordings from tetrodotoxin-treated CA3 neurons revealed a strong postsynaptic component of endogenous adenosinergic inhibition. Both 1,3-dipropyl-8-cyclopentylxanthine and the adenosine-degrading enzyme adenosine deaminase produced an apparently irreversible depolarization of the membrane potential by about 20 mV. Sometimes, this depolarization attained the threshold for the generation of putative calcium spikes, but no potential changes resembling paroxysmal depolarization shift-like events were observed

Characteristics of long-duration inhibitory postsynaptic potentials in rat neocortical neurons in vitro

1. The characteristics of long-duration inhibitory postsynaptic potentials (l-IPSPs) which are evoked in rat frontal neocortical neurons by local electrical stimulation were investigated with intracellular recordings from anin vitro slice preparation. 2. Stimulation with suprathreshold intensities evoked l-IPSPs with typical durations of 600–900 msec at resting membrane potential. Conductance increases of 15–60% were measured at the peak amplitude of l-IPSPs (150–250 msec poststimulus). 3. The duration of the conductance increases during l-IPSPs displayed a significant voltage dependence, decreasing as the membrance potential was depolarized and increasing with hyperpolarization. 4. The reversal potential of l-IPSPs is significantly altered by reductions in the extracellular potassium concentration. Therefore it is concluded that l-IPSPs in rat neocortical neurons are generated by the activation of a potassium conductance. 5. l-IPSPs exhibit stimulation fatigue. Stimulation with a frequency of 1 Hz produces a complete fatigue of the conductance increases during l-IPSPs after approximately 20 consecutive stimuli. Recovery from this fatigue requires minutes. 6. l-IPSPs are not blocked by bicuculline but are blocked by baclofen

Transcriptional Mechanisms of Proneural Factors and REST in Regulating Neuronal Reprogramming of Astrocytes

© 2015 Elsevier Inc. Direct lineage reprogramming induces dramatic shifts in cellular identity, employing poorly understood mechanisms. Recently, we demonstrated that expression of Neurog2 or Ascl1 in postnatal mouse astrocytes generates glutamatergic or GABAergic neurons. Here, we take advantage of this model to study dynamics of neuronal cell fate acquisition at the transcriptional level. We found that Neurog2 and Ascl1 rapidly elicited distinct neurogenic programs with only a small subset of shared target genes. Within this subset, only NeuroD4 could by itself induce neuronal reprogramming in both mouse and human astrocytes, while co-expression with Insm1 was required for glutamatergic maturation. Cultured astrocytes gradually became refractory to reprogramming, in part by the repressor REST preventing Neurog2 from binding to the NeuroD4 promoter. Notably, in astrocytes refractory to Neurog2 activation, the underlying neurogenic program remained amenable to reprogramming by exogenous NeuroD4. Our findings support a model of temporal hierarchy for cell fate change during neuronal reprogramming