The α2A -adrenoceptor suppresses excitatory synaptic transmission to both excitatory and inhibitory neurons in layer 4 barrel cortex.

KEY POINTS: The effects of noradrenaline on excitatory synaptic transmission to regular spiking (excitatory) cells as well as regular spiking non-pyramidal and fast spiking (both inhibitory) cells in cortical layer 4 were studied in thalamocortical slice preparations, focusing on vertical input from thalamus and layer 2/3 in the mouse barrel cortex. Excitatory synaptic responses were suppressed by noradrenaline. However, currents induced by iontophoretically applied glutamate were not suppressed. Further, paired pulse ratio and coefficient of variation analysis indicated the site of action was presynaptic. Pharmacological studies indicated that the suppression was mediated by the α2- adrenoceptor. Consistent with this, involvement of α2A -adrenoceptor activation in the synaptic suppression in excitatory and inhibitory cells was confirmed by the use of α2A -adrenoceptor knockout mice. ABSTRACT: The mammalian neocortex is widely innervated by noradrenergic (NA) fibres from the locus coeruleus. To determine the effects of NA on vertical synaptic inputs to layer 4 (L4) cells from the ventrobasal thalamus and layer 2/3 (L2/3), thalamocortical slices were prepared and whole-cell recordings were made from L4 cells. Excitatory synaptic responses were evoked by electrical stimulation of the thalamus or L2/3 immediately above. Recorded cells were identified as regular spiking, regular spiking non-pyramidal or fast spiking cells through their firing patterns in response to current injections. NA suppressed (∼50% of control) excitatory vertical inputs to all cell types in a dose-dependent manner. The presynaptic site of action of NA was suggested by three independent studies. First, responses caused by iontophoretically applied glutamate were not suppressed by NA. Second, the paired pulse ratio was increased during NA suppression. Finally, a coefficient of variation (CV) analysis was performed and the resultant diagonal alignment of the ratio of CV-2 plotted against the ratio of the amplitude of postsynaptic responses suggests a presynaptic mechanism for the suppression. Experiments with phenylephrine (an α1 -agonist), prazosin (an α1 -antagonist), yohimbine (an α2 -antagonist) and propranolol (a ß-antagonist) indicated that suppression was mediated by the α2 -adrenoceptor. To determine whether the α2A -adrenoceptor subtype was involved, α2A -adrenoceptor knockout mice were used. NA failed to suppress EPSCs in all cell types, suggesting an involvement of the α2A -adrenoceptor. Altogether, we concluded that NA suppresses vertical excitatory synaptic connections in L4 excitatory and inhibitory cells through the presynaptic α2A -adrenoceptor.

Fast activation of feedforward inhibitory neurons from thalamic input and its relevance to the regulation of spike sequences in the barrel cortex.

Thalamocortical afferents innervate both excitatory and inhibitory cells, the latter in turn producing disynaptic feedforward inhibition, thus creating fast excitation-inhibition sequences in the cortical cells. Since this inhibition is disynaptic, the time lag of the excitation-inhibition sequence could be approximately 2-3 ms, while it is often as short as only slightly above 1 ms; the mechanism and function of such fast IPSPs are not fully understood. Here we show that thalamic activation of inhibitory neurons precedes that of excitatory neurons, due to increased conduction velocity of thalamic axons innervating inhibitory cells. Developmentally, such latency differences were seen only after the end of the second postnatal week, prior to the completion of myelination of the thalamocortical afferent. Furthermore, destroying myelination failed to extinguish the latency difference. Instead, axons innervating inhibitory cells had consistently lower threshold, indicating they had larger diameter, which is likely to underlie the differential conduction velocity. Since faster activation of GABAergic neurons from the thalamus can not only curtail monosynaptic EPSPs but also make disynaptic ISPSs precede disynaptic EPSPs, such suppression theoretically enables a temporal separation of thalamically driven mono- and disynaptic EPSPs, resulting in spike sequences of 'L4 leading L2/3'. By recording L4 and L2/3 cells simultaneously, we found that suppression of IPSPs could lead to deterioration of spike sequences. Thus, from the end of the second postnatal week, by activating GABAergic neurons prior to excitatory neurons from the thalamus, fast feedforward disynaptic suppression on postsynaptic cells may play a role in establishing the spike sequences of 'L4 leading L2/3 cells'.

Electroacupuncture induces the expression of Fos in rat dorsal horn via capsaicin-insensitive afferents.

Fos is expressed in rat dorsal horn neurons after electroacupuncture (E-acupuncture), but it is unclear which types of afferent fibers are involved in the expression. It is thought that the Fos expression is induced via Adelta afferents rather than C afferents, since the threshold of Adelta afferents to electrical stimulation is much lower than that of unmyelinated ones. Therefore, neonatally capsaicin treated rats lacking many C afferents were examined to clarify this. Fos expression in the dorsal horn after injection of formalin into the hindpaw was severely attenuated by neonatal capsaicin treatment. However, Fos expression after E-acupuncture to the pads of the hindpaw was unaffected by the same treatment. These results suggest that E-acupuncture induces the expression of Fos in the dorsal horn neurons via capsaicin-insensitive afferents, presumably Adelta afferents.