2-amino-5-phosphono-3-pentenoic-acid and biocytin

GABAergic interneurons play central roles in the regulation of neuronal activity in the basolateral nucleus of the amygdala (BLA). They are also suggested to be the principal targets of the brainstem noradrenergic afferents which are involved in the enhancement of the BLA-related memory. In addition, behavioral stress has been shown to impair noradrenergic facilitation of GABAergic transmission. However, the noradrenaline (NA) effects in the BLA have not been differentiated among medium- to large-sized GABAergic neurons and principal cells, and remain to be elucidated in terms of their underlying mechanisms. Glutamate decarboxylase 67 (GAD67) is a biosynthetic enzyme of GABA and is specifically expressed in GABAergic neurons. To facilitate the study of the NA effects on GABAergic neurons in live preparations, we generated GAD67-green fluorescent protein (GFP) knock-in mice, in which GFP was expressed under the control of an endogenous GAD67 gene promoter. Here, we show that GFP was specifically expressed in GABAergic neurons in the BLA of this GAD67-GFP knock-in mouse. Under whole-cell patch-clamp recordings in vitro, we identified a certain subpopulation of GABAergic neurons in the BLA chiefly on the basis of the electrophysiological properties. When depolarized by a current injection, these neurons, which are referred to as type A, generated action potentials at relatively low frequency. We found that NA directly excited type-A cells via alpha1-adrenoceptors, whereas its effects on the other types of neurons were negligible. Two ionic mechanisms were involved in this excitability: the activation of nonselective cationic conductance and the suppression of the resting K+ conductance. NA also increased the frequency of spontaneous IPSCs in the principal cells of the BLA. It is suggested that the NA-dependent excitation of type-A cells attenuates the BLA output for a certain period.

Topics: 2-Amino-5-phosphonovalerate; Action Potentials; Adrenergic Agents; Amygdala; Anesthetics, Local; Animals; Biophysical Phenomena; Dose-Response Relationship, Drug; Electric Conductivity; Electric Stimulation; Excitatory Amino Acid Antagonists; GABA Antagonists; gamma-Aminobutyric Acid; Glutamate Decarboxylase; Green Fluorescent Proteins; In Vitro Techniques; Lysine; Membrane Potentials; Mice; Mice, Inbred C57BL; Mice, Inbred ICR; Mice, Transgenic; Neurons; Norepinephrine; Patch-Clamp Techniques; Phosphinic Acids; Potassium; Propanolamines; Tetrodotoxin