Yasuhiro Morita

Localization of GABAA-receptor α1 subunit mRNA-containing neurons in the lower brainstem of the rat

The localization of gamma-aminobutyric acid-A (GABAA) receptors (GABAA-R) in the lower brainstem of the rat was examined by means of in situ hybridization histochemistry using an oligonucleotide probe to the sequence of the alpha 1 subunit (GABAA-R alpha 1). Strongly labeled neurons were found in the cranial motor nuclei, the dorsal motor nucleus of the vagus, reticular formation (large neurons), lateral vestibular nucleus, dorsal nucleus of the lateral lemniscus, central nucleus of the inferior colliculus, intermediate and white layers of the superior colliculus, red nucleus and substantia nigra. In addition, moderately labeled cells were abundant in the nucleus of the solitary tract, medial and inferior vestibular nuclei, parabrachial area, dorsal and ventral tegmental nuclei of Gudden, central gray matter, ventral nucleus of the lateral lemniscus, and reticular formation (small neurons). This study has therefore revealed some of the target neurons of GABA-containing fibers in the lower brainstem.

Different ontogenetic profiles of cells expressing prepro-neurotensin/neuromedin N mRNA in the rat posterior cingulate cortex and the hippocampal formation

The ontogeny of the expression of prepro-neurotensin/neuromedin N messenger RNA (prepro-NT/NN mRNA) in the rat posterior cingulate cortex (retrosplenial cortex) and the hippocampal formation was investigated using in situ hybridization histochemistry. In the primordium of the posterior cingulate cortex and the hippocampal formation, prepro-NT/NN mRNA was first expressed on embryonic day 17, and was found in the subiculum, layers II-III in areas 29a and 29b, and layer VI in the posterior cingulate cortex at birth. Expression was also observed in the CA1 field. In the adult rat, the expression of prepro-NT/NN mRNA was reduced in the posterior cingulate cortex, and only a few positive cells were seen here. However, the CA1 field and the subiculum still contained numerous positive cells.

Localization and ontogeny of cells expressing preprodynorphin mRNA in the rat cerebral cortex

The regional distribution of preprodynorphin (PPD) mRNA-containing cells (PPD cells) in the rat cerebral cortex was investigated by in situ hybridization histochemistry using a synthetic oligonucleotide probe. In the isocortex, PPD cells were small or medium-sized and were mainly located in layer V. While they were less numerous in the allocortex than in the isocortex. Only a few labeled cells were seen in the piriform and entorhinal cortices. In the hippocampal formation, labeled cells were observed in the granular layer of the dentate gyrus. An ontogenetic study revealed that PPD mRNA-containing cells appeared on postnatal day 7 in the isocortex and the allocortex and on day 14 in the dentate gyrus. Thereafter, they increased in number and signal intensity to reach a plateau on postnatal day 14 in both the isocortex and the allocortex and on day 35 in the dentate gyrus. The time-course of development of PPD mRNA-containing neurons in the cerebral cortex suggested that PPD-derived peptide has a neuromodulator and/or neurotransmitter role in these regions of the brain.

Possible regulation of the cytosolic-free calcium concentration by Na+ spikes in immature cerebellar Purkinje cells

Isolated immature cerebellar Purkinje cell bodies often exhibit the spontaneous fluctuations of a cytosolic free Ca concentration ([ Ca]in), which was measured by fura-2 microfluorimetry. In silent cells, however, the increase of [Ca]in could be triggered by pharmacological depolarizations (adding veratridine or tetraethylammonium). In any case, these [Ca]in changes were sensitive to voltage-dependent channel blockers (tetrodotoxin and Ca channel blockers) and to an inhibitory transmitter (GABA). The results suggest that the spontaneous activation of voltage-dependent Na+ and Ca2+ channels might be involved in the increase of [Ca]in.

GABA inhibits the rise in cytosolic free calcium concentration in depolarized immature cerebellar Purkinje cells

gamma-Aminobutyric acid (GABA) reduced the peak rise or slowed the rate of rise in cytosolic free calcium concentration ([Ca]in) induced by quisqualate, kainate, or high KCl in immature cerebellar Purkinje cell bodies. The sustained rise or repeated transient of [Ca]in induced by tetraethylammonium, veratridine, or Bay-K-8644 was lowered to the basal level by adding GABA, although the inhibition by GABA of Bay-K-8644-induced rise in [Ca]in was only slight and transient in some cells. These findings indicate that GABA inhibits the rise in [Ca]in by hyperpolarizing the membrane potentials at Purkinje cell bodies.

N-Methyl-d-aspartate and aspartate raise the cytosolic freee calcium concentration by acting upon receptors transiently expressed on immature cerebellar Purkinje cells

N-Methyl-D-aspartate (NMDA) or aspartate (Asp) increased the cytosolic free calcium concentration ([Ca]in) in some populations of Purkinje cells dissociated from immature rat cerebellum. The NMDA- and Asp-induced rise in [Ca]in was affected only a little by adding glycine or NMDA antagonists, but was reduced either by adding Mg2+, Gallopamil hydrochloride (D-600) and gamma-amino-butyric acid, or by removing external Na+. The results suggest that stimulation of the NMDA-sensitive receptors transiently expressed on immature Purkinje cell soma results in a rise in [Ca]in through the activation of voltage-dependent Ca2+ channels.

Regulation of the cytosolic free calcium concentration by Na+ spikes in immature cerebellar neurons with N-methyl-d-aspartate receptors

N-Methyl-D-aspartate and glycine increased the cytosolic free calcium concentration ([Ca]in) in medium-sized cerebellar neurons. Spontaneous changes in [Ca]in were occasionally observed in NMDA-responsive cells, but large increases in [Ca]in were triggered only through depolarizations by adding veratridine or K+ channel blockers in every cell examined. The [Ca]in increase was suppressed by voltage-dependent Na+ and Ca2+ channel blockers and by an inhibitory transmitter (GABA), suggesting that the generation of Na+ spikes is involved in the increase in [Ca]in.