Kunio Murakami

An ultrastructural study of the neural circuit between the prefrontal cortex and the mediodorsal nucleus of the thalamus

Synaptic connectivity between the prefrontal cortex (PFC) and the mediodorsal thalamic nucleus (MD) of the rat has been investigated with the electron microscope after labeling both the pre- and postsynaptic elements. Prefrontal corticothalamic fibers end exclusively as small axon terminals with round synaptic vesicles (SR boutons), which make asymmetrical synaptic contacts with distal dendritic segments of MD neurons. Thalamocortical terminals from MD in PFC are also of the SR type and form asymmetrical synaptic contacts predominantly with dendritic spines arising from the apical or basal dendrites of pyramidal cells whose somata reside in layers III, V and VI. At least some pyramidal cells in layer III that receive MD afferents are callosal cells, whereas deep layer pyramidal cells projecting to MD receive directly some of the thalamocortical terminations from MD, suggesting that the recurrent loop to MD is monosynaptically mediated. Thus, taken together with recent evidence that both the PFC-MD and MD-PFC pathways are glutamatergic and excitatory, the cortical excitation exerted by afferent fibers from MD is transferred, not only back to MD itself through deep pyramidal cells, but also the contralateral prefrontal cortex via pyramidal cells in layer III of the ipsilateral prefrontal cortex. Concerning modulatory and inhibitory inputs, fibers to MD from the ventral pallidum and substantia nigra pars reticulata have been shown to be inhibitory and GABAergic. In addition, fibers from the ventral tegmental area preferentially make symmetrical membrane thickenings (i.e. inhibitory synapses) on deep pyramidal cells in PFC that receive synaptic endings from MD. From these morphological grounds, therefore, cells in the ventral pallidum, the substantia nigra pars reticulata and the ventral tegmental area may mediate, to some extent, an inhibitory effect on the reverberatory excitation between PFC and MD.

The Convergence of Axon Terminals from the Mediodorsal Thalamic Nucleus and Ventral Tegmental Area on Pyramidal Cells in Layer V of the Rat Prelimbic Cortex

We investigated the ultrastructural basis of the synaptic convergence of afferent fibres from the mediodorsal thalamic nucleus (MD) and the ventral tegmental area (VTA) on the prefrontal cortical neurons of the rat by examining the synaptic relationships between thalamocortical or tegmentocortical terminals labelled with anterograde markers [lesion‐induced degeneration or transport of wheat germ agglutinin conjugated to horseradish peroxidase (WGA—HRP)] and randomly selected unlabelled apical dendrites of layer V pyramidal cells in the prelimbic cortex. WGA—HRP‐labelled terminals from the VTA ranged in diameter from 0.7 to 2.8 μm and established synaptic contacts with large dendritic profiles, i.e. proximal segments of apical dendritic shafts and spines from layer V pyramidal cells. Symmetrical synapses, i.e. inhibitory synapses, were more often seen than asymmetrical ones. Degenerating terminals from the MD formed asymmetrical synapses on dendritic spines or occasionally on small dendritic shafts of apical dendrites from layer V pyramidal cells, which received tegmentocortical synapses, mostly within layer III. Thalamocortical synapses were more distally distributed over common apical dendrites than tegmentocortical synapses, although some of them overlapped. The numerical density of direct synaptic inputs from the MD and VTA was low. These results suggest that fibres from the VTA exert their inhibitory effects directly on pyramidal cells in layer V via synaptic junctions with apical dendrites of these pyramidal cells, and that the tegmentocortical fibres are in an ideal anatomical position to modulate the reverberatory circuits between the MD and the prelimbic cortex.

Direct synaptic connections between thalamocortical axon terminals from the mediodorsal thalamic nucleus (MD) and corticothalamic neurons to MD in the prefrontal cortex.

A combined anterograde axonal degeneration with ibotenic acid and wheat germ agglutinin-horseradish peroxidase (WGA-HRP) retrograde tracing study revealed that some degenerating thalamocortical axon terminals from the mediodorsal thalamic nucleus (MD) directly formed asymmetrical synaptic contacts predominantly with dendritic spines of apical dendrites of WGA-HRP-labeled corticothalamic projection neurons to MD in the prelimbic cortex of the rat. This result suggests that there is a monosynaptic feedback loop from and to MD via deeper layer neurons in the prelimbic cortex.

Dual termination modes of corticothalamic fibers originating from pyramids of layers 5 and 6 in cat visual cortical area 17

Terminal morphology of corticothalamic fibers originating from cat area 17 was examined. Injections of an anterograde axonal tracer, phaseolus vulgaris leucoagglutinin (PHA-L), in area 17 resulted in labeling of small boutons in the dorsal lateral geniculate, perigeniculate, and thalamic reticular nuclei and in labeling of large boutons in the lateral nucleus of lateral posterior-pulvinar, ventral lateral geniculate, and pulvinar nuclei. Since it is well known that the dorsal geniculate nucleus is a major corticothalamic target for layer 6 pyramids and the lateral posterior-pulvinar complex is that for layer 5 pyramids, the findings indicate that layer 5 pyramids in cat area 17 project axons ending with large boutons, while layer 6 pyramids project those ending with small boutons.

Distribution of the piriform cortical terminals to cells in the central segment of the mediodorsal thalamic nucleus of the rat

A Golgi electron microscopic study was undertaken to investigate the distribution of terminals from the piriform cortex that synapse on identified dendrites of neurons in the central segment of the mediodorsal thalamic nucleus of the rat. The piriform cortical terminals were identified as degenerating terminals following lesions in the cortex. They consisted of two types, i.e., large (LR type) and small (SR type) presynaptic terminals, both of which had round synaptic vesicles and formed asymmetric synaptic contacts. SR boutons terminated preferentially onto distal dendrites and never synapsed on primary dendrites. LR terminals synapsed preferentially on proximal dendrites, but were also found on more distal dendritic segments.

Quantitative analysis of axon collaterals of single cells in layer III of the piriform cortex of the guinea pig

Recent physiological and morphological studies suggest that the piriform cortex (PC) functions like the association areas of the neocortex rather than the typical primary sensory area as was previously assumed. The axon connection patterns of single cells are important for understanding the functional organization of the PC. The axon collaterals of three single pyramidal cells and one spiny multipolar cell in layer III of the PC were labeled and quantitatively analyzed by intracellular injections of biocytin in guinea pigs. The individual pyramidal and spiny multipolar cells have highly distributed axon collaterals, which display little tendency for patchy concentrations, within the PC and multiple higher order behavior/reward/contextual‐related areas, such as the prefrontal cortex, amygdaloid nuclei, and entorhinal cortex. For the pyramidal cells, the average length of axonal collaterals is 143 mm; the average number of boutons is 12,930. For the spiny multipolar cell, the length of the axonal collaterals is 88 mm; the number of boutons is 7,052. The pyramidal cells in the anterior subdivision of the PC (APC) have both rostrally and caudally directed intrinsic association fibers, whereas the pyramidal and spiny multipolar cells in the posterior subdivision (PPC) have predominantly caudally directed intrinsic association fibers in the PC. Our results reveal that the connection patterns of single cells in layer III resemble those of pyramidal cells in layer II, suggesting that the PC performs correlative functions analogous to those in the association area of other sensory systems. The rostrally‐to‐caudally directed connections in the APC provide a substrate for the recurrent process, whereas largely caudally directed connections in the PPC suggest the dominance of the feed‐forward process. J. Comp. Neurol. 465:455–465, 2003.

Electron microscopic evidence that axon terminals from the mediodorsal thalamic nucleus make direct synaptic contacts with callosal cells in the prelimbic cortex of the rat

A combined study of anterograde axonal degeneration and HRP retrograde labeling has shown that there exist monosynaptic connections between afferent fibers from the mediodorsal thalamic nucleus (MD) and callosal cells in the prelimbic cortex of the rat. Degenerating axon terminals from MD made asymmetrical synaptic contacts with dendritic spines from apical dendrites of layer III pyramidal cells that were retrogradely labeled with HRP after its injection into the prelimbic cortex contralateral to MD lesions.

Synaptic relationships between axon terminals from the mediodorsal thalamic nucleus and layer III pyramidal cells in the prelimbic cortex of the rat.

A combined study of anterograde axonal degeneration and Golgi electron microscopic technique was designed to examine the distribution and density of axon terminals from the mediodorsal thalamic nucleus (MD) over layer III pyramidal cells in the prelimbic cortex of the rat. The reconstructive analysis of serial ultrathin sections of gold-toned apical and basal dendrites of layer III pyramidal cells showed that degenerating thalamocortical axon terminals from MD formed asymmetrical synaptic contacts predominantly with dendritic spines of the identified basal dendrites as well as apical dendrites. There was little difference in the numerical density of thalamocortical synapses from MD per unit length of both apical and basal dendrites.

Localization of α7 nicotinic acetylcholine receptor immunoreactivity on GABAergic interneurons in layers I–III of the rat retrosplenial granular cortex

The rat retrosplenial granular cortex (RSG) receives cholinergic input from the medial septum-diagonal band (MS-DB) of the cholinergic basal forebrain (CBF), with projections terminating in layers I-III of RSG. The modulatory effects of acetylcholine (ACh) on cortical GABAergic interneurons in these layers are mediated by α7 nicotinic acetylcholine receptors (α7nAChRs). α7nAChRs are most abundant in the cerebral cortex and are largely localized to GABAergic interneurons. However, the CBF projection to the RSG has not been studied in detail, and the cellular or subcellular distribution of α7nAChRs in the rat RSG remains unclear. The main objective of this study was to test that α7nAChRs reside on GABAergic interneurons in CBF terminal fields of the rat RSG. First, we set out to define the characteristics of CBF projections from the MS-DB to layers of the RSG using anterograde neural tracing and immunohistochemical labeling with cholinergic markers. These results revealed that the pattern of axon terminal labeling in layer Ia, as well as layer II/III of the RSG is remarkably similar to the pattern of cholinergic axons in the RSG. Next, we investigated the relationship between α7nAChRs, labeled using either α-bungarotoxin or α7nAChR antibody, and the local neurochemical environment by labeling surrounding cells with antibodies against glutamic acid decarboxylase (GAD), parvalbumin (PV) and reelin (a marker of the ionotropic serotonin receptor-expressing GABAergic interneurons). α7nAChRs were found to be localized on both somatodendritic and neuronal elements within subpopulations of GABAergic PV-, reelin-stained and non PV-stained neurons in layers I-III of the RSG. Finally, electron microscopy revealed that α7nAChRs are GAD- and PV-positive cytoplasmic and neuronal elements. These results strongly suggest that ACh released from CBF afferents is transmitted via α7nAChR to GAD-, PV-, and reelin-positive GABAergic interneurons in layers I-III of the RSG.

Anatomical architecture of the brachial plexus in the common hippopotamus (Hippopotamus amphibius) with special reference to the derivation and course of its unique branches.

The anatomy of the brachial plexus in the common hippopotamus (Hippopotamus amphibius), which has not been previously reported, was first examined bilaterally in a newborn hippopotamus. Our observations clarified the following: (1) the brachial plexus comprises the fifth cervical (C5) to first thoracic (T1) nerves. These formed two trunks, C5–C6 and C7–T1; in addition, the axillary artery passed in between C6 and C7, (2) unique branches to the brachialis muscle and those of the lateral cutaneous antebrachii nerves ramified from the median nerve, (3) nerve fibre analysis revealed that these unique nerve branches from the median nerve were closely related and structurally similar to the musculocutaneous (MC) nerve; however, they had changed course from the MC to the median nerve, and (4) this unique branching pattern is likely to be a common morphological feature of the brachial plexus in amphibians, reptiles and certain mammals.