Johji Kohno

Two distinct strio-nigral substance P pathways in the rat: An experimental immunohistochemical study

Re-examination of the strio-nigral substance P (SP) tract by means of an experimental immunohistochemical method in the rats demonstrated the presence of two distinct pathways from the nucleus caudatus putamen (CP) to the substantia nigra (SN). Destruction of the posterior portion of the CP resulted in the disappearance of SP-positive fibers in the SN pars lateralis but not in the SN pars compacta or reticulata. On the other hand, destruction of the ventrolateral portion of the anterior portion of the CP caused the disappearance of SP-positive fibers in the SN pars compacta and pars reticulata but not in the SN pars lateralis. In addition, destruction of the dorsal portion of the anterior portion of the CP, where 3-6 cell islets of SP-positive cells are located, failed to decrease SP-positive fibers in any of the subdivisions of the SN. These findings strongly suggest that SP-positive neurons in the posterior portion of the CP project to the SN pars lateralis (posterior strio-nigral SP tract), SP-positive cells in the lateroventral part of the anterior portion of the CP extend to the SN pars compacta and pars reticulata, but SP-positive cells in the dorsal part of the anterior portion of the CP do not innervate the SN.

Enkephalin fibers synapse on cholinergic neurons in the rat sacral intermediolateral nucleus: A double-immunostaining at the light and electron microscopic levels

Relationships between leucine-enkephalin fibers and cholinergic neurons in the rat sacral intermediolateral nucleus were examined by light and electron microscopy using double-immunostaining method. Cholinergic neurons in the sacral intermediolateral nucleus were labeled by a rat-mouse monoclonal antibody to choline acetyltransferase and stained bluish green with 5-bromo-4-chloro-3-indolyl-beta-D- galactoside reaction products using beta-galactosidase as a marker. On the same sections, leucine-enkephalin fibers were labeled by a rabbit polyclonal antiserum to leucine-enkephalin and stained brown by diaminobenzidine reaction products using peroxidase as a marker. After embedding in Epon, the sections were examined in light and electron microscopes. In the light microscope, choline acetyltransferase-like immunoreactive cells were seen in the sacral intermediolateral nucleus. In the same region, leucine-enkephalin-like immunoreactive cells. In the electron microscope, 5-bromo-4-chloro-3-indolyl-beta-D-galactoside reaction products were in the form of coarse electron dense deposits in the choline acetyltransferase-like immunoreactive structures and could be distinguished from the much finer grained diaminobenzidine reaction products. Choline acetyltransferase-like immunoreactive neurons received synaptic inputs from leucine-enkephalin fibers-like immunoreactive terminals. These findings suggest that leucine-enkephalin fibers may affect the activity of cholinergic parasympathetic preganglionic neurons.

Direct adrenergic inputs to sacral autonomic neurons: using a double-immunostaining method at the light and electron microscopic levels

Relationships between adrenergic fibers and cholinergic neurons were examined in the rat sacral intermediolateral nucleus using a double-immunostaining method at light and electron microscopic levels. Adrenergic fibers were immunohistochemically stained brown by peroxidase reaction, and cholinergic neurons in the same sections were stained bluish green by beta-galactosidase reaction. In the light microscope, many cholinergic neurons were seen in the intermediolateral nucleus and some of them were surrounded by adrenergic fibers. In the electron microscope, adrenergic fibers made synapses with the cholinergic neurons in the intermediolateral nucleus. These results suggest that adrenergic fibers directly influence the activity of the sacral parasympathetic preganglionic neurons.