2002;283:C126CC134. CVOs. Besides affecting glial-neural functions in the CVOs, the astrocytes found in the white matter may affect saltatory nerve conduction, serving as a sodium buffer. The ENaC -expressing astrocytes of the ventral medulla send processes into the raphe pallidus which intermingle with the serotoninergic (5-HT) neurons found in this region as well as with the other nearby 5-HT neurons distributed along ventral medullary surface. hybridization studies (Miller, 2013). The immunostaining pattern obtained with the ENaC -subunit antibody was similar to the results found for the anti-sera that was generated against ENaC -subunit. In contrast, the ENaC -immunostaining pattern was different, yielding very weak neuronal staining in the three sensory CVOs: AP, SFO, and OVLT. An example of the weak ENaC- immunoreactive neurons is presented in Figure 1. Unlike the immunostaining resulting from the antibodies directed against the ENaC – and -subunits, the ENaC -subunit resulted in robust staining of the astrocytes that border these three CVOs (Figs. 1C5). In addition, ENaC -immunoreactive astrocytes were also prominent in white matter, such as the optic chiasm (Figs. 1 and ?and2)2) and pyramidal tract (Fig.6) as well as with the pia mater (Figs. 1, ?,2,2, ?,4,4, ?,5,5, ?,6).6). The pia mater was also Tegafur immunostained with antibodies directed against ENaC -and -subunits. ENaC -subunit immunoreactivity was colocalized in GFAP pial materials (Figs. 2A and ?and5A).5A). In addition, all three antibodies resulted in immunostaining of the ependymal lining of the brain, but, as expected, no GFAP immunoreactivity was found in this tissue. Open in a separate window Number 1 Distribution of ENaC -subunit immunoreactivity as shown inside a parasagittal section of the rat forebrain. ENaC -subunit immunoreactivity was present in the lamina terminalis (LT), pia mater, and strongly indicated in the neurons of the nucleus of the diagonal band (NDB). Weak ENaC -manifestation was observed in the neurons of the dorsal cap and Tegafur lateral zone of the organum vasculosum of the lamina terminalis (OVLT), median preoptic nucleus (MnPO), and periventricular hypothalamic region (Pe). In the optic chiasm (OX), very strong ENaC -manifestation was recognized Tegafur in astrocytes that lay in the border of this dietary fiber bundle; this region is designated by asterisks (*). Notice the absence of ENaC -subunit immunoreactive staining in the astrocytes in the brain appropriate. The ENaC -subunit expressing astrocytes are distributed in select mind Tegafur regions, including in the zone between the NDB and LT. III, third ventricle; or, optic recess. Open in a separate window Number 2 A. Brightfield preparation of a transverse section through the forebrain showing dense ENaC -subunit expressing astrocyte plexus that forms the lamina terminalis (LT). This plexus is definitely continuous with the pia mater that invests the ventral mind surface and surrounds the optic chiasm (OX). The ENaC -positive glial materials format the optic recess (or) which lies in probably the most rostral part of the third ventricle. A’ Two times immunofluorescence preparation to demonstrate co-expression of glial fibrillary acidic protein (GFAP) and ENaC -subunit protein is seen in the pia. Lower part of this number shows enlarged photoimages to show overlap ENaC -subunit protein and GFAP. B. ENaC -subunit expressing neurons were present in the dorsal cap and lateral zone of the OVLT. Dorsal cap OVLT neurons were more ENaC -subunit immunoreactive than the lateral zone OVLT neurons. B’. Double-immunostained preparation to show the ENaC -subunit immunoreactive in the lateral zone OVLT neurons also communicate NeuN C a neuronal nuclear marker. Related results were found for the dorsal cap OVLT neurons. Open in FKBP4 a separate window Number 4 Transverse section of the dorsomedial medulla to show the distribution of ENaC -immunoreactivity in neurons and astrocytes..