Elsevier

Brain Research

Volume 475, Issue 1, 13 December 1988, Pages 91-102
Brain Research

Aromaticl-amino acid decar☐ylase-immunoreactive neurons in and around the cerebrospinal fluid-contacting neurons of the central canal do not contain dopamine or serotonin in the mouse and rat spinal cord

https://doi.org/10.1016/0006-8993(88)90202-8Get rights and content

Abstract

Using a specific antibody against each of the 4 catecholamine-synthesizing enzymes, dopamine or serotonin, we compared the localization of these substances in rat and mouse spinal cord with two kinds of fixative, such as paraformaldehyde and glutaraldehyde, and two types of sectioning techniqye, i.e. cryostat and microslicer. In this study, we demonstrated aromaticl-amino acid decar☐ylase (AADC)-like immunoreactivity (LI) in cerebrospinal fluid (CSF)-contacting neurons for the first time in the mouse. These cells and some AADC-immunoreactive cells localized outside the subependymal layer of mouse and rat spinal cord did not show tyrosine hydroxylase-, serotonin- and dopamine-LI with and without colchicine treatment.

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      Coronal sections (40 μm in thickness) were cut on a cryostat and collected in PBS. IHC was performed by incubation overnight with TH (1:50,000) (Nagatsu et al., 1979) or AADC (1:20,000) antibodies (Nagatsu et al., 1988). The slides were washed, incubated for 2 h with a biotinylated anti-rabbit IgG secondary antibody (1:200, Vector Laboratories, Burlingame, CA, USA), and visualized using the VECTASTAIN Elite ABC Kit and Peroxidase Substrate DAB Kit (both from Vector Laboratories, Burlingame, CA, USA).

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