| Popis: |
GAD, the GABA biosynthetic enzyme, and CSD, the enzyme which converts cysteic acid to taurine, a putative inhibitory transmitter, and cysteine sulfinate to hypotaurine, the proposed precursor of taurine, were partially copurified from rat brain. Polyvalent antisera against this preparation were raised in sheep. GAD/CSD was localized on two-dimensional electrophoresis and line immunoelectrophoresis (LIE) using the radioactive suicide substrate 2- 3 H-γ-Acetylenic GABA. LIE with less purified GAD/CSD and a polyvalent antiserum yielded three precipitin lines. An antiserum (1440) to the radioactively labelled precipitin line precipitated both GAD and CSD activity. On crossed immunoelectrophoresis with an intermediate gel containing antiserum 1440, the mobility of one antigen in brain homogenate supernatant was altered. Immunocytochemistry using antiserum 1440 and the indirect “PAP” method of Sternberger yielded apparently specific staining in the cerebellum and olfactory bulb. |
