Journal Article

Histamine Monoclonal Antibody for Brain Immunocytochemistry

Kunio Fujiwara, Yukinobu Masuyama, Shiroki Yagisawa, Toshio Tanabe, Masahiko Yabuuchi and Daisuke Tsuru

in The Journal of Biochemistry

Published on behalf of The Japanese Biochemical Society

Volume 126, issue 3, pages 503-509
Published in print September 1999 | ISSN: 0021-924X
Published online September 1999 | e-ISSN: 1756-2651 | DOI: http://dx.doi.org/10.1093/oxfordjournals.jbchem.a022479
Histamine Monoclonal Antibody for Brain Immunocytochemistry

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Among five monoclonal antibodies (AHA-1 to 5 mAbs) prepared against glutaraldehyde (GA)-conjugated histamine (HA) in our previous study, only mAb AHA-2 was found to detect HA specifically in rat brain neurons by an immunocytochemistry method (ICC) using GA as a tissue fixative. All the other mAbs, except for AHA-5, reacted with HA in the enterochromaflln-like cells (ECL cells) of rat stomach [Fujiwara et at. (1997) Histochem. Cell Biol 107, 39–45]. Enzyme-linked immunosorbent assay (ELISA) binding and inhibition tests demonstrated that AHA-2 is specific for HA, with almost no detectable cross-reaction with any other established or putative amino acid neurotransmitters, LH-RH, TRH, or peptides with N-terminal histidines. ELJSA assays also suggested that the AHA-2 mAb recognizes a HA epitope structure different from the one recognized by the AHA-1 mAb. The immunostaining patterns with AHA-2 mAb, as seen in the five subgroups of the tuberomammillary nuclei in the rat posterior hypothalamus, were very similar to those described by Inagaki et al. [(1988) Brain Res. 439, 402–405; (1990) Exp. Brain Res. 80, 374–380] and Panula et al. [(1984) Proc. Natl. Acad. Sci. USA 81, 2572–2576; (1988) J. Histochem. Cytochem. 36, 259–269] using polyclonal anti-HA serum. However, it was also noted that moderate numbers of immunoreactive nerve fibers projected into the median eminence. The present HA ICC method using AHA-2 mAb allows highly sensitive HA detection in brain, and thus might permit detailed studies of HA localization hitherto impossible using previously available anti-HA polyclonal antibodies produced against carbodiimide-conjugated HA.

Keywords: glutaraldehyde; histamine; immunocytochemistry; monoclonal antibody; neuron

Journal Article.  0 words. 

Subjects: Biochemistry

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