Summary
L-homocysteic acid (HCA) and other amino acids were conjugated to rat brain material (extracted rat brain protein) with glutaraldehyde to form HCA- and amino acids-brain material conjugates. The specificity of monoclonal antibody (McAb) was tested on serialdilution test and absorption test on enzyme-linked immunosorbent assay (ELISA) using these conjugates as antigens instead of amino acids-BSA (bovine serum albumin) conjugates used previously. The characterized McAb was applied for immunohistochemical staining using PAP (peroxidase antiperoxidase) technique in combination with silver enhancement of diamino-benzene (DAB) products. The results indicated that McAb to L-HCA reacted with L-HCA-brain material conjugates, but not with other amino acids-brain material conjugates so far tested. McAb absorbed with L-HCA-brain material abolished or decreased immunoreactivity of L-HCA-brain material with McAb. The antibody selectively stained subpopulation of cells and processes in the hippocampus fixed with glutaradehyde. Absorption of McAb with L-HCA-brain material abolished immunohistochemical staining. These results suggested that McAb was specific for L-HCA-brain materials and could be used for imunohistocytochemistry. This would provide a new tool for immunohistochemical visualization and localization of L-HCA in the nervous system.
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References
Fonnum F. Glutamate: a neurotransmitter in mammalian brain. J Neurochem, 1984, 43: 1
Watkins J C, Evance R H. Excitatory amino acid transmitters. Annu Rev Pharmacol Toxicol, 1981, 21: 165
Cuenod M, Do K Q, Vollenweider Fet al. The puzzle of the transmitters in the climbing fibers. In: Experimental Brain Research Series (Symposium on “The olivo-cerebellar system in motor control”, Turin). Berlin, Heidelberg, New York: Springer, 1988
Neal M J, Cunningham J R. L-homocysteic acid—a possible bipolar cell transmitter in the rabbit retina. Neurosci Lett, 1989, 102(1): 114
Do K Q, Mattenberger M, Streit Pet al. (1986b) In vitro release of endogenous excitatory sulfur-containing amino acids from various rat brain regions. J Neurochem, 1986, 46: 779
Do K Q, Herrling P L, Streit Pet al. In vitro release and electrophysiological effects in situ of homocysteic acid, an endogenous N-methyl-(D)-aspartic acid agonist, in the mammalian striatum. J Neurosci, 1986, 6: 2226
Steinbush H W M, Verhofstad A A, Joosten H W J. Localization of serotonin in the central nervous system by immunohistochemistry; description of a specific and sensitive technique and some applications. Neuroscience, 1978, 3: 811
Sterm-Mathisen J, Leknes A K, Bore A Tet al. First visualization of glutamate and GABA in neurons by immunocytochemistry. Nature, 1983, 301: 517
Liu C J, Li A, Hu X Wet al. Preparation and characterization of monoclonal antibody to L-homocysteic acid. Acta Univ Med Tongji (Chineses), 2001, 30(5): 410
Liu C J, Grandes P, Matute Cet al. (1989) Glutamate-like immunoreactivity revealed in rat olfactory bulb, hippocampus and cerebellum by monoclonal antibody and sensitive staining method. Histoche, 1989, 90: 427
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LIU Changjin, female, born in 1953, Professor
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Changjin, L., Ai, L., Xinwu, H. et al. Characterization of monoclonal antibody to L-homocysteic acid and its immunohistochemistry in rat hippocampus. Current Medical Science 25, 1–4 (2005). https://doi.org/10.1007/BF02831371
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DOI: https://doi.org/10.1007/BF02831371