Antibody Markers for Studying Neurodegeneration in the Nothobranchius Central Nervous System

Abstract

Fish of the genus Nothobranchius have a naturally short lifespan in which they develop several neurological biomarkers of aging. This short lifespan (several months compared to several years in mice and zebrafish) make these fish a promising alternative in aging research. Resveratrol dosing retarded the development of the neurological biomarkers of aging identified in N. furzeri. Resveratrol's method of action was not determined due to the lack of celltype specific markers of degenerative processes. Further research of the Nothobranchius CNS and its degeneration is hampered by the lack of cell-type specific antibodies and other cellular markers of degenerative processes. In this article the authors present a molecular toolbox of five antibodies and a lectin which can be used to study the Nothobranchius central nervous system and its degeneration. These antibodies were tested against western blots of rat, zebrafish and Nothobranchius guentheri brain homogenates as well as frozen sections. Antibodies shown to reliably bind with N. guentheri proteins were: SMI31 which labeled axons and dendrites; mouse anti-human GFAP and rabbit anti-cow GFAP which labeled astroglia; mouse anti-rat tenascin-R which labeled oligodendrocytes; and E587 anti-goldfish L1 which labeled neuronal and astrocytic cell bodies and processes. The BS-I isolectin B4 labeled microglia in the optic nerve and retina. This toolbox will enable researchers to study the aging brain of Nothobranchius as well as its normal development at a cellular level.