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Journal of Neuroscience, Vol 12, 2130-2143, Copyright © 1992 by Society for Neuroscience
Characterization of a novel synapse-specific protein. I. Developmental expression and cellular localization of the F1-20 protein and mRNA
R Sousa, NH Tannery, S Zhou and EM Lafer
Department of Biological Sciences, University of Pittsburgh, Pennsylvania 15260.
A molecular description of the nerve terminal will be required to
understand synaptic function fully. The goals of this study were to
contribute toward such a description by characterizing a novel synapse-
specific protein. A monoclonal antibody library was screened for antibodies
to synaptic proteins. The antibodies were then used to isolate cDNA clones
by expression screening. Here we report a detailed characterization of the
protein reactive with monoclonal antibody F1- 20. Immunohistochemical and
biochemical analyses revealed that the F1- 20 protein is synapse
associated. Western blot analyses revealed that the F1-20 protein is a
brain-specific polypeptide with an apparent molecular weight on SDS-PAGE of
190,000 Da. Northern blot analyses indicated that probes generated from an
F1-20 cDNA clone hybridize to a single brain-specific mRNA of approximately
4.8 kilobases. In situ hybridization experiments demonstrated that F1-20
mRNA expression is neuronal specific. Northern and Western blot analyses
indicated that F1- 20 mRNA levels increase abruptly at postnatal day 4 and
protein levels increase abruptly at postnatal day 7. This corresponds to a
period of active synaptogenesis and synaptic maturation in the mouse CNS.
We characterized the neuroanatomical distribution of the F1-20 protein by
immunohistochemistry, and of the F1-20 mRNA by in situ hybridization. We
found that the F1-20 mRNA and protein are expressed nonuniformly in brain.
Variation in the expression of the F1-20 protein is complex and reveals
patterns also exhibited by probes directed against other synapse-associated
molecules. The highest levels of F1-20 protein are found in the cortically
organized regions of the brain. The highest levels of F1-20 mRNA are found
in long-distance projection neurons. There is also variation in the
expression of F1-20 mRNA between different classes of large output neuron,
as well as extensive variation in the expression of F1-20 mRNA between
different nuclear groups.
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