THE ISOLATION AND CHARACTERIZATION OF NOVEL POSTSYNAPTIC ANTIBODIES FOR DANIO RERIO
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The proteins and molecular agents that govern synapse formation in developing organisms are essential for cognitive and neural maturation. Mutations or disruptions in proteins that aid in the formation of a mature synapse have been implicated in developmental and neural disorders such as autism, mental retardation, and schizophrenia. The current body of knowledge on synapses lacks great detail about the processes that occur following the release of neurotransmitters into the synapse, including the events occurring in the synaptic cleft and postsynaptically. The general aim of this project was to generate antibodies that would specifically recognize synaptic targets in order to better characterize the various processes of synaptogenesis, including the establishment of the postsynaptic apparatus and the recruitment of assorted structural proteins and neurotransmitter receptors. Clonal hybridomas were generated by injecting mice with purified fractions of synaptoneurosomes and postsynaptic density. A total of 96 clonal hybridomas were screened for the desired characteristics by enzyme-linked immunosorbent assay, immunolabeling, Western blotting, and immunoprecipitation. A number of hybridomas recognized targets with high affinity and exhibited specificity for neuronal and postsynaptic targets. Notably, a number of subclonal hybridomas labeled neuronal targets strongly in whole-mount immunolabeling. Additionally, immunoprecipitation showed that selected subclonal hybridomas could precipitate antigens from solution, which will allow the isolated antigens to be identified.