Establecimiento de Protocolos de Inmunotinción para los Antígenos GFAP, NeuN Y BrdU en Cerebro de Rata
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In the Neuroscience Research Center (CIN) at the University of Costa Rica, research projects are performed in which behavioral, neurochemical and genetic variables involving albino rats as experimental subjects are analyzed. The development of techniques and immunohistochemical protocols is essential for the study, research and careful analysis of the cellular and molecular components of rodents’ brain, so that they complement other techniques existing and successful. This allows us to understand the functional and structural changes in them under certain conditions and enhances the comparison between different experimental groups under various conditions. The objective of this research was to establish protocols for immunostaining with fluorochromes to study GFAP, NeuN and BrdU antigens in the hippocampus of rat brains. A protocol for the identification and quantification of glial cells was developed and also a protocol for the identification and quantification of neuronal cells was implemented. It was evident that there is no ideal protocol for immunofluorescent detection of BrdU, and rather, there are multiple tactics used by scientists to detect this marker; so, it is recommended to change the synthetic nucleoside BrdU by the nucleoside EdU with Alexa Fluor 555 to achieve the implementation of the protocol to determine cell proliferation in tissue sections of rat brain and to study neuronal plasticity indirectly.
Proyecto de Graduación (Licenciatura en Ingeniería en Biotecnología) Instituto Tecnológico de Costa Rica, Escuela de Biología, 2016.