Hippocampal levels and activity of the sodium/potassium transporting ATPase subunit alpha-3 (AT1A3) are paralleling memory training in the multiple T-Maze in the C57BL/6J mouse

摘要

Although the sodium/potassium transporting ATPase subunit alpha-3 (AT1A3) has been linked to memory mechanisms in rodents, regulation of this ATPase in terms of activity and complex levels by memory performance in a land maze has not been shown so far. It was therefore the aim of the study to link memory retrieval in the multiple T-Maze (MTM) to AT1A3 protein levels and activity. C57BL/6J mice were trained in the MTM and euthanized 6 h following memory retrieval. Hippocampal membrane proteins were prepared by ultracentrifugation and run on blue native gel electrophoresis (BN-PAGE). Enzyme activity was evaluated using an in-gel method. AT1A3 protein was characterized using mass spectrometry (nano-LC-ESI-MS/MS). On BN-PAGE a single band was observed at 240 kDa, which corresponds to the dimeric form of the enzyme. Higher levels of AT1A3 complex were seen in trained mice. Also ATPase activity was higher in trained mice, and was observed both at 110 and at 240 kDa. Mass spectrometry unambiguously identified AT1A3 with 98.91% sequence coverage. A series of novel AT1A3 phosphorylation sites were detected. Taken together, it was shown that increased AT1A3 protein levels for the dimer as well as AT1A3 activity represented by the monomer and the dimer were paralleling memory training in the MTM. This may be relevant for understanding the role of the catalytic hydrolysis of ATP coupled with the exchange of sodium and potassium ions across the plasma membrane that generates the electrochemical gradient of sodium and potassium ions. Herein, we provide evidence for a possible role of AT1A3 in memory mechanisms and support previous findings using different animal models for memory formation.