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Because of their ability to maintain and retrieve memories encoding spatial information, hippocampal place cells have been widely studied in experimental models of memory and have emerged as a model system for understanding normal memory function in healthy animals and the processes that fail during memory impairment. Life-long spatial learning in mice requires repeated (10-20 day) exposures to a novel environment to form a stable hippocampal map, from which memory can then be retrieved upon re-exposure25,26,27,28,29,30,31,32,33. This protocol, which we use in the current study, uses the virtual reality system OctaStage (Richmond, UK) in which mice are either removed from their home cage and placed in a novel arena or stay in their home cage while being exposed to the virtual environment. Virtual reality tasks can be completed in as little as 40 minutes and are therefore experimentally convenient for creating large numbers of data points. To validate the current experimental paradigm, we first studied the same mice using a parallel approach in which they were placed in a familiar cage for 20 days while being exposed to the same virtual environment. For these familiar animals, we recorded 425 place cells that were identified to have the same properties as previously reported34. Each neuron tracked the mouse’s progress through the maze via the system’s position sensor. We found that similar numbers of place cells were identified in both groups of mice (371 cells vs 430 cells; group difference, p=0.14). Moreover, 82.6% of cells encoded spatial position and the remaining 17.4% encoded cell-specific altered place fields. Finally, some cells encoded these position- and cell-specific altered places fields across days in the familiar environment (58.9% of cells vs 54.6% of cells; p=0.16). This validation suggests that the current experimental paradigm yields place cells with the same types of responses seen in previous work, but under a lower level of experimental demand. The ability to record spatial information under strong behavioural demand will be important for future studies in which the molecular basis of memory consolidation and retrieval processes can be studied under the same conditions and analyzed using the same statistics as place cells. a3f8a02ae1