Nature research paper: Entorhinal cortex directs learning-related changes in CA1 representations
= 44 mice, Jackson Laboratories) mice of either sex by an experimenter who was not blind to the experimental conditions. Animals were housed under an inverse 12-h dark/12-h light cycle in the Magee laboratory satellite facility with the temperature and humidity controlled. All surgical procedures were performed under deep isoflurane anaesthesia. After locally applying topical anaesthetics, the scalp was removed, and the skull was cleaned.
For the experiments with GCaMP6f and tdTomato expression in EC3 or ArchT or tdTomato expression in EC3 and GCaMP6f expression in CA1, the hippocampal window surgery was preceded in the pOxr1–Cre mice by ipsilateral virus injections using the coordinates stated above. Notably, the pOxr1–Cre mouse line expresses Cre recombinase predominantly in the medial entorhinal cortex. For the virus injections, we first made a small craniotomy. This was followed by injecting a small volume of one of the following mixtures : AAV1.Syn.GCaMP6f.WPRE.SV40 and AAVrg.Syn.Flex.ArchT–tdTomato.WPRE.SV40 into area CA1 ; AAV1.Syn.GCaMP6f.WPRE.SV40 and AAVrg.Syn.Flex.tdTomato.WPRE.SV40 into area CA1 ; AAV1.Syn.Flex.
At 5–7 days after the optical window implantation, running wheels were added to the home cages, and mice were placed on water restriction . After both training and recording sessions, mice were supplemented with additional water to guarantee a 1.5 ml per day water intake. After 5–6 days of familiarizing the animals with the experimenter, mice were trained to run head-fixed on the linear treadmill for 3–5 days.
To record neuronal activity and study the development of CA1 representations as mice learned to navigate in a new environment, we exposed the animals to two different environments . Environment A consisted of a belt enriched with three different visual and tactile cues , which covered the entire length of the belt
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