Data from: Glutamate sensor and calcium signals in dopamine neurons and dopamine release
Data files
Dec 21, 2023 version files 3.34 GB
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Ap_DA_362_session1.mat
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Ap_DA_362_session2.mat
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Ap_DA_362_session3.mat
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Ap_DA_456_session1.mat
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Ap_DA_456_session2.mat
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Ap_DA_456_session3.mat
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Ap_GCaMP_582_session1.mat
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Ap_GCaMP_582_session2.mat
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Ap_GCaMP_582_session3.mat
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Ap_GCaMP_583_session1.mat
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Ap_GCaMP_583_session2.mat
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Ap_GCaMP_583_session3.mat
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Ap_GCaMP_585_session1.mat
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Ap_GCaMP_585_session2.mat
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Ap_GCaMP_585_session3.mat
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Ap_GCaMP_586_session1.mat
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Ap_GCaMP_586_session2.mat
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Ap_GCaMP_586_session3.mat
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Ap_GCaMP_588_session1.mat
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Ap_GCaMP_588_session2.mat
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Ap_GCaMP_588_session3.mat
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Ap_GCaMP_597_session1.mat
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Ap_GCaMP_597_session2.mat
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Ap_GCaMP_597_session3.mat
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Ap_GCaMP_599_session1.mat
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Ap_GCaMP_599_session2.mat
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Ap_GCaMP_599_session3.mat
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Ap_GCaMP_600_session1.mat
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Ap_GCaMP_600_session2.mat
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Ap_GCaMP_600_session3.mat
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Ap_GCaMP_602_session1.mat
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Ap_GCaMP_602_session2.mat
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Ap_GCaMP_602_session3.mat
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Ap_Glu_455_session1.mat
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Ap_Glu_455_session2.mat
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Ap_Glu_455_session3.mat
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Ap_Glu_578_session1.mat
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Ap_Glu_578_session2.mat
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Ap_Glu_587_session1.mat
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Ap_Glu_587_session2.mat
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Ap_Glu_587_session3.mat
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Ap_GluDA_361_session1.mat
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Ap_GluDA_361_session3.mat
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Ap_GluDA_363_session1.mat
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Ap_GluDA_363_session3.mat
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Ap_GluDA_457_session1.mat
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Ap_GluDA_457_session2.mat
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Ap_GluDA_457_session3.mat
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Ap_GluDA_524_session1.mat
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Ap_GluDA_524_session2.mat
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Ap_GluDA_524_session3.mat
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Ap_GluDA_537_session1.mat
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Ap_GluDA_537_session2.mat
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Ap_GluDA_537_session3.mat
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ApOpi_GluDA_638_session1_post.mat
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ApOpi_GluDA_638_session1_pre.mat
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ApOpi_GluDA_638_session2_post.mat
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ApOpi_GluDA_638_session2_pre.mat
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ApOpi_GluDA_639_session1_post.mat
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ApOpi_GluDA_640_session1_post.mat
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ApOpi_GluDA_640_session1_pre.mat
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ApOpi_GluDA_640_session2_post.mat
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ApOpi_GluDA_640_session2_pre.mat
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ApOpi_GluDA_640_session3_pre.mat
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ApOpi_GluDA_641_session1_post.mat
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ApOpi_GluDA_643_session1_post.mat
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ApOpi_GluDA_645_session1_post.mat
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ApOpi_GluDA_646_session3_pre.mat
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ApOpi_GluDA_646_session4_pre.mat
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ApSal_GluDA_638_session1_post.mat
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ApSal_GluDA_638_session1_pre.mat
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ApSal_GluDA_638_session2_post.mat
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ApSal_GluDA_638_session2_pre.mat
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ApSal_GluDA_639_session1_post.mat
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ApSal_GluDA_640_session1_post.mat
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ApSal_GluDA_640_session1_pre.mat
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ApSal_GluDA_640_session2_postmat.mat
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ApSal_GluDA_640_session2_pre.mat
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ApSal_GluDA_641_session1_post.mat
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ApSal_GluDA_641_session2_post.mat
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ApSal_GluDA_641_session3_pre.mat
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ApSal_GluDA_642_session1_post.mat
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ApSal_GluDA_642_session1_pre.mat
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ApSal_GluDA_642_session2_post.mat
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ApSal_GluDA_642_session2_pre.mat
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ApSal_GluDA_642_session3_post.mat
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ApSal_GluDA_642_session3_pre.mat
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ApSal_GluDA_642_session4_post.mat
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ApSal_GluDA_642_session4_pre.mat
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ApSal_GluDA_643_session1_post.mat
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ApSal_GluDA_643_session1_pre.mat
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ApSal_GluDA_643_session2_post.mat
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ApSal_GluDA_643_session2_pre.mat
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ApSal_GluDA_643_session3_post.mat
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ApSal_GluDA_643_session3_pre.mat
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ApSal_GluDA_643_session4_post.mat
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ApSal_GluDA_643_session4_pre.mat
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ApSal_GluDA_645_session1_post.mat
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ApSal_GluDA_645_session1_pre.mat
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ApSal_GluDA_645_session2_post.mat
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ApSal_GluDA_645_session3_pre.mat
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ApSal_GluDA_645_session4_post.mat
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ApSal_GluDA_646_session1_post.mat
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ApSal_GluDA_646_session1_pre.mat
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ApSal_GluDA_646_session2_post.mat
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ApSal_GluDA_646_session2_pre.mat
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ApSal_GluDA_646_session3_post.mat
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ApSal_GluDA_646_session3_pre.mat
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ApSal_GluDA_646_session4_post.mat
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ApSal_GluDA_646_session4_pre.mat
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CC_DA_437_session1.mat
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CC_DA_437_session2.mat
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CC_DA_437_session3.mat
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CC_DA_438_session1.mat
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CC_DA_438_session2.mat
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CC_DA_438_session3.mat
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CC_DA_439_session1.mat
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CC_DA_440_session1.mat
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CC_DA_444_session1.mat
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CC_DA_445_session1.mat
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CC_DA_446_session1.mat
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CC_DA_516_session1.mat
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CC_DA_516_session3.mat
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CC_DA_524_session1.mat
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CC_DA_524_session2.mat
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CC_DA_524_session3.mat
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CC_DA_537_session1.mat
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CC_DA_537_session2.mat
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CC_DA_537_session3.mat
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CC_DA_541_session1.mat
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CC_DA_541_session2.mat
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CC_DA_541_session3.mat
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CC_GCaMP_582_session1.mat
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CC_GCaMP_582_session2.mat
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CC_GCaMP_582_session3.mat
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CC_GCaMP_583_session1.mat
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CC_GCaMP_583_session2.mat
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CC_GCaMP_583_session3.mat
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CC_GCaMP_585_session1.mat
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CC_GCaMP_585_session2.mat
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CC_GCaMP_585_session3.mat
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CC_GCaMP_586_session1.mat
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CC_GCaMP_586_session2.mat
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CC_GCaMP_586_session3.mat
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CC_GCaMP_588_session1.mat
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CC_GCaMP_588_session2.mat
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CC_GCaMP_588_session3.mat
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CC_GCaMP_597_session1.mat
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CC_GCaMP_597_session2.mat
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CC_GCaMP_597_session3.mat
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CC_GCaMP_599_session1.mat
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CC_GCaMP_599_session2.mat
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CC_GCaMP_599_session3.mat
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CC_GCaMP_600_session1.mat
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CC_GCaMP_600_session2.mat
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CC_GCaMP_600_session3.mat
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CC_GCaMP_601_session1.mat
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CC_GCaMP_601_session2.mat
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CC_GCaMP_601_session3.mat
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CC_GCaMP_602_session1.mat
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CC_GCaMP_602_session2.mat
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CC_GCaMP_602_session3.mat
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CC_GCaMP_612_session1.mat
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CC_GCaMP_612_session2.mat
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CC_GCaMP_612_session3.mat
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CC_Glu_18_session1.mat
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CC_Glu_18_session2.mat
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CC_Glu_18_session3.mat
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CC_Glu_346_session1.mat
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CC_Glu_346_session2.mat
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CC_Glu_346_session3.mat
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CC_Glu_362_session1.mat
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CC_Glu_362_session2.mat
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CC_Glu_362_session3.mat
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CC_Glu_364_session1.mat
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CC_Glu_364_session2.mat
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CC_Glu_364_session3.mat
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CC_Glu_436_session1.mat
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CC_Glu_436_session2.mat
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CC_Glu_436_session3.mat
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CC_Glu_455_session1.mat
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CC_Glu_455_session2.mat
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CC_Glu_455_session3.mat
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CC_Glu_516_session1.mat
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CC_Glu_516_session2.mat
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CC_Glu_516_session3.mat
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CC_Glu_524_session1.mat
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CC_Glu_524_session2.mat
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CC_Glu_524_session3.mat
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CC_Glu_537_session1.mat
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CC_Glu_537_session2.mat
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CC_Glu_537_session3.mat
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CC_Glu_541_session1.mat
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CC_Glu_541_session2.mat
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CC_Glu_541_session3.mat
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CC_Glu_577_session1.mat
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CC_Glu_577_session2.mat
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CC_Glu_577_session3.mat
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CC_Glu_578_session1.mat
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CC_Glu_578_session2.mat
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CC_Glu_578_session3.mat
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CC_Glu_580_session1.mat
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CC_Glu_580_session2.mat
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CC_Glu_580_session3.mat
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CC_Glu_581_session1.mat
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CC_Glu_581_session2.mat
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CC_Glu_581_session3.mat
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CC_Glu_587_session1.mat
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CC_Glu_587_session2.mat
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CC_Glu_587_session3.mat
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OptoChr_DA_589_session1.mat
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OptoChr_DA_589_session2.mat
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OptoChr_DA_589_session3.mat
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OptoChr_DA_592_session1.mat
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OptoChr_DA_592_session2.mat
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OptoChr_DA_592_session3.mat
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OptoChr_DA_593_session1.mat
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OptoChr_DA_593_session2.mat
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OptoChr_DA_608_session1.mat
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OptoChr_DA_608_session2.mat
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OptoChr_Glu_590_session1.mat
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OptoChr_Glu_606_session1.mat
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OptoChr_Glu_606_session2.mat
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OptoChr_Glu_606_session3.mat
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OptoChr_Glu_613_session1.mat
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OptoChr_Glu_614_session1.mat
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OptoCtrl_DA_594_session1.mat
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OptoCtrl_DA_609_session1.mat
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OptoCtrl_Glu_591_session1.mat
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OptoCtrl_Glu_607_session1.mat
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OptoCtrl_Glu_611_session1.mat
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README.md
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SeqC_GluDA_431_session1.mat
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SeqC_GluDA_431_session2.mat
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SeqC_GluDA_432_session1.mat
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SeqC_GluDA_432_session2.mat
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SeqC_GluDA_433_session1.mat
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SeqC_GluDA_456_session1.mat
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Abstract
We recorded glutamate input to dopamine neurons in the ventral tegmental area (VTA) with iGluSnFR(A184S), a glutamate sensor, somatodendritic dopamine neuron activity in the VTA with jGCaMP7f, a calcium indicator, and dopamine release at the ventral striatum (VS) with GrabDA2m, a dopamine sensor, using fiber fluorometry. Recordings were performed in two types of reward association tasks (classical conditioning and sequential conditioning), with presentation of aversive airpuff to the eye with or without buprenorphine treatment, and with optogenetic activation of glutamate input axons at the VTA.
README: Glutamate sensor and calcium signals in dopamine neurons and dopamine release
The recording data for glutamate input to dopamine, somatodendritic dopamine activity, and dopamine release at the ventral striatum. The datasets are divided into 5 different task conditions: ‘Classical_conditioning’, ‘Sequential conditioning’, ‘Airpuff_response’, ‘Airpuff_with_Buprenorphine’, and ‘Optogenetic_stimulation’.
Description of the data and file structure
File names
Data of each recording session is stored in each “.mat” file (_******.mat).
Task type is indicated at the beginning of the file name (‘task type’******.mat).
The mouse ID number is indicated in the middle of the file name (_’mouse ID’**.mat).
The session number is indicated at the end of the file name (__**_session’session number’.mat). Task structures were exactly the same in each session, and each session was performed on different days.
The block name is indicated at the end of the file name after the session number in some files (see below).
* *
Classical_conditioning
Files start from “CC_Glu_”
Activity of glutamate inputs to dopamine neurons in VTA was recorded using a glutamate sensor, SF-iGluSnFR(A184S), during classical conditioning tasks. Animals were trained for more than nine days before neural recording.
“dFoverFGCaMP2” is the motion-corrected and z-scored glutamate signal (see Methods for the details). Each row corresponds to each trial (-1 sec to 8 sec from water onset for free reward trial, and -1 sec to 8 sec from odor onset for other trial types) in the order in the session, and each column corresponds to signals in 1 ms.
“lickmatrix” is the lick event stamp (1: lick, 0: no lick) for the same time window as photometry data.
“TrialType” is the list of trial types in the session.
Trial types for the task are as follows.
Mouse ID: 18, 436, 455, 516, 524, 537, 541, 577, 578, 580, 581, 587
1: free reward trial (reward at 1001)
9: 80% reward-predicting cue followed by reward (cue at 1001 and reward at 4001)
10: 80% reward-predicting cue followed by reward omission (cue at 1001)
11: 40% reward-predicting cue followed by reward (cue at 1001 and reward at 4001)
12: 40% reward-predicting cue followed by reward omission (cue at 1001)
14: 0% reward (nothing)-predicting cue (cue at 1001)
Mouse ID: 346, 362, 364
1: free reward trial (reward at 1001)
4: free airpuff trial (airpuff at 1001)
5: 80% airpuff predicting cue followed by airpuff (cue at 1001 and airpuff at 4001)
9: 80% reward predicting cue followed by reward (cue at 1001 and reward at 4001)
10: 80% reward predicting cue followed by reward omission (cue at 1001)
11: 40% reward predicting cue followed by reward (cue at 1001 and reward at 4001)
12: 40% reward predicting cue followed by reward omission (cue at 1001)
14: 0% reward (nothing) predicting cue (cue at 1001)
16: 80% airpuff predicting cue followed by airpuff omission (cue at 1001)
Files start from “CC_DA_”
Dopamine sensor signal data recorded from VS neurons during classical conditioning are stored in these files.
“dFoverFGCaMP2” is the z-scored dopamine sensor signal for each trial (-1 sec to 8 sec from water onset for free reward trial, and -1 sec to 8 sec from odor onset for other trial types) in a single session.
“lickmatrix” is the lick event stamp (1: lick, 0: no lick) for the same time window as photometry data.
“TrialType” is the list of trial type order in the session.
Trial types for the task are as follows.
All mice
1: free reward trial (reward at 1001)
9: 80% reward predicting cue reward (cue at 1001 and reward at 4001)
10: 80% reward predicting cue followed by reward omission (cue at 1001)
11: 40% reward predicting cue followed by reward (cue at 1001 and reward at 4001)
12: 40% reward predicting cue followed by reward omission (cue at 1001)
14: 0% reward (nothing) predicting cue (cue at 1001)
Files start from “CC_GCaMP_”
Calcium sensor signal data recorded from VTA dopamine neurons during classical conditioning are stored in these files.
“dFoverFGCaMP2” is the z-scored dopamine sensor signal for each trial (-1 sec to 8 sec from water onset for free reward trial, and -1 sec to 8 sec from odor onset for other trial types) in a single session.
“lickmatrix” is the lick event stamp (1: lick, 0: no lick) for the same time window as photometry data.
“TrialType” is the list of trial type order in the session.
Trial types for the task are as follows.
All mice
1: Free reward trial (reward at 1001)
9: 80% reward predicting cue followed by reward (cue at 1001 and reward at 4001)
10: 80% reward predicting cue followed by reward omission (cue at 1001)
11: 40% reward predicting cue followed by reward (cue at 1001 and reward at 4001)
12: 40% reward predicting cue followed by reward omission (cue at 1001)
14: 0% reward (nothing) predicting cue (cue at 1001)
Sequential_conditioning
Files start from “SeqC_GluDA_”
Simultaneously recorded glutamate sensor signals from VS neurons and dopamine sensor signals from VTA dopamine neurons during sequential conditioning are stored in these files.
“dFoverFGCaMP2” is the motion-corrected and z-scored glutamate sensor signal, and “dFoverFGCaMP” is the z-scored dopamine sensor signal for each trial in a single session. -6 sec to 8 sec from water onset for free reward trial (trial type 1), -4sec to 10 sec from proximal cue onset for unexpected proximal cue trial (trial types 9 and 14), and -1 sec to 13 sec from distal odor onset for other trial types (trial types 18, 19, 20, and 21).
“lickmatrix” is the lick event stamp (1: lick, 0: no lick) for the same time window as photometry data.
“TrialType” is the list of trial type order in the session.
Trial types for the task are as follows.
All mice
1: Free reward trial (reward at 6001)
9: Unexpected 100% reward predicting proximal cue at 4001 and reward at 6001
14: Unexpected 0% reward (nothing) predicting proximal cue at 4001
18: Distal cue 100% predicting 100% reward predicting proximal cue at 1001/ 100% reward predicting proximal cue at 4001/ reward at 6001
19: Distal cue 50% predicting 100% reward or 0% reward predicting proximal cue at 1001/ 100% reward predicting proximal cue at 4001/ reward at 6001
20: Distal cue 50% predicting 100% reward or 0% reward predicting proximal cue at 1001/ 0% reward predicting proximal cue at 4001
21: Distal cue 100% predicting 0% reward (nothing) predicting proximal cue at 1001/ 0% reward predicting proximal cue at 4001
Airpuff_response
Files start from “Ap_GluDA_”
Mouse ID: 361, 363, 457, 524, and 537
Simultaneously recorded glutamate sensor signals from VTA dopamine neurons and dopamine sensor signals from VS neurons during free airpuff and free reward trials are stored in these files.
“dFoverFGCaMP2” is the z-scored and motion-corrected glutamate signal, and “dFoverFGCaMP” is the z-scored dopamine sensor signal for each trial in a single session. -1 sec to 8 sec from water or airpuff onset.
“lickmatrix” is the lick event stamp (1: lick, 0: no lick) for the same time window as photometry data.
“TrialType” is the list of trial type order in the session.
Trial types for the task are as follows.
Mouse ID: 457 and 537
1: Free reward trial (reward at 1001)
4: Free airpuff trial (airpuff at 1001)
Mouse ID: 361, 363, and 524
1: Free reward trial (reward at 1001)
4: Free airpuff trial (airpuff at 1001)
5: 80% airpuff predicting cue followed by airpuff (cue at 1001 and airpuff at 4001)
9: 80% reward predicting cue followed by reward (cue at 1001 and reward at 4001)
10: 80% reward predicting cue followed by reward omission (cue at 1001)
11: 40% reward predicting cue followed by reward (cue at 1001 and reward at 4001)
12: 40% reward predicting cue followed by reward omission (cue at 1001)
14: 0% reward (nothing) predicting cue (cue at 1001)
16: 80% airpuff predicting cue followed by airpuff omission (cue at 1001)
Files start from “Ap_Glu_”
Mouse ID: 455, 578, and 587
Glutamate sensor signals from VTA dopamine neurons at free airpuff and free reward trials are stored in these files.
“dFoverFGCaMP2” is the z-scored and motion-corrected glutamate signal for each trial in a single session. -1 sec to 8 sec from water or airpuff onset.
“lickmatrix” is the lick event stamp (1: lick, 0: no lick) for the same time window as photometry data.
“TrialType” is the list of trial type order in the session.
Trial types for the task are as follows.
1: Free reward trial (reward at 1001)
4: Free airpuff trial (airpuff at 1001)
Files start from “Ap_DA_”
Mouse ID: 362 and 456
Dopamine sensor signals from VS neurons during free airpuff and free reward trials are stored in these files.
“dFoverFGCaMP2” is the z-scored dopamine signal for each trial in a single session. -1 sec to 8 sec from water or airpuff onset.
“lickmatrix” is the lick event stamp (1: lick, 0: no lick) for the same time window as photometry data.
“TrialType” is the list of trial type order in the session.
Trial types for the task are as follows.
1: Free reward trial (reward at 1001)
4: Free airpuff trial (airpuff at 1001)
Files start from “Ap_GCaMP_”
Mouse ID: 582, 583, 585, 586, 588, 597, 599, 600, and 602
Calcium sensor signal data during free airpuff and free reward trials are stored in these files.
“dFoverFGCaMP2” is the z-scored dopamine signal for each trial in a single session. -1 sec to 8 sec from water or airpuff onset.
“lickmatrix” is the lick event stamp (1: lick, 0: no lick) for the same time window as photometry data.
“TrialType” is the list of trial type order in the session.
Trial types for the task are as follows.
1: Free reward trial (reward at 1001)
4: Free airpuff trial (airpuff at 1001)
Airpuff_with_Buprenorphine
(Correspond to Figure 8)
Files start from “ApOpi_GluDA_”
Simultaneously recorded glutamate sensor signals from VTA dopamine neurons and dopamine sensor signals from VS neurons for free airpuff and free reward responses before and after buprenorphine treatment are stored in these files.
The treatment (“_pre”: before buprenorphine injection, and “_post”: after buprenorphine treatment) is indicated at the end of the file name.
“dFoverFGCaMP2” is the z-scored and motion-corrected glutamate signal, and “dFoverFGCaMP” is the z-scored dopamine sensor signal for each trial in a single session. -1 sec to 8 sec from water or airpuff onset.
“lickmatrix” is the lick event stamp (1: lick, 0: no lick) for the same time window as photometry data.
“TrialType” is the list of trial type order in the session.
Trial types for the task are as follows.
All mice
1: Free reward trial (reward at 1001)
4: Free airpuff trial (airpuff at 1001)
Files start from “ApSal_GluDA_”
Simultaneously recorded glutamate sensor signal and dopamine sensor signal data for free airpuff and free reward responses before and after control saline treatment are stored in these files.
The treatment (“_pre”: before saline injection, and “_post”: after saline injection) is indicated at the end of the file name.
“dFoverFGCaMP2” is the z-scored and motion-corrected glutamate signal, and “dFoverFGCaMP” is the z-scored dopamine sensor signal for each trial in a single session. -1 sec to 8 sec from water or airpuff onset.
“lickmatrix” is the lick event stamp (1: lick, 0: no lick) for the same time window as photometry data.
“TrialType” is the list of trial type order in the session.
Trial types for the task are as follows.
All mice
1: Free reward trial (reward at 1001)
4: Free airpuff trial (airpuff at 1001)
Optogenetic_stimulation
(Correspond to Figure 1)
Files start from “OptoChr_Glu_”
Mouse ID: 590, 606, 613, and 614
Glutamate sensor signal data during free light stimulation (single 20, 50, or 250 ms pulse) and free reward trials in the mice expressing ChrimsonR in glutamate input to the VTA are stored in these files.
“dFoverFGCaMP2” is the z-scored glutamate signal for each trial in a single session. -1 sec to 8 sec from water or airpuff onset.
“lickmatrix” is the lick event stamp (1: lick, 0: no lick) for the same time window as photometry data.
“TrialType” is the list of trial type order in the session.
Trial types for the task are as follows.
All mice
1: Free reward trial (reward at 1001)
23: Light stimulation (20 ms pulse stimulation at 1001)
25: Light stimulation (50 ms pulse stimulation at 1001)
26: Light stimulation (250 ms pulse stimulation at 1001)
Files start from “OptoCtrl_Glu_”
Mouse ID: 591, 607, and 611
Glutamate sensor signal data during free light stimulation (single 20, 50, or 250 ms pulse) and free reward trials in the mice expressing tdTomato in glutamate input to the VTA are stored in these files.
“dFoverFGCaMP2” is the z-scored glutamate signal for each trial in a single session. -1 sec to 8 sec from water or airpuff onset.
“lickmatrix” is the lick event stamp (1: lick, 0: no lick) for the same time window as photometry data.
“TrialType” is the list of trial type order in the session.
Trial types for the task are as follows.
All mice
1: Free reward trial (reward at 1001)
23: Light stimulation (20 ms pulse stimulation at 1001)
25: Light stimulation (50 ms pulse stimulation at 1001)
26: Light stimulation (250 ms pulse stimulation at 1001)
Files start from “OptoChr_DA_”
Mouse ID: 589, 692, 693, and 608
Dopamine sensor signal data during free light stimulation (single 20, 50, or 250 ms pulse) and free reward trials in the mice expressing ChrimsonR in glutamate input to the VTA are stored in these files
“dFoverFGCaMP2” is the z-scored dopamine sensor signal for each trial in a single session. -1 sec to 8 sec from water or airpuff onset.
“lickmatrix” is the lick event stamp (1: lick, 0: no lick) for the same time window as photometry data.
“TrialType” is the list of trial type order in the session.
Trial types for the task are as follows.
All mice
1: Free reward trial (reward at 1001)
23: Light stimulation (20 ms pulse stimulation at 1001)
25: Light stimulation (50 ms pulse stimulation at 1001)
26: Light stimulation (250 ms pulse stimulation at 1001)
Files start from “OptoCtrl_DA_”
Mouse ID: 591, 607, and 611
Dopamine sensor signals from VTA neurons during free light stimulation (single 20, 50, or 250 ms pulse) and free reward trials in the mice expressing tdTomato in glutamate input to the VTA are stored in these files.
“dFoverFGCaMP2” is the z-scored dopamine sensor signal for each trial in a single session. -1 sec to 8 sec from water or airpuff onset.
“lickmatrix” is the lick event stamp (1: lick, 0: no lick) for the same time window as photometry data.
“TrialType” is the list of trial type order in the session.
Trial types for the task are as follows.
All mice
1: Free reward trial (reward at 1001)
23: Light stimulation (20 ms pulse stimulation at 1001)
25: Light stimulation (50 ms pulse stimulation at 1001)
26: Light stimulation (250 ms pulse stimulation at 1001)
Methods
Behavior
After 1 week of recovery from surgery, mice were water-restricted in their cages. All conditioning tasks were controlled by a NIDAQ board and LabVIEW. Mice were handled for 2 days, acclimated to the experimental setup for 1-2 days including consumption of water from the tube, and head-fixed with random interval water for 1-3 days until mice showed reliable water consumption. For odor-based classical conditioning, all mice were head-fixed, and the volume of water reward was constant for all reward trials (predicted or unpredicted) in all conditions (6 ml). Some sessions included mild air puff trials, directed at one of the eyes and the intensity of air puff was constant for all air puff trials (predicted or unpredicted; 2.5 psi). In classical conditioning, each association trial began with an odor cue (lasting 1 s) followed by a 2 s delay, and then an outcome (either water, nothing, or air puff) was delivered. In sequential conditioning, each association trial began with a distal odor cue (lasting 1 s) followed by a 2 s delay, and then a distal odor cue (lasting 1 s) followed by 1 s delay, and then an outcome (either water, or nothing) was delivered. Some trial types began with a proximal odor cue (lasting 1 s) followed by 1 s delay, and then an outcome (either water, or nothing) was delivered. Odors were delivered using a custom olfactometer. Each odor was dissolved in mineral oil at 1:10 dilution and 30 ml of diluted odor solution was applied to the syringe filter (2.7mm pore, 13mm; Whatman, 6823-1327). Odorized air was further diluted with filtered air by 1:8 to produce a 900 ml/min total flow rate. Different sets of odors (Ethyl butyrate, p-Cymene, Isoamyl acetate, Isobutyl propionate, 1-Butanol, 4-Methylanisole, Caproic acid, Eugenol, and 1-Hexanol) were selected for each animal. Some of the animals shared the same odor set (4 animals and 3 animals for DA sensor classical conditioning; 3 animals and 2 animals for sequential conditioning)
A variable inter-trial interval (ITI) of flat hazard function (minimum 10s, mean 13s, truncated at 20s) was placed between trials. Each session was composed of multiple blocks (12-24 trial/block) and all trial types were pseudorandomized in each block. Each day, the mice did about 70-350 trials over the course of 20-75 min, and with constant excitation from the laser and continuous recording in recording sessions.
Training for classical conditioning used 4 types of trials; odor cue predicting 100% water, odor cue predicting 40% water/60% no outcome (nothing), odor cue predicting nothing (29.4% of all trials for each odor), and water without cue (free water) (11.8%) for 7-10 days, and then odor cue predicting 80% water/20% nothing, odor cue predicting 40% water/60% nothing, odor cue predicting nothing (29.4% each), and free water (11.8%) for more than 2 days of training followed by recording sessions. The first 170 trials or trials before the animal stops licking for each session are used for analysis. 3 animals used for glutamate sensor recording (Figure 2) were trained with classical conditioning with air puff trial; odor cue predicting 100% water, odor cue predicting 100% air puff, odor cue predicting 40% water/60% no outcome (nothing), odor cue predicting nothing (20.8 % of all trials for each odor), and water without cue (free water), air puff without cue (free air puff) (8.3 % of all trials for each stimulus) for 8-9 days, and then odor cue predicting 80% water/20% nothing, odor cue predicting 80% air puff/20% nothing, odor cue predicting 40% water/60% nothing, odor cue predicting nothing (20.8 % each), and water without cue (free water), air puff without cue (free air puff) (8.3% each) for more than 7 days of training followed by recording sessions. The first 192 trials or trials before the animal stops licking for each session are used for analysis. Of note these 3 animals are used only in the analysis for glutamate sensor activity pattern, and not included for comparison between different sensor recordings.
For sequential conditioning step1, mice were trained with proximal odor for 3-7 days, using 3 types of trials; proximal odor cue predicting 100% water, proximal odor cue predicting nothing (45.8 % of all trials for each odor), and water without cue (free water) (8.3 %). Then, for sequential conditioning step2, mice were further trained with distal odor and proximal odor using 6 types of trials; distal odor cue predicting proximal odor cue predicting 100% water (25 %), distal odor cue predicting 50% proximal odor cue predicting water/50% proximal odor cue predicting nothing (25 %), distal odor cue predicting proximal odor cue predicting nothing (25 %), proximal odor cue predicting 100% water (8.3 %), proximal odor cue predicting nothing (8.3 %), and water without cue (free water) (8.3 %). Sensor signals were recorded after 8-12 days of step2 training. The first 192 trials of each session are used for analysis.
For buprenorphine treatment, the mouse received unexpected air puff and water with a variable ITI of the flat hazard function (minimum 10s, mean 13s, truncated at 20s). Animals were first acclimated to head-fixation with water reward for 2 days. Then the animals were habituated to a test procedure for two days, which was composed of 40-70 trials of air puff and water presented in pseudorandomized order (the order of trials is pseudorandomized within each block of 10 trials composed of the same number of air puff trials and reward trials) with subcutaneous injection of saline (10 ml/g in body weights) at the end of sessions. After habituation sessions, buprenorphine sessions and control saline sessions were performed in pseudorandom order. Each session was separated into pre-injection sub-session and post-injection sub-session. The sub-session was composed of 48-75 trials with the air puff and water trials in the pseudorandom order. After pre-injection sub-sessions, either buprenorphine (Buprenorphine hydrochloride, Par Pharmaceutical: diluted in saline 0.03 mg/ml) or control saline was subcutaneously injected (10 ml/g; 0.3 mg/g buprenorphine or corresponding volume of saline). Post-injection sub-session started 5 min after injection. To prevent acute adverse effects, we used a smaller dose of buprenorphine (0.15 mg/g) in the first session and this session is not included in the analysis.
Optogenetic stimulation
Red light from 625 nm LED light (M625F2, Thorlab) was applied through an optical patch cord (400 mm, 0.39 NA, Thorlab). 12 mW single block pulse light of 20, 50, and 250ms duration was triggered pseudorandom order through custom software written in LabVIEW (National Instruments) via a NIDAQ board (PCI-e6321, National Instruments). A variable inter-trial interval (ITI) of flat hazard function (minimum 10s, mean 13s, truncated at 20s) was placed between trials.
Data analysis
Fiber-fluorometry
The noise from the power line in the voltage signal was cleaned by removing 58-62Hz signals through a band stop filter. The global change within a session was normalized using a moving median of 100 s. Then, the correlation between green and red signals during ITI was examined by linear regression when the red signal data was available. If the correlation is significant (p ≤ 0.05), fitted tdTomato signals were subtracted from green signals. Responses were calculated by subtracting the average baseline activity from the average activity of the target window. Z-scores of the signals were obtained using mean and standard deviation of signals in all ITI (from 5 s before odor onset to odor onset for classical conditioning, from 5 s before distal odor onset to odor onset for sequential conditioning) in each animal.
Licking
Licking from a water spout was detected by a photoelectric sensor that produces a change in voltage when the light path is broken. The timing of each lick was detected at the peak of the voltage signal above a threshold.