DOI: 10.5061/dryad.18931zd7t
title: Data from: Norepinephrine signals through astrocytes to modulate synapses

description: Locus coeruleus (LC)-derived norepinephrine (NE) drives network and
 behavioral adaptations to environmental saliencies by reconfiguring
 circuit functional connectivity, but the underlying synapse-level
 mechanisms are elusive. Here, we show that NE remodeling of synaptic
 function is independent from its binding on neuronal receptors. Instead,
 astrocytic adrenergic receptors and Ca2+ dynamics fully gate the effect of
 NE on synapses as the astrocyte-specific deletion of adrenergic receptors
 and three independent astrocyte-silencing approaches all render synapses
 insensitive to NE. Additionally, we find that NE suppression of synaptic
 strength results from an ATP-derived and adenosine A1 receptor-mediated
 control of presynaptic efficacy. An accompanying study from Chen et al.
 reveals the existence of an analogous pathway in the larval zebrafish and
 highlights its importance to behavioral state transitions. Together, these
 findings support a new model wherein astrocytes are a core component of
 neuromodulatory systems and the circuit effector through which
 norepinephrine produces network and behavioral adaptations.
description: # Data from: Norepinephrine signals through astrocytes to modulate
 synapses Dataset DOI: [10.5061/dryad.18931zd7t](10.5061/dryad.18931zd7t)
 ## Description of the data and file structure #### File:
 Lefton_et_al_Data.xlsx **Description:** The data file contains the raw and
 analyzed data for each figure (and supplemental figures) of the
 manuscript. Each tab corresponds to a separate figure and in each tab the
 data are organized by figure panel. The main metric, variable, or value
 measured, and their unit, are clearly stated (in bold) and reflect the
 corresponding figure panel. The numerical values listed represent the raw
 data from which the figures were compiled. **Figure1**: **NE and LC-NE
 activity inhibit presynaptic efficacy:** (A) Schematic of the recording
 conditions. (B) Left: time course of the effect of 20 μM NE applied at
 time t = 0 (gray area) on fEPSP slope and PPF. Right: representative
 traces showing the effect of NE on the slope of the first fEPSP (top) and
 on the PPF of the second fEPSP (bottom) from the same recording.
 Stimulation artifacts were cropped for clarity. (C) Pairwise
 quantification of the effect of NE on fEPSP slope and PPF for the
 experiments shown in (B). (D) Correlation between NE-induced fEPSP
 decrease and PPF increase in experiments shown in (B) and (C). (E)
 Representative recording of EPSC amplitude in response to minimal
 stimulations, showing failures (gray) and successes (black). (F) Left:
 averaged time course (per minute) of minimal-stimulation experiments
 showing the effect of 20 μM NE on synaptic efficacy (gray) and potency
 (black). Right: representative traces illustrating the effect of NE on
 efficacy (gray traces, failures; black traces, successes) and potency
 (average of successes) over 1-min epochs. (G) Quantification and pairwise
 comparison of the effect of NE on synaptic efficacy, potency, and strength
 for individual experiments shown in (F). (H) Schematic illustration of the
 procedure for expressing ChR2 in LC-NE fibers. (I) Immunohistochemistry
 (IHC) images of ChR2-EYFP expression in the LC (left) and hippocampal CA1
 (right) at 12 weeks, and quantification of efficiency and specificity of
 ChR2-EYFP expression in the LC (center). (J) Schematic of the recording
 and optogenetic stimulation conditions. (K) Time course of the effect of
 the optical stimulation of LC-NE fibers on fEPSPs and PPFs and
 representative traces. (L) Pairwise quantification of the effect of light
 (1 Hz, 10 min) on fEPSP slope and PPF for the experiments shown in (K) and
 in EYFP-control slices. (M) Plot summarizing the effect of light in
 ChR2-positive slices, EYFP-control slices, and ChR2-positive slices in the
 presence of silodosin (silo.) (50 nM). (N) Plot summarizing the effect of
 NE on synaptic strength in the presence of blockers of α2-AR (yohimbine,
 500 nM), β-AR (propranolol, 1 μM), α1B-AR (LY746-314, 1 μM), α1-AR
 (prazosin, 1 μM), α1A-AR (silodosin, 50 nM), or the α1A-AR agonist A61603
 (70 nM). n = 5 to 10 mice. (O and P) Time course of the effect of 20 μM NE
 on fEPSP and PPF in the presence of silodosin (50 nM), representative
 traces, and pairwise quantification. **Figure 2: Astrocyte Ca2+ dynamics
 gate the effect of NE on synapses:** (A) Approach for astrocyte Ca2+
 silencing with iβARK. (B) Representative IHC images of iβARK-mCherry
 expression in the hippocampal CA1, along with quantification of efficiency
 and specificity (n = 5 sections). (C) Plot of the stimulation
 intensity/fEPSP slope relationship (left, unpaired Student’s t test on
 slope/stimulation ratio) and summary bar graphs of PPF values (right) in
 RFP-control and iβARK slices at baseline. (D) Kymograph, in which each row
 shows the average fluorescence across regions of activity (ROAs) of an
 individual astrocyte, and five representative ΔF/F0 traces (from
 individual ROAs) of spontaneous Ca2+ transients in RFP-control and iβARK
 slices. Horizontal time axis applies to the kymograph and representative
 traces. (E) Plots of the peak amplitude, frequency, and kinetics of
 spontaneous Ca2+ transients in RFP-control and iβARK slices. (F)
 Kymographs, in which each row shows the whole-cell fluorescence of an
 individual astrocyte, and average ΔF/F0 traces (±SEM) across all
 astrocytes in response to 20 μM NE application in RFP-control and iβARK
 slices. (G) Plot of the peak ΔF/F0 response in RFP-control and iβARK
 conditions for experiments shown in (F). (H and I) Time courses of the
 effect of 20 μM NE on fEPSP slope and PPF and representative traces in
 RFP-control and iβARK slices. (J) Pairwise quantifications of the effect
 of NE on fEPSP slope and PPF for the experiments shown in (H) and (I). (K)
 Plots summarizing the effect of NE on fEPSP slope in RFP-control and iβARK
 slices. (L) Correlation between the effect of 20 μM NE on astrocyte peak
 Ca2+ responses and fEPSP slope across three methods of astrocyte silencing
 and RFP controls (figs. S4 and S5 and table S1).  **Figure 3: Astrocytic,
 but not neuronal, α1A-ARs are required for NE to affect synapses:** (A)
 Approach for the neuronal deletion of Adra1a. (B) Representative IHC
 images of Cre-GFP expression in hippocampal CA1 and CA3 neurons, along
 with quantification of efficiency and specificity (n = 4 sections from 2
 mice). (C) Quantification of bulk Adra1a gDNA polymerase chain reaction
 (PCR) band intensity, normalized to Atcb (β-actin), in Cre-injected
 Adra1a+/+ and Adra1afl/fl mice. (D) Quantification of Adra1a gDNA,
 normalized to Atcb (β-actin), in GFP-positive and GFP-negative cells from
 Cre-injected Adra1afl/fl mice (n = 5) relative to Adra1a+/+ mice (n = 5)
 (left), and representative PCR gels (right). (E) Time course of the effect
 of 20 μM NE on fEPSPs and PPFs in N-Adra1aKO slices and representative
 traces. (F and G) Pairwise quantification of the effect of NE on fEPSP
 slope and PPF in N-Adra1aKO and N-Adra1aCre-GFP control slices and summary
 plot of the effect of NE on fEPSPs in both conditions. (H) Approach for
 the astrocytic deletion of Adra1a. (I) Representative IHC images of
 Cre-GFP expression in hippocampal CA1 astrocytes, along with
 quantification of efficiency and specificity (n = 6 sections from 2 mice).
 (J) Quantification of bulk Adra1a gDNA levels, normalized to Actb
 (β-actin), in Cre-injected Adra1afl/fl mice and Cre-injected Adra1a+/+
 controls (left) and quantification of Adra1a gDNA, normalized to Atcb
 (β-actin), in GFP-positive and GFP-negative cells from Cre-injected
 Adra1afl/fl mice (n = 4) relative to Adra1a+/+ mice (n = 3) (right).
 Representative PCR gels are shown. (K and L) Kymographs, in which each row
 shows the whole-cell fluorescence of an individual astrocyte; average
 ΔF/F0 traces (± SEM) across all astrocytes; and quantification of the peak
 Ca2+ signal in response to 20 μM NE application in A-Adra1aCre-GFP and
 A-Adra1aKO slices. (M to P) Time courses of the effect of NE on fEPSPs and
 PPFs and representative traces in A-Adra1acontrol (M) and A-Adra1aKO
 slices (N), pairwise quantification of the effect of NE on fEPSP slope and
 PPF (O), and summary plot of the effect of NE in both conditions (P). (Q)
 Summary plot of the inhibitory effect of NE on fEPSPs in A-Adra1aKO and
 N-Adra1aKO slices relative to their respective controls. **Figure 4: NE
 leverages ATP-adenosine-A1R signaling to modulate synaptic efficacy:** (A)
 Left: plot summarizing the effect of 20 μM NE in the presence of A1R
 antagonists (CPT, 200 nM, or DPCPX, 100 nM), an adenosine scavenger (ADA,
 1U/ml), a cocktail of P2X/P2Y (PPADS, 10 μM), A2A (ZM241385, 50 nM) and
 A2B (PSB603 50 nM) receptor antagonists, a cocktail of mGluR inhibitors
 (CPPG, 5 μM; MPEP, 3.6 μM; YM298198, 2 μM), or in slices from NT5eKO mice
 (n = 4 to 9 mice). Right: schematic of the ATP-adenosine-A1R pathways
 showing different points of genetic or pharmacological intervention. (B)
 Time course of the effect of NE on fEPSPs and PPFs in the presence of the
 A1R antagonist CPT and representative traces. (C) Pairwise quantification
 for the experiments shown in (B). (D) Approach for the deletion of Adora1
 in CA3 or CA1 neurons. (E) Representative IHC images of Cre-GFP expression
 in CA1- and CA3-injected animals, along with quantification of regional
 specificity (n = 4 sections from 2 mice). (F and G) Time course,
 representative traces, and quantification of the effect of 5 μM adenosine
 on fEPSPs in CA1-Ado1KD and CA3-Ado1KD slices. (H to J) Time course,
 representative traces, pairwise quantification, and summary plot of the
 effect of 20 μM NE on fEPSPs in CA1-Ado1KD and CA3-Ado1KD slices. (K and
 L) Time course, representative traces, and summary plot of the effect of
 20 μM NE on fEPSPs and PPFs in NT5eKO slices. (M and N) Time course,
 representative traces, and quantification of the effect of adenosine on
 fEPSPs in NT5eKO and control slices.  **Figure S1: Effect of NE on the
 AMPAR-mediated fEPSP:** (A) Pairwise quantification of the effect of the
 AMPAR antagonist NBQX (10μM) on fEPSPs and representative traces. n = 2
 mice. (B) Dose-response curve of the inhibitory effect of NE on fEPSP
 slope (n = 5 to 7 slices per dose). A 4-parameter logistic non-linear
 symmetrical sigmoid fit was used to determine the IC50. (C) Coefficient of
 variation analysis showing 1/CV2 of the slope of fEPSPs in the NE
 condition against the mean, normalized to baseline. All but one recording
 give rise to a data point below the diagonal line (slope = 1), and the
 slope of the line from the baseline coordinate (1,1) to the average
 coordinate in NE (0.73,0.44) is greater than 1, indicative of a
 presynaptic locus of action. (D) Time course and pairwise quantification
 of the effect of NE on fEPSP and PPF with stimulations of Schaffer
 collaterals paused for five minutes at the onset of NE application. (E)
 Pairwise quantification of the effect of 20μM NE on fEPSP and PPF in the
 presence of the NMDAR antagonist D-AP5 (50μM, n = 4 mice). (F) Plots of
 the inhibitory effect of NE as a function of the initial fEPSPs slope
 (left) or initial PPF (right) across control conditions. The naïve and
 vehicle condition (DMSO, 0.01%) are denoted separately, but the linear
 regression, correlation coefficient and p-values are shown for the control
 condition as a whole. (G) Quantification and representative images of
 ChR2-eYFP expression of LC-NE projections in the hippocampal CA1 at 7, 10,
 11, 12 and 13 weeks post-injection (n = 6-12 slices, from 1-2 animals).
 (H) Time course and representative traces showing the effect of the
 optical stimulation of LC-NE fibers on fEPSPs and PPF in EYFP-control
 slices. (I,J) Time course, representative traces and pairwise
 quantification showing the effect of the optical stimulation of LC-NE
 fibers on fEPSPs and PPF in the presence of silodosin (50nM). (K-Q)
 Pairwise quantification of the effect of 20 μM NE on fEPSP slope and PPF
 in prazosin (α1-AR antagonist, (K,L)), yohimbine (α2-AR antagonist,
 (M,N)), propranolol (β antagonist, (O)), yohimbine and propranolol (P),
 and LY746-314 (α1B-AR antagonist, (Q)) at indicated concentrations. n =
 5-10 mice. (R,S) Pairwise quantification showing the inhibitory effect of
 the α1A-AR agonist A61603 (70nM) on fEPSP and PPF (R) and its blockade by
 silodosin (S). n = 9-3 mice respectively. (T) Pairwise quantification
 showing the potentiating effect of silodosin perfusion (50nM) on fEPSP and
 inhibitory effect on PPF (n = 3 mice). **Figure S2: Presynaptic Gq
 signaling enhances presynaptic efficacy and synaptic strength:**  (A)
 Schematic of micro-injections to express the excitatory hM3D(Gq) DREADD
 actuator in CA3 (presynaptic) neurons. (B) IHC of DREADD expression (in
 green) in CA3 but not CA1 neurons (NeuN, in red) and quantification of
 regional specificity. (C) Time courses and representative traces of the
 effect of 10μM of the hM3D(Gq) agonist CNO, on fEPSP and PPF in control
 slices. (D) Time courses and representative traces of the effect of 10μM
 CNO, on fEPSP and PPF in slices expressing hM3D(Gq) in the CA3. Note the
 increase in synaptic strength (fEPSP) and release probability (decreased
 PPF). (E) Pairwise quantification of the effect of CNO on fEPSP and PPF in
 DREADD and control slices. (F) Summary plots comparing the effect of NE on
 fEPSP and PPF in DREADD and control slices. n=3 biological replicates
 each. **Figure S3: Imaging and analysis of astrocyte Ca2+ responses to NE
 and α1A-AR pharmacology:**  (A) Schematic of micro-injections for
 lck-GCaMP6f expression in astrocytes in the s. radiatum of the CA1. (B)
 IHC of lck-GCaMP6f expression in CA1 astrocytes and quantification of
 efficiency and specificity. (C) Schematic of the 2-PSLM conditions for
 astrocyte lck-GCaMP6f imaging in hippocampal slices. (D) Overview of the
 STARDUST analysis workflow for ROA and cell-based timeseries analysis. (E)
 Left, Kymographs of pseudo-colored astrocyte Ca2+ signals (each row
 represents an individual cell) and average ΔF/F0 traces (± s.e.m.) across
 all astrocytes, in responses to the application of NE at indicated
 concentrations. Right, Dose-response curve showing the peak amplitude of
 the astrocyte Ca2+ response as a function of NE concentration. (F) Left,
 Kymograph of pseudo-colored astrocyte Ca2+ signals (each row represents a
 single cell) and average ΔF/F0 traces (± s.e.m.) across all astrocytes, in
 response to the application of 20μM NE in control conditions and in the
 presence of silodosin (50nM, α1A-AR antagonist). Right, Quantification of
 the peak astrocyte Ca2+ response to 20μM NE in control and silodosin. (G)
 Kymographs (each row shows the average fluorescence across ROAs of a
 single astrocyte) and 5 representative ΔF/F0 traces (from individual ROAs)
 showing spontaneous astrocyte Ca2+ transients in control and silodosin
 conditions. (H) Quantification of the peak amplitude, frequency, and
 kinetics of spontaneous astrocyte Ca2+ transients in control conditions
 and in the presence of silodosin.  **Figure S4: CalEx blocks the effect of
 NE on astrocyte Ca2+ and synapses:**  (A) Schematic of micro-injections to
 express the CalEx actuator PMCA2 (plasma membrane Ca2+ pump) in CA1
 astrocytes. (B) IHC of CalEx expression in CA1 s. radiatum astrocytes and
 quantification of specificity and efficiency. (C) Plot of the stimulation
 intensity/fEPSP slope relationship (left, unpaired Student’s t-test on
 slope/stim ratio) and summary bar graphs of PPF values (right) in
 RFP-control and CalEx slices at baseline. (D,E) Kymograph (each row shows
 the average fluorescence across ROAs of an individual astrocyte), 5
 representative ΔF/F0 traces (from individual ROAs), and quantification of
 the peak amplitude, frequency, and kinetics of spontaneous Ca2+ transients
 in RFP-control and CalEx slices. (F,G) Kymographs (each row represents an
 individual cell), average ΔF/F0 traces (± s.e.m.) across all astrocytes,
 and quantification of the peak amplitude, in response to 20μM NE
 application in RFP-control and CalEx slices. (H) Time course and
 representative traces of the effect of adenosine on fEPSP and PPF in CalEx
 slices. (I) Pairwise quantification of the effect of adenosine on fEPSP
 and PPF in CalEx slices. (J,K) Time course, representative traces, and
 pairwise quantifications of the effect of NE on fEPSP and PPF in CalEx
 slices. (L) Summary bar graphs showing the inhibitory effect of NE and
 adenosine on fEPSP in RFP-control and CalEx slices. The p-values in (L)
 are from ANOVAs across iβark, CalEx, and RFP-control conditions followed
 by Tukey’s post-hoc test, reflecting the fact that the RFP-control
 condition for the experiments shown in (L) is the same across iβark
 (Fig.2) and CalEx (this figure) experiments. **Figure  S5: The SERCA pump
 inhibitor thapsigargin blocks the effect of NE on astrocyte Ca2+ and
 synapses:** (A,B) Kymograph (each row shows the average fluorescence
 across ROAs of an individual astrocyte), 5 representative ΔF/F0 traces
 (from individual ROAs), and quantification of the peak amplitude,
 frequency, and kinetics of spontaneous Ca2+ transients in TTX alone
 (RFP-control) and TTX + thapsigargin (1μM) conditions. All slices were
 obtained from animals with RFP-transduced astrocytes for cell-segmentation
 purposes. Thapsigargin was bath applied 20min prior to the start of
 recording. (C,D) Kymographs (each row represents a cell), average ΔF/F0
 traces (± s.e.m.) across all astrocytes, and quantification of the peak
 amplitude, in response to 20μM NE application in RFP-control and
 thapsigargin conditions. (E,F) Time course, representative traces, and
 pairwise quantification of the effect of 5μM adenosine on fEPSP and PPF in
 thapsigargin-treated slices. (G,H) Time course, representative traces, and
 pairwise quantifications of the effect of 20μM NE on fEPSP and PPF in
 thapsigargin-treated slices. (I) Summary of the inhibitory effect of NE
 and adenosine on fEPSP in control and thapsigargin-treated slices.
 **Figure S6: Ca2+ and synaptic recordings in A- AdraKO and N-AdraKO
 slices:**  (A) Schematic of the experimental design and qPCR analysis of
 the expression of astrocyte markers (Kcnj10: Kir4.1 and Slc1a2: GLT-1) and
 neuronal markers (Snap25: SNAP25 and Dlg4: PSD95) in GFP-sorted cells from
 AAV5-gfaABC1D::Cre-GFP (n = 9 mice) and AAV5-hSyn::Cre- GFP injected mice
 (n = 12 mice). Statistical analysis was done by z-scoring ΔΔCt values and
 averaging them for a ‘marker type’ (i.e. astrocyte, or neuron) within
 samples, followed by a paired Student’s t-test. (B) Adra1a mRNA levels
 measured by qPCR in GFP-positive and GFP-negative cells from hSyn::Cre-GFP
 injected Adra1a+/+ (n = 4) and Adra1afl/fl mice (n = 3). (C) Same as (B)
 in gfaABC1D::Cre-GFP injected mice (n=3 biological replicates each). (D)
 Schematic of microinjections of AAV5-hSyn::Cre or AAV5-gfaABC1D::Cre in
 LSL-tdTomato mice (Ai14 mice), and strategy for validation of
 cell-specificity. (E) IHC of Cre-dependent tdTomato expression in neurons
 (NeuN), oligodendrocytes (Olig2), microglia (Iba1), and astrocytes (GFAP)
 in the hippocampus of Cre-injected Ai14 mice. Cell markers are shown in
 green and tdTomato in red. Owing to the CAG promoter, expression of
 tdTomato appears stronger in recombined neurons than in recombined
 astrocytes. Scale bar: 40μm. (F) Quantification of tdTomato expression in
 Creinjected Ai14 mice (n = 3 mice, 4 sections/mouse), in % of
 maker-positive cells that are tdTomatopositive and marker-positive.
 Numbers indicate the number tdTomato-positive cells over the total number
 of cells counted for each cell-type. (G) Same as (A) for tdTomato-positive
 and negative cells from Cre-injected Ai14 mice (n = 3 mice). (H) Pairwise
 quantifications of the effect of 20μM NE on the holding current in
 whole-cell patch-clamp recordings (V-clamp) of CA1 pyramidal neurons in
 slices from control mice, in the absence or presence of the α1A-AR
 antagonist silodosin (100nM), and in slices from N-AdraKO mice
 (GFP-positive neurons), and illustrative traces. Note the inward current
 (downward shift) caused by NE, which is completely absent in the presence
 of silodosin and in N-AdraKO neurons. Numbers in grey in the traces
 indicate the holding current (I(h)) at the start and end of the recording.
 (I) Summary plot of the effect of NE on the change in holding current in
 control conditions, in the presence of silodosin and in GFP-positive
 N-AdraKO neurons. n = 2-3 mice. (J) Plot of the stimulation
 intensity/fEPSP slope relationship (left, unpaired Student’s t-test on
 slope/stim ratio) and summary bar graphs of PPF values (right) in N-AdraKO
 and N-AdraCre-GFP slices at baseline. (K) Time course and representative
 traces of the effect of 20μM NE on fEPSP and PPF in N-AdraCre-GFP slices.
 (L) Plot of the stimulation intensity/fEPSP slope relationship (left,
 unpaired Student’s t-test on slope/stim ratio) and summary bar graphs of
 PPF values (right) in A-AdraKO and A-AdraCre-GFP slices at baseline. (M,N)
 Kymograph (each row shows the average fluorescence across ROAs of a single
 astrocyte), 5 representative ΔF/F0 traces (from individual ROAs), and
 quantification of the peak amplitude, frequency, and kinetics of
 spontaneous Ca2+ transients in A-AdraKO and A-AdraCre-GFP slices. (O) Time
 course and representative traces of the effect of 5μM adenosine on fEPSP
 and PPF in A-AdraKO slices. (P) Pairwise quantification of the effect of
 5μM adenosine on fEPSP and PPF in A-AdraKO and AAdraCre-GFP slices. (Q)
 Summary plot of the inhibitory effect of adenosine on fEPSP in A-AdraKO
 and A-AdraCre-GFP slices. (R) Summary plot of the inhibitory effect of
 adenosine on fEPSP in NAdraKO and N-AdraCre-GFP slices.  **Figure
 S7: Adenosine signaling in control conditions, in NT5eKO animals and
 across astrocyte Ca2+ interventions:** (A,B) Time course, representative
 traces, and pairwise quantifications of the effect of 5μM adenosine on
 fEPSP and PPF in slices from naïve animals. (C) Time courses and
 representative traces of the effect of 5μM adenosine on fEPSP and PPF in
 slices in which astrocytes express RFP (RFP-control). (D-E) Pairwise
 quantification of the effect of 20μM NE on fEPSP and PPF in DPCPX (100nM,
 D), and a cocktail of P2X/P2Y (PPADS, 10μM), A2A (ZM241385, 50nM) and A2B
 (PSB603, 50nM) receptor antagonists (E). (F) Time courses and
 representative traces of the effect of 20μM NE and 5μM adenosine on fEPSP
 and PPF in slices from CA3-Ado1KD that received 2 bilateral
 AAV5-hSyn::Cre-GFP injections in the CA3 (4 total) (G) Pairwise
 quantification of the effect of 20μM NE and 5μM adenosine on fEPSP
 experiments shown on (F). n = 2 mice. (H) Pairwise quantification of the
 effect of 20μM NE on fEPSP and PPF in the presence of a cocktail of mGluR
 antagonists (CPPG, 5μM; MPEP, 3.6μM and YM298198, 2μM). n = 4 mice. (I)
 Pairwise quantification of the effect of 20μM NE on fEPSP and PPF in
 adenosine deaminase (ADA, 1U/mL, n = 9 mice). (J,K) Time course,
 representative traces, and pairwise quantification of the effect of 5μM
 adenosine on fEPSP and PPF in NT5eKO slices (paired Student’s t-tests).
 (L) Time courses and representative traces of the effect of 5μM adenosine
 on fEPSP and PPF in iβARK slices. (M) Pairwise quantification of the
 effect of adenosine on fEPSP and PPF in iβARK and RFP-control slices (N)
 Summary plot of the inhibitory effect of adenosine on fEPSP in iβARK and
 RFP-control slices. The p-value in (N) is from an ANOVA across iβARK,
 CalEx and RFP-control conditions followed by Tukey’s post-hoc test,
 reflecting the fact that the RFP-control condition for fEPSP recordings
 experiments is common to CalEx (SupFig.4) and iβark (Fig.2). #####
 Variables * fEPSP slope: slope of the field excitatory post-synaptic
 potential. Either raw value (mV/ms) or normalized * PPF: paired-pulse
 facilitation. Either raw value (no dimension) or normalized * PPR:
 paired-pulse ratio. Either raw value (no dimension) or normalized *
 Efficacy: rate of synaptic transmission successes (percentage or
 probability) * Potency: amplitude (in pA) of successful synaptic events *
 Strength: a product of potency and efficacy (in pA) * Number of cells
 counted * Efficiency: percent of a cell type expressing the genetic
 reporter * Specificity: precent of reporter-expressing cells that are of
 desired cell-type * Spontaneous calcium activity (dF/F0). Activity
 observed in the absence of stimulation * Evoked calcium activity (dF/F0).
 Activity observed in response to stimulation (e.g., NE application) *
 Amplitude of calcium events. Peak amplitude (dF/F0) of calcium events *
 Frequency of calcium events (transients/cell/min) * Duration of calcium
 events. Time (in s) from event onset to return to baseline * Rise time of
 calcium event. Time (in s) from 10% or peak amplitude to 90% of peak
 amplitude * gDNA levels. Genomic DNA intensity band  * mRNA levels. Log2
 ratio of mRNA expression relative to that of housekeeping gene ##
 Code/software N/A ## Access information Other publicly accessible
 locations of the data: * None Data was derived from the following sources:
 *
 [https://www.biorxiv.org/content/10.1101/2024.05.21.595135v1](https://www.biorxiv.org/content/10.1101/2024.05.21.595135v1)