We blocked gap junctions via application of the gap junction blocker MFA (50 µM, Sigma-Aldrich). We blocked the nicotinic acetylcholine signaling pathway via application of the broad nicotinic receptor antagonists hexamethonium (Hex, 100 µM, Sigma-Aldrich) and epibatidine (EPB, 10 nM, Sigma-Aldrich) as well as the specific antagonist dihydro-ß-erythroidine hydrobromide (DHßE, 8 µM, Sigma-Aldrich).
The following procedure was used for all pharmacology experiments: We recorded baseline activity in ACSF for 8 min before pharmacological agents were applied to the perfusion system. We then waited 15–30 min for the agents to take effect before acquiring another 8 min recording session.
To attempt to assay off-target effects of MFA, we used whole-cell voltage-clamp recordings to compare voltage-gated ion channels on RGCs in E16–18 retina but found inconsistent results (Figure 2—figure supplement 1 ). We associate this high variance with a rapid changing complement of ion channels during development and the quick washout of these conductances during whole-cell recordings.
The following procedure was used for all pharmacology experiments: We recorded baseline activity in ACSF for 8 min before pharmacological agents were applied to the perfusion system. We then waited 15–30 min for the agents to take effect before acquiring another 8 min recording session.
To attempt to assay off-target effects of MFA, we used whole-cell voltage-clamp recordings to compare voltage-gated ion channels on RGCs in E16–18 retina but found inconsistent results (
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