Three weeks after the AAV injections, mice were anaesthetized with urethane (i.p. 1.9 g/Kg) at two estrous stages (diestrus or proestrus) and two different time points ZT4-ZT5 (in vitro recording at ZT6-ZT10) or ZT9-ZT10 (in vitro recording at ZT11-ZT14, time of the preovulatory LH surge at the proestrus stage). An intracardic perfusion was performed with oxygenated iced-cold sucrose-artificial cerebrospinal fluid (sucrose-aCSF, containing 248 mM sucrose, 11 mM glucose, 26 mM NaHCO3, 2 mM KCl, 1.25 mM KH2PO4, 2 mM CaCl2, 1.3 mM MgSO4, 5 mM kynurenic acid). The solution was continuously bubbled with 5% CO2 and 95% O2. Brains were quickly removed and 300 μm thick coronal slices were cut at the level of the DMH using a vibratome (Leica VT1200S). Slices were then transferred in artificial cerebrospinal fluid (aCSF) containing 126 mM NaCl, 26 mM NaHCO3, 2.5 mM KCl, 1.25 mM NaH2PO4, 2 mM CaCl2, 2 mM MgCl2, 10 mM glucose at room temperature for 1 hour before starting recordings.

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