4.8. Electrophysiological Recording
This protocol is extracted from research article:
A Role for Xanthurenic Acid in the Control of Brain Dopaminergic Activity
Int J Mol Sci, Jun 28, 2021; DOI: 10.3390/ijms22136974

Animals (6 rats per condition) were treated during 21 consecutive days with Xanthurenic acid (XA; 100 mg/kg/day; dissolved in 1% bicarbonate solution). The drug was i.p injected in a 1 mL volume. The XA effect was compared to that of the neuroleptic haloperidol (0.5 mg/kg i.p in 1 mL) given daily for 21 days. Control animals were daily i.p injected with 1 mL of vehicle for the same period.

After the treatment schedule (i.e., on day 22) spontaneously firing dopaminergic neurons within the SN and the VTA nuclei were counted.

The animals were anesthetized with ketamine (100 mg/kg i.p) and mounted on a stereotaxic apparatus (TSE system, Germany). During the experiment supplemental doses of anesthetic were administered if necessary. The skull was revealed by incision of the scalp and remaining flesh removed using cotton sticks. Left and right skull areas overlying the SN and VTA were removed. The following stereotaxic coordinates were used, relative to bregma and cortical surface: SN: AP = −5.4 to −5.8, ML = 1.8 to 2.5, DV = −7.0 to −8.0 and VTA: AP = −5.4 to −5.8, ML = 0.4 to 0.8, DV = −7.0 to −8.5 [43]. Nine electrode tracks (separated from each other by 200 µm), with a constant stereotaxic coordinates pattern for all animals, were made in each recorded region. Extracellular recordings were performed using a single-barrel glass micropipette that was filled with 2 M NaCl (in vitro resistance 4–7 MΩ). Electrical signal of spontaneous spiking activity was measured using a high input impedance amplifier (IsoDAM-8A; WPI) and a 10–3000 Hz bandwidth filtering. The signal was digitized with a digidata 1224 (Axon Instruments) and acquired using the Pclamp software (Axon Instrument). Dopaminergic neurons were identified according to previously established electrophysiological characteristics [44]. The data are expressed as the mean active neurons per track (±S.E.M.).

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