Donald-iScience

Published: 10 December 2020| Version 2 | DOI: 10.17632/r94jkwmd5z.2
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Donald Joseph

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Using conventional cre-recombination techniques under the control of the parvalbumin (PV) promoter, we successfully ablated the aristaless (Arx) transcription factor in postnatal PV interneurons. To investigate the postnatal function of Arx in these cells, we first used intracranial EEG recordings to measure network properties and seizures in Arx conditional knockout (CKO) mice. We also measured adaptive behavioral properties using a battery of well-established behavioral paradigms. To determine the molecular mechanisms of postnatal Arx signaling in PV interneurons, we obtained RNA seq data from dissociated parvalbumin interneurons isolated from control and conditional Arx knockout mice at P35-40. Standard sequencing techniques were employed to prepare RNA libraries and validation. Specifically, real time PCR was used to ascertain the validity of the RNA seq data set and immunohistochemistry was used to assess the integrity of the perineuronal net around PV interneurons given that the dysregulated genes in the RNAseq data were primarily associated with the extracellular matrix. To determine the cellular basis of network and behavioral abnormalities, we characterized the electrophysiological properties of PV interneurons. Electrophysiological data were obtained in the current- and voltage clamp modes from PV interneurons in the first region of the cornu Ammonis in the hippocampus. Intrinsic membrane and synaptic properties of these cells were measured using well-established protocols. Finally, Sholl analysis was used to determine dendritic complexity Arx CKO PV interneurons.