Inhibitory collaterals in genetically identified medium spiny neurons in mouse primary corticostriatal cultures. Issue 6 (24th November 2013)

• Record Type: Journal Article
• Authors: Lalchandani, Rupa R.; Vicini, Stefano
• Is Part Of: Physiological reports. Volume 1:Issue 6(2013:Nov.)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2013-11-24
• Subjects: Dissociated culture -- dopamine receptors -- inhibitory collaterals -- striatum -- synaptic physiology; Physiology -- Periodicals; 571
• DOI: 10.1002/phy2.164
• Languages: English
• ISSNs: 2051-817X

Abstract

Abstract: Inhibitory collaterals between striatal medium spiny neuron (MSN) subtypes have been shown to critically influence striatal output. However, the low rate of inhibitory collateral detection between striatal MSNs in conventional ex vivo slice recordings has made the study of these connections challenging. Furthermore, most studies on MSN collaterals have been conducted either blind or in models, in which only one MSN subtype can be distinguished. Here, we describe a dissociated culture system using striatal and cortical neurons harvested from genetically modified mice at postnatal day 0. These mice express tdTomato and enhanced green fluorescent protein (EGFP) downstream of the dopamine D1 and D2 receptor promoters, respectively, allowing for simultaneous distinction between the two major subtypes of MSNs. In vitro, these neurons develop spines, hyperpolarized resting membrane potentials and exhibit up‐and‐down states, while also maintaining expression of both fluorophores through time. Using paired whole‐cell patch‐clamp recordings from identified MSNs at 14 days in vitro, we are able to detect a much higher rate of inhibitory functional synapses than what has been previously reported in slice recordings. These collateral synapses release γ‐Aminobutyric acid (GABA) and shape the firing patters of other MSNs. Although reduced in vitro models have a number of inherent limitations, the cultures described here provide a unique opportunity to study frequently observed functional collaterals between identifiable MSNs. Additionally, cultured neurons allow for control of the extracellular environment, with the potential to investigate pharmacological regulation of inhibitory MSNs collaterals. Abstract : e00164 D1 and D2 MSNs extend inhibitory collaterals that shape neuron firing and striatal output. Using BAC transgenics and paired whole‐cell recordings, we describe a paradigm that allows for the simultaneous identification of both MSN subtypes and the targeted study of MSN collaterals.