PT  - JOURNAL ARTICLE
AU  - Shen, Jun
AU  - Blute, Todd A.
AU  - Liberti, William A.
AU  - Yen, William
AU  - Liberti, Derek C.
AU  - Kotten, Darrell N.
AU  - Cruz-Martín, Alberto
AU  - Gardner, Timothy J.
TI  - Songbird organotypic culture as an <em>in vitro</em> model for interrogating sparse sequencing networks
AID  - 10.1101/164228
DP  - 2017 Jan 01
TA  - bioRxiv
PG  - 164228
4099  - http://biorxiv.org/content/early/2017/07/17/164228.short
4100  - http://biorxiv.org/content/early/2017/07/17/164228.full
AB  - Sparse sequences of neuronal activity are fundamental features of neural circuit computation; however, the underlying homeostatic mechanisms remain poorly understood. To approach these questions, we have developed a method for cellular-resolution imaging in organotypic cultures of the adult zebra finch brain, including portions of the intact song circuit. These in vitro networks can survive for weeks, and display mature neuron morphologies. Neurons within the organotypic slices exhibit a diversity of spontaneous and pharmacologically induced activity that can be easily monitored using the genetically encoded calcium indicator GCaMP6. In this study, we primarily focus on the classic song sequence generator HVC and the surrounding areas. We describe proof of concept experiments including physiological, optical, and pharmacological manipulation of these exposed networks. This method may allow the cellular rules underlying sparse, stereotyped neural sequencing to be examined with new degrees of experimental control.HighlightsOrganotypic brain slices from adult zebra finch (Taeniopygia guttata), expressing the calcium indicator GCaMP6, can be cultured and maintained for at least several weeks and display spontaneous and evoked calcium transients.