Three-photon in vivo imaging of neurons and glia in the medial prefrontal cortex with sub-cellular resolution
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Abstract
The medial prefrontal cortex (mPFC) is important for higher cognitive functions, including working memory, decision making, and emotional control. In vivo recordings of neuronal activity in the mPFC have been achieved via invasive electrical and optical approaches. Here we apply low invasive three-photon in vivo imaging in the mPFC of the mouse at unprecedented depth. Specifically, we measure neuronal and astrocytic Ca 2+ -transient parameters in awake head-fixed mice up to a depth of 1700 µm. Furthermore, we longitudinally record dendritic spine density (0.41 ±0.07 µm -1 ) deeper than 1 mm for a week. Using 1650 nm wavelength to excite red fluorescent microglia, we quantify their processes’ motility (58.9 ±2% turnover rate) at previously unreachable depths (1100 µm). We establish three-photon imaging of the mPFC enabling neuronal and glial recordings with subcellular resolution that will pave the way for novel discoveries in this brain region.
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3P-imaging of astrocytic Ca2+-activity in mPFC in vivo
Very cool section! A couple of questions:
Would you expect that astrocyte calcium activity in deeper layers in mPFC—either in somata or microdomains—would be similar to deep layers in primary sensory areas, which I believe you cite in your comparisons. If they are indeed similar in the features you describe here, does that give specific hints about functions of astrocyte across cortical areas with different architecture.
There are several indications recently that rather than number/amplitude/duration of astrocyte calcium events being the most salient, their propagation features can distinguish differential astrocyte activity. Have you looked at that for this comparison? I might temper the last sentence of this section more without a propagative analysis included.
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