Figure 5.

Transduction of the cortex with SuperClomeleon. (A) Coronal slab of brain with bilateral injections of SuperClomeleon AAV into the rostral gyrus, 5 weeks after injection and 5 h after unilateral cortical impact (right hemisphere). As expected, the cortical impact site is moderately swollen with subarachnoid (marked red in inset) and intraparenchymal hemorrhages (marked yellow in inset) and the injection site visible (blue dye). On the left cortex, the injection site (blue dye) remains pristine following AAV injection and after 2 h of 2P imaging. (B) Schematic demonstrating the distribution of SuperClomeleon⁺ neurons in the rostral gyrus down to the sulci. SuperClomeleon⁺ neurons were dense in Layers 2/3 of the gray matter, were dispersed through all layers to Layer 6, and were in the gyral white matter, perhaps expressed by interstitial neurons in pig. This was pig 86.027 (Supplemental Fig. 1), where conditions were optimized and repeated for several pigs and results consistent. (C) Low power (10×) photomicrograph of SuperClomeleon⁺ neurons (green) with nuclei (DAPI, blue; scale bar = 100 μM). SuperClomeleon⁺ neurons are dense in Layer 2/3²⁰ and dispersed in Layer 4. (D) Higher power photomicrograph (20×; scale bar = 50 μM) of Layer 2/3 in Panel (C) (white box) demonstrates SuperClomeleon expression in pyramidal neuron cell bodies and neurites.