
Methods and Protocols, Journal Year: 2025, Volume and Issue: 8(2), P. 31 - 31
Published: March 13, 2025
Butterfly larvae display intricate cognitive capacities and behaviors, but relatively little is known about how those behaviors alter their brains at the molecular level. Here, we optimized a hybridization chain reaction 3.0 (HCR v3.0) protocol to visualize expression of multiple RNA molecules in fixed larval African butterfly Bicyclus anynana. We polyacrylamide gel mounting, fixation, sample permeabilization steps, mapped domains ten genes whole brain tissue single-cell resolution. The included optomotor blind (omb), yellow-like, zinc finger protein SNAI2-like (SNAI2), weary (wry), extradenticle (exd), Synapsin, Distal-less (Dll), bric-à-brac 1 (bab1), dachshund (dac), acetyl coenzyme A acetyltransferase B (AcatB). This method can be used alongside sequencing spatial location cells that change gene or splicing patterns response specific experiences.
Language: Английский