Banerjee Tirtha Das, Zhang Linwan, Monteiro Antónia
Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543, Singapore.
Methods Protoc. 2025 Mar 13;8(2):31. doi: 10.3390/mps8020031.
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 the expression of multiple RNA molecules in fixed larval brains of the African butterfly . We optimized the polyacrylamide gel mounting, fixation, and sample permeabilization steps, and mapped the expression domains of ten genes in whole larval brain tissue at single-cell resolution. The genes included (), , (), (), (), , (), (), (), and (). This method can be used alongside single-cell sequencing to visualize the spatial location of brain cells that change in gene expression or splicing patterns in response to specific behaviors or cognitive experiences.
蝴蝶幼虫表现出复杂的认知能力和行为，但对于这些行为如何在分子水平上改变它们的大脑，我们所知相对较少。在这里，我们优化了一种杂交链式反应3.0（HCR v3.0）方案，以可视化非洲蝴蝶固定幼虫大脑中多个RNA分子的表达。我们优化了聚丙烯酰胺凝胶固定、固定和样品通透步骤，并以单细胞分辨率绘制了全幼虫脑组织中十个基因的表达域。这些基因包括（）、、（）、（）、（）、、（）、（）、（）和（）。该方法可与单细胞测序一起使用，以可视化因特定行为或认知体验而在基因表达或剪接模式上发生变化的脑细胞的空间位置。
