Work

Optimized protocol for nuclei isolation from aquatic fish brain tissue for single-cell Genomic assays

Year: 2024

Type: article

Abstract: Single-cell sequencing technology has become increasingly prominent in aquaculture for investigating complex biological processes , such as development, reproduction, and disease resistance in aquatic species like Nile tilapia , an economically important fish. This technology reveals cellular heterogeneity across various biological functions . However, traditional enzymatic dissociation methods often fail to generate high-quality, intact single-cell suspensions from complex brain tissue, limiting single-cell research in this area. This study aims to optimize the isolation of single-cell nuclei from tilapia ( Oreochromis niloticus ) brain tissue. We systematically fine-tuned the parameters of homogenization buffer, washing buffer, and centrifugation conditions, identifying the optimal method for nuclei isolation from tilapia brain. We developed a streamlined, rapid, and efficient protocol that eliminates the need for costly equipment, such as flow cytometry and ultracentrifugation . As a proof of principle, we successfully identified the cellular heterogeneity of tilapia brain tissue using single-nucleus RNA-seq analysis. The method we developed yields intact nuclei with minimal background contamination and a broad representation of cell types, making it a reliable tool for single-cell genomic assays in aquatic fish species.

Source: Aquaculture and Fisheries

DOI: 10.1016/j.aaf.2024.12.001