Analysis of Translation in the Developing Mouse Brain using Polysome Profiling

The proper development of the mammalian brain relies on a fine balance of neural stem cell proliferation and differentiation into different neural cell types. This balance is tightly controlled by gene expression that is fine-tuned at multiple levels, including transcription, post-transcription and translation. In this regard, a growing body of evidence highlights a critical role of translational regulation in coordinating neural stem cell fate decisions. Polysome fractionation is a powerful tool for the assessment of mRNA translational status at both global and individual gene levels. Here, we present an in-house polysome profiling pipeline to assess translational efficiency in cells from the developing mouse cerebral cortex. We describe the protocols for sucrose gradient preparation, tissue lysis, ultracentrifugation and fractionation-based analysis of mRNA translational status.

Automated summary

The proper development of the mammalian brain relies on a fine balance of neural stem cell proliferation and differentiation into different neural cell types, which is controlled by gene expression at multiple levels including transcription, post-transcription, and translation. Translational regulation plays a critical role in coordinating neural stem cell fate decisions, and polysome fractionation is a powerful tool for evaluating mRNA translational status. The polysome profiling pipeline presented in this study includes sucrose gradient preparation, tissue lysis, ultracentrifugation, and fractionation-based analysis of mRNA translational status.