Protocol for single-molecule FISH in the developing mouse retinal vasculature.

The developing vasculature of the post-natal mouse retina is a powerful model to discover mechanisms of vessel formation and to test modulators of neovascularization. We present a protocol for single-molecule fluorescent in situ hybridization (smFISH) in whole-mount mouse retinas enabling the detection of individual mRNAs in vascular endothelial cells. We describe procedures from initial retina preparation to smFISH and detection.

Insights into the regulation of mRNA translation by scaffolding proteins.

Regionalisation of molecular mechanisms allows cells to fine-tune their responses to dynamic environments. In this context, scaffolds are well-known mediators of localised protein activity. These phenomenal proteins act as docking sites where pathway components are brought together to ensure efficient and reliable flow of information within the cell.

The scaffolding protein AKAP12 regulates mRNA localization and translation.

Regulation of subcellular messenger (m)RNA localization is a fundamental biological mechanism, which adds a spatial dimension to the diverse layers of post-transcriptional control of gene expression. The cellular compartment in which mRNAs are located may define distinct aspects of the encoded proteins, ranging from production rate and complex formation to localized activity. Despite the detailed roles of localized mRNAs that have emerged over the past decades, the identity of factors anchoring mRNAs to subcellular domains remains ill-defined.

RNA-binding proteins and translation control in angiogenesis.

Tissue vascularization through the process of angiogenesis ensures adequate oxygen and nutrient supply during development and regeneration. The complex morphogenetic events involved in new blood vessel formation are orchestrated by a tightly regulated crosstalk between extra and intracellular factors. In this context, RNA-binding protein (RBP) activity and protein translation play fundamental roles during the cellular responses triggered by particular environmental cues.