An RNA-immunoprecipitation protocol to identify RNAs associated with RNA-binding proteins in cytoplasmic and nuclear head fractions.

RNA-binding proteins (RBPs) are multifunctional proteins that shuttle between the nucleus and the cytoplasm where they assemble with target RNAs to form multi-molecular complexes. Here, we describe a protocol to selectively identify RNAs associated with RBPs of interest in the cytoplasmic and nuclear compartments of adult brain cells. Cytoplasmic and nuclear fractions are differentially collected and used for immunoprecipitation-based purification of GFP-tagged RBPs.

Expression of the Human Serotonin 5-HT Receptor Rescues Phenotype Profile and Restores Dysregulated Biomarkers in a Glioma Model.

Gliomas are the most common primary brain tumors in adults. Significant progress has been made in recent years in identifying the molecular alterations involved in gliomas. Among them, an amplification/overexpression of the EGFR (Epidermal Growth Factor Receptor) proto-oncogene and its associated signaling pathways have been widely described.

LINGO family receptors are differentially expressed in the mouse brain and form native multimeric complexes.

Leucine-rich repeat and immunoglobin-domain containing (LRRIG) proteins that are commonly involved in protein-protein interactions play important roles in nervous system development and maintenance. LINGO-1, one of this family members, is characterized as a negative regulator of neuronal survival, axonal regeneration, and oligodendrocyte precursor cell (OPC) differentiation into mature myelinating oligodendrocytes. Three LINGO-1 homologs named LINGO-2, LINGO-3, and LINGO-4 have been described.

Bioluminescence Resonance Energy Transfer as a Method to Study Protein-Protein Interactions: Application to G Protein Coupled Receptor Biology.

The bioluminescence resonance energy transfer (BRET) approach involves resonance energy transfer between a light-emitting enzyme and fluorescent acceptors. The major advantage of this technique over biochemical methods is that protein-protein interactions (PPI) can be monitored without disrupting the natural environment, frequently altered by detergents and membrane preparations. Thus, it is considered as one of the most versatile technique for studying molecular interactions in living cells at "physiological" expression levels.