ABSTRACT
Recent years have seen both considerable progress and controversy in the Alzheimer's disease (AD) field. After decades of slow to negligible movement towards the development of disease modifying therapies, promising outcomes in recent clinical trials with several monoclonal antibodies targeting various forms of the amyloid-ß (Aß) peptide have at last opened a possible way forward. In fact, at this point multiple anti-Aß therapeutics are close to receiving (or have already received) regulatory approval. Although these outcomes are not without some degree of divisiveness, the fact remains that targeting amyloid for removal has finally shown at least modest efficacy in slowing the otherwise relentless progression of the disease. Although the validation of the long standing amyloid cascade hypothesis would seem to be at hand, what remains is the puzzling issue of why - if Aß indeed causes AD - does removing it from the brain not stop the disease entirely.
Subject(s)
Alzheimer Disease , Amyloid beta-Peptides , Humans , Amyloid beta-Peptides/metabolism , Alzheimer Disease/drug therapy , Brain/metabolism , Antibodies, Monoclonal/therapeutic use , CognitionABSTRACT
The dialdehyde glyoxal is an alternative chemical fixative that cross-links tissues faster than formaldehyde, retains higher antigenicity, and is less hazardous than either formaldehyde or glutaraldehyde. Here we present a glyoxal-based fixation protocol for use with Drosophila embryos. We describe steps to prepare acid-free glyoxal, fix embryos, and then stain with antibodies for immunofluorescence (IF). We also describe methods for RNA fluorescence in situ hybridization (FISH) and FISH plus IF (FISH-IF) using glyoxal-fixed embryos. This protocol was adapted for Drosophila embryos from the methods of Bussolati et al.1 and Richter et al.2.
