Short communicationNew methods for investigation of neuronal migration in embryonic brain explants

- New immunocytochemistry (ICC) protocol for staining neurons migrating out of the MGE explant embedded in Matrigel was developed in this work.
- Our new protocol transfects MGE explants.
- Our microfluidic chambers allow visualization of the vectorial migration of individual neuronsfrom explants, and tracking of cellular organelles.
- Our methods provide new paradigms to study neuronal migration in real-time.

BackgroundProper migration of neurons is essential for the formation and normal functioning of the nervous system. Defects in neuronal migration underlie a number of neurologic diseases in humans. Although cell migration is crucial for neural development, molecular mechanisms guiding neuronal migration remain to be elucidated fully. Newborn neurons from the embryonic medial ganglionic eminence (MGE) migrate a long distance dorsally in the developing brain, giving rise to several types of interneurons in the neocortex.New methodIn this study, we developed an immunocytochemistry (ICC) protocol to stain neurons migrating out of the MGE explant embedded in Matrigel. We also established a protocol to efficiently transfect cells in MGE explants, achieving a transduction efficiency of more than 30%.Comparison with existing methodIn addition, we developed microfluidic chambers for explants that allow visualization of the vectorial migration of individual neurons from mouse embryonic MGE explants. Our microfluidic system allows monitoring of the distribution of cellular organelles (e.g. Golgi) within migrating neurons which have been stained with commercial molecular dyes or transfected with adeno-associated virus (AAV) expressing reporter proteins.ConclusionThese methods provide new paradigms to study neuronal migration in real-time.