title: New methods for investigation of neuronal migration in embryonic brain explants.
abstract: BACKGROUND
  Proper 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.
  
  
  METHODS
  In 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%.
  
  
  CONCLUSIONS
  In 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.
  
  
  CONCLUSIONS
  These methods provide new paradigms to study neuronal migration in real-time.
authors.affiliation: Department of Neurology and Radiology, Massachusetts General Hospital and Program in Neuroscience, Harvard Medical School, Boston, MA 02114, USA. Electronic address: neryfc@yahoo.com.
authors.forename: Flávia C
authors.initials: FC
authors.lastname: Nery
authors.affiliation: Department of Neurology and Radiology, Massachusetts General Hospital and Program in Neuroscience, Harvard Medical School, Boston, MA 02114, USA.
authors.forename: Cintia C
authors.initials: CC
authors.lastname: da Hora
authors.affiliation: Department of Neurology and Radiology, Massachusetts General Hospital and Program in Neuroscience, Harvard Medical School, Boston, MA 02114, USA.
authors.forename: Uzma
authors.initials: U
authors.lastname: Yaqub
authors.affiliation: Department of Neurology and Radiology, Massachusetts General Hospital and Program in Neuroscience, Harvard Medical School, Boston, MA 02114, USA.
authors.forename: Xuan
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Neurology and Radiology, Massachusetts General Hospital and Program in Neuroscience, Harvard Medical School, Boston, MA 02114, USA; Center for Brain Repair, Department of Biomedical Sciences, Florida State University College of Medicine, Tallahassee, FL 32306, USA.
authors.forename: Deirdre M
authors.initials: DM
authors.lastname: McCarthy
authors.affiliation: Department of Neurology and Radiology, Massachusetts General Hospital and Program in Neuroscience, Harvard Medical School, Boston, MA 02114, USA; Center for Brain Repair, Department of Biomedical Sciences, Florida State University College of Medicine, Tallahassee, FL 32306, USA.
authors.forename: Pradeep G
authors.initials: PG
authors.lastname: Bhide
authors.affiliation: BioMEMS Resource Center, Department of Surgery, Massachusetts General Hospital, Charlestown, MA 02129 USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Irimia
authors.affiliation: Department of Neurology and Radiology, Massachusetts General Hospital and Program in Neuroscience, Harvard Medical School, Boston, MA 02114, USA.
authors.forename: Xandra O
authors.initials: XO
authors.lastname: Breakefield
mesh_terms: D000818:Animals; D000941:Antigens / Q000378:metabolism; D002465:Cell Movement / Q000502:physiology*; D000229:Dependovirus / Q000235:genetics; D004622:Embryo, Mammalian; D005260:Female; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D066298:In Vitro Techniques; D008297:Male; D008473:Median Eminence / Q000166:cytology*; D051379:Mice; D008810:Mice, Inbred C57BL; D046210:Microfluidic Analytical Techniques; D009474:Neurons / Q000502:physiology*; D009924:Organ Culture Techniques; D014161:Transduction, Genetic; D014404:Tubulin / Q000378:metabolism
chemical_list: D000941:Antigens; D014404:Tubulin; C462941:beta3 tubulin, mouse; C086178:pericentrin; D049452:Green Fluorescent Proteins
keywords: MGE explant; Microfluidic device; Neuron migration; Transduction