A simple slice culture system for the imaging of nerve development in embryonic mouse.
Newborn neurons elaborate an axon that undertakes a complicated journey to find its ultimate target in the brain or periphery. Although major progress in the study of this process has been made by analysis of dissociated neurons in vitro, one would like to observe and manipulate axonal outgrowth and pathfinding as it occurs in situ, as fasciculated nerves growing within the tissue itself. Here, we present a simple technique to do this, through cultivation of embryonic mouse slices expressing enhanced green fluorescent protein (EGFP) specifically in newborn neurons. This system allows for imaging of outgrowth of peripheral nerves into structures such as the developing limb. We demonstrate a reproduction of normal innervation patterns by spinal nerves derived from spinal cord motor neurons and sensory neurons of the dorsal root ganglia. The slices can be manipulated pharmacologically as well as genetically, by crossing the EGFP-expressing line with lines containing targeted mutations in genes of interest.
Brachmann
Isabel
I
Interdisciplinary Center for Neurosciences, Department of Neuroanatomy, University of Heidelberg, Heidelberg, Germany.
Jakubick
Vera Catherine
VC
Shakèd
Maya
M
Unsicker
Klaus
K
Tucker
Kerry Lee
KL
D000818:Animals; D000831:Animals, Newborn; D019521:Body Patterning; D016923:Cell Death; D005260:Female; D005727:Ganglia, Spinal / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D009046:Motor Neurons / Q000166:cytology / Q000378:metabolism; D009420:Nervous System / Q000166:cytology / Q000196:embryology* / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D009475:Neurons, Afferent / Q000166:cytology / Q000378:metabolism; D009924:Organ Culture Techniques; D011247:Pregnancy; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D040121:Semaphorin-3A / Q000172:deficiency / Q000235:genetics; D013116:Spinal Cord / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D013127:Spinal Nerves / Q000166:cytology / Q000196:embryology / Q000378:metabolism
D011994:Recombinant Proteins; C488217:Sema3a protein, mouse; D040121:Semaphorin-3A; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins