Abstract：
:This protocol describes an optimized method for direct in vitro monitoring of homo- and heterotypic axon-axon interactions involved in the developmental assembly of neural circuits. The assay exploits a classical example of heterotypic axonal interactions by modeling the sequential extension of spinal motor and somatosensory neuron axons, but the procedure should be readily adaptable to other neuron types. The protocol is based on the rapid isolation and primary culture of genetically identified motor neurons combined with straightforward vital dye labeling and culture of dorsal root ganglion sensory neurons. Subsequently, axonal interactions are directly monitored via live fluorescence microscopy, whereas axon type identities can be unambiguously delineated throughout the experiments. Through chemical compound application or by using neurons derived from genetically engineered mice, the protocol facilitates the dissection of molecular pathways driving the axonal interactions that are crucial for neural pathway and circuit assembly. The whole procedure can be completed in 3 d.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Wang L,Marquardt Tdoi
10.1038/nprot.2011.442subject
Has Abstractpub_date
2012-01-26 00:00:00pages
351-63issue
2eissn
1754-2189issn
1750-2799pii
nprot.2011.442journal_volume
7pub_type
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