Short communicationMuscle specific nucleus ambiguus neurons isolation and culturing

- The development of a new protocol for specific brainstem motoneuron isolation from postnatal rats.
- Abductor and adductor motoneurons were isolated in separate plates.
- The utility of neural tracers for a long term neuron culture isolation was addressed.
- The best period for cranial motoneuron isolation and culture in postnatal rats was determined.

BackgroundPeripheral nerve injury leads to a regenerative state. However, the reinnervation process is highly non-selective. Growing axons are often misrouted and establish aberrant synapsis to abductor or adductor muscles. Determining the complex properties of abductor and adductor motoneurons in a neuron culture, may lay the groundwork for future studies on axon guidance, leading to a clinical treatment for a selective reinnervation.New methodIn the present study we develop a neuron culture protocol to isolate recurrent laryngeal nerve abductor and adductor motoneurons in order to study their unique properties.Comparison with existing methods the best period to perform the present protocol for postnatal rat cranial motoneurons isolation was determined. In addition, the method allows identification of specific motoneurons from other primary motoneurons and interneurons within brainstem.ConclusionThe present protocol will allow investigators to perform targeted and novel studies of the mechanisms of peripheral nerve regeneration.