Abstract
The purpose of this study is to determine whether synapses can be formed between spinal motor neurons (SMNs) and major pelvic ganglion (MPG) neurons of a rat in vitro. The green fluorescent protein (GFP)-labelled MPG cells were cultured together with SMNs in a specific medium. The synaptic-like contacts established between SMNs and MPG neurons were studied in co-cultures using morphologic and immunocytochemistry approaches. Phase-contrast observation of co-cultures showed apparent SMNs-MPG neurons contacts as early as three or four days in vitro. We demonstrate some evidence of synaptic contacts between SMNs and MPG neurons in vitro by immunostaining with antibody directed against postsynaptic density protein 95 (PSD-95). We describe the development process of a defined SMNs-MPG neurons co-culture system. The results suggest that the hybrid synapse formation that may occur between SMNs and MPG neurons in vitro played an essential role in the mechanisms of a regenerated bladder with an artificial somatic-autonomic reflex arc.
Highlights
There has been evidence that indicates synaptogenesis can occur between somatic and autonomic neurons in vivo
To explore whether synapses would form between spinal motor neurons (SMNs) and major pelvic ganglion (MPG) neurons of the rat, we developed a system for the co-culture of SMNs from a newborn rat with MPG neurons from an adult rat
After MPG cells were transfected with Ad-green fluorescent protein (GFP) vector, the intensity of fluorescence gradually increased over time in each GFP-positive cell
Summary
There has been evidence that indicates synaptogenesis can occur between somatic and autonomic neurons in vivo. Previous studies show that functional synapses have been formed in mammalian sympathetic ganglia re-innervated with somatic nerves[1]. Nurse et al[6] described the formation of cholinergic synapses between dissociated sympathetic neurons and skeletal myotubes of the rat in cell culture. Belenky et al[7] developed a system for long-term co-culturing of explants of sympathetic chain ganglia and dorsal root ganglia from newborn rats. They found that some apparently specific sympathetic-sensory contacts could be observed, suggesting that a functional interaction may develop between sympathetic axons and sensory neurons in vitro
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