IADR Abstract Archives
DPSC Derived Neurons Functionally Express Senory Neuron Specific Receptor (MRGX1).
Objectives:
To use human dental pulp stem cells (hDPSCs) to derive functionally active nerve cells as an alternative, more directly human relevant in vitro model system. This system will be used to determine functional expression of sensory neuron specific receptor MRGX1.
Methods: Neurons were characterised by expression of specific neuronal markers using immunohistochemistry (IHC) and functionally with patch clamp recordings. Protein and gene expression of TRP channels and MRGX1 in neurons were determined using IHC and quantitative polymerase chain reaction (qPCR) respectively. Receptor-channel functionality was assessed using calcium imaging.
Results: IHC showed that both TRP channels and MRGX1 co-localise at protein level. qPCR confirmed gene expression of both TRP channels and MRGX1 in neurons, although MRGX1 expression was relatively low. Calcium imaging demonstrated TRP channels to be functionally active in neurons in response to full agonists. MRGX1 was also shown to be functional in response to enkephalin fragment – bovine adrenal medulla (BAM8-22).
Conclusions: TRP channels and MRGX1 are expressed and functional in neurons derived from hDPSCs at a molecular level. Future work will determine if their expression and function are affected by injury and inflammation.
To use human dental pulp stem cells (hDPSCs) to derive functionally active nerve cells as an alternative, more directly human relevant in vitro model system. This system will be used to determine functional expression of sensory neuron specific receptor MRGX1.
Methods: Neurons were characterised by expression of specific neuronal markers using immunohistochemistry (IHC) and functionally with patch clamp recordings. Protein and gene expression of TRP channels and MRGX1 in neurons were determined using IHC and quantitative polymerase chain reaction (qPCR) respectively. Receptor-channel functionality was assessed using calcium imaging.
Results: IHC showed that both TRP channels and MRGX1 co-localise at protein level. qPCR confirmed gene expression of both TRP channels and MRGX1 in neurons, although MRGX1 expression was relatively low. Calcium imaging demonstrated TRP channels to be functionally active in neurons in response to full agonists. MRGX1 was also shown to be functional in response to enkephalin fragment – bovine adrenal medulla (BAM8-22).
Conclusions: TRP channels and MRGX1 are expressed and functional in neurons derived from hDPSCs at a molecular level. Future work will determine if their expression and function are affected by injury and inflammation.