Abstract
Today we have a nice technique to look at deeper tissue or cells, called two-photon laser scanning microscope (2PLSM). We are taking up some topics here related to this sexy technique. I will give a talk about in vivo two-photon targeted patch (TPTP) method which is an example of combination 2PLSM and electrophysiological technique.First we made a shRNA against the voltage gated sodium channel subunits, Nav1.1, 1.2, 1.3, which are reported to be mainly expressing in the dendrites. We packed it into a self-inactivated lentiviral vector with GFP as an indicator. Then we injected it in the barrel cortex before critical period of it at P8 old. Ten days after injection, we patched the cell which expressed the shRNA and GFP under the 2PLSM to investigate the property of somatosensory receptive field. We could reviel that the reduction of somatosensory responses of neurons in layer 2/3 of the barrel cortex with TPTP. This result suggest that the dendritic sodium channel played a critical role for maturation of sensory receptive field during its critical period, which could be accomplished via a form of neural plasticity, such as spike timing dependent plasticity.Now we are standing at the starting point with this result, because we have many molecules, brain areas and speices which have to be done with this method, TPTP. [J Physiol Sci. 2007;57 Suppl:S27]
