The Roles Of Cystic Fibrosis Transmembrane Conductance Regulator In Prefrontal Cognitive Functions The Application Of Chirped Pulse Amplification Laser-mediated Ablation In The Neurology

Cystic fibrosis transmembrane conductance regulator(CFTR),an ATP-binding chloride channel,is regulated by intracellular cAMP/PKA phosphorylation.It is widely expressed in mammalian peripheral tissues and organs and contributed to chloride balance and cell homeostasis.Preliminary work has found rich expression of CFTR in the central nervous system(CNS).However,whether the CFTR channel also affects the neural network of the prefrontal cortex by calibrating the intracellular chloride level of neurons is still lacking.Here,we used immunohistochemical staining,chloride imaging,patch-clamp recording,and two-photon imaging to comprehensively analyze the specific functions of the CFTR channel in the prefrontal cortex.We found that regulation of CFTR channel activity can alter the structural complexity of cultured prefrontal cortical neurons,intracellular chloride levels;the resting membranes potential and evoked action potentials of layer 5 pyramidal neurons in brain slices;it also produces significant changes in inhibitory synaptic transmission.Furthermore,we found that inhibition of CFTR channels increased calcium activity of apical dendrites in the primary motor cortex in vivo and promoted motor learning.These results implicate a prominent role of CFTR in modulating Clhomeostasis and neuronal excitability in the prefrontal cortex(PFC),which may promote to understand the roles of the chloride channel in PFC related cognitive functions.Targeted cell ablation is an important strategy for dissecting the function of individual cells within complicated cellular networks.To perform targeted cell ablation rapidly and precisely in the brain,we developed an chirped pulse amplification laser-coupled two-photon microscope(CPA-TPM)system that allows instantaneous and targeted ablation of individual cells and real-time monitoring of neural network changes in the living mouse cortex.Through precise and iterative control of the laser power and position,we show that individual cells could be ablated by a single pulse with minimum collateral damage to surrounding neuropils.We further show that ablation of individual somatostatin(SST)-expressing interneuron increases the activity of nearby neurons in layer 2/3 of the primary motor cortex during motor skill learning.Furthermore,by performing precise dendrotomy of layer5 pyramidal neurons,we reveal that different apical dendritic branches of these cells are structurally and functionally independent.By rapid deactivating individual cells and their processes in a spatiotemporally specific manner,the CPA-TPM system could serve as an effective means for understanding the functions of cells within the complex neuronal network.