Photodynamic Modification of Native HCN Channels Expressed in Thalamocortical Neurons
| Autor: | Qinglian Liu, Priyodarshan Goswamee, Fusheng Wei, Jizhong Han, Adam Rory McQuiston, Ankush Gupta, Qiang Wang, Lei Zhou |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Patch-Clamp Techniques
Physiology Cognitive Neuroscience Cyclic Nucleotide-Gated Cation Channels chemistry.chemical_element Biochemistry Oxygen Article Membrane Potentials Mice 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Thalamus Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels HCN channel medicine Animals Photosensitizer 030304 developmental biology Cerebral Cortex Neurons Membrane potential 0303 health sciences biology Chemistry Singlet oxygen Depolarization Cell Biology General Medicine Mice Inbred C57BL Ventral tegmental area medicine.anatomical_structure biology.protein Biophysics Neuron 030217 neurology & neurosurgery |
| Zdroj: | ACS Chem Neurosci |
| ISSN: | 1948-7193 |
| DOI: | 10.1021/acschemneuro.9b00475 |
| Popis: | The photodynamic process requires three elements: light, oxygen, and photosensitizer, and involves the formation of singlet oxygen, the molecular oxygen in excited electronic states. Previously, we reported that heterologously expressed hyperpolarization-activated cAMP-gated (HCN) channels in excised membrane patches are sensitive to photodynamic modification (PDM). Here we extend this study to native HCN channels expressed in thalamocortical (TC) neurons in the ventrobasal (VB) complex of the thalamus and dopaminergic neurons (DA) of the ventral tegmental area (VTA). To do this, we introduced the photosensitizer FITC-cAMP into TCs or DAs of rodent brain slices via a whole-cell patch-clamp recording pipette. After illumination with blue light pulses, we observed an increase in the voltage-insensitive, instantaneous I(inst) component, accompanied by a long-lasting decrease in the hyperpolarization-dependent I(h) component. Both I(h) and the increased I(inst) after PDM could be blocked by the HCN blockers Cs(+) and ZD7288. When FITC and cAMP were dissociated and loaded into neurons as two separate chemicals, light application did not result in any long-lasting changes of the HCN currents. In contrast, light pulses applied to HCN2−/− neurons loaded with FITC-cAMP generated a much greater reduction in the I(inst) component compared to that of WT neurons. Next, we investigated the impact of the long-lasting increases in I(inst) after PDM on the cellular physiology of VB neurons. Consistent with an upregulation of HCN channel function, PDM elicited a depolarization of the resting membrane potential (RMP). Importantly, Trolox-C, an effective quencher for singlet oxygen, could block the PDM-dependent increase in I(inst) and depolarization of the RMP. We propose that PDM of native HCN channels under physiological conditions may provide a photodynamic approach to alleviate HCN channelopathy in certain pathological conditions. |
| Databáze: | OpenAIRE |
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