Sediqi, H.; Mathews, J.; de Nola, G.; Lytton-Jean, A. K. R.; Levin, M.

While bioelectricity is increasingly recognized as an important regulator of cell function and morphogenesis, the field has almost exclusively focused on plasma membrane states. Voltage across the inner nuclear membrane (INM) has been proposed as a potential regulator of nuclear function, but how it responds to extracellular ionic perturbations and whether it relates to chromatin organization remain unclear. Here, we targeted the ratiometric genetically encoded voltage indicator ASAP3-R3 to SUN2-associated nuclear membranes in intact NRK cells and combined INM voltage measurements with Gray-Level Co-Occurrence Matrix (GLCM)-based chromatin texture analysis. Reporter localization was confirmed by fluorescence imaging and electron microscopy, and functional validation in isolated nuclei showed that sodium-potassium pump inhibition produced INM depolarization consistent with Goldman-Hodgkin-Katz (GHK)-based prediction. We then used our validated construct to determine the response of Vnuc and chromatin texture to changing ionic conditions via two exposure methods, gradual (ramped) exposure or direct application. In intact cells, ramping different sets of ionic solutions of decreasing sodium/increasing potassium, decreasing sodium, increasing potassium, or decreasing chloride induced INM hyperpolarization and coordinated changes in chromatin texture, including increased contrast and entropy, reduced homogeneity, and reduced nuclear area. These effects were strongly path-dependent, with nuclear responses shaped by the history and order of ionic exposure: sodium and potassium responses emerged most clearly during ramping exposure, whereas reducing chloride by direct exposure showed a more pronounced response profile. Direct changes in sodium exposure produced limited electrical and chromatin-texture effects, while direct potassium exposure altered chromatin texture and nuclear area without significantly changing VNuc. Importantly, shifting baseline chromatin state in either direction, through Trichostatin-A (TSA)-induced chromatin relaxation or sodium azide/2-deoxy-D-glucose-induced compaction, blunted ion-associated Vnuc and chromatin responses across sodium, potassium, and chloride conditions. Together, these findings identify the nucleus as a dynamic, ion-responsive electro-structural system in which INM voltage and chromatin organization are functionally coupled, and in which both ionic trajectory and pre-existing chromatin state shape the magnitude of the nuclear response.