The hypertonic inner medulla poses challenges to the cells that inhabit this area of the nephron. We employed discovery tools including proteomics and genomics to identify proteins that subserve the adaptive response. The gamma subunit of the Na/K-ATPase is critical to the survival of cells in hypertonic conditions, as silencing it increases osmosensitvity, and overexpression increases osmotolerance. The inner medullary collecting duct (IMCD) has high transepithelial resistance (TER). Proteins responsible for tight junction integrity are upregulated in hypertonic states. Multi PDZ protein 1 (MUPP1), a PDZ scaffolding protein, targets Claudin 4 to the tight junction. The silencing of either of these proteins decreases TER and renders the epithelium leaky. The accumulation of inert osmolytes is integral to the adaptive response. The genes involved are regulated by the transcription factor Tonicity Enhancer Binding Protein. An osmoregulated nuclear protein Nup88 is critical to the retention of this transcription factor in the nucleus and to the generation of the osmolytes. In summary, IMCD cells bring forth a coordinated response to hypertoncity that is necessary for cell survival and function of these cells in anisotonic conditions.