Collective Cell Migration Drives Morphogenesis of the Kidney Nephron

Tissue organization in epithelial organs is achieved during development by the combined processes of cell differentiation and morphogenetic cell movements. In the kidney, the nephron is the functional organ unit. Each nephron is an epithelial tubule that is subdivided into discrete segments with specific transport functions. Little is known about how nephron segments are defined or how segments acquire their distinctive morphology and cell shape. Using live, in vivo cell imaging of the forming zebrafish pronephric nephron, we found that the migration of fully differentiated epithelial cells accounts for both the final position of nephron segment boundaries and the characteristic convolution of the proximal tubule. Pronephric cells maintain adherens junctions and polarized apical brush border membranes while they migrate collectively. Individual tubule cells exhibit basal membrane protrusions in the direction of movement and appear to establish transient, phosphorylated Focal Adhesion Kinase–positive adhesions to the basement membrane. Cell migration continued in the presence of camptothecin, indicating that cell division does not drive migration. Lengthening of the nephron was, however, accompanied by an increase in tubule cell number, specifically in the most distal, ret1-positive nephron segment. The initiation of cell migration coincided with the onset of fluid flow in the pronephros. Complete blockade of pronephric fluid flow prevented cell migration and proximal nephron convolution. Selective blockade of proximal, filtration-driven fluid flow shifted the position of tubule convolution distally and revealed a role for cilia-driven fluid flow in persistent migration of distal nephron cells. We conclude that nephron morphogenesis is driven by fluid flow–dependent, collective epithelial cell migration within the confines of the tubule basement membrane. Our results establish intimate links between nephron function, fluid flow, and morphogenesis. : The kidney's job is to maintain blood ion and metabolite concentrations in a narrow range that supports the function of all other organs. Blood is filtered and essential solutes are recovered in a structure called the nephron. Human kidneys have one million nephrons, while simpler kidneys like the zebrafish larval kidney have only two. Nephrons are segmented epithelial tubules; each segment takes on a particular shape (such as convoluted, straight, or U-shaped) and plays a specific role in recovering filtered solutes. How the nephron is proportioned into segments and how some tubule segments become convoluted is not known. This work takes advantage of the simple zebrafish kidney to image living cells during nephron formation. Unexpectedly, we found that nephron cells are actively migrating “upstream” toward the filtering end of the nephron. The cells remain connected to each other and migrate as an intact tube. This is similar to a process called “collective cell migration.” We find that collective cell migration establishes the final position of nephron segment boundaries and drives convolution of the tubule. We also find that cell migration is dependent on fluid flow in the tubules, supporting the idea that organ function is important in defining its final form. Epithelial cell shape, tubule convolution, and segment boundary position along the kidney nephron unexpectedly involve the migration of fully differentiated epithelial cells against the flow of lumenal fluid.