Basolateral Mg2+ Extrusion via CNNM4 Mediates Transcellular Mg2+ Transport across Epithelia: A Mouse Model

Abstract

Transcellular Mg²⁺ transport across epithelia, involving both apical entry and basolateral extrusion, is essential for magnesium homeostasis, but molecules involved in basolateral extrusion have not yet been identified. Here, we show that CNNM4 is the basolaterally located Mg²⁺ extrusion molecule. CNNM4 is strongly expressed in intestinal epithelia and localizes to their basolateral membrane. CNNM4-knockout mice showed hypomagnesemia due to the intestinal malabsorption of magnesium, suggesting its role in Mg²⁺ extrusion to the inner parts of body. Imaging analyses revealed that CNNM4 can extrude Mg²⁺ by exchanging intracellular Mg²⁺ with extracellular Na⁺. Furthermore, CNNM4 mutations cause Jalili syndrome, characterized by recessive amelogenesis imperfecta with cone-rod dystrophy. CNNM4-knockout mice showed defective amelogenesis, and CNNM4 again localizes to the basolateral membrane of ameloblasts, the enamel-forming epithelial cells. Missense point mutations associated with the disease abolish the Mg²⁺ extrusion activity. These results demonstrate the crucial importance of Mg²⁺ extrusion by CNNM4 in organismal and topical regulation of magnesium. Magnesium is an essential element for living organisms. Its absorption occurs at the intestine through the barrier comprised of epithelial cells. In this process, transcellular Mg²⁺ transport across epithelia, involving both entry from one side and extrusion from the other side, is important. Previous studies have revealed the role of Mg²⁺-permeable channel protein in Mg²⁺ entry into the epithelial cells. However, the identity of proteins involved in Mg²⁺ extrusion to the inner parts of body has remained unknown. Mice genetically engineered not to express CNNM4, which localizes to the epithelial membrane facing to the inner parts of body, show hypomagnesemia due to the defect in magnesium absorption. Functional analyses using culture cells directly reveal that CNNM4 can extrude intracellular Mg²⁺ to the outside of cells. These results indicate that CNNM4 mediates transcellular Mg²⁺ transport across the intestinal epithelia. Furthermore, we also show that these CNNM4-lacking mice also have a defect in amelogenesis, which is consistent with the disease symptoms of Jalili syndrome that is known to be caused by mutations in the CNNM4 gene.