|Klotho in Mouse Kidney. Klotho was detected in perfusion fixed frozen sections of mouse kidney using 1.7 µg/mL Mouse Klotho Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1819) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.|
Klotho, also called Klotho-alpha, is the founding member of the Klotho family within the glycosidase-1 superfamily. Klotho is expressed in areas concerned with calcium regulation, predominantly in the kidney distal convoluted tubules, but also in the brain choroid plexus (which produces cerebrospinal fluid) and the parathyroid. The 1014 amino acid (aa) type I transmembrane protein contains a 34 aa signal sequence, a 948 aa extracellular domain (ECD) containing two extracellular glycosidase-like domains, a 21 aa transmembrane domain and an 11 aa intracellular domain. Within the ECD, mouse Klotho shares 95%, 87%, and 87% aa identity with rat, human, and equine Klotho, respectively. Although a truncated 554 aa isoform predicts a soluble 70 kDa form, the soluble form found in plasma and cerebrospinal fluid is a 130 kDa form produced by proteolytic cleavage of the glycosylated 135 kDa full-length Klotho. A prominent intracellular 120 kDa form of Klotho is localized to endoplasmic reticulum and Golgi membranes. Klotho is named for the Greek goddess who spins the thread of life. The phenotype of Klotho-deficient mice resembles premature aging, including arteriosclerosis, osteoporosis, skin atrophy, infertility, emphysema, and premature death. Conversely, excess Klotho extends lifespan. Klotho acts as a cofactor for interaction of FGF-23 with FGF R1. This interaction negatively regulates 1 alpha -hydroxylase, the rate-limiting enzyme in the synthesis of 1,25(OH)2D3 (vitamin D). Klotho-deficient mice show severe hyperphosphatemia and ectopic calcification of soft tissues due to excess vitamin D. Both Klotho and beta ‑Klotho are co-factors for FGF-19 binding. Klotho also shows glucuronidase activity which activates the renal ion channel TRPV5 to reabsorb urinary calcium. Klotho has been reported to downregulate insulin or IGF-I signaling in adipocytes, to bind and antagonize Wnt molecules, and to facilitate release of parathyroid hormone.
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