Supplemental materials for Altered expression of several molecular mediators of cerebral spinal fluid production in Hyp mice

X-Linked Hypophosphatemia (XLH) is a genetic disease causing life-long hypophosphatemia due to overproduction of FGF23. XLH is associated with Chiari malformations, cranial synostosis, and syringomyelia. FGF23 signals through FGFR1c and requires a co-receptor, a-Klotho, which is expressed in the renal distal convoluted tubules and the choroid plexus (ChP). In the ChP, a-Klotho participates in regulating cerebral spinal fluid (CSF) production by shuttling the sodium/potassium adenosine triphosphatase (Na+/K+-ATPase) to the luminal membrane. The sodium/potassium/chloride cotransporter 1 (NKCC1) also makes a substantial contribution to CSF production. Real-time PCR (RT-PCR) demonstrated significant upregulation of Klotho transcripts in the fourth ventricle but a trend toward reduced expression in the lateral ventricles of Hyp mice compared to controls. Transcript levels for Fgfr1c were unchanged in Hyp mice. Na+/K+-ATPase alpha-1 subunit transcripts were downregulated in all three ventricles, reaching significance in the third and lateral ventricles. Expression levels of the Slc12a2 transcript (which encodes NKCC1) were unchanged in Hyp mice compared to controls. In situ hybridization (ISH) was used to obtain anatomical corelates to the changes observed by RT-PCR. ISH confirmed the presence of all four transcripts in the lateral ventricle ChP of both WT and Hyp mice. This is the first study to document a significant change in the level of expression of the molecular machinery required for CSF production in Hyp mice. Whether similar changes occur in patients with XLH, potentially contributing to the cranial and spinal cord abnormalities frequently seen in XLH remains to be determined.