Conjunctive effects of fibroblast growth factor and glycosaminoglycan on bone metabolism in neonatal bartter syndrome.
The calciotropic activity of urine from a subject with neonatal Bartter syndrome (NBS) has been partially purified using ion-exchange and gel chromatographic techniques. A bioassay using bone disks from rat calvaria was used to estimate calciotropic activity, which in the urine of the subject with NBS appears to be due to basic fibroblast growth factor (bFGF) bound to a glycosaminoglycan susceptible to heparitinase digestion. The calciotropic activity is eluted from DEAE-Sephacel and Sepharose CL-6B in a narrow band in association with metachromatic material and is destroyed by heparitinase and blocked by an antibody to bFGF. After treatment of purified preparations with heparitinase, a component that is inactive alone but develops calciotropic activity in association with heparin can be isolated by affinity chromatography on heparin-Sepharose columns. This component is recovered from the column at NaCl concentrations expected to elute bFGF and is inactivated by antibodies to bFGF. No calciotropic activity can be shown in glycosaminoglycan-containing fractions from urine from a normal boy or a normal man, but such fractions exhibit calciotropic activity if bFGF is added to the assay system. When bFGF is added to urine from either normal subject followed by ion-exchange chromatography on DEAE-Sephacel, calciotropic activity is eluted at NaCl concentrations closely similar to those found to elute calciotropic activity from the urine of the NBS subject. It appears that the abnormal findings in NBS urine are due to excess bFGF, although they could be due to some abnormality of the glycosaminoglycan component.
Williams
W J
WJ
Department of Pediatrics, SUNY Health Science Center, Syracuse 13210, USA.
Shoemaker
L R
LR
Schurman
S J
SJ
Welch
T R
TR
Bergstrom
W H
WH
D000368:Aged; D000818:Animals; D001477:Bartter Syndrome / Q000151:congenital / Q000652:urine*; D001681:Biological Assay; D002118:Calcium / Q000097:blood; D002648:Child; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D016222:Fibroblast Growth Factor 2 / Q000302:isolation & purification / Q000494:pharmacology / Q000652:urine*; D006025:Glycosaminoglycans / Q000302:isolation & purification / Q000494:pharmacology / Q000652:urine*; D006801:Humans; D008297:Male; D051381:Rats; D012016:Reference Values; D012886:Skull / Q000187:drug effects / Q000502:physiology*
D006025:Glycosaminoglycans; D016222:Fibroblast Growth Factor 2; D002118:Calcium