Impaired calcification around matrix vesicles of growth plate and bone in alkaline phosphatase-deficient mice.
The presence of skeletal hypomineralization was confirmed in mice lacking the gene for bone alkaline phosphatase, ie, the tissue-non-specific isozyme of alkaline phosphatase (TNAP). In this study, a detailed characterization of the ultrastructural localization, the relative amount and ultrastructural morphology of bone mineral was carried out in tibial growth plates and in subjacent metaphyseal bone of 10-day-old TNAP knockout mice. Alizarin red staining, microcomputerized tomography (micro CT), and FTIR imaging spectroscopy (FT-IRIS) confirmed a significant overall decrease of mineral density in the cartilage and bone matrix of TNAP-deficient mice. Transmission electron microscopy (TEM) showed diminished mineral in growth plate cartilage and in newly formed bone matrix. High resolution TEM indicated that mineral crystals were initiated, as is normal, within matrix vesicles (MVs) of the growth plate and bone of TNAP-deficient mice. However, mineral crystal proliferation and growth was inhibited in the matrix surrounding MVs, as is the case in the hereditary human disease hypophosphatasia. These data suggest that hypomineralization in TNAP-deficient mice results primarily from an inability of initial mineral crystals within MVs to self-nucleate and to proliferate beyond the protective confines of the MV membrane. This failure of the second stage of mineral formation may be caused by an excess of the mineral inhibitor pyrophosphate (PPi) in the extracellular fluid around MVs. In normal circumstances, PPi is hydrolyzed by the TNAP of MVs' outer membrane yielding monophosphate ions (Pi) for incorporation into bone mineral. Thus, with TNAP deficiency a buildup of mineral-inhibiting PPi would be expected at the perimeter of MVs.
Department of Pathology and Laboratory Medicine, University of Kansas Medical Center, Kansas City, Kansas 66160, USA. handerso@kumc.edu
H Clarke
HC
Anderson
Joseph B
JB
Sipe
Lovisa
L
Hessle
Rama
R
Dhanyamraju
Elisa
E
Atti
Nancy P
NP
Camacho
José Luis
JL
Millán
Rama
R
Dhamyamraju
D000469:Alkaline Phosphatase / Q000172:deficiency*; D000818:Animals; D000880:Anthraquinones; D015519:Bone Density; D001857:Bone Matrix / Q000502:physiology / Q000648:ultrastructure; D002113:Calcification, Physiologic / Q000502:physiology*; D006132:Growth Plate / Q000502:physiology* / Q000648:ultrastructure*; D051379:Mice; D018345:Mice, Knockout; D008854:Microscopy, Electron; D017550:Spectroscopy, Fourier Transform Infrared; D014057:Tomography, X-Ray Computed
D000880:Anthraquinones; C010078:alizarin; D000469:Alkaline Phosphatase