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Sex-Linked Skeletal Phenotype of Lysyl Oxidase Like-1 Mutant Mice

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Abstract

Lysyl oxidases are required for collagen and elastin cross-linking and extracellular matrix maturation including in bone. The lysyl oxidase family consists of lysyl oxidase (LOX) and 4 isoforms (LOXL1-4). Here we investigate whether deletion of LOXL1, which has been linked primarily to elastin maturation, leads to skeletal abnormalities. Left femurs (n = 8), L5 vertebrae (n = 8), and tibiae (n = 8) were analyzed by micro-computed tomography in 13-week-old wild-type (WT) and LOXL1−/− male and female mice. Right femurs (n = 8) were subjected to immunohistochemistry for LOXL1, and histochemical/histology analyses of osteoclasts and growth plates. Sera from all mice were analyzed for bone turnover markers. Results indicate strong expression of LOXL1 in wild-type growth plates in femurs. Significant deterioration of trabecular bone structure in long bones and vertebrae from female was observed but not from male, mutant mice compared with WT. Decreases in BV/TV, Conn.D, trabecular thickness, and number in the femoral distal metaphysis were observed in female, but not in male, mutant mice. Trabecular spacing was increased significantly in femurs of female mutant mice. Findings were similar in trabeculae of L5 vertebrae from female mutant mice. The number of TRAP positive osteoclasts at the trabecular bone surface was increased in female mutant mice compared with WT females, consistent with increased serum RANKL and decreased OPG levels. Analysis of bone turnover markers confirmed increased bone resorption as indicated by significantly elevated CTX-1 in the serum of female LOXL1−/− mice compared to their wild-type counterparts, as well as decreased bone formation as measured by decreased serum levels of PINP. Picrosirius red staining revealed a loss of heterogeneity in collagen organization in female LOXL1−/− mice only, with little to no yellow and orange birefringence. Organization was also impaired in chondrocyte columns in both female and male LOXL1−/− mice, but to a greater extent in females. Data indicate that LOXL1−/− mutant mice develop appendicular and axial skeletal phenotypes characterized by decreased bone volume fraction and compromised trabecular microstructure, predominantly in females.

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Acknowledgments

This study was supported by DE14066 (PCT), and AG028048 (RAW).

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    Authors and Affiliations

    1. Department of Molecular and Cell Biology, Henry M. Goldman School of Dental Medicine, Boston University, 700 Albany Street, W-201, Boston, MA, 02118, USA

      Loai Alsofi, Eileen Daley & Philip C. Trackman

    2. Department of Endodontics, Faculty of Dentistry, King Abdulaziz University, Jeddah, Saudi Arabia

      Loai Alsofi

    3. Division of Dermatology, Washington University School of Medicine, Saint Louis, MO, USA

      Ian Hornstra

    4. Department of Mechanical Engineering, Boston University, 110 Cummington Mall, Boston, MA, 02215, USA

      Elise F. Morgan & Zachary D. Mason

    5. Department of Obstetrics and Gynecology, University of Texas Southwestern Medical Center, Dallas, TX, 75390, USA

      Jesus F. Acevedo & R. Ann Word

    6. Department of Orthopaedic Surgery, Boston University School of Medicine, Boston, MA, 02118, USA

      Louis C. Gerstenfeld

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    Correspondence to Philip C. Trackman.

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    Loai Alsofi, Eileen Daley, Ian Hornstra, Elise F. Morgan, Zachary D. Mason, Jesus F. Acevedo, R. Ann Word, Louis C. Gerstenfeld, and Philip C. Trackman declare no conflicts of interest.

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    The Institutional Animal Care and Use Committees of Boston University Medical Center and of the University of Texas Southwestern Medical Center approved the protocols regarding the use of mice in the reported experiments.

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    Loai Alsofi and Eileen Daley have contributed equally to this study.

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    Alsofi, L., Daley, E., Hornstra, I. et al. Sex-Linked Skeletal Phenotype of Lysyl Oxidase Like-1 Mutant Mice. Calcif Tissue Int 98, 172–185 (2016). https://doi.org/10.1007/s00223-015-0076-4

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