Depletion of annexin A5, annexin A6, and collagen X causes no gross changes in matrix vesicle-mediated mineralization, but lack of collagen X affects hematopoiesis and the Th1/Th2 response.
Numerous biochemical studies have pointed to an essential role of annexin A5 (AnxA5), annexin A6 (AnxA6), and collagen X in matrix vesicle-mediated biomineralization during endochondral ossification and in osteoarthritis. By binding to the extracellular matrix protein collagen X and matrix vesicles, annexins were proposed to anchor matrix vesicles in the extracellular space of hypertrophic chondrocytes to initiate the calcification of cartilage. However, mineralization appears to be normal in mice lacking AnxA5 and AnxA6, whereas collagen X-deficient mice show only subtle alterations in the growth plate organization. We hypothesized that the simultaneous lack of AnxA5, AnxA6, and collagen X in vivo induces more pronounced changes in the growth plate development and the initiation of mineralization. In this study, we generated and analyzed mice deficient for AnxA5, AnxA6, and collagen X. Surprisingly, mice were viable, fertile, and showed no obvious abnormalities. Assessment of growth plate development indicated that the hypertrophic zone was expanded in Col10a1(-/-) and AnxA5(-/-) AnxA6(-/-) Col10a1(-/-) newborns, whereas endochondral ossification and mineralization were not affected in 13-day- and 1-month-old mutants. In peripheral quantitative computed tomography, no changes in the degree of biomineralization were found in femora of 1-month- and 1-year-old mutants even though the diaphyseal circumference was reduced in Col10a1(-/-) and AnxA5(-/-) AnxA6(-/-) Col10a1(-/-) mice. The percentage of naive immature IgM(+) /IgM(+) B cells and peripheral T-helper cells were increased in Col10a1(-/-) and AnxA5(-/-) AnxA6(-/-) Col10a1(-/-) mutants, and activated splenic T cells isolated from Col10a1(-/-) mice secreted elevated levels of IL-4 and GM-CSF. Hence, collagen X is needed for hematopoiesis during endochondral ossification and for the immune response, but the interaction of annexin A5, annexin A6, and collagen X is not essential for physiological calcification of growth plate cartilage. Therefore, annexins and collagen X may rather fulfill functions in growth plate cartilage not directly linked to the mineralization process.
Center for Biochemistry, Medical Faculty, University of Cologne, Cologne, Germany.
Ivan
I
Grskovic
Anna
A
Kutsch
Christian
C
Frie
Gergely
G
Groma
Jacek
J
Stermann
Ursula
U
Schlötzer-Schrehardt
Anja
A
Niehoff
Stephen E
SE
Moss
Sabrina
S
Rosenbaum
Ernst
E
Pöschl
Markus
M
Chmielewski
Gunter
G
Rappl
Hinrich
H
Abken
John F
JF
Bateman
Kathryn Se
KS
Cheah
Mats
M
Paulsson
Bent
B
Brachvogel
D000818:Animals; D000831:Animals, Newborn; D017304:Annexin A5 / Q000172:deficiency* / Q000378:metabolism; D017317:Annexin A6 / Q000172:deficiency* / Q000378:metabolism; D001842:Bone and Bones / Q000473:pathology; D002113:Calcification, Physiologic; D024121:Collagen Type X / Q000172:deficiency* / Q000378:metabolism; D003433:Crosses, Genetic; D005109:Extracellular Matrix / Q000378:metabolism*; D005260:Female; D006132:Growth Plate / Q000378:metabolism / Q000473:pathology; D006410:Hematopoiesis; D006984:Hypertrophy; D008213:Lymphocyte Activation / Q000276:immunology; D008297:Male; D051379:Mice; D008817:Mice, Mutant Strains; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology*; D018417:Th1 Cells / Q000276:immunology; D018418:Th2 Cells / Q000276:immunology
D017304:Annexin A5; D017317:Annexin A6; D024121:Collagen Type X