(A) Scleraxis expression is localized to tibiotalar
ligament, patellar and Achilles tendons as observed by ex vivo fluorescence in
Scx-GFP transgenic mice. Spontaneous ossification of tibiotalar ligament,
patellar tendon and Achilles tendon progresses slowly in
Scx-Cre:ACVR1R206H mice from 4
through 18 wks, with rapid progression in
Scx-Cre:ACVR1Q207D-Tg
mice from 4 through 8 weeks (B) Micro-CT of
Scx-Cre:ACVR1R206H mice reveals
distinct ossification of the tibiotalar ligament, Achilles tendon, and
periarticular ossification, but no intramuscular ossification.
(C-D)
Scx-Cre:ACVR1Q207D-Tg:Rosa26-YFP
mice injected with CTX (P21, gastrocnemius) develop typical tendon and ligament
ossification, as seen by x-ray (P42) corresponding to areas of
Scleraxis expression via
Scx+YFP fluorescence but
lack evidence of any intramuscular ossification (0/5 transgenic mice injected).
(E) Cryosections of fibular head counterstained for CD45
(magenta) reveal Scleraxis expression (green) at ligamentous insertions (white
arrow) but not in adjacent skeletal muscle (red arrow) in
Scx-GFP transgenic mice. The HO lesions infiltrating the
(F) talonavicular ligament, (G) talonavicular
ligament in higher magnification, and (H) patellar tendon in
Scx-Cre:ACVR1Q207D-Tg;Rosa26-YFP
mice reflect the contribution of Scx+YFP+
cells to nearly all Alcian Blue (AB)-stained hypertrophic chondrocytes and
heterotopic cartilage (white arrows) in these lesions, but essentially no
contribution to osteocytes or mineralized matrix (red arrows) stained with
Alizarin Red (AR), with DAPI as a nuclear counter-stain.