Aria Myers
Research Mentor: Yuji Mishina
Mentor Department: Biological and Material Sciences, Dentistry
Author(s): Aria Myers, Mylene Pighini, Yuji Mishina
Session: Session 4 (1:00 PM – 1:50 PM)
Presentation Type: Poster 120
Abstract
Bone development and remodeling rely on the coordinated actions of osteoblasts and osteoclasts: osteoclasts resorb bone, and osteoblasts form it. Their direct interaction is critical for sustaining osseous tissue, as demonstrated in studies of long bones. In the tibia, ongoing osteogenesis throughout development supports its roles in leg stabilization and movement, as well as serving as a muscle attachment point. However, the mechanisms by which osteoclasts and osteoblasts communicate to maintain bone tissue in physiological conditions remain unclear. To explore this, we designed a cross-sectional study using tibiae from Col1a1-GFP; LysMCre; Rosa26-tdTomato reporter mice, in which osteoblasts express GFP (green) and osteoclasts express tdTomato (red). We selected two time points—postnatal day 23 and 2 months—to compare the phase of active bone growth with that of established remodeling. After dissection, tibiae were fixed overnight in 4% paraformaldehyde (PFA) at 4°C, cryoprotected in 30% sucrose, embedded in OCT, and cryosectioned into 10 µm-thick cross sections. Confocal microscopy enabled the identification of osteoblasts and osteoclasts. To quantify these cells at the metaphysis-diaphysis junction, we used three approaches: local cell counting, cell density assessment, and spatial distribution analysis. Together, these analyses provide insight into the spatial localization and potential communication of osteoblast and osteoclast populations. Understanding these interactions under physiological conditions will advance our understanding of long-bone homeostasis and development.
