From gene activity to 3D cell structure and protein-protein interactions in the joint interzone cells

Joints are essential for the integrity and movement of our skeleton.

During embryological development the formation of the joint starts with the interzone, which, together with the surrounding tissue, forms all components of the adult joint.

Molecular markers of the early interzone formation are associated with congenital and age-related joint diseases.In this project I will compare three models of zebrafish joint development: disrupted joint development resulting in joint loss, caused by mutation in nkx3.2 gene; a delayed joint development, which is later recovered, caused by a deletion of the regulatory sequence of nkx3.2 gene; and normal joint development.

I will investigate heterogeneity in the joint interzone cell layers by combining single-cell RNA sequencing and spatial analysis of interzone cells.

I will further identify key molecular differences during interzone formation between my three models. 3D cell organization, 3D morphology, and subcellular structures of the identified interzone cell clusters will be investigated by using revolutionary genetically-tagged enzymes in combination with X-ray synchrotron microtomography and advanced electron microscopy imaging.

I will also investigate interacting proteins and signalling pathways in identified interzone cell clusters.This project will generate new ground-breaking discoveries of the cellular and molecular mechanisms during normal and altered joint development that have important biomedical implications.