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doi: 10.1242/10.1242/dev.00186

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Impaired intervertebral disc formation in the absence of Jun

Axel Behrens^1,*, Jody Haigh^1,, Fatima Mechta-Grigoriou², Andras Nagy³, Moshe Yaniv² and Erwin F. Wagner^1,

¹ Research Institute of Molecular Pathology (IMP), Dr Bohr-Gasse 7, A-1030 Vienna, Austria
² Institut Pasteur, Unite des Virus Oncogenes, URA1644 du CNRS, 25, Rue du Dr Roux, 75724 Paris Cedex 15, France
³ Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Canada
^* Present address: Mammalian Genetics Laboratory, Cancer Research UK, London Research Institute, Lincoln's Inn Fields Laboratories, 44, Lincoln's Inn Fields, London WC2A 3PX, UK
Present address: Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Canada

View larger version (98K): [in a new window]   Fig. 1. Jun is required for axial skeletogenesis. (A) Skeletal phenotype of Bal1;Jun/ mice. Photograph of an adult Bal1;Jun/ and Junf/f control littermate. Arrowhead indicates scoliosis. (B,C) Alcian Blue/Alizarin Red skeletal preparations of E18.5 Bal1;Jun/-mutant (C) and Junf/f control (B) embryos. Arrow in C indicates fused atlas and axis. (D,E) Dorsal view of Alcian Blue stained E16.5 Bal1;Jun/-mutant (E) and Junf/f control (D) mice. The epidermis and the neural tube were removed to visualise vertebral cartilage. Arrowheads in E indicate fusions of Alcian Blue-positive domains.

View larger version (92K): [in a new window]   Fig. 2. Jun is dispensable for somitogenesis. (A,B) Expression of Jun is detected in the most recently formed somite. (A) Wild-type foetus with four somites; (B) wild-type foetus with seven somites. Arrowheads indicate Jun expression in the somite; arrow in B indicates midline expression of Jun. (C-L) Sox9 (C,D), Col2a1 (E,F), Uncx4.1 (G,H), Myod1 (I,J) and Pax1 (K,L) expression detected by whole-mount in situ hybridisation is comparable between Jun+/+ control (C,E,G,I,K) and Jun-/- (D,F,H,J,L) E12.5 embryos.

View larger version (53K): [in a new window]   Fig. 3. Jun expression in the sclerotome and notochord is required for skeletogenesis. (A,B) Skeletal X-ray analysis of adult human Col2a1;Jun/ (B) and Junf/f control (A) mice. Arrowheads indicate scoliosis. (C,D) Alcian Blue/Alizarin Red skeletal preparations of the thoraxic region of adult Col2a1;Jun/ (D) and Junf/f control (C) mice. (E-H) mRNA in situ analysis of E14.5 Col2a1;Jun/ (F,H) and Junf/f control foetuses (E,G) using a Col2a1 (E,F) and a collagen X (G,H) antisense probe.

View larger version (156K): [in a new window]   Fig. 4. Jun regulates the viability of notochordal cells. (A-D) Histology of sagittal sections of human Col2a1;Jun/ (B,D) and Junf/f control foetuses (A,C) at E13.5 (A,B) and E14.5 (C,D). Arrowheads in D indicate the nucleus pulposus; nc, notochord; pv, prevertebrae; im, intervertebral mesenchyme; vc, vertebral cartilage; np, nucleus pulposus. (E,F) Cell proliferation measured by BrdU incorporation in E13.5 Col2a1;Jun/ (F) and Junf/f control foetuses (E). (G,H) Apoptotic cell death measured by TUNEL staining (green) in E13.5 Col2a1;Jun/ (G) and Junf/f control foetuses (H). Sections were counterstained with propidium iodide (red). Arrows indicate TUNEL-positive cells.