Expression of genes encoding the collagen-binding heat shock protein (Hsp47) and type II collagen in developing...

Hsp47 is a heat-shock protein which interacts with newly synthesize procollagen chains in the endoplasmic reticulum (ER) of collagen-secreting cells and is thought to assist in procollagen triple helix assembly and subsequent transport to the cis-Golgi. This is supported by studies which have reported that genes encoding collagen and Hsp47 are subject to co-ordinate increases and decreases in expression in cultured cells. However, limited information is available regarding hsp47 expression in vivo, particularly during early embryonic development when a variety of collagen genes are expressed. Here we show that the zebrafish hsp47 gene is expressed in a dynamic spatiotemporal pattern in developing embryos. Strong expression of hsp47 mRNA is co-incident predominantly with expression of the type II collagen gene (col2a1) in a number of chondrogenic and non-chondrogenic tissues including the notochord, otic vesicle and developing fins. Notochordal expression of both genes is disrupted in floating head (flh) and no tail (ntl) embryos, which lack properly differentiated notochords. Surprisingly, no hsp47 mRNA is detectable in the strongly col2a1-expressing cells of the floor plate and hypochord, indicating that the two genes are not strictly co-regulated. Finally, Northern blot analysis revealed two alternative transcripts of col2a1 which are expressed in distinct temporal patterns. Appearance of the larger transcript occurs following somitogenesis, a time which coincides with the co-activation of hsp47 and col2a1 gene expression in tissues outside of the notochord.