Restricted expression of a novel retinoic acid responsive gene during limb bud dorsoventral patterning and endochondral ossification.

Abstract

Using a differential substractive hybridization cloning procedure we have recently identified Stra6 as a novel retinoic acid-induced gene in murine P19 embryonal carcinoma cells. The putative amino acid sequence of Stra6 shows no similarity with previously characterised proteins. We report here the pattern of expression of Stra6 transcripts during mouse limb development as revealed by in situ hybridization. In 8.5-9.0 days post-coitum (dpc) embryos, Stra6 was expressed in the lateral plate mesenchyme prior to limb bud outgrowth. By 9.5 dpc, expression was restricted to the proximal and dorsal forelimb bud mesoderm. Over the next 2 gestational days, Stra6 expression was specific of the dorsal mesoderm of the undifferentiated forelimb and hindlimb buds with the exception of their distal-most region or progress zone. A novel proximal-ventral expression domain appeared, however, by 11.0-11.5 dpc. Stra6 also remained expressed in the flank mesoderm. From 11.5-13.5 dpc, Stra6 expression was restricted to the superficial mesenchyme surrounding the chondrogenic blastemas, and progressively extended until the distal extremities of the limbs upon disappearance of the progress zone. Progressive restriction of Stra6 expression to perichondrium and developing muscles was seen at 13.5-14.5 dpc. Upon the initiation of endochondral ossification (15.5-16.5 dpc), Stra6 expression was limited to the area of perichondrium opposing cells of high metabolic and proliferative activity (the elongation zone). We suggest that Stra6 may play a role in early dorsoventral limb patterning and later in the control of endochondral ossification.