In situ expression of collagen and proteoglycan genes in notochord and during skeletal development and growth

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Abstract
Cartilage is an important tissue in skeletogenesis, in the growth of long bones, and as a flexible component of the mature skeleton. The extracellular matrix proteins type II collagen and aggrecan comprise 90% of the matrix and are characteristic of cartilage. Type II collagen provides structural integrity to the tissue, while aggrecan confers resiliency. The quantity of type II procollagen is controlled at the level of transcription of mRNA from the COL2A1 gene. In addition, type II procollagen can be expressed in two isoforms by differential splicing of the primary gene transcript, a post-transcript, a post-transcriptional control mechanism. The two mRNAs either include exon 2 (type IIA) or exclude exon 2 (type IIB) which encodes the major portion of the amino (NH2)-propeptide [Ryan and Sandell (1990), J. Biol. Chem., 265:10334–10339]. The aggrecan gene also encodes alternative splice forms that may be developmentally expressed. The regulation of aggrecan splicing or transcription has not been studied in detail. To determine the spatial and temporal patterns of expression of extracellular matrix in the development of cartilage, we have examined the expression of type II collagen and aggrecan during chondrogenesis in the vertebral column and during elongation of a newborn growth plate. Our results indicate that there is a developmental sequence of type II collagen splice form expression during chondrogenesis with type IIA expressed in prechondrocytes and type IIB expressed in chondrocytes. During elongation of the growth plate, mature chondrocytes express type IIB procollagen and then differentiate into hypertrophic chondrocytes and initiate expression of type X collagen. In all cases, aggrecan was coordinately expressed with type IIB procollagen. As cartilage-like proteins have been observed in more primitive structures such as notochord, the expression of type II collagen mRNAs was also examined in the notochordal remnants of the vertebral column. In the notochord, the predominant collagen expressed was the type IIA collagen prechondrocyte isoform. Notochordal cells also expressed mRNAs more characteristic of fibroblasts such as versican and decorin: low expression of type I collagen, type IIB collagen, and aggrecan were observed. © 1994 Wiley-Liss, Inc 1 This Article is a US Government work and, as such, is in the public domain in the United States of America. .

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