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A large-scale analysis of mRNAs expressed by primary mesenchyme cells of the sea urchin embryo

Xiaodong Zhu1,2, Gregory Mahairas3,*, Michele Illies1, R. Andrew Cameron4, Eric H. Davidson4 and Charles A. Ettensohn1,{ddagger}

1 Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, PA 15213, USA
2 Stowers Institute for Medical Research, Kansas City, MO 64110, USA
3 Department of Molecular Biotechnology, University of Washington School of Medicine, Seattle, WA 98195, USA
4 Division of Biology, California Institute of Technology, Pasadena, CA 91124, USA
* Present address: Institute for Systems Biology, Seattle, WA 98105, USA



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  		Fig. 1. Histograms of maximum ORF lengths. (A) Black bars are plots of the lengths of the longest ORFs represented in 120 segments (average length=800 nucleotides) of sea urchin 3'-UTRs. The segments were obtained from all sea urchin genes in GenBank as of 7/11/2000 for which 3'-UTR sequences >600 nucleotides were available (39 genes). Most of the ORFs are short and none was >350 nucleotides in length. White bars are of the lengths of the longest ORFs in a random sample of 120 ESTs (average length=794 nucleotides) that yielded strong BLAST matches (P<10-7). There is little overlap between this distribution and the one derived from 3'-UTR sequences. (B) Plot of maximum ORF lengths in 120 randomly selected ESTs from the ‘no match’ category (average length=796 nucleotides). Because the cDNA library was directionally cloned, only the longest ORFs in the three positive reading frames (+1, +2, +3) were considered. 20/120 clones (16.7%) contained an ORF equal to or longer than 350 nucleotides.

 


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  		Fig. 2. Whole-mount in situ hybridization analysis. (A) Aristaless. (B) Fibrinogen-related protein (FRP). (C) MSP130-like. (D) C-lectin. (E) NET-7. (F) ERG. (G) Fibronectin. (H) SM50-related. Expression of aristaless, MSP130-like, SM50-related and C-lectin is restricted to PMCs at the mesenchyme blastula stage. FRP is expressed by PMCs and by ectoderm cells at the animal pole. ERG mRNA is expressed by PMCs and non-skeletogenic mesoderm cells in the vegetal plate. At the late gastrula stage, fibronectin mRNA is enriched in PMCs (arrow, G).

 




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