Gaillard C et al. (1999), Differential expression of two skeletal muscle ...

XB-ART-13066

Int J Dev Biol 1999 Mar 01;432:175-8.

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Differential expression of two skeletal muscle beta-tropomyosin mRNAs during Xenopus laevis development.

Gaillard C , Lerivray H , Thézé N , Cooper B , Lepetit D , Mohun T , Thiébaud P .

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A cDNA clone for a Xenopus laevis skeletal muscle beta-tropomyosin (beta-TMad) isoform was isolated from an adult skeletal muscle cDNA library. Sequence analysis revealed that this clone corresponded to a second beta-tropomyosin mRNA distinct from the one that was previously characterized (beta-TMemb). The two skeletal beta-TM mRNAs originate from distinct genes and are differentially expressed during development. Beta-TMemb mRNA is expressed only in the somites of the early embryo while beta-TMad mRNA is expressed in pre-metamorphic tadpoles and adult skeletal muscles. We have isolated the promoter region of the beta-TMemb gene and shown that a DNA construct containing 2.9 kb of promoter region is properly expressed after injection in the embryo.

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Species referenced: Xenopus laevis
Genes referenced: cat.2 mt-tr tpm2 trna

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	Fig. 2. Expression of b-TMemb and b-TMad transcripts in embryos and adult tissues. (A) The distribution of b-TMemb and b-TMad transcripts was analyzed by RT-PCR. RNA was from eggs (Eg), different staged embryos (22-48), body (B) and tail (T) muscles of pre-metamorphic stage 57 tadpoles, adult skeletal muscle (Sk) and stomach (St). tRNA (-) was used as control. (B)Whole-mount in situ analysis of b-TMemb transcripts. Embryos were stained at stage 25, 28 and 35. (C) In situ hybridization on transverse sections of a stage 25 (upper) and stage 35 (lower) embryo.
	tpm2 (tropomyosin 2 (beta)) gene expression in Xenopus laevis embryos, NF stage 35, as assayed by in situ hybridization. Lateral view: anterior left, dorsal up.
	Fig. 3. Xenopus laevis b-TMemb gene 5' structure and transcriptional activation of the promoter region in the embryo. (A) Schematic representation of the 13 kb genomic clone. Exons are figured by black boxes and 5'UTR by an open box. Major restriction enzymes sites are BamHI (B), EcoRI (E), HindIII (H) and KpnI (K). (B) RNase mapping of the transcription start site. Total RNA from eggs (Eg), skeletal muscle (Sk), stomach (St), oviduct (Ov), embryo (Emb) or tRNA (-) were analyzed as described in Experimental Procedures. Similar size major products of 69 and 74-77 nt are detected in embryo and smooth muscle tissues. (C) CAT activity detection in Xenopus oocytes and embryos injected with the 2.9 kb b-TM/CAT plasmid construction. Embryos are from stage 12 (12), 18 (18), 24 (24) 28 (28), and ventral (V) or dorsal (D) part of a stage 28 embryo. The promoterless plasmid PBCOCAT was used as control (C). The migration of the acetylated forms of chloramphenicol (CM) is indicated by arrows.