Lunardi A and Dente L (2002), Molecular cloning and expression analysis of dy...

XB-ART-4619

Mech Dev 2002 Dec 01;119 Suppl 1:S49-54. doi: 10.1016/s0925-4773(03)00091-1.

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Molecular cloning and expression analysis of dystroglycan during Xenopus laevis embryogenesis.

Lunardi A , Dente L .

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Dystroglycan is a transmembrane receptor protein that provides a structural linkage between extracellular matrix components and cytoskeletal proteins. It was originally characterized as a member of dystrophin associated protein complex in muscle but, unlike other proteins of this complex, mutations in the dystroglycan gene have not been implicated as a cause of muscular dystrophies. Indeed, dystroglycan is an essential gene, expressed early in development that, if removed in knockout mice, provokes lethal defects before the onset of myogenesis. Dystroglycan is synthesized as a precursor propeptide that is post-translationally cleaved and glycosylated to yield alpha and beta subunits. We have cloned and characterized a cDNA clone, containing the complete coding region of the dystroglycan precursor, from a Xenopus laevis cDNA library. We have performed a spatial and temporal analysis of its expression in X. laevis embryos, using whole-mount in situ hybridization and reverse transcription-polymerase chain reaction analysis. Early expression of dystroglycan in a variety of tissues of different embryological derivation suggests a crucial role in morphogenetic events, especially during central nervous system differentiation.

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Species referenced: Xenopus laevis
Genes referenced: dag1 dmd dmd.2 rho
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	Fig. 3. Expression of X-dg during Xenopus development (A). Whole-mount in situ hybridization was carried out using digoxigenin-labeled cRNA probe. Nieuwkoop-Faber stages of embryogenesis are indicated in higher left corners. Dorsal views, anterior down (A, B, E); postero-lateral view (C); anterior views (D, F, H); lateral views, anterior on the right (G, I). Transverse section of stage 22 (J): arrow head indicates sensorial layer of the ectoderm; parasagittal section of stage 22 (K) shows strong expression in the eye anlage and in the diencephalon/mesencephalon regions. X-dg expression sites at stage 33 (arrow heads in I): central nervous system (black), heart (orange), lens epithelium (turquoise), mandibular, hyoid and branchial arches (yellow and green respectively), myotomes (purple), otic vesicle (red), pronephron (blue). Parasagittal and longitudinal sections of stage 33 showing X-dg transcripts (L) and protein localization (M) in the myotomes. Transverse section of adult skeletal muscle reveals protein localization in the sarcolemma (N).
	Fig. 4. (A) Antero-posterior transverse sections of central nervous system at stage 33. Eye expression at level of ganglion cells layer in the neural retina (G); and in the optic stalk (H). Abbreviations: die, diencephalon; ep, epiphysis; gcl, ganglion cells layer; h, hypothalamus; mes, mesencephalon; os, optic stalk; rho, rhombencephalon; st, stomodeum.
	Fig. 5. (A) Expression of X-dg as detected in dissected brains of stage 48 embryos by whole-mount in situ hybridization. Lateral view, anterior right (A); ventral (B); and dorsal (C) view, anterior up.