Mouse muscle identity: The position‐dependent and fast fiber‐specific expression of a transgene in limb muscles is methylation‐independent and cell‐autonomous

• Published in: Developmental dynamics Vol. 228; no. 4; pp. 594 - 605
• Main Authors: Ngô‐Muller, Valerie, Bertrand, Anne, Concordet, Jean‐Paul, Daegelen, Dominique
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 01.12.2003; Wiley
• Subjects: aldolase A; Amino Acid Motifs; Animals; Cells, Cultured; Chromatin - metabolism; CpG Islands; DNA Methylation; Extremities - physiology; Female; Fructose-Bisphosphate Aldolase - metabolism; Hindlimb - metabolism; Life Sciences; Methylation; Mice; Mice, Transgenic; Muscle Fibers, Fast-Twitch - physiology; muscle identity; Muscles - metabolism; Muscles - physiology; Mutation; positional information; Promoter Regions, Genetic; Tissue Transplantation; Transcriptional Activation; Transfection; Transgenes; muscle identity positional information aldolase A; positional information; aldolase A; muscle identity
• ISSN: 1058-8388; 1097-0177
• DOI: 10.1002/dvdy.10402

We previously characterised transgenic mice in which fast‐muscle‐specific regulatory sequences from the human aldolase A pM promoter drive the chloramphenicol acetyltransferase gene expression. Mutation of a NF1/MEF2 binding site (M2 motif) in this promoter does not affect fibre‐type specificity of the transgene but modifies its expression in a subset of fast‐twitch fibres at the limb level, preferentially affecting distal limb muscles. We investigated the molecular and cellular bases of this peculiar expression pattern that provided an adequate model to characterise the mechanisms responsible for muscle positional information. By direct electrotransfer of mutated M2 construct in adult muscle, we demonstrate that positional differences in mutated M2 transgene expression are not observed when the transgene is not integrated into chromatin. Also, this transgene expression pattern does not seem to be correlated with the extent of CpG methylation in its promoter sequence. Finally, we show that positional values reflected by CAT levels are maintained in primary cultures established from different adult limb muscles, as well as in heterotopically transplanted muscles. Our results suggest that mutation of the M2 site contributes to reveal a molecular memory of fibre fate that would be set up on pM promoter during development and persist into adulthood possibly through a chromatin imprint maintained in satellite cells associated with various limb muscles. Developmental Dynamics 228:594–605, 2003. © 2003 Wiley‐Liss, Inc.