An axial gradient of transgene methylation in murine skeletal muscle

Genomic imprint of rostrocaudal position

Maria J. Donoghue, Bruce Patton, Joshua R. Sanes, John P. Merlie

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

We previously used mice bearing a myosin light chain-chloramphenicol acetyltransferase (MLC1-CAT) transgene to show that adult muscle cells bear a heritable, cell autonomous memory of their rostrocaudal position. CAT mRNA and protein are expressed in a > 100-fold rostrocaudal gradient in skeletal muscles of developing and adult MLC1-CAT mice (Donoghue, M. J., Merlie, J. P., Rosenthal, N. and Sanes, J. R. (1991). Proc. Natl. Acad. Sci. USA 88, 5847-5851; Donoghue, M. J., Alvarez, J. D., Merlie, J. P. and Sanes, J. R. (1991). J. Cell Biol. 115, 423-434). Moreover, both in primary cultures and in myogenic cell lines prepared from individual muscles of these mice, CAT levels reflect the body position from which the myoblasts were derived (Donoghue, M.J., Morris-Valero, R., Johnson, Y.R., Merlie, J.P. and Sanes, J. R. (1992). Cell 69, 67-77). Here, we show that the methylation state of the MLC1-CAT transgene in skeletal muscles is also graded along the rostrocaudal axis: methylation levels decrease and expression levels increase in the order, jaw → neck → chest and forelimb → hindlimb. Methylation levels are also approx. 10-fold higher in rostrally derived than in caudally derived myogenic cell lines, which express low and high levels of CAT, respectively. Within each cell line, undifferentiated cells (myoblasts), which do not express the transgene, and differentiated cells (myotubes), which do, are indistinguishable in methylation state. Thus, differentiation-related changes in transgene expression do not affect position-related levels of transgene methylation. On the other hand, treatment of rostrally derived lines with the demethylating agent, 5-azacytidine, decreases methylation and increases expression of the transgene. Thus, perturbation of methylation affects expression. Taken together, these results suggest that methylation provides a genomic imprint of rostrocaudal body position that may serve as a component of the positional memory that mammalian cells retain into adulthood.

Original languageEnglish (US)
Pages (from-to)1101-1112
Number of pages12
JournalDevelopment
Volume116
Issue number4
StatePublished - Dec 1992
Externally publishedYes

Fingerprint

Transgenes
Methylation
Skeletal Muscle
Myoblasts
Cell Line
Azacitidine
Myosin Light Chains
Chloramphenicol O-Acetyltransferase
Forelimb
Skeletal Muscle Fibers
Hindlimb
Jaw
Muscle Cells
Thorax
Muscles
Messenger RNA
Proteins

Keywords

  • Methylation
  • Muscle
  • Myosin light chain
  • Positional information
  • Rostrocaudal gradient
  • Transgene

ASJC Scopus subject areas

  • Anatomy
  • Cell Biology

Cite this

Donoghue, M. J., Patton, B., Sanes, J. R., & Merlie, J. P. (1992). An axial gradient of transgene methylation in murine skeletal muscle: Genomic imprint of rostrocaudal position. Development, 116(4), 1101-1112.

An axial gradient of transgene methylation in murine skeletal muscle : Genomic imprint of rostrocaudal position. / Donoghue, Maria J.; Patton, Bruce; Sanes, Joshua R.; Merlie, John P.

In: Development, Vol. 116, No. 4, 12.1992, p. 1101-1112.

Research output: Contribution to journalArticle

Donoghue, MJ, Patton, B, Sanes, JR & Merlie, JP 1992, 'An axial gradient of transgene methylation in murine skeletal muscle: Genomic imprint of rostrocaudal position', Development, vol. 116, no. 4, pp. 1101-1112.
Donoghue, Maria J. ; Patton, Bruce ; Sanes, Joshua R. ; Merlie, John P. / An axial gradient of transgene methylation in murine skeletal muscle : Genomic imprint of rostrocaudal position. In: Development. 1992 ; Vol. 116, No. 4. pp. 1101-1112.
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