TY - JOUR
T1 - Distribution of myosin heavy chain mRNA in embryonic muscle tissue visualized by ultrastructural in situ hybridization
AU - Pomeroy, Marsha E.
AU - Lawrence, Jeanne Bentley
AU - Singer, Robert H.
AU - Billings-Gagliardi, Susan
N1 - Funding Information:
We thank Gary Langevin, John McNeil, and Carol Johnson for their generous technical assistance. Gary Bassell, Constance Cardasis, Ger-aldine Gauthier, and Fred Silva provided encouraging and helpful discussions of this work. The MHC clone was obtained from Jeff Robbins. Supported by Grant NS 11425f rom the National Institutes of Health to S. Billings-Gagliardi and M. K. Wolf, and by HD 18066 to R. H. Singer and J. B. Lawrence.
PY - 1991/1
Y1 - 1991/1
N2 - We have localized myosin heavy chain (MHC) mRNAs in cells of intact embryonic chick muscle using high resolution in situ hybridization. Blocks of muscle were aldehyde-fixed prior to detergent treatment and hybridized with a biotinated cDNA probe, followed by colloidal gold-labeled antibodies, before embedment. Labeling was determined to represent MHC mRNA by extensive quantitative comparisons of electron micrographs from experimental and four different types of control samples. MHC mRNA was localized primarily to peripheral regions of 14-day chick pectoral muscle cells, where the majority of developing myofibrils were found. MHC mRNAs were consistently associated with the nonmyofibrillar cytoskeletal filaments which had diameters ranging from 4 to 10 nm. They were often oriented parallel to the longitudinal axis of the cell. The resolution of the ultrastructural approach allowed us to demonstrate that the mRNA molecules visualized were not directly associated with myofilaments, suggesting that nascent chains read from those messages do not assemble directly into myofilaments simultaneous with translation.
AB - We have localized myosin heavy chain (MHC) mRNAs in cells of intact embryonic chick muscle using high resolution in situ hybridization. Blocks of muscle were aldehyde-fixed prior to detergent treatment and hybridized with a biotinated cDNA probe, followed by colloidal gold-labeled antibodies, before embedment. Labeling was determined to represent MHC mRNA by extensive quantitative comparisons of electron micrographs from experimental and four different types of control samples. MHC mRNA was localized primarily to peripheral regions of 14-day chick pectoral muscle cells, where the majority of developing myofibrils were found. MHC mRNAs were consistently associated with the nonmyofibrillar cytoskeletal filaments which had diameters ranging from 4 to 10 nm. They were often oriented parallel to the longitudinal axis of the cell. The resolution of the ultrastructural approach allowed us to demonstrate that the mRNA molecules visualized were not directly associated with myofilaments, suggesting that nascent chains read from those messages do not assemble directly into myofilaments simultaneous with translation.
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U2 - 10.1016/0012-1606(91)90054-7
DO - 10.1016/0012-1606(91)90054-7
M3 - Article
C2 - 1985024
AN - SCOPUS:0025957356
SN - 0012-1606
VL - 143
SP - 58
EP - 67
JO - Developmental Biology
JF - Developmental Biology
IS - 1
ER -