Transcription occurs in pulses in muscle fibers

Newlands, Sarah; Levitt, Linda K.; Robinson, Charles Alexander; Karpf, A.B. Carmen; Hodgson, Vanessa R.M.; Wade, Robert; Hardeman, Edna C.

doi:10.1101/gad.12.17.2748

Cited by 182 publications

(177 citation statements)

References 31 publications

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“…Non-periodic bursts in expression are predicted from mathematical models of transcription together with stationary distributions obtained by flow cytometry (30) and microscopy (31)(32)(33), and are observed in living systems including bacteria (34), during differentiation in Dictyostelium (35), and following DNA damage in mammalian cells (36). Protein levels within a single cell can fluctuate as a result of the stochastic nature of reactions that rely on components present in low copy numbers.…”

Section: Resultsmentioning

confidence: 99%

Tracking lineages of single cells in lines using a microfluidic device

Rowat

Bird²,

Agresti³

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

114

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Cells within a genetically identical population exhibit phenotypic variation that in some cases can persist across multiple generations. However, information about the temporal variation and familial dependence of protein levels remains hidden when studying the population as an ensemble. To correlate phenotypes with the age and genealogy of single cells over time, we developed a microfluidic device that enables us to track multiple lineages in parallel by trapping single cells and constraining them to grow in lines for as many as 8 divisions. To illustrate the utility of this method, we investigate lineages of cells expressing one of 3 naturally regulated proteins, each with a different representative expression behavior. Within lineages deriving from single cells, we observe genealogically related clusters of cells with similar phenotype; cluster sizes vary markedly among the 3 proteins, suggesting that the time scale of phenotypic persistence is protein-specific. Growing lines of cells also allows us to dynamically track temporal fluctuations in protein levels at the same time as pedigree relationships among the cells as they divide in the chambers. We observe bursts in expression levels of the heat shock protein Hsp12-GFP that occur simultaneously in mother and daughter cells. In contrast, the ribosomal protein Rps8b-GFP shows relatively constant levels of expression over time. This method is an essential step toward understanding the time scales of phenotypic variation and correlations in phenotype among single cells within a population.epigenetics ͉ protein expression ͉ single cell assay ͉ yeast

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Section: Resultsmentioning

confidence: 99%

Tracking lineages of single cells in lines using a microfluidic device

Rowat

Bird²,

Agresti³

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

114

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“…The complexity of myonuclear organization is compounded by the pulsatile alternating pattern of transcriptional activity (Newlands et al, 1998). It has been suggested that the organization of myonuclei provides more efficient coverage of the fibre volume (Bruusgaard et al, 2003).…”

Section: Nearest Neighbour Value (Nn)mentioning

confidence: 99%

Effects of aging and gender on the spatial organization of nuclei in single human skeletal muscle cells

et al. 2010

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SummaryThe skeletal muscle fibre is a syncitium where each myonucleus regulates the gene products in a finite volume of the cytoplasm, i.e., the myonuclear domain (MND). We analysed aging-and gender-related effects on myonuclei organization and the MND size in single muscle fibres from six young (21-31 years) and nine old men (72-96 years), and from six young (24-32 years) and nine old women (65-96 years), using a novel image analysis algorithm applied to confocal images. Muscle fibres were classified according to myosin heavy chain (MyHC) isoform expression. Our image analysis algorithm was effective in determining the spatial organization of myonuclei and the distribution of individual MNDs along the single fibre segments. Significant linear relations were observed between MND size and fibre size, irrespective age, gender and MyHC isoform expression. The spatial organization of individual myonuclei, calculated as the distribution of nearest neighbour distances in 3D, and MND size were affected in old age, but changes were dependent on MyHC isoform expression. In type I muscle fibres, average NN-values were lower and showed an increased variability in old age, reflecting an aggregation of myonuclei in old age. Average MND size did not change in old age, but there was an increased MND size variability. In type IIa fibres, average NN-values and MND sizes were lower in old age, reflecting the smaller size of these muscle fibres in old age. It is suggested that these changes have a significant impact on protein synthesis and degradation during the aging process.

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“…However, the cells showed a variable compartmental staining that was not related to the penetration of the Bluo-gal solution. It has been shown in skeletal muscle that the proportion of positive nuclei changes during development and regeneration, and declines upon muscle maturation (Newlands et al, 1998).…”

Section: Discussionmentioning

confidence: 99%

Changing intracellular compartmentalization of β‐galactosidase in the ROSA26 reporter mouse during embryonic development: A light‐ and electron‐microscopic study

Aoyama¹,

Molin²,

Mentink³

et al. 2004

Anat. Rec.

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The beta-geo (LacZ) reporter gene encodes for ␤-galactosidase (␤-gal) in all cells of the ROSA26 mouse during embryonic development. As such, ␤-gal activity constitutes an excellent marker for in situ labeling of expressing cells. However, the intracellular distribution of ␤-gal differs between cells, and changes during embryonic development. Therefore, we studied LacZ-encoded ␤-gal using light and electron microscopy in the heart, lung, liver, and small intestine on days 13 and 16 of gestation, and the kidney on day 16 of gestation in ROSA26 mice. The Bluo-gal method was carried out under standardized conditions, including fixation, washing, and incubation procedures. Intracellular ␤-gal staining is encountered in a combination of membranous compartments, including the nuclear envelope, the endoplasmic reticulum, and the plasma membrane. Its exact localization depends on the cell type and is regulated during development. Therefore, one must take the compartmental transition of intracellular ␤-gal staining into consideration when interpreting results obtained from experiments using ROSA26 mice.

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Transcription occurs in pulses in muscle fibers

Cited by 182 publications

References 31 publications

Tracking lineages of single cells in lines using a microfluidic device

Tracking lineages of single cells in lines using a microfluidic device

Effects of aging and gender on the spatial organization of nuclei in single human skeletal muscle cells

Changing intracellular compartmentalization of β‐galactosidase in the ROSA26 reporter mouse during embryonic development: A light‐ and electron‐microscopic study

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