Muscle gene activation by induction and the nonrequirement for cell division

Development ◽  
1986 ◽  
Vol 97 (Supplement) ◽  
pp. 75-84
Author(s):  
J. B. Gurdon ◽  
S. Fairman
Keyword(s):  
Gene Activation ◽  
Cell Types ◽  
Northern Analysis ◽  
Muscle Actin ◽  
S1 Nuclease ◽  
Cardiac Actin ◽  
Gene Transcripts ◽  
Cloned Cdna ◽  
Muscle Gene ◽  
Neurula Stage

In amphibia, as in many other animals with free-swimming larvae, muscle is one of the first differentiated cell types to be formed in early development. In Xenopus, the first contractions of axial body muscle take place about 30 h after fertilization, but genes required to form muscle are activated long before this, during gastrulation. Muscle actin proteins are first seen to be synthesized at the early neurula stage (Sturgess et al. 1980). More recently Mohun et al. (1984), using cloned cDNA probes, have found that cardiac actin, the type of muscle actin characteristic of adult heart, is a major component of the larval axial muscle. Xenopus cardiac actin gene transcripts are detected by S1 nuclease and Northern analysis at the early neurula stage (Mohun et al. 1984), and the use of SP6 probes on poly(A)+ RNA enables cardiac actin transcripts to be seen as early as the midgastrula stage (Cascio & Gurdon, 1986).

scholarly journals A Novel Complex Regulates cardiac actin Gene Expression through Interaction of Emb, a Class VI POU Domain Protein, MEF2D, and the Histone Transacetylase p300

2004 ◽  
Vol 24 (7) ◽  
pp. 2944-2957 ◽  
Author(s):  
S. Molinari ◽  
F. Relaix ◽  
M. Lemonnier ◽  
B. Kirschbaum ◽  
B. Schäfer ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Gene Activation ◽  
Distal Enhancer ◽  
Cardiac Actin ◽  
Pou Domain ◽  
Cell Extracts ◽  
E Box ◽  
Novel Complex ◽  
Domain Protein ◽  
Muscle Gene

ABSTRACT Expression of the mouse cardiac actin gene depends on a distal enhancer (−7 kbp) which has been shown, in transgenic mice, to direct expression to embryonic skeletal muscle. The presence of this distal sequence is also associated with reproducible expression of cardiac actin transgenes. In differentiated skeletal muscle cells, activity of the enhancer is driven by an E box, binding MyoD family members, and by a 3′ AT-rich sequence which is in the location of a DNase I-hypersensitive site. This sequence does not bind MEF2 proteins, or other known muscle transcription factors, directly. Oct1 and Emb, a class VI POU domain protein, bind to consensus sites on the DNA, and it is the binding of Emb which is important for activity. Emb binds as a major complex with MEF2D and the histone transacetylase p300. The form of Emb present in this complex and as a major form in muscle cell extracts is longer (80 kDa) than that previously described. These results demonstrate the importance of this novel complex in the transcriptional regulation of the cardiac actin gene and suggest a potential role in chromatin remodeling associated with muscle gene activation.

Regulation of a human cardiac actin gene introduced into rat L6 myoblasts suggests a defect in their myogenic program

1986 ◽  
Vol 6 (9) ◽  
pp. 3287-3290
Author(s):  
R Hickey ◽  
A Skoultchi ◽  
P Gunning ◽  
L Kedes
Keyword(s):  
Gene Expression ◽  
Cell Line ◽  
Human Gene ◽  
Actin Gene ◽  
Cardiac Actin ◽  
Gene Transcripts ◽  
Actin Mrna ◽  
Muscle Gene ◽  
Alpha Actin ◽  
Myogenic Program

The rat myogenic cell line L6E9 induces skeletal but not cardiac alpha-actin mRNA upon fusion to form myotubes. However, when a human cardiac alpha-actin gene was introduced into L6E9 myoblasts, differentiation of the cells led to the accumulation of human gene transcripts in parallel with those derived from the endogenous skeletal alpha-actin gene. This result demonstrates that factors which direct rat myogenesis can regulate a muscle gene from another species and that the L6E9 cells may have a defect in their ability to activate endogenous cardiac actin gene expression.

scholarly journals Regulation of a human cardiac actin gene introduced into rat L6 myoblasts suggests a defect in their myogenic program.

1986 ◽  
Vol 6 (9) ◽  
pp. 3287-3290 ◽  
Author(s):  
R Hickey ◽  
A Skoultchi ◽  
P Gunning ◽  
L Kedes
Keyword(s):  
Gene Expression ◽  
Cell Line ◽  
Human Gene ◽  
Actin Gene ◽  
Cardiac Actin ◽  
Gene Transcripts ◽  
Actin Mrna ◽  
Muscle Gene ◽  
Alpha Actin ◽  
Myogenic Program

The rat myogenic cell line L6E9 induces skeletal but not cardiac alpha-actin mRNA upon fusion to form myotubes. However, when a human cardiac alpha-actin gene was introduced into L6E9 myoblasts, differentiation of the cells led to the accumulation of human gene transcripts in parallel with those derived from the endogenous skeletal alpha-actin gene. This result demonstrates that factors which direct rat myogenesis can regulate a muscle gene from another species and that the L6E9 cells may have a defect in their ability to activate endogenous cardiac actin gene expression.

Alternative Splicing Is Responsible for the Presence of Two Tissue Factor mRNA Species in LPS Stimulated Human Monocytes

1992 ◽  
Vol 67 (02) ◽  
pp. 272-276 ◽  
Author(s):  
C Paul ◽  
E van der Logt ◽  
Pieter H Reitsma ◽  
Rogier M Bertina
Keyword(s):  
Alternative Splicing ◽  
Tissue Factor ◽  
Stop Codon ◽  
Cell Types ◽  
Northern Analysis ◽  
Human Monocytes ◽  
Mrna Species ◽  
Protein Levels ◽  
Intron 1 ◽  
Tf Gene

SummaryAlthough normally absent from the surface of all circulating cell types, tissue factor (TF) can be induced to appear on circulating monocytes by stimulants like bacterial lipopolysaccharide (LPS) and phorbolesters. Northern analysis of RNA isolated from LPS stimulated human monocytes demonstrates the presence of 2.2 kb and 3.1 kb TF mRNA species. The 2.2 kb message codes for the TF protein. As demonstrated by Northern blot analysis with a variety of TF gene probes, the 3.1 kb message arises from an alternative splicing process which fails to remove 955 bp from intron 1. Because of a stop codon in intron 1 no TF protein is produced from the 3.1 kb transcript. This larger transcript should therefore not be taken into account when comparing TF gene transcription and TF protein levels.

Muscle gene activation in Xenopus requires intercellular communication during gastrula as well as blastula stages

Development ◽  
1992 ◽  
Vol 116 (Supplement) ◽  
pp. 137-142 ◽  
Author(s):  
J. B. Gurdon ◽  
K. Kao ◽  
K. Kato ◽  
N. D. Hopwood
Keyword(s):  
Protein Synthesis ◽  
Developmental Stage ◽  
Cell Transplantation ◽  
Intercellular Communication ◽  
Gene Activation ◽  
Gastrula Stage ◽  
Blastula Stage ◽  
Morphological Marker ◽  
Muscle Gene ◽  
Early Gastrula

In Xenopus an early morphological marker of mesodermal induction is the elongation of the mesoderm at the early gastrula stage (Symes and Smith, 1987). We show here that the elongation of equatorial (marginal) tissue is dependent on protein synthesis in a mid blastula, but has become independent of it by the late blastula stage. In animal caps induced to become mesoderm, the time when protein synthesis is required for subsequent elongation immediately follows the time of induction, and is not related to developmental stage. For elongation, intercellular communication during the blastula stage is of primary importance. Current experiments involving cell transplantation indicate a need for further celhcell interactions during gastrulation, and therefore after the vegetal-animal induction during blastula stages. These secondary cell interactions are believed to take place among cells that have already received a vegetal induction, and may facilitate some of the later intracellular events known to accompany muscle gene activation.

The Florey Lecture, 1988 - From egg to embryo: the initiation of cell differentiation in Amphibia

1989 ◽  
Vol 237 (1286) ◽  
pp. 11-25 ◽  
Keyword(s):  
Muscle Cell ◽  
Gene Activation ◽  
Cell Formation ◽  
Gene Promoter ◽  
Cell Types ◽  
Specific Gene ◽  
Embryonic Induction ◽  
Differentiated Cells ◽  
Wide Range ◽  
Muscle Genes

Some of the principles by which different cell types first arise at the beginning of animal development are illustrated by muscle cell formation in Amphibia. If the nucleus of a differentiated muscle cell is transplanted to an enucleated egg, some of the resulting embryos develop into tadpoles with a wide range of normally differentiated cells. These experiments show that genes undergo major changes in activity as a response to components of egg cytoplasm. Two fundamental mechanisms account for the regional activation of genes in early embryos. One involves the effect of localized ‘determinants’ in egg cytoplasm, and the other concerns cell interactions or embryonic induction. Both these mechanisms seem to be responsible for muscle cell formation in amphibian development. The old problem of embryonic induction has recently become accessible to analysis at the molecular level, especially in the case of the mesoderm or muscle-forming induction. This has been greatly facilitated by using a sensitive and quantitative assay to detect the first transcripts of muscle genes a few hours after the start of induction. The role of early events and of interactions among like cells during response to induction is discussed. In analysing specific gene activation following induction, DNA injection into fertilized eggs has shown that a very small part of the cardiac actin gene promoter is sufficient to enable it to respond to induction. Although the experimental work summarized here has been done on amphibian embryos, which are more suitable than other embryos for embryological manipulation, the conclusions reached are believed to be generally applicable to the development of other organisms.

scholarly journals Nonviral gene editing via CRISPR/Cas9 delivery by membrane-disruptive and endosomolytic helical polypeptide

2018 ◽  
Vol 115 (19) ◽  
pp. 4903-4908 ◽  
Author(s):  
Hong-Xia Wang ◽  
Ziyuan Song ◽  
Yeh-Hsing Lao ◽  
Xin Xu ◽  
Jing Gong ◽  
...  
Keyword(s):  
Gene Editing ◽  
Transfection Efficiency ◽  
Gene Activation ◽  
Cell Types ◽  
Biological Research ◽  
Knock Out ◽  
Safe Delivery ◽  
Pegylated Nanoparticles

Effective and safe delivery of the CRISPR/Cas9 gene-editing elements remains a challenge. Here we report the development of PEGylated nanoparticles (named P-HNPs) based on the cationic α-helical polypeptide poly(γ-4-((2-(piperidin-1-yl)ethyl)aminomethyl)benzyl-l-glutamate) for the delivery of Cas9 expression plasmid and sgRNA to various cell types and gene-editing scenarios. The cell-penetrating α-helical polypeptide enhanced cellular uptake and promoted escape of pCas9 and/or sgRNA from the endosome and transport into the nucleus. The colloidally stable P-HNPs achieved a Cas9 transfection efficiency up to 60% and sgRNA uptake efficiency of 67.4%, representing an improvement over existing polycation-based gene delivery systems. After performing single or multiplex gene editing with an efficiency up to 47.3% in vitro, we demonstrated that P-HNPs delivering Cas9 plasmid/sgRNA targeting the polo-like kinase 1 (Plk1) gene achieved 35% gene deletion in HeLa tumor tissue to reduce the Plk1 protein level by 66.7%, thereby suppressing the tumor growth by >71% and prolonging the animal survival rate to 60% within 60 days. Capable of delivering Cas9 plasmids to various cell types to achieve multiplex gene knock-out, gene knock-in, and gene activation in vitro and in vivo, the P-HNP system offers a versatile gene-editing platform for biological research and therapeutic applications.

Actin genes in Xenopus and their developmental control

Development ◽  
1985 ◽  
Vol 89 (Supplement) ◽  
pp. 125-136
Author(s):  
J. B. Gurdon ◽  
T. J. Mohun ◽  
S. Brennan ◽  
S. Cascio
Keyword(s):  
Skeletal Muscle ◽  
Early Development ◽  
Actin Gene ◽  
Muscle Actin ◽  
Cell Type ◽  
Developmental Control ◽  
Normal Contact ◽  
Actin Genes ◽  
Cardiac Actin ◽  
Cell Type Specific

The results summarized here have established the temporal and regional activation of three kinds of Xenopus actin genes. The cardiac and skeletal muscle actin genes are among the first cell-type-specific genes to be expressed in early development. The first transcripts to be synthesized by these genes appear to be correctly initiated, spliced, and at once translated into proteins. Both cardiac and skeletal actin genes are strongly transcribed in the axial skeletal muscle of embryos. The mechanism by which the cardiac actin gene is first transcribed in only the somite region of an embryo depends, at least in part, on materials already localized in the subequatorial region of a fertilized but uncleaved egg. Cells which acquire this material seem able to activate their cardiac actin genes without requiring normal contact with other cells.

A role for cytoplasmic determinants in mesoderm patterning: cell-autonomous activation of the goosecoid and Xwnt-8 genes along the dorsoventral axis of early Xenopus embryos

Development ◽  
1994 ◽  
Vol 120 (5) ◽  
pp. 1191-1199 ◽  
Author(s):  
P. Lemaire ◽  
J.B. Gurdon
Keyword(s):  
Gene Activation ◽  
Cell Communication ◽  
Cell Types ◽  
Mesoderm Induction ◽  
Mesoderm Cell ◽  
Xenopus Embryos ◽  
Cell Dispersion ◽  
Embryo Axes ◽  
Alpha Actin ◽  
Cytoplasmic Determinants

Although an induction event is required for the formation of mesoderm in Xenopus embryos, it is not clear that this induction is wholly sufficient to give rise to a correctly patterned mesodermal layer. We have studied the expression of the two genes, goosecoid and Xwnt-8, in Xenopus gastrulae in which cell-cell communication, and therefore mesoderm induction, has been prevented by frequent cell dispersion. Although neither the early panmesodermal marker Xbra nor the muscle-specific alpha-actin gene were activated under these conditions, goosecoid and Xwnt-8 were activated in cells of dorsal and ventrolateral origin respectively, thus correctly reflecting their distribution during normal development. We also show that the spatial pattern of expression of these two genes along the animal-vegetal axis is similar in normal and in dissociated early gastrulae: goosecoid is mainly expressed in future mesoderm while the domain of expression of Xwnt-8 spans the mesoderm-endoderm boundary. These results show that, during the blastula and early gastrula stages, gene activation can be controlled cell-autonomously along both the animal-vegetal and dorsoventral embryo axes. This suggests that the inheritance of localised maternal cytoplasmic determinants is a key event for the patterning of mesoderm. We present a modified model of mesoderm formation in which the different mesoderm cell types are produced as a result of cooperation between induction-dependent and induction-independent immediate-early genes.

Renal expression of the gene for atrial natriuretic factor

1992 ◽  
Vol 263 (5) ◽  
pp. F974-F978
Author(s):  
J. E. Greenwald ◽  
D. Ritter ◽  
E. Tetens ◽  
P. S. Rotwein
Keyword(s):  
Epithelial Cells ◽  
Atrial Natriuretic Factor ◽  
Rat Kidney ◽  
Pcr Amplification ◽  
Northern Analysis ◽  
Mammalian Kidney ◽  
Natriuretic Factor ◽  
Normal Rat ◽  
Gene Transcripts ◽  
Atrial Natriuretic

To date, atrial natriuretic factor (ANF) mRNA has eluded detection in the mammalian kidney, although we and others have identified ANF protein in the kidney using immunohistochemical and immunoassay techniques. Furthermore, we have demonstrated the synthesis and secretion of the ANF prohormone in the distal cortical nephron of the intact rat kidney and from rat primary cultured renal distal cortical tubular epithelial cells. In the present study, we show that the ANF gene is expressed in the kidney. Amplification of RNA isolated from rat distal cortical tubular epithelial cultures using ANF specific primers produced a 213-bp fragment that specifically hybridized to a 32P-labeled ANF cDNA. We had previously demonstrated these cultures to be enriched for the renal ANF synthetic and secretory cell type. However, we were unable to detect an ANF gene transcript in total rat kidney RNA using the above-mentioned polymerase chain reaction (PCR) conditions. Reanalysis of normal rat kidney PCR products by a second round of PCR amplification using nested primers successfully identified ANF mRNA. Similar to cultured kidney epithelial cells, normal rat kidney expresses ANF mRNA, but at a very low abundance, thus necessitating two rounds of PCR amplification. Further characterization of rat cortical distal tubular epithelia poly(A)+ RNA by Northern analysis revealed two ANF gene transcripts. A 1.0-kb message that comigrated with rat atrial ANF mRNA, and a second larger 1.4-kb transcript. These studies further substantiate the synthesis of ANF in the mammalian kidney. Unlike the mammalian heart, the kidney contains two ANF gene transcripts.

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