Induction of avian cardiac myogenesis by anterior endoderm

Development ◽  
1995 ◽  
Vol 121 (12) ◽  
pp. 4203-4214 ◽  
Author(s):  
T.M. Schultheiss ◽  
S. Xydas ◽  
A.B. Lassar
Keyword(s):  
Cardiac Muscle ◽  
Cardiac Myocytes ◽  
Cardiac Myocyte ◽  
Sequence Similarity ◽  
Experimental System ◽  
Primitive Streak ◽  
Fate Map ◽  
Cardiac Myogenesis ◽  
Stage 4

An experimental system was devised to study the mechanisms by which cells become committed to the cardiac myocyte lineage during avian development. Chick tissues from outside the fate map of the heart (in the posterior primitive streak (PPS) of a Hamburger & Hamilton stage 4 embryo) were combined with potential inducing tissues from quail embryos and cultured in vitro. Species-specific RT-PCR was employed to detect the appearance of the cardiac muscle markers chick Nkx-2.5 (cNkx-2.5), cardiac troponin C and ventricular myosin heavy chain in the chick responder tissues. Using this procedure, we found that stage 4–5 anterior lateral (AL) endoderm and anterior central (AC) mesendoderm, but not AL mesoderm or posterior lateral mesendoderm, induced cells of the PPS to differentiate as cardiac myocytes. Induction of cardiogenesis was accompanied by a marked decrease in the expression of rho-globin, implying that PPS cells were being induced by anterior endoderm to become cardiac myocytes instead of blood-forming tissue. These results suggest that anterior endoderm contains signaling molecules that can induce cardiac myocyte specification of early primitive streak cells. One of the cardiac muscle markers induced by anterior endoderm, cNkx-2.5, is here described for the first time. cNkx-2.5 is a chick homeobox-containing gene that shares extensive sequence similarity with the Drosophila gene tinman, which is required for Drosophila heart formation. The mesodermal component of cNkx-2.5 expression from stage 5 onward, as determined by in situ hybridization, is strikingly in accord with the fate map of the avian heart. By the time the myocardium and endocardium form distinct layers, cNkx-2.5 is found only in the myocardium. cNkx-2.5 thus appears to be the earliest described marker of avian mesoderm fated to give rise to cardiac muscle.

scholarly journals Tissue engineering of functional cardiac muscle: molecular, structural, and electrophysiological studies

2001 ◽  
Vol 280 (1) ◽  
pp. H168-H178 ◽  
Author(s):  
M. Papadaki ◽  
N. Bursac ◽  
R. Langer ◽  
J. Merok ◽  
G. Vunjak-Novakovic ◽  
...  
Keyword(s):  
Cardiac Muscle ◽  
Cardiac Myocytes ◽  
Conduction Velocity ◽  
Capture Rate ◽  
Signal Amplitude ◽  
Model System ◽  
Neonatal Rat ◽  
Excitation Threshold ◽  
Electrophysiological Properties

The primary aim of this study was to relate molecular and structural properties of in vitro reconstructed cardiac muscle with its electrophysiological function using an in vitro model system based on neonatal rat cardiac myocytes, three-dimensional polymeric scaffolds, and bioreactors. After 1 wk of cultivation, we found that engineered cardiac muscle contained a 120- to 160-μm-thick peripheral region with cardiac myocytes that were electrically connected through gap junctions and sustained macroscopically continuous impulse propagation over a distance of 5 mm. Molecular, structural, and electrophysiological properties were found to be interrelated and depended on specific model system parameters such as the tissue culture substrate, bioreactor, and culture medium. Native tissue and the best experimental group (engineered cardiac muscle cultivated using laminin-coated scaffolds, rotating bioreactors, and low-serum medium) were comparable with respect to the conduction velocity of propagated electrical impulses and spatial distribution of connexin43. Furthermore, the structural and electrophysiological properties of the engineered cardiac muscle, such as cellularity, conduction velocity, maximum signal amplitude, capture rate, and excitation threshold, were significantly improved compared with our previous studies.

Induction of cardiac myogenesis in avian pregastrula epiblast: the role of the hypoblast and activin

Development ◽  
1997 ◽  
Vol 124 (13) ◽  
pp. 2561-2570 ◽  
Author(s):  
T.A. Yatskievych ◽  
A.N. Ladd ◽  
P.B. Antin
Keyword(s):  
Heart Development ◽  
Heart Muscle ◽  
Primitive Streak ◽  
Myocardial Cell ◽  
Posterior Region ◽  
Cell Specification ◽  
Potent Inducer ◽  
Vitro Assay ◽  
Cardiac Myogenesis ◽  
Stage 4

An in vitro assay has been developed to investigate tissue interactions regulating myocardial cell specification in birds. Explants from the posterior region of stage XI-XIV blastulas were found to form heart muscle at high frequency with a timing that corresponded to onset of cardiac myocyte differentiation in vivo. Isolation and recombination experiments demonstrated that a signal from the hypoblast was required to induce cardiac myogenesis in the epiblast, and regional differences in epiblast responsiveness and hypoblast inductiveness restrict appearance of cardiac myocytes to the posterior region. Explantation studies provided evidence that myocardial cell specification is underway by stage 3, indicating that the hypoblast-derived signal occurs shortly before specification is detected. Recombinations were also performed to compare cardiac-inducing capacities of pregastrula hypoblast and stage 5 anterior lateral endoderm. The hypoblast possessed broad capacity to induce heart muscle cells in pregastrula and mid-gastrula epiblast, and modest ability to induce cardiac myogenesis in stage 4 posterior primitive streak. Stage 5 anterior lateral endoderm, in contrast, showed no ability to induce heart development in epiblast cells but was a potent inducer of cardiac myogenesis in cells from stage 4 posterior primitive streak. These findings suggest that the hypoblast-derived signal likely acts upstream of proposed heart-inducing signals provided by anterior lateral endoderm. Experiments were also performed to investigate whether activin, or an activin-like molecule, is involved in regulating cardiac myogenesis. Follistatin blocked cardiac myogenesis in stage XI-XIV posterior region explants and activin induced cardiac myogenesis in a dose-dependent fashion in posterior epiblast. These findings indicate that activin, or an activin-like molecule, is required for and is sufficient to stimulate cardiac myogenesis in posterior region pregastrula epiblast. Three models are presented to explain these results.

scholarly journals Cellular Physiology of Rat Cardiac Myocytes in Cardiac Fibrosis: In Vitro Simulation Using the Cardiac Myocyte/Cardiac Non-Myocyte Co-Culture System

2008 ◽  
Vol 31 (4) ◽  
pp. 693-706 ◽  
Author(s):  
Keiichi Ikeda ◽  
Katsuyoshi Tojo ◽  
Takashi Udagawa ◽  
Chikara Otsubo ◽  
Masahiro Ishikawa ◽  
...  
Keyword(s):  
Cardiac Myocytes ◽  
Culture System ◽  
Cardiac Fibrosis ◽  
Cardiac Myocyte ◽  
Cellular Physiology ◽  
In Vitro Simulation ◽  
Rat Cardiac Myocytes

Confocal and Electron Microscopy of cultured cardiac myocytes

1993 ◽  
Vol 51 ◽  
pp. 364-365
Author(s):  
D.G. Simpson ◽  
R.L. Price ◽  
M. Terracio ◽  
L. Terracio ◽  
T.K. Borg
Keyword(s):  
Cardiac Myocytes ◽  
Heart Development ◽  
Cardiac Myocyte ◽  
Striated Muscle ◽  
Intercellular Junctions ◽  
Membrane Receptors ◽  
Cell Junctions ◽  
The Many

Early in heart development cardiac myocytes are spherical in shape, intercellular junctions are distributed at irregular intervals around the periphery of the cell, and myofibrillar organization is essentially random. As myocytes mature, they undergo extensive morphogenesis during which the phenotype changes to a tubular rodlike shape, cell junctions congregate at the distal ends of cells to form intercalated disks, and myofibrils become organized in parallel arrays typical of striated muscle. Although not fully understood, it is known that these changes are a result of interactive processes between intracellular components of the cytoskeleton, integrin membrane receptors, and the extracellular matrix (ECM).In vivo studies on the process of cardiac myocyte maturation and myofibrillogenesis are difficult because of the complex biochemical environment of the intact animal and the many extra- and intracellular interactions which are required for proper development and myofibrillogenesis. Unfortunately, in previously available in vitro modelling systems, isolated myocytes spread out over the culture substratum, assume a stellate nonpolar shape, and myofibril organization remains essentially random.

Abstract 127: Phosphorylation of Serum Responsive Factor by p90 Ribosomal S6 Kinase 3 Promotes Concentric Growth of Cardiac Myocytes

2016 ◽  
Vol 119 (suppl_1) ◽  
Author(s):  
Jinliang Li ◽  
Eliana C Martinez Valencia ◽  
Catherine L Passariello ◽  
Hrishikesh Thakur ◽  
Michael Kapiloff
Keyword(s):  
Cardiac Myocytes ◽  
Cardiac Myocyte ◽  
Pressure Overload ◽  
Post Translational Modification ◽  
S6 Kinase ◽  
Knock Out ◽  
Gene Therapy Vector ◽  
P90 Ribosomal S6 Kinase ◽  
Ribosomal S6 Kinase

p90 Ribosomal S6 Kinase 3 (RSK3) is required for the induction of concentric hypertrophy in hearts subjected to pressure overload and catecholamine infusion, as well as in a model for Noonan Syndrome-associated hypertrophic cardiomyopathy. Serving important roles in both cardiac development and adult function, the transcription factor Serum Responsive Factor (SRF) regulates genes expression involved in myocyte growth and sarcomeric assembly. It has been reported that SRF can be phosphorylated by RSK protein kinases on residue Ser-103, but the function of that post-translational modification has remained uncertain. We now show that SRF is a substrate for RSK3 associated with muscle A-Kinase-Anchoring-Protein (mAKAP) scaffold in cardiac myocytes, contributing to the regulation of concentric myocyte growth. By co-immunoprecipitation assay, SRF and RSK3 were both associated with the mAKAP scaffold in heart extracts, such that a ternary complex could be detected when recombinant proteins were expressed in cells. Silencing RSK3 or mAKAP expression by siRNA transfection reduced phenylephrine-induced SRF(S103) phosphorylation in neonatal cardiac myocytes. Similar results were acquired following expression of a peptide comprising the mAKAP RSK3-binding domain (RBD) that competed the binding of the kinase to the scaffold. Overexpression of either a SRF S103A phosphoablative mutant or the RBD peptide diminished the increase in width of adult cardiac myocytes stimulated by phenylephrine. In contrast, overexpression of RSK3 or a SRF S103D phosphomimetic mutant promoted adult cardiac myocyte concentric hypertorphy in vitro. While baseline SRF S103 phosphorylation in the heart was inhibited by mAKAP or RSK3 gene knock-out in mice, acute transverse aortic constriction significantly increased SRF S103 phosphorylation in the heart. Based upon these results, we porpose that RSK3 phosphorylation of SRF at mAKAP signalosomes is an important regulator of concentric cardiac myocyte growth. These results are consistent with additional findings that an adeno-associated virus gene therapy vector that expresses RBD in the cardiac mycoyte attenuates pathological hypertrophy and prevents heart failure in response to pressure overload.

scholarly journals Involvement of Cardiotrophin-1 in Cardiac Myocyte-Nonmyocyte Interactions During Hypertrophy of Rat Cardiac Myocytes In Vitro

Circulation ◽  
1999 ◽  
Vol 100 (10) ◽  
pp. 1116-1124 ◽  
Author(s):  
Koichiro Kuwahara ◽  
Yoshihiko Saito ◽  
Masaki Harada ◽  
Masahiro Ishikawa ◽  
Emiko Ogawa ◽  
...  
Keyword(s):  
Cardiac Myocytes ◽  
Cardiac Myocyte ◽  
Rat Cardiac Myocytes

The formation of mesodermal tissues in the mouse embryo during gastrulation and early organogenesis

Development ◽  
1987 ◽  
Vol 99 (1) ◽  
pp. 109-126 ◽  
Author(s):  
P.P. Tam ◽  
R.S. Beddington
Keyword(s):  
Anterior Part ◽  
Primitive Streak ◽  
Paraxial Mesoderm ◽  
Substantial Contribution ◽  
Fate Map ◽  
Extraembryonic Mesoderm ◽  
Lateral Mesoderm ◽  
Embryonic Ectoderm ◽  
Specific Labelling

Orthotopic grafts of [3H]thymidine-labelled cells have been used to demonstrate differences in the normal fate of tissue located adjacent to and in different regions of the primitive streak of 8th day mouse embryos developing in vitro. The posterior streak produces predominantly extraembryonic mesoderm, while the middle portion gives rise to lateral mesoderm and the anterior region generates mostly paraxial mesoderm, gut and notochord. Embryonic ectoderm adjacent to the anterior part of the streak contributes mainly to paraxial mesoderm and neurectoderm. This pattern of colonization is similar to the fate map constructed in primitive-streak-stage chick embryos. Similar grafts between early-somite-stage (9th day) embryos have established that the older primitive streak continues to generate embryonic mesoderm and endoderm, but ceases to make a substantial contribution to extraembryonic mesoderm. Orthotopic grafts and specific labelling of ectodermal cells with wheat germ agglutinin conjugated to colloidal gold (WGA-Au) have been used to analyse the recruitment of cells into the paraxial mesoderm of 8th and 9th day embryos. The continuous addition of primitive-streak-derived cells to the paraxial mesoderm is confirmed and the distribution of labelled cells along the craniocaudal sequence of somites is consistent with some cell mixing occurring within the presomitic mesoderm.

A correlation between the capacity of cavity formation and the subsequent differentiation of teratocarcinoma embryoid body lines

Development ◽  
1982 ◽  
Vol 69 (1) ◽  
pp. 127-140
Author(s):  
Kazuko Uno
Keyword(s):  
Cardiac Muscle ◽  
Embryoid Body ◽  
High Capacity ◽  
Primitive Streak ◽  
Muscle Differentiation ◽  
Cavity Formation ◽  
Short Term ◽  
Mouse Teratocarcinoma

Seven different embryoid body (EB) lines of mouse teratocarcinoma were isolated from a single EB. With regard to each of the lines, a comparison was made of the following developmental properties, including potentiality: (1) cavity formation in a short term intraperitoneal passage, (2) growth in vivo, (3) cardiac muscle differentiation in vitro following intraperitoneal passage and (4) differentiation of solid tumours in vivo. These lines could be divided into three distinct groups withrespect to their capacity for cardiac muscle differentiation. It has been shown that a high capacity for celldifferentiation in vitro correlates well with the capacity for cavity formation of an EB during the in vivo period. This cavity formation was followed by the appearance of primitive-streak-like structures, from which mesodermal cells were subsequently formed.

scholarly journals Pre-Conditioning with CDP-Choline Attenuates Oxidative Stress-Induced Cardiac Myocyte Death in a Hypoxia/Reperfusion Model

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Héctor González-Pacheco ◽  
Aurelio Méndez-Domínguez ◽  
Salomón Hernández ◽  
Rebeca López-Marure ◽  
Maria J. Vazquez-Mellado ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Reperfusion Injury ◽  
Cardiac Myocytes ◽  
Cardiac Myocyte ◽  
Cell Signalling ◽  
Annexin V ◽  
Neonatal Rat ◽  
Ischaemia Reperfusion Injury ◽  
Ischaemia Reperfusion

Background. CDP-choline is a key intermediate in the biosynthesis of phosphatidylcholine, which is an essential component of cellular membranes, and a cell signalling mediator. CDP-choline has been used for the treatment of cerebral ischaemia, showing beneficial effects. However, its potential benefit for the treatment of myocardial ischaemia has not been explored yet.Aim. In the present work, we aimed to evaluate the potential use of CDP-choline as a cardioprotector in anin vitromodel of ischaemia/reperfusion injury.Methods. Neonatal rat cardiac myocytes were isolated and subjected to hypoxia/reperfusion using the coverslip hypoxia model. To evaluate the effect of CDP-choline on oxidative stress-induced reperfusion injury, the cells were incubated with H2O2during reperfusion. The effect of CDP-choline pre- and postconditioning was evaluated using the cell viability MTT assay, and the proportion of apoptotic and necrotic cells was analyzed using the Annexin V determination by flow cytometry.Results. Pre- and postconditioning with 50 mg/mL of CDP-choline induced a significant reduction of cells undergoing apoptosis after hypoxia/reperfusion. Preconditioning with CDP-choline attenuated postreperfusion cell death induced by oxidative stress.Conclusion. CDP-choline administration reduces cell apoptosis induced by oxidative stress after hypoxia/reperfusion of cardiac myocytes. Thus, it has a potential as cardioprotector in ischaemia/reperfusion-injured cardiomyocytes.

A differential display strategy identifies Cryptic, a novel EGF-related gene expressed in the axial and lateral mesoderm during mouse gastrulation

Development ◽  
1997 ◽  
Vol 124 (2) ◽  
pp. 429-442 ◽  
Author(s):  
M.M. Shen ◽  
H. Wang ◽  
P. Leder
Keyword(s):  
Growth Factor ◽  
Differential Display ◽  
Sequence Similarity ◽  
Es Cells ◽  
Embryonic Stem ◽  
Primitive Streak ◽  
Lateral Plate ◽  
New Family ◽  
Lateral Mesoderm

We have developed a differential display screening approach to identify mesoderm-specific genes, relying upon the differentiation of embryonic stem (ES) cells in vitro. Using this strategy, we have isolated a novel murine gene that encodes a secreted molecule containing a variant epidermal growth factor-like (EGF) motif. We named this gene Cryptic, based on its predicted protein sequence similarity with Cripto, which encodes an EGF-related growth factor. Based on their strong sequence similarities, we propose that Cryptic, Cripto, and the Xenopus FRL-1 gene define a new family of growth factor-like molecules, which we name the ‘CFC’ (Cripto, Frl-1, and Cryptic) family. Analysis of Cryptic expression by in situ hybridization shows that it is expressed during gastrulation in two spatial domains that correspond to the axial and lateral mesoderm. In the first domain of expression, Cryptic expression is progressively localized to the anterior primitive streak, the head process, and the node and notochordal plate. In the second domain, Cryptic expression is initially concentrated in the lateral region of the egg cylinder, and is later found circumferentially in the intermediate and lateral plate mesoderm. Furthermore, Cryptic expression can also be detected at the early head-fold stage in the midline neuroectoderm, and consequently is an early marker for the prospective floor plate of the neural tube. Expression of Cryptic ceases at the end of gastrulation, and has not been observed in later embryonic stages or in adult tissues. Thus, Cryptic encodes a putative signaling molecule whose expression suggests potential roles in mesoderm and/or neural patterning during gastrulation.

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