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Early heart development: dynamics of endocardial cell sorting suggests a common origin with cardiomyocytes.

The myocardial and endocardial cell sorting out processes take place primarily between 19 and 29 hr of development in the avian embryo. This occurs in an apparent rostral to caudal wave through the heart forming region. During heart development considerable uncertainty exists regarding the processes that regulate cell commitments, progressive aggregation, and sorting out of the different precardiac cell populations. The question addressed in this report is whether endocardial and myocardial cells have a common origin or do the endocardial cells arise from a distinct population of cells from within the precardiac mesoderm. These cells then migrate to become localized between the developing myocardium above and the endoderm below. The distribution of preendocardial cells and premyocardial cells has been followed immunohistochemically in quail heart-forming region mesoderm explants from embryos approximately 18 hr in development and incubated for a 24-hr period. Differentiating myocardiocytes were immunostained with anti-N-cadherin and endocardiocytes with QH-1, a monoclonal antibody that recognizes an antigenic determinant on quail endothelial cells. Sparsely localized QH-1 labeled endothelial cells are localized in the stage 5 heart-forming region. These cells are often arranged in a columnar fashion in the mesoderm explants 6 hr after explantation. By 15-22 hr large patches of QH-1 expressing cells are interspersed with the N-cadherin expressing myocardiocytes. A subpopulation of cells express both N-cadherin and QH-1 antigen suggesting that endocardial and myocardial cells may arise from a common precursor population and that N-cadherin regulation may be a mechanism underlying specific cell sorting of these two cell populations during heart development.

Animals↗

Temporally and spatially restricted expression of apolipoprotein J in the developing heart defines discrete stages of valve morphogenesis.

During cardiac valve morphogenesis, a series of interactions between the mesodermal-derived myocardium and the overlying endothelium lead to condensed leaflet structure formation. At the atrioventricular (AV) canal, endocardial cells are transformed by specialized underlying myocardial cells into endocardial cushions, and then remodeled into mitral and tricuspid valves. Aortic and pulmonary valves develop by a similar mechanism in the primitive outflow tract. Few genes exhibit restricted spatiotemporal expression in these critical embryonic structures, thus limiting the clues to the sequence of molecular events necessary for valvulogenesis. Apolipoprotein J (ApoJ), a secreted glycoprotein expressed in a variety of cell types at tissue interfaces, exhibits a highly restricted and dynamic expression pattern in the developing heart. ApoJ transcripts were detected in mice at day 9.0 of gestation in the wall of the developing truncus arteriosus. By day 10, intense signal occurred in a thin layer of myocardial cells adjacent to developing endocardial cushions of both atrioventricular canal and truncus arteriosus. No apoJ mRNA was present in the overlying endocardial cushions until day 13.5 when prevalvular condensation begins. Intense expression occurred in the stromal connective tissue throughout leaflet formation. The highly restricted spatiotemporal expression pattern of apoJ in the developing heart implicates its role in the morphogenesis of the AV canal and outflow tract into cardiac valves.

Animals↗

Fibronectin distribution during early chick embryo heart development.

The distribution of fibronectin (FN) during early stages of chick embryo heart development has been studied by indirect immunofluorescence methods. The cardiac extracellular matrix (cardiac jelly) was almost devoid of FN-positive material throughout the period studied (stages 8-18). Intensely extracellular fluorescent material was only demonstrated at the heart midline and in the dorsal mesocardium. Fluorescence associated with the basal surface of the myocardium was demonstrated first at the time of fusion of the two heart tubes. While the heart remains attached to the embryonic trunk by the dorsal mesocardium, two different myocardial basal zones can be distinguished according to the intensity of fluorescence: an intensely stained dorsal zone and a much less fluorescent ventral zone. The endocardium did not present a strongly fluorescent basement membrane until stage 13. The intensity of fluorescence of the endocardial basal surface varied according to the rostrocaudal levels of the heart and also to the development stage of the embryo. The levels of fluorescence increased in myocardium and endocardium at the onset of trabeculation but decreased as trabeculation was completed. The quantitative and qualitative variations of FN distribution have been associated with a number of developmental events.

Animals↗

The orphan nuclear receptor COUP-TFII is required for angiogenesis and heart development.

The embryonic expression of COUP-TFII, an orphan nuclear receptor, suggests that it may participate in mesenchymal-epithelial interactions required for organogenesis. Targeted deletion of the COUP-TFII gene results in embryonic lethality with defects in angiogenesis and heart development. COUP-TFII mutants are defective in remodeling the primitive capillary plexus into large and small microcapillaries. In the COUP-TFII mutant heart, the atria and sinus venosus fail to develop past the primitive tube stage. Reciprocal interactions between the endothelium and the mesenchyme in the vascular system and heart are essential for normal development of these systems. In fact, the expression of Angiopoietin-1, a proangiogenic soluble factor thought to mediate the mesenchymal-endothelial interactions during heart development and vascular remodeling, is down-regulated in COUP-TFII mutants. This down-regulation suggests that COUP-TFII may be required for bidirectional signaling between the endothelial and mesenchymal compartments essential for proper angiogenesis and heart development.

Animals↗

Coxsackievirus B3 infection in pregnancy and its influence on foetal heart development.

Infection of mice on the 12th or 14th day of pregnancy with Coxsackievirus B3 resulted in the birth of growth-retarded young which died soon after birth and exhibited an abnormal heart development. The ratio of heart weight to body weight in these offspring was higher than normal. The auricles were prominent and the ventricles developed such that the heart apex was bifid in appearance. This anomalous cardiac development may have been due to a direct viral pathogenicity in the developing tissue or, as seems more likely, resulted from a generalized disturbance in foetal growth attributable to a virus-induced pancreatic insufficiency in the mother. Retarded development in the pulmonary system also resulting from aberrations in foetal growth may have been contributory to impaired postnatal cardiac growth.

Animals↗

Inhibitory effects of ouabain on early heart development and cardiomyogenesis in the chick embryo.

Pericardial cavity formation and epithelialization of the cardiac precursor cell population constitute a critical developmental period that precedes stable cardiac cell commitment and differentiation. These events delineate the myocardial and endocardial precursor population in the embryo. Restriction of Na/K-ATPase (the sodium pump) expression to the pre-cardiomyocyte lateral membranes coincides with these events. Na/K-ATPase has been implicated developmentally in cavitation and in maintaining membrane potential. Experiments were undertaken to determine if the effects of perturbing sodium pump activity will affect pericardial cavity formation and, in turn, whether heart formation and/or cardiac cell commitment will be affected. We incubated whole chick embryos in vitro between stages 5 to 8 in the presence of the highly specific Na/K-ATPase inhibitor ouabain. Exposure of whole embryos to 10 microM ouabain (10(-5) M) demonstrated that heart development and precardiomyocyte differentiation are inhibited principally between stage 5 through stage 7. In each stage the degree of inhibition follows a rostrocaudal gradient as development proceeds along the anterior to posterior axis. After stage 8 ouabain no longer affects heart development or cardiomyogenesis. The inhibition is concentration- and developmental stage-dependent. The inhibition is reversible by elevating the outside potassium ion concentration [Ko] in the culture medium or by transferring the embryos into normal medium minus ouabain even after 20 hr of ouabain exposure. The results also suggest that the regulation of the formation of the three-dimensional organ is independent from regulation of myogenesis.

Animals↗

Biophysics of the developing heart.

The force-interval relationship was evaluated in the developing heart of the lamb. The qualitative characteristics of the relationship were the same in isolated muscle (from 93 to 141 days' gestation to 1 year old) as in the chronically instrumented in utero fetus (122 to 141 days' gestation): (1) contractility, e.g., the maximum rate of rise of force, increased monotonically from a small value immediately after a contraction to a plateau, and (2) postextrasystolic potentiation was present in all preparations. In the intact animal, postextrasystolic potentiation depended on the basic pacing interval, t0, and the timing of the extrasystole, t1: when t0 was held constant and t1 was increased, potentiation decreased; when t1 was held constant and t0 was increased, potentiation increased. The qualitative characteristics of the relationship, and so the underlying myocardial basis, were unchanged over the developmental period studied.

Animals↗

Heart-targeted overexpression of Nip3a in zebrafish embryos causes abnormal heart development and cardiac dysfunction.

We transiently expressed a proapoptotic protein, Nip3a, by a heart-specific BMP4 promoter in zebrafish embryos and generated two variants of embryos with abnormal heart phenotypes (A and B). Embryos with phenotype A heart defects showed hypoplastic or elongated ventricles, elongated or enlarged atriums with no normal cardiac looping resulting a significant longer SV-BA distance, and bradycardia at 48 h post-fertilization (hpf). Embryos with phenotype B heart defects showed an enlarged fluid-filled pericardium, severe hypoplasia, non-contracting ventricles, and elongated or enlarged slowly beating atriums with no normal looping. Histological sections further revealed the absence of a proper atrioventricular boundary and no endocardial cells lining this region in both 48- and 72-hpf Nip3a-overexpressing embryos, implicating defective endocardial cushion formation. These phenotypes are reminiscent of atrioventricular canal defects in humans. In addition, induced apoptotic myocardium cells were clustered in the presumptive atrioventricular boundary as well as in the adjacent ventricle and atrium of 48- and 72-hpf Nip3a-overexpressing embryos. Nip3a expression was readily detected in 80% epiboly BMP4-Nip3a-injected embryos, and defects in heart development were observed in both the linear heart tube and subsequent chamber formation stages. These results showed that myocyte apoptosis is a universal pathogenic factor for congenital heart failure using zebrafish as a model organism.

Animals↗

Regulating heart development: the role of Nf1.

Neurofibromatosis type 1 (NF1) is one of the most common human genetic disorders and is associated with significant morbidity and mortality. The gene responsible for this disorder, NF1, encodes neurofibromin, which can function to down-regulate ras activity. Mutations that inactivate NF7 result in elevated levels of ras signaling and increased cell proliferation in some tissues. NF7 functions as a tumor suppressor gene; patients inherit one mutated copy and are believed to acquire a "second hit" in tissues that go on to form benign or malignant tumors. NF7 is expressed widely, yet certain tissues are more susceptible to growth dysregulation in NF1 patients. Cardiovascular defects also contribute to NF1, though the cause remains unclear. In a recent study, we used tissue-specific gene inactivation in mice to study the role of neurofibromin in heart development. A further understanding of neurofibromin function will help to elucidate the pathophysiology of NF1 and will also lead to a better understanding of cell cycle regulation and ras pathways in specific cell types. Finally, we comment on how similar genetic strategies can be used in mice to study the role of additional signaling pathways involved in heart development.

Animals↗

Elfin is expressed during early heart development.

Elfin (previously named CLIM1) is a protein that possesses an N-terminal PDZ domain and a C-terminal LIM domain. It belongs to the family of Enigma proteins. Enigma proteins are a family of cytoplasmic proteins that contain an N-terminal PDZ domain and a series of C-terminal LIM domains. By virtue of these two protein interacting domains, Enigma proteins are capable of protein-protein interactions. It has been proposed that Enigma proteins may act as adapters between kinases and the cytoskeleton. We have previously shown that Elfin is most abundantly expressed in the heart and it colocalizes with alpha-actinin 2 at the Z-disks of the myocardium. In this report, Elfin was shown to localize at the actin stress fibers of myoblasts, as revealed by green fluorescent protein (GFP) tagging. In situ hybridization and immunostaining showed that Elfin expression begins at an early stage in mouse development and is present throughout the developing heart. Taken together, our experimental results suggest that Elfin may play an important role in myofibrillogenesis and heart development.

Actinin↗

[Urinary albumin excretion increases during an acute myocardial infarct especially in patients who develop heart failure].

AIM OF THE STUDY: To evaluate the profile of albumin excretion rate (AER) in the first days of acute myocardial infarction (AMI), its relationship with serum enzymes and the presence of heart failure, and the effect of thrombolytic therapy. METHODS: Two hundred and thirty-one consecutive patients admitted to coronary care unit for suspected AMI were examined. Patients with diabetes mellitus, urinary tract infections or proteinuric diseases were excluded. In 135 patients (95 males, 40 females) AMI diagnosis was confirmed. The remaining 96 (56 males, 40 females) were considered as controls. AER was measured by radioimmunoassay in 24-hour urine samples at the first, third and seventh day after admission and expressed as mg/24h. Statistical analysis was performed after AER logarithmic transformation using repeated measure ANOVA: RESULTS: Mean age was 66.9 +/- 12.2 years (range = 35 -91) in the AMI group and 63.2 +/- 12.3 years (range = 33-91) in the controls (p = 0.023) Age-adjusted blood pressure was lower in the AMI group than in the controls (p < 0.0001 for both systolic and diastolic), while no difference was found in heart rate. Plasma cholesterol, triglycerides, creatinine and uric acid were similar in the 2 groups. Mean AER was 43.4 +/- 64.8, 26.9 +/- 51.2 and 23.9 +/- 52.7 mg/24h at 1st, 3rd and 7th day respectively in the AMI group and 24.9 +/- 58.2, 13.7 +/- 25.8 and 17.9 +/- 44.1 mg/24h respectively in the controls (p = 0.014). In the AMI group, first day AER significantly and positively correlated with CPK (r = 0.287, p = 0.001), CPK-MB (r = 0.239, p = 0.007) and GOT (r = 0.300, p = 0.001). Within the patients with AMI, those who developed heart failure (n = 57), had higher AER (48.6 +/- 68.4, 29.7 +/- 54.9 and 28.1 +/- 55.8 mg/24h at 1st, 3rd and 7th day in patients with mild heart failure -2nd Killip Class- and 100.0 +/- 141.7, 50.3 +/- 66.4 and 64.2 +/- 74.4 mg/24h in those with severe heart failure -3rd and 4th Killip Class-) than those who did not (31.0 +/- 41.7, 19.6 +/- 45.6 and 16.5 +/- 45.7 mg/24h respectively) (p = 0.004). In a multiple linear regression model AER was significantly related to peak values of GOT (1st day) and CPK (3rd day) and to presence of heart failure (3rd and 7th day). Thrombolytic therapy (n = 48) did not influence AER. CONCLUSIONS: The results of the present study show that AER increases following AMI, chiefly in the subjects who develop heart failure. AER correlates with serum enzymes peak levels at 1st and 3rd day and with presence of heart failure at 3rd and 7th day after admission, and is not influenced by thrombolytic therapy. These data suggest that in AMI the initial increase in AER is due to the inflammatory process which accompanies cardiac necrosis, while in a later phase its rise is mainly due to the increased intraglomerular capillary pressure consequent to heart failure.

Adult↗

GATA factors in vertebrate heart development and disease.

Vertebrate heart formation is dependent upon complex hierarchical gene regulatory networks, which effect both the specification and differentiation of cardiomyocytes and subsequently cardiac morphogenesis. GATA-4, -5 and -6 comprise an evolutionarily conserved subfamily of transcription factors, which are expressed within the precardiac mesoderm from early stages in its specification and continue to be expressed within the adult heart. We review here the functional roles of individual GATA transcription factors in cardiac development, normal homeostasis and disease. We also review the cellular mechanisms employed to regulate the expression and downstream targets of the different GATA factors.

Animals↗

A novel role for cardiac neural crest in heart development.

Ablation of premigratory cardiac neural crest results in defective development of the cardiac outflow tract. The purpose of the present study was to correlate the earliest functional and morphological changes in heart development after cardiac neural crest ablation. Within 24 hours after neural crest ablation, the external morphology of the hearts showed straight outflow limbs, tighter heart loops, and variable dilations. Incorporation of bromodeoxyuridine in myocytes, an indication of proliferation, was doubled after cardiac neural crest ablation. The myocardial calcium transients, which are a measure of excitation-contraction coupling, were depressed by 50% in both the inflow and outflow portions of the looped heart tube. The myocardial transients could be rescued by replacing the cardiac neural crest. The cardiac jelly produced by the myocardium was distributed in an uneven, rather than uniform, pattern. An extreme variability in external morphology could be attributed to the uneven distribution of cardiac jelly. In the absence of cardiac neural crest, the myocardium was characterized by somewhat disorganized myofibrils that may be a result of abnormally elevated proliferation. In contrast, endocardial development appeared normal, as evidenced by normal expression of fibrillin-2 protein (JB3 antigen) and normal formation of cushion mesenchyme and trabeculae. The signs of abnormal myocardial development coincident with normal endocardium suggest that the presence of cardiac neural crest cells is necessary for normal differentiation and function of the myocardium during early heart development. These results indicate a novel role for neural crest cells in myocardial maturation.

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Cardiac imaging to identify patients at risk for developing heart failure after myocardial infarction.

The development of heart failure (HF) after acute myocardial infarction (MI) is recognized as a major complication that leads to a significant increase in morbidity and mortality. Given the availability of effective treatments for improving both quality of life and survival for patients at increased risk for developing HF after MI, early identification of these individuals is critical. Noninvasive cardiac imaging offers a detailed characterization of two important pathophysiological processes related to the development of HF post-MI: left ventricular (LV) remodeling and LV functional recovery. Cardiovascular MRI has recently emerged as the preferred noninvasive imaging modality because of its ability to provide the most comprehensive and informative evaluation of these processes. In addition to allowing for an accurate and reproducible longitudinal follow-up of LV volumes and mass, MRI also offers information on infarct size, the presence of microvascular obstruction, and the transmural extent of infarct scar, all of which are valuable parameters that can assist in identifying patients at risk for developing HF after MI.

Heart Failure↗

Microarray analysis of Tbx5-induced genes expressed in the developing heart.

Tbx5 is a member of the T-box family of transcription factors and is associated with Holt-Oram syndrome (HOS), a congenital disorder characterized by heart and limb defects. Although implicated in several processes during development, only a few genes regulated by Tbx5 have been reported. To identify candidate genes regulated by Tbx5 during heart development, a microarray approach was used. A cardiac-derived mouse cell line (1H) was infected with adenoviruses expressing Tbx5 or beta-galactosidase and RNA was isolated for analysis using an Affymetrix gene chip representing over 39,000 transcripts. Real-time reverse transcriptase-polymerase chain reaction confirmed Tbx5 induction of a subset of the genes, including nppa, photoreceptor cadherin, brain creatine kinase, hairy/enhancer-of-split related 2, and gelsolin. In situ hybridization analysis indicated overlapping expression of these genes with tbx5 in the embryonic mouse heart. In addition, the effect of HOS-associated mutations on the ability of Tbx5 to induce target gene expression was evaluated. Together, these data identify several genes induced by Tbx5 that are potentially important during cardiac development. These genes represent new candidate gene targets of Tbx5 that may be related to congenital heart malformations associated with HOS.

Adenoviridae↗

NFATc1 expression in the developing heart valves is responsive to the RANKL pathway and is required for endocardial expression of cathepsin K.

NFATc1 is necessary for remodeling endocardial cushions into mature heart valve leaflets and is also an essential effector of receptor activator of NFkappaB ligand (RANKL) signaling required for transcriptional activation of bone matrix remodeling enzymes during osteoclast differentiation. Therefore, developing heart valves were examined to determine if NFATc1 functions in the RANKL pathway during leaflet remodeling. Key components of RANKL signal transduction including RANKL, its receptor RANK, and the downstream remodeling enzyme cathepsin K (Ctsk) are expressed in the heart during valve remodeling and colocalize with NFATc1 in developing valve endocardium. However, the absence of tartrate-resistant acid phosphatase (TRAP) activity and the lack of F4/80-positive macrophage lineage contribution to the remodeling valves demonstrate that certain aspects of osteoclast RANKL function are not shared during valve formation. Analysis of NFATc1-/- mouse embryos shows that NFATc1 is specifically required for endocardial expression of RANKL and Ctsk during valve formation. In addition, RANKL treatment augments expression of NFATc1 and Ctsk in embryonic heart cultures, and the RANKL-mediated increase in Ctsk expression is dependent on NFATc1. Together, these results support a role for RANKL signaling during heart valve development and suggest that valve leaflet morphogenesis involves NFATc1-dependent expression of remodeling enzymes including Ctsk.

Animals↗