Initiation Of Heartbeat Research Articles

A novel conceptual model of heart rate autonomic modulation based on a small-world modular structure of the sinoatrial node.

The present view on heartbeat initiation is that a primary pacemaker cell or a group of cells in the sinoatrial node (SAN) center paces the rest of the SAN and the atria. However, recent high-resolution imaging studies show a more complex paradigm of SAN function that emerges from heterogeneous signaling, mimicking brain cytoarchitecture and function. Here, we developed and tested a new conceptual numerical model of SAN organized similarly to brain networks featuring a modular structure with small-world topology. In our model, a lower rate module leads action potential (AP) firing in the basal state and during parasympathetic stimulation, whereas a higher rate module leads during β-adrenergic stimulation. Such a system reproduces the respective shift of the leading pacemaker site observed experimentally and a wide range of rate modulation and robust function while conserving energy. Since experimental studies found functional modules at different scales, from a few cells up to the highest scale of the superior and inferior SAN, the SAN appears to feature hierarchical modularity, i.e., within each module, there is a set of sub-modules, like in the brain, exhibiting greater robustness, adaptivity, and evolvability of network function. In this perspective, our model offers a new mainframe for interpreting new data on heterogeneous signaling in the SAN at different scales, providing new insights into cardiac pacemaker function and SAN-related cardiac arrhythmias in aging and disease.

Ultrastructural Assessment and Proteomic Analysis in Myofibrillogenesis in the Heart Primordium After Heartbeat Initiation in Rats.

Myofibrillogenesis is an essential process for cardiogenesis and is closely related to excitation-contraction coupling and the maintenance of heartbeat. It remains unclear whether the formation of myofibrils and sarcomeres is associated with heartbeat initiation in the early embryonic heart development. Here, we investigated the association between the ultrastructure of myofibrils assessed by transmission electron microscopy and their proteomic profiling assessed by data-independent acquisition mass spectrometry (DIA-MS) in the rat heart primordia before and after heartbeat initiation at embryonic day 10.0, when heartbeat begins in rats, and in the primitive heart tube at embryonic day 11.0. Bundles of myofilaments were scattered in a few cells of the heart primordium after heartbeat initiation, whereas there were no typical sarcomeres in the heart primordia both before and after heartbeat initiation. Sarcomeres with Z-lines were identified in cells of the primitive heart tube, though myofilaments were not aligned. DIA-MS proteome analysis revealed that only 43 proteins were significantly upregulated by more than 2.0 fold among a total of 7,762 detected proteins in the heart primordium after heartbeat initiation compared with that before heartbeat initiation. Indeed, of those upregulated proteins, 12 (27.9%) were constituent proteins of myofibrils and 10 (23.3%) were proteins that were accessories and regulators for myofibrillogenesis, suggesting that upregulated proteins that are associated with heartbeat initiation were enriched in myofibrillogenesis. Collectively, our results suggest that the establishment of heartbeat is induced by development of bundles of myofilaments with upregulated proteins associated with myofibrillogensis, whereas sarcomeres are not required for the initial heartbeat.

Enhanced glucose metabolism through activation of HIF-1\u03b1 covers the energy demand in a rat embryonic heart primordium after heartbeat initiation

The initiation of heartbeat is an essential step in cardiogenesis in the heart primordium, but it remains unclear how intracellular metabolism responds to increased energy demands after heartbeat initiation. In this study, embryos in Wistar rats at embryonic day 10, at which heartbeat begins in rats, were divided into two groups by the heart primordium before and after heartbeat initiation and their metabolic characteristics were assessed. Metabolome analysis revealed that increased levels of ATP, a main product of glucose catabolism, and reduced glutathione, a by-product of the pentose phosphate pathway, were the major determinants in the heart primordium after heartbeat initiation. Glycolytic capacity and ATP synthesis-linked mitochondrial respiration were significantly increased, but subunits in complexes of mitochondrial oxidative phosphorylation were not upregulated in the heart primordium after heartbeat initiation. Hypoxia-inducible factor (HIF)-1α was activated and a glucose transporter and rate-limiting enzymes of the glycolytic and pentose phosphate pathways, which are HIF-1α-downstream targets, were upregulated in the heart primordium after heartbeat initiation. These results suggest that the HIF-1α-mediated enhancement of glycolysis with activation of the pentose phosphate pathway, potentially leading to antioxidant defense and nucleotide biosynthesis, covers the increased energy demand in the beating and developing heart primordium.

PAH-pollution effects on sensitive and resistant embryos: Integrating structure and function with gene expression.

Polycyclic aromatic hydrocarbons (PAHs) are among the most widespread natural and anthropogenic pollutants, and some PAHs are proven developmental toxicants. We chemically characterized clean and heavily polluted sites and exposed fish embryos to PAH polluted sediment extracts during four critical developmental stages. Embryos were collected from Fundulus heteroclitus populations inhabiting the clean and heavily polluted Superfund estuary. Embryos of parents from the clean sites are sensitive to PAH pollutants while those of parents from the heavily polluted site are resistant. Chemical analysis of embryos suggests PAH accumulation and pollution-induced toxicity among sensitive embryos during development that ultimately kills all sensitive embryos before hatching, while remarkably, the resistant embryos develop normally. The adverse effects on sensitive embryos are manifested as developmental delays, reduced heart rates, and severe heart, liver, and kidney morphological abnormalities. Gene expression analysis of early somitogenesis, heartbeat initiation, late organogenesis, and pre-hatching developmental stages reveals genes whose expression significantly differs between sensitive and resistant embryo populations and helps to explain mechanisms of sensitivity and resistance to polluted environments during vertebrate animal development.

Energy Metabolism in Rat Fetal Hearts Just after the Initiation of Heartbeat is Altered via Increased Glycolytic Flux and Activated Mitochondrial Function

Energy Metabolism in Rat Fetal Hearts Just after the Initiation of Heartbeat is Altered via Increased Glycolytic Flux and Activated Mitochondrial Function

The expression dynamics of mechanosensitive genes in extra-embryonic vasculature after heart starts to beat in chick embryo.

Fluid flow plays an important role in vascular development. However, the detailed mechanisms, particularly the link between flow and modulation of gene expression during vascular development, remain unexplored. In chick embryo, the key events of vascular development from initiation of heart beat to establishment of effective blood flow occur between the stages HH10 and HH13. Therefore, we propose a novel in vivo model to study the flow experienced by developing endothelium. Using this model, we aimed to capture the transcriptome dynamics of the pre- and post-flow conditions. RNA was isolated from extra embryonic area vasculosa (EE-AV) pooled from three chick embryos between HH10-HH13 and RNA sequencing was performed. The whole transcriptome sequencing of chick identified up-regulation of some of the previously well-known mechanosensitive genes including NFR2, HAND1, CTGF and KDR. GO analyses of the up-regulated genes revealed enrichment of several biological processes including heart development, extracellular matrix organization, cell-matrix adhesion, cell migration, blood vessel development, patterning of blood vessels, collagen fibril organization. Genes encoding for gap junctions proteins which are involved in vascular remodeling and arterial-venous differentiation, and genes involved in cell-cell adhesion, and ECM interactions were significantly up-regulated. Validation of selected genes through semi quantitative PCR was performed. The study indicates that shear stress plays a major role in development. Through appropriate validation, this platform can serve as an in vivo model to study conditions of disturbed flow in pathology as well as normal flow during development.

Regular heartbeat rhythm at the heartbeat initiation stage is essential for normal cardiogenesis at low temperature

BackgroundThe development of blood flow in the heart is crucial for heart function and embryonic survival. Recent studies have revealed the importance of the extracellular matrix and the mechanical stress applied to the valve cushion that controls blood flow to the formation of the cardiac valve during embryogenesis. However, the events that trigger such valve formation and mechanical stress, and their temperature dependence have not been explained completely. Medaka (Oryzias latipes) inhabits a wide range of East Asia and adapts to a wide range of climates. We used medaka embryos from different genomic backgrounds and analyzed heartbeat characteristics including back-and-forth blood flow and bradyarrhythmia in embryos incubated at low temperature. We also used high-speed imaging analysis to examine the heartbeat of these animals after transient exposure to low temperature.ResultsEmbryos of the Hd-rR medaka strain exhibited back-and-forth blood flow in the heart (blood regurgitation) after incubation at 15°C. This regurgitation was induced by exposure to low temperature around the heartbeat initiation period and was related to abnormalities in the maintenance or pattern of contraction of the atrium or the atrioventricular canal. The Odate strain from the northern Japanese group exhibited normal blood flow after incubation at 15°C. High-speed time-lapse analysis of the heartbeat revealed that bradyarrhythmia occurred only in Hd-rR embryos incubated at 15°C. The coefficient of contraction, defined as the quotient of the length of the atrium at systole divided by its length at diastole, was not affected in either strain. The average heart rate after removing the effect of arrhythmia did not differ significantly between the two strains, suggesting that the mechanical stress of individual myocardial contractions and the total mechanical stress could be equivalent, regardless of the presence of arrhythmia or the heart rate. Test-cross experiments suggested that this circulation phenotype was caused by a single major genomic locus.ConclusionsThese results suggest that cardiogenesis at low temperature requires a constant heartbeat. Abnormal contraction rhythms at the stage of heartbeat initiation may cause regurgitation at later stages. From the evolutionary viewpoint, strains that exhibit normal cardiogenesis during development at low temperature inhabit northern environments.

Expression of L-Type Calcium Channel Cav1.3 and Sarcomeric-Related Proteins in the Period of Heart-Beat Initiation in the Early Rat Embryo

Expression of L-Type Calcium Channel Cav1.3 and Sarcomeric-Related Proteins in the Period of Heart-Beat Initiation in the Early Rat Embryo

Hematopoietic Stem Cell Development Is Dependent on Blood Flow and Nitric Oxide Signaling

During vertebrate embryogenesis, definitive hematopoietic stem cells (HSC) arise in the aorta-gonads-mesonephros (AGM). Based on the functional conservation of AGM hematopoiesis from fish to man, an evolutionary advantage for the production of stem cells within the aorta must exist. The identification of the signals that induce HSCs at this developmental stage is of significant interest. Through a chemical genetic screen in zebrafish, a diverse group of compounds that regulate blood flow were found to affect the production of runx1/cmyb+ HSCs. These compounds represented modulators of the adrenergic and renin/angiotensin pathways, and Ca+, Na+ and nitric oxide (NO) signaling. In general, we determined that compounds which increased blood flow enhanced HSC number, whereas chemicals that decreased blood flow diminished runx1/cmyb expression. The conserved physiological mechanism of action of each compound on the vasculature was confirmed in vivo by confocal microscopy of transgenic fli1:GFP reporter fish. In the zebrafish, the step-wise initiation of heartbeat, establishment of vigorous circulation and onset of definitive hematopoiesis in the aorta-gonad-mesonephros region (AGM) suggests that blood flow may trigger HSC formation. silent heart (sih) embryos that lack a heartbeat and fail to establish blood circulation exhibit severely reduced numbers of runx1+ HSCs in the AGM. Blood flow modifying agents primarily exerted their effects after the onset of the heartbeat (>24 hpf), however, only compounds that increase NO production (L-Arginine, S-nitroso-N-acetyl-penicillamine (SNAP)) could induce HSC formation prior to the initiation of circulation (5 somites to 22 hpf). Furthermore, SNAP rescued HSC production in sih mutant zebrafish, whereas other drugs that increased blood flow could not. Treatment with the NO synthase (NOS) inhibitor, N-nitro-L-arginine methyl ester (L-NAME), and morpholino-oligonucleotide (MO)-knockdown of nos1 (nnos/enos) blocked HSC development. Additionally, modulation of downstream components of the NO pathway affected HSC production in the zebrafish embryo. Together these data indicate that NO signaling is the downstream effector of blood flow on AGM HSC induction. To document that NO-mediated regulation of HSC formation was conserved across vertebrate species, we examined definitive HSC production in the murine AGM. Nos3 (eNos) was found to be expressed in the AGM endothelium and aortic hematopoietic clusters. Additionally, Nos3 expression specifically marks the population of HSCs with long-term adult bone marrow repopulating activity. Intrauterine NOS inhibition with L-NAME resulted in a lack of hematopoietic clusters in the AGM and a failure to generate transplantable hematopoietic progenitors. Our work provides a direct link between the initiation of circulation and the onset of AGM hematopoiesis, and identifies NO signaling as a conserved downstream regulator of HSC development. ^TEN and WG contributed equally to this work

A Reduction of Tropomyosin Limits Development of Sarcomeric Structures in Cardiac Mutant Hearts of the Mexican Axolotl

The cardiac lethal mutation in Mexican axolotl (Ambystoma mexicanum) results in a lack of contractions in the ventricle of mutant embryos. Previous studies have demonstrated that tropomyosin, a component of thin filaments, is greatly reduced in mutant hearts lacking myofibril organization. Confocal microscopy was used to examine the structure and comparative amount of tropomyosin at heartbeat initiation and at a later stage. The formation of functional sarcomeres coincided with contractions in normal hearts at stage 35. A-bands and I-bands were formed at stage 35 and did not change at stage 39. The widening of Z-bodies into z-lines was the main developmental difference between stage 35 and 39 normal hearts. Relative to normal hearts, a reduction of sarcomeric protein levels in mutant hearts at stage 35 was found, and a greater reduction occurred at later stages. The lower level of tropomyosin limited the areas where organized myofibrils formed in the mutant. The areas that had tropomyosin staining also had staining for alpha-actinin and myosin. Early myofibrils formed in these areas but the A-bands and I-bands were shorter than normal. At a later stage in the mutant, A-bands and I-bands remained shorter and importantly the Z-bodies also did not form wider z-lines.

Morphometry and estimated bulk oxygen diffusion in larvae of Xenopus laevis under chronic carbon monoxide exposure.

To understand the mechanisms that allow tadpoles of the African clawed frog Xenopus laevis to develop under conditions of impaired convective transport (hemoglobin poisoning with carbon monoxide), whole animal surface area and volume were measured and bulk oxygen diffusion was modeled at four developmental stages (from initiation of heartbeat to pre-metamorphic climax). Surface area [8.5 mm2 at stages Nieuwkoop-Faber (NF) 33-34 to 70.2 mm2 at stages NF 50-51] and volume (1.8 mm3 at stages NF 33-34 to 35.7 mm3 at stages NF 50-51) measured from volumetric analysis from dual plane images of each animal were not significantly different between treatments. Bulk oxygen radial diffusion was estimated by modeling the larvae as a set of adjacent cylinders with different radii. The model was used to predict the oxygen tension at the water-skin interface at which the oxygen tension in the center of the animal is nil (0.7 kPa at stage NF 33-34 and 14.0 kPa at stage NF 50-51), suggesting that bulk oxygen diffusion is sufficient to meet the metabolic demand up to stages NF 46-47 irrespective of the oxygen tension at the water-skin interface. At NF 50-51 an anoxic core in the animal would appear if bulk oxygen diffusion were the only means of oxygen transport at oxygen tensions below 15 kPa. However, the relative volume of the anoxic core would only exceed 10% of the total volume of the animal only at oxygen tensions below 5 kPa. Therefore, the ten-fold increase in mass between NF 50-51 and metamorphosis would prove insufficient for embryonic oxygen requirements via simple diffusion, and therefore would require additional transport mechanisms.

Thiobencarb-induced embryotoxicity in medaka ( Oryzias latipes): stage-specific toxicity and the protective role of chorion

Thiobencarb ( S-(4-chlorobenzyl)- N, N-diethyl thiol carbamate) has been one of the herbicides previously associated with fish kills in agricultural drains near the Sacramento/San Joaquin rivers and their Delta. This area is an important spawning ground for fish, and thus there are concerns over possible toxic effects on early life stages of fishes. To define targets of thiobencarb embryotoxicity and to determine the degree of protection afforded by the chorion, medaka ( Oryzias latipes) embryos were exposed under static nonrenewal conditions. Responses to exposures initiated at blastula or at initiation of heart beat (stages 10 and 23, respectively) were assessed. In addition, enzymatically dechorionated embryos (stage 13, gastrula) were exposed and compared to responses in embryos with intact chorions. Embryos were observed daily for development and for gross abnormalities including: bradycardia, pericardial edema, hemostasis, poor yolk resorption, cephalic and spinal deformities, and abnormal hatching. A subset was also evaluated for histologic alterations. Based on gross abnormalities, the concentration of thiobencarb affecting 50% (EC 50) of embryos exposed at blastula was 3.6 mg/l, while the putative no observable effect concentration (NOEC) was 1.0 mg/l. For embryos exposed at onset of heart beat (stage 23), these values were 4.1 and 2.5 mg/l, respectively. Dechorionated embryos tended to be more sensitive than their chorionated cohorts (LC 50=2.5 vs. 1.0 mg/l). Liver histologic alterations were seen in chorionated embryos at EC 50 levels and higher. Stage-specific toxicity was evident; nevertheless, the EC 50 and NOEC values for embryos treated at stage 10 and stage 23 were similar.

Diluted rotifer, Brachionus plicatilis (Muller), extract as culture medium stimulates hatching of red sea bream, Pagrus major (Temminck & Schlegel)

Hatching rates and hatching times (50% hatch) of red sea bream, Pagrus major (Temminck & Schlegel), eggs in culture media that were prepared to dilute rotifer Brachionus plicatilis Muller extract at 4000-, 6000-. 8000- and 10 000-fold with artificial sea water (σ15, = 24.0) were investigated. Although differences in hatching rate between the culture media and control artificial sea water were negligible, hatching time was significantly shorter in media at 4000- and 6000-fold dilutions than in media at 8000- and 10 000-fold dilutions and in the control artificial sea water, when the eggs immediately post spawning were incubated. The hatching rates of eggs at three developmental stages, immediately post-spawning, gastrula stage and heart-beat initiation stage, incubated respectively in the medium at 6000-fold dilution and the control, did not differ either, yet the hatching times at each egg stage in the medium were significantly shorter than that in the control. These results suggest that hatching enzyme secretion from the hatching glands is promoted to perceive chemical stimuli in the water-soluble fraction of rotifer extract via the embryo's chemoreceptors, from the heart-beat initiation stage to immediately before hatching, thus shortening the hatching time.

Extracellular matrix of the developing heart in normal and cardiac lethal mutant axolotls, Ambystoma mexicanum

As part of an ongoing study of heart development in normal and cardiac lethal mutant axolotls (Mexican salamanders) we examined the extracellular matrix (ECM) by microscopical methods. With scanning electron microscopy we are unable to detect ECM on the apical surface of cells of the early cardiogenic mesoderm. During the period of lateral plate migration, which coincides with the period of cardiogenic induction of mesoderm by anterior endoderm, there is little ECM, aside from some microfibrils, on the basal surface of the endoderm or mesoderm of the pharyngeal region. Later, a basal lamina (BL) is found on the endoderm and along portions of the developing endocardial and myocardial tubes. By the time of heartbeat initiation the BLs are complete and invested with striated collagen-like fibrils that are sparsely distributed in the "cardiac jelly" of normal and mutant hearts. Striated fibril deposition, which increases with time, is generally random in orientation, with the exception of some regions where there is a preferred directionality. During the post-hatching period striated fibrils appear in the subepicardial space. In addition, branching fibers that are probably elastin appear in the bulbus arteriosus. In these later stages the density of fibrils in the cardiac lethal mutant heart is much less than normal. Indirect immunofluorescent microscopy reveals laminin and fibronectin in the basal laminae of the endocardial and myocardial tubes of both normal and cardiac lethal mutant hearts. In addition, punctate and fibrillar staining for fibronectin, and punctate staining for laminin are found in the cardiac jelly. These matrix proteins are not abundant at the apical (exterior) surface of the myocardium until the epicardium appears.(ABSTRACT TRUNCATED AT 250 WORDS)

The specification of heart mesoderm occurs during gastrulation in Xenopus laevis.

The establishment of heart mesoderm during Xenopus development has been examined using an assay for heart differentiation in explants and explant combinations in culture. Previous studies using urodele embryos have shown that the heart mesoderm is induced by the prospective pharyngeal endoderm during neurula and postneurula stages. In this study, we find that the specification of heart mesoderm must begin well before the end of gastrulation in Xenopus embryos. Explants of prospective heart mesoderm isolated from mid- or late neurula stages were capable of heart formation in nearly 100% of cases, indicating that the specification of heart mesoderm is complete by midneurula stages. Moreover, inclusion of pharyngeal endoderm had no statistically significant effect upon either the frequency of heart formation or the timing of the initiation of heartbeat in explants of prospective heart mesoderm isolated after the end of gastrulation. When the superficial pharyngeal endoderm was removed at the beginning of gastrulation, experimental embryos formed hearts, as did explants of prospective heart mesoderm from such embryos. These results indicate that the inductive interactions responsible for the establishment of heart mesoderm occur prior to the end of gastrulation and do not require the participation of the superficial pharyngeal endoderm.

Quantitative analysis of cardiac arrhythmias.

Quantitative analysis of cardiac arrhythmias has been the subject of intensive research during the last 10 years. Several systems have been designed to help in the processing of cardiac signals: single or multiple lead electrocardiograms (ECG), electrograms from intracardiac catheters, esophageal recordings, etc. The main objective of these developments was oriented toward positive identification of arrhythmias or rhythm-disturbance counts to imitate the cardiologist's interpretation in contexts such as routing ECG and ambulatory recordings. However, these systems were mainly measurement tools aimed at extracting auricular and ventricular depolarization timings plus gross morphology description. The domain of morphology analysis of beat-to-beat auricular depolarization on ECG has never been highly active due to poor signal conditions. For routine ECG, automatic interpretation was set as an objective to complement computer-assisted ECG interpretation of conduction problems (i.e., morphology analysis of a representative beat extracted or averaged from the dominant rhythm). The limitations of rhythm interpretation in this context are well known. In the ambulatory ECG context, the analysis procedures are relatively simple and are often summarized as trivial counts describing, most exclusively, the ventricular arrhythmic behavior of the heart over a relatively long duration. Waveform-detection and measurement have been the bottleneck of advancement in arrhythmia analysis since highly reliable detection of events on a beat-to-beat basis are necessary to perform a valid analysis. Rare approaches have proposed probabilistic definition of event detection. The present review puts emphasis on the potential of several methods which have been demonstrated as powerful in identifying short- or long-duration heartbeat patterns, mode of heartbeat initiation, mode of heartbeat coupling, etc. Globally, these methods are referred to as time series analysis, modeling of rhythm patterns, simulation, and pattern recognition. A delay in the advancement of the study of arrhythmogenesis and limiting the analysis of arrhythmias to textbook descriptions is not justified when put in perspective of the potential of implementing powerful techniques which have been more or less neglected or used in a narrow way.

Precardiac mesoderm differentiation in vitro: Actin-isotype synthetic transitions, myofibrillogenesis, initiation of heartbeat, and the possible involvement of collagen

The differentiation of precardiac mesoderm into beating heart tissue was examined during explant culture. Explanted tissue forms tubular heart-like vesicles and initiates rhythmic contractility within 18–24 h in vitro, a developmental time-course approximating that observed during in vivo development. Electron-microscopic observations reveal that beating heart cells are rich in cytoplasmic myofibrils in varying degrees of order, with some regions containing highly organized myofibrillar arrays. The analysis of actin-isotype biosynthesis, using metabolic labeling with [ 35S]-methionine and isoelectric-focusing resolution of the synthesized radioactive polypeptides, demonstrates that the initiation of cardiac α-actin synthesis and the pattern of transition in the synthesis of α-, β-, and γ-actin isotypes is equivalent to the initiation time and pattern observed in vivo. A possible collagen involvement in the differentiation process was investigated by assessing the effects of collagen-synthesis inhibitors on the development of the explant cultures. Two different agents, α,α′-dipyridyl and L-azetidine-2-carboxylic acid, exhibited a dose-dependent ability to inhibit the formation of beating heart tissue. When examined by electron microscopy, the nonbeating tissue exhibited a drastic depression of myofibrillogenesis, but otherwise appeared healthy. Further examination of the effect of L-azetidine-2-carboxylic acid demonstrated that the inhibition of myofibril formation and heartbeat was correlated with a 60% inhibition of native collagen synthesis; however, the time-course and pattern of actinisotype biosynthesis was completely unaffected. The data suggest a possible collagen involvement in heart differentiation that is necessary for either the synthesis of non-actin cardiac contractile proteins or the assembly of cardiac contractile proteins into myofibrils.

Optical signals from early embryonic chick heart stained with potential sensitive dyes: evidence for electrical activity.

1. Using an optical method for monitoring membrane potential, spontaneous electrical activity in the very early embryonic chick heart at the 7-9 somite stages was measured. 2. Spontaneous absorption signals from the 7-8 somite embryonic chick hearts stained with a potential sensitive merocyanine-oxazolone dye were demonstrated. The signals were observed also when a merocyanine-rhodanine dye was used. These signals were identified as spontaneous electrical activity in the embryonic heart cells. 3. The action spectrum in absorption of the merocyanine-oxazolone dye was triphasic in early embryonic chick heart with an increase in transmittance from 750 to 700, a decrease from 700 to 600, and an increase from 600 to 525 nm. 4. The magnitude of the signal was about 10(-3) of the resting intensity at 675 nm, with the merocyanine-oxazolone dye. The spontaneous absorption signals had a signal-to-noise ratio of about 10, respectively. 5. The absorption signals were markedly depressed by a higher external K+-concentration, however, were not affected by tetrodotoxin (TTX). 6. The results indicate that spontaneous electrical activity is generated at the 7-9 somite developmental stage before the initiation of heartbeat.

Location of pacemaker in chick embryo heart at the time of initiation of heartbeat.

Location of pacemaker in chick embryo heart at the time of initiation of heartbeat.

Cardia bifida and the development of pacemaker function in the early chick heart

Double-hearted or “cardia bifida” embryos were produced by making a mid-line cut through the tissues of the anterior intestinal portal of the 1–3 somite chick embryo, at a time when the foregut is a shallow crescent and the cardiac primordia have not yet fused in the bulboventricular region. Each lateral primordium in the operated chicks forms a separately beating tubular heart. From the time of initiation of heart beat (at 10–11 somites) to the stage of 18–19 somites, the left heart beats faster than does the right in the great majority of embryos. This observation is in consonance with the concept of “left dominance” as applied to early embryonic development. As development progresses, however, the left heart declines in rate while the right remains stable or increases in rate. By the 21–24 somite stage, the right heart usually has a faster rate than the left. It is at 19–20 somites that the sinus venosus is just forming in the heart. It is concluded that: (1) material from the left cardiac primordium is predominant in many respects over that from the right during early embryonic development and acts as pacemaker for the entire embryonic heart up to the stage of 18–19 somites. (2) At about the time the sinus venosus forms, this dominance relation reverses, material from the right primordium taking over the pacemaker function. Whether this change represents a collapse of some left-right gradient of metabolic activity, or the overriding of such a gradient by specific sinoatrial cells from the right side, is not known. It is suggested that the present results provide evidence that the definitive sinoatrial pacemaker of the adult heart is derived wholly or in the main from the right cardiac primordium.