Hemolymph Flow Research Articles

Roles of Keber's valve and foot chamber for foot manipulation in the mussel Nodularia douglasiae.

ABSTRACTIn order to analyse the roles of Keber's valve for foot manipulation in the mussel Nodularia douglasiae, the anatomy and haemolymph flow in the cardiovascular system were detected by magnetic resonance imaging. The superficial layer of the foot was covered by a dense muscle layer, which extended to the dorsal side and connected with the shell. This closed space, the foot chamber, had an inlet (anterior aorta) and an outlet (Keber's valve). At rest, in the beginning of the systolic phase, flows in the anterior aorta and the pedal artery increased, followed by the pedal and visceral sinuses. Then these flows ceased at the end of the systolic phase, followed by inflow to the ventricle in the diastolic phase; therefore, the compliance of the foot chamber is low enough to transfer pressure pulses to the visceral sinus. Extension of the foot started with relaxation of the foot muscle, so the compliance of the foot chamber increased. Then, Keber's valve closed so that the haemolymph filled the foot haemocoel. Retraction of the foot is initiated by the opening of Keber's valve. Judging from these results Keber's valve and the foot chamber are essential for circulation at rest, foot extension and retraction.This article has an associated First Person interview with the first author of the paper.

Hemolymph circulation in insect flight appendages: physiology of the wing heart and circulatory flow in the wings of the mosquito Anopheles gambiae.

The wings of insects are composed of membranes supported by interconnected veins. Within these veins are epithelial cells, nerves and tracheae, and their maintenance requires the flow of hemolymph. For this purpose, insects employ accessory pulsatile organs (auxiliary hearts) that circulate hemolymph throughout the wings. Here, we used correlative approaches to determine the functional mechanics of hemolymph circulation in the wings of the malaria mosquito Anopheles gambiae Examination of sectioned tissues and intravital videos showed that the wing heart is located underneath the scutellum and is separate from the dorsal vessel. It is composed of a single pulsatile diaphragm (indicating that it is unpaired) that contracts at 3 Hz and circulates hemolymph throughout both wings. The wing heart contracts significantly faster than the dorsal vessel, and there is no correlation between the contractions of these two pulsatile organs. The wing heart functions by aspirating hemolymph out of the posterior wing veins, which forces hemolymph into the wings via anterior veins. By tracking the movement of fluorescent microspheres, we show that the flow diameter of the wing circulatory circuit is less than 1 µm, and we present a spatial map detailing the flow of hemolymph across all the wing veins, including the costa, sub-costa, ambient costa, radius, media, cubitus anterior, anal vein and crossveins. We also quantified the movement of hemolymph within the radius and within the ambient costa, and show that hemolymph velocity and maximum acceleration are higher when hemolymph is exiting the wing.

Mosquito hemocytes preferentially aggregate and phagocytose pathogens in the periostial regions of the heart that experience the most hemolymph flow

When a mosquito acquires an infection in the hemocoel, dedicated immune cells called hemocytes aggregate around the valves of the heart. These sessile hemocytes are called periostial hemocytes. In the present study we scrutinized the immune response mounted by the periostial hemocytes of the malaria mosquito, Anopheles gambiae, against bacterial pathogens, and tested the relationship between periostial hemocyte aggregation, immune activity, and hemolymph flow. Initially, we quantified the process of periostial hemocyte aggregation and found that hemocytes migrate to the periostial regions in response to infection with Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, and Micrococcus luteus (all infections tested). Then, we investigated whether the periostial hemocytes are evenly distributed along the six periostial regions of the heart, and found that they preferentially aggregate in the periostial regions of the mid-abdominal segments (4, 5 and 6). This distribution perfectly correlates with the spatial distribution of phagocytic activity along the surface of the heart, and to a lesser extent, with the distribution of melanin deposits. Finally, experiments measuring circulatory physiology found that the majority of hemolymph enters the heart through the ostia located in the periostial regions of abdominal segments 4, 5, and 6. These data show that periostial hemocytes aggregate on the surface of the heart in response to diverse foreign stimuli, and that both hemocytes and immune activity preferentially occur in the regions that experience the swiftest hemolymph flow. Thus, these data show that two major organ systems – the immune and circulatory systems – interact to control infections.

Respiratory and Metabolic Impacts of Crustacean Immunity: Are there Implications for the Insects?

Extensive similarities in the molecular architecture of the crustacean immune system to that of insects give credence to the current view that the Hexapoda, including Insecta, arose within the clade Pancrustacea. The crustacean immune system is mediated largely by hemocytes, relying on suites of pattern recognition receptors, effector functions, and signaling pathways that parallel those of insects. In crustaceans, as in insects, the cardiovascular system facilitates movement of hemocytes and delivery of soluble immune factors, thereby supporting immune surveillance and defense along with other physiological functions such as transport of nutrients, wastes, and hormones. Crustaceans also rely heavily on their cardiovascular systems to mediate gas exchange; insects are less reliant on internal circulation for this function. Among the largest crustaceans, the decapods have developed a condensed heart and a highly arteriolized cardiovascular system that supports the metabolic demands of their often large body size. However, recent studies indicate that mounting an immune response can impair gas exchange and metabolism in their highly developed vascular system. When circulating hemocytes detect the presence of potential pathogens, they aggregate rapidly with each other and with the pathogen. These growing aggregates can become trapped in the microvasculature of the gill where they are melanized and may be eliminated at the next molt. Prior to molting, trapped aggregates of hemocytes also can impair hemolymph flow and oxygenation at the gill. Small shifts to anaerobic metabolism only partially compensate for this decrease in oxygen uptake. The resulting metabolic depression is likely to impact other energy-expensive cellular processes and whole-animal performance. For crustaceans that often live in microbially-rich, but oxygen-poor aquatic environments, there appear to be distinct tradeoffs, based on the gill's multiple roles in respiration and immunity. Insects have developed a separate tracheal system for the delivery of oxygen to tissues, so this particular tradeoff between oxygen transport and immune function is avoided. Few studies in crustaceans or insects have tested whether mounting an immune response might impact other functions of the cardiovascular system or alter integrity of the gut, respiratory, and reproductive epithelia where processes of the attack on pathogens, defense by the host, and physiological functions play out. Such tradeoffs might be fruitfully addressed by capitalizing on the ease of molecular and genetic manipulation in insects. Given the extensive similarities between the insect and the crustacean immune systems, such models of epithelial infection could benefit our understanding of the physiological consequences of immune defense in all of the Pancrustacea.

Integrated Immune and Cardiovascular Function in Pancrustacea: Lessons from the Insects.

When pathogens invade the insect hemocoel (body cavity) they immediately confront two major forces: immune-responses and circulatory currents. The immune response is mediated by circulating and sessile hemocytes, the fat body, the midgut, and the salivary glands. These tissues drive cellular and humoral immune processes that kill pathogens via phagocytosis, melanization, lysis, encapsulation, and nodulation. Moreover, immune-responses take place within a three-dimensional and dynamic space that is governed by the forces of the circulatory system. The circulation of hemolymph (insect blood) is primarily controlled by the wave-like contraction of a dorsal vessel, which is a muscular tube that extends the length of the insect and is divided into a thoracic aorta and an abdominal heart. Distributed along the heart are valves, called ostia, that allow hemolymph to enter the vessel. Once inside the heart, hemolymph is sequentially propelled to the anterior and to the posterior of the body. During an infection, circulatory currents sweep small pathogens to all regions of the body. As they circulate, pathogens encounter immune factors of the insect that range from soluble cytotoxic peptides to phagocytic hemocytes. A prominent location for these encounters is the surface of the heart. Specifically, periostial hemocytes aggregate in the extracardiac regions that flank the heart's ostia (the periostial regions) and phagocytoze pathogens in areas of high flow of hemolymph. This review summarizes the biology of the immune and circulatory systems of insects, including how these two systems have co-adapted to fight infection. This review also compares the immune and circulatory systems of insects to that of crustaceans, and details how attachment of hemocytes to cardiac tissues and the biology of the lymphoid organ demonstrate that dynamic interactions between the immune and circulatory systems also occur in lineages of crustaceans.

Effect of temperature on leg kinematics in sprinting tarantulas (Aphonopelma hentzi): high speed may limit hydraulic joint actuation.

Tarantulas extend the femur-patella (proximal) and tibia-metatarsal (distal) joints of their legs hydraulically. Because these two hydraulically actuated joints are positioned in series, hemolymph flow within each leg is expected to mechanically couple the movement of the joints. In the current study, we tested two hypotheses: (1) at lower temperatures, movement of the two in-series hydraulic joints within a leg will be less coupled because of increased hemolymph viscosity slowing hemolymph flow; and (2) at higher temperatures, movement of the two in-series hydraulic joints will be less coupled because the higher stride frequencies limit the time available for hemolymph flow. We elicited maximal running speeds at four ecologically relevant temperatures (15, 24, 31 and 40°C) in Texas Brown tarantulas (Aphonopelma hentzi). The spiders increased sprint speed 2.5-fold over the temperature range by changing their stride frequency but not stride length. The coefficient of determination for linear regression (R(2)) of the proximal and distal joint angles was used as the measure of the degree of coupling between the two joints. This coupling coefficient between the proximal and distal joint angles, for both forelegs and hind-legs, was significantly lowest at the highest temperature at which the animals ran the fastest with the highest stride frequencies. The coordination of multiple, in-series hydraulically actuated joints may be limited by operating speed.

Effects of salinity on the accumulation of hemocyte aggregates and bacteria in the gills of Callinectes sapidus, the Atlantic blue crab, injected with Vibrio campbellii

In addition to respiration and ion regulation, crustacean gills accumulate and eliminate injected particles, along with hemocyte aggregates that form in response to those particles. Here we report that the dose of Vibrio campbellii previously shown to induce a decrease in respiration and hemolymph flow across the gill in the Atlantic blue crab, Callinectes sapidus, also triggered the formation of aggregates containing four or more hemocytes in the gills, compared with saline-injected controls. More bacteria were trapped and rendered non-culturable per unit weight by anterior respiratory gills than posterior gills specialized for ion regulation. Further, more bacteria accumulated in the anterior gills of animals held at 30ppt than those at 10ppt. Thus, the role of the gills in immune defense comes at an energetic cost to this and likely to other crustaceans; this cost is influenced by acclimation salinity and the position and specialized function of individual gills.

Spatial and temporal in vivo analysis of circulating and sessile immune cells in mosquitoes: hemocyte mitosis following infection

BackgroundMosquitoes respond to infection by mounting immune responses. The primary regulators of these immune responses are cells called hemocytes, which kill pathogens via phagocytosis and via the production of soluble antimicrobial factors. Mosquito hemocytes are circulated throughout the hemocoel (body cavity) by the swift flow of hemolymph (blood), and data show that some hemocytes also exist as sessile cells that are attached to tissues. The purpose of this study was to create a quantitative physical map of hemocyte distribution in the mosquito, Anopheles gambiae, and to describe the cellular immune response in an organismal context.ResultsUsing correlative imaging methods we found that the number of hemocytes in a mosquito decreases with age, but that regardless of age, approximately 75% of the hemocytes occur in circulation and 25% occur as sessile cells. Infection induces an increase in the number of hemocytes, and tubulin and nuclear staining showed that this increase is primarily due to mitosis and, more specifically, autonomous cell division, by circulating granulocytes. The majority of sessile hemocytes are present on the abdominal wall, although significant numbers of hemocytes are also present in the thorax, head, and several of the appendages. Within the abdominal wall, the areas of highest hemocyte density are the periostial regions (regions surrounding the valves of the heart, or ostia), which are ideal locations for pathogen capture as these are areas of high hemolymph flow.ConclusionsThese data describe the spatial and temporal distribution of mosquito hemocytes, and map the cellular response to infection throughout the hemocoel.

Disseminated neoplasia causes changes in ploidy and apoptosis frequency in cockles Cerastoderma edule

A proliferative disease, usually referred as disseminated neoplasia (DN), shows high prevalence in some cockle Cerastoderma edule beds of Galicia (NW Spain). Chromosome counts, examination of chromosome morphology, DNA quantification by flow cytometry and estimation of apoptosis frequency by TUNEL assay and flow cytometry were performed in cockles with different DN severity. Metaphases obtained from gills of DN-affected cockles displayed a chromosome number ranging from 41 to 145, while normal number is 38; changes in chromosome morphology were also evident, with numerous microchromosomes occurring. Haemolymph flow cytometry analysis revealed difference in DNA content between healthy and DN-affected cockles. Aneuploid peaks ranged from 1.3n to 8.9n. Apoptosis frequency was determined on histological sections (TUNEL assay) and haemolymph samples (flow cytometry). Both techniques revealed neoplastic cells in apoptosis. The higher DN severity, the lower the percentage of apoptotic cells. According to flow cytometry results, the negative association between DN severity and apoptosis frequency only affected the neoplastic cells, whereas DN did not significantly affect the percentage of apoptotic hyalinocytes or apoptotic granulocytes.

Cardioacceleratory function of the neurohormone CCAP in the mosquito Anopheles gambiae

Crustacean cardioactive peptide (CCAP) is a highly conserved arthropod neurohormone that is involved in ecdysis, hormone release and the modulation of muscle contractions. Here, we determined the CCAP gene structure in the malaria mosquito Anopheles gambiae, assessed the developmental expression of CCAP and its receptor and determined the role that CCAP plays in regulating mosquito cardiac function. RACE sequencing revealed that the A. gambiae CCAP gene encodes a neuropeptide that shares 100% amino acid identity with all sequenced CCAP peptides, with the exception of Daphnia pulex. Quantitative RT-PCR showed that expression of CCAP and the CCAP receptor displays a bimodal distribution, with peak mRNA levels in second instar larvae and pupae. Injection of CCAP revealed that augmenting hemocoelic CCAP levels in adult mosquitoes increases the anterograde and retrograde heart contraction rates by up to 28%, and increases intracardiac hemolymph flow velocities by up to 33%. Partial CCAP knockdown by RNAi had the opposite effect, decreasing the mosquito heart rate by 6%. Quantitative RT-PCR experiments showed that CCAP mRNA is enriched in the head region, and immunohistochemical experiments in newly eclosed mosquitoes detected CCAP in abdominal neurons and projections, some of which innervated the heart, but failed to detect CCAP in the abdomens of older mosquitoes. Instead, in older mosquitoes CCAP was detected in the pars lateralis, the subesophageal ganglion and the corpora cardiaca. In conclusion, CCAP has a potent effect on mosquito circulatory physiology, and thus heart physiology in this dipteran insect is under partial neuronal control.

In vivo imaging of Drosophila wing heart development during pupal stages

Wing hearts are small pumping organs that maintain the flow of hemolymph through the wing veins of insects. In Drosophila, these organs consist of parallel oriented muscle cells and a simple epithelium of connective tissue. Both tissues originate from eight embryonic wing heart progenitors (WHPs), which remain dormant until late larval stages. Most of the differentiation and maturation takes place during the pupal stage following head eversion. In this study, we have used the tissue specific expression of Gal4 enhancer lines, in combination with the live cell markers GFP and DsRed to investigate pupal wing heart development in conjunction with the surrounding tissues. We found that WHPs interact with the tracheal system and specific expression domains of the adult epidermis. Additionally, wing heart development occurs simultaneously with the remodeling of the dorso-lateral epidermis into the scutellum and the scutellar arms. Myogenesis in wing hearts comprises known processes such as founder cell specification, but also new features like removal of growing myotubes, and nuclei movement. Wing heart epithelium development is accomplished by the mesenchymal-epithelial transition of WHPs and occurs slightly delayed to muscle development. The epithelium represents a novel mesodermally derived secondary epithelium. Moreover, we have identified a nerve that runs along the epithelium and innervates the wing heart muscle cells.

Antimicrobial properties of Anopheles albimanus pericardial cells

Insect pericardial cells (PCs) are strategically located along the dorsal vessel where they encounter a high hemolymph flow enabling them to undertake their osmoregulatory, detoxifying, and scavenging functions. In this location, PCs also encounter foreign molecules and microorganisms. The response of PCs of the mosquito Anopheles albimanus, one of the most important Plasmodium vivax vectors in Mexico and Latin America, to Saccharomyces cerevisiae was analyzed by using biochemical, cellular, ultrastructural, and bioinformatics approaches. Immune gene transcripts were identified in the PC transcriptome of A. albimanus. PCs responded to the presence of yeast and zymosan with increased lysosomal and phosphatase activities and produced lytic activity against bacteria. Our results indicate that mosquito PCs play a key role in the neutralization and elimination of pathogens.

Anopheles gambiae corazonin: gene structure, expression and effect on mosquito heart physiology

Haemolymph flow in mosquitoes is primarily driven by the contraction of a dorsal vessel that is subdivided into an abdominal heart and a thoracic aorta. The factors that regulate mosquito heart contractions are not understood, but in other insects heart physiology is partially controlled by several neurohormones. One of these is corazonin, a neuropeptide initially discovered because of its cardioacceleratory activity in the cockroach Periplaneta americana. In the present study, we describe the corazonin gene and transcript structure in the mosquito Anopheles gambiae, characterize its developmental expression, and test its role in modulating heart physiology. We show that the A. gambiae corazonin gene encodes the most common form of the corazonin peptide ([Arg(7) ]-corazonin) and that it is alternatively spliced, with the only difference between the transcripts occurring in the 5' untranslated region. Analysis of the developmental expression of corazonin and the corazonin receptor revealed that transcription of both follows a bimodal distribution, with highest mRNA levels in 2nd instar larvae and during the pupa to adult transition. Finally, experiments where mosquitoes were injected with various doses of corazonin and experiments where the transcription of corazonin and the corazonin receptor were reduced by RNA interference failed to detect a significant role for this neuropeptide in modulating mosquito heart physiology.

Properties of Bacteria That Trigger Hemocytopenia in the Atlantic Blue Crab, Callinectes sapidus

In the blue crab Callinectes sapidus, injection with the bacterial pathogen Vibrio campbellii causes a decrease in oxygen consumption. Histological and physiological evidence suggests that the physical obstruction of hemolymph flow through the gill vasculature, caused by aggregations of bacteria and hemocytes, underlies the decrease in aerobic function associated with bacterial infection. We sought to elucidate the bacterial properties sufficient to induce a decrease in circulating hemocytes (hemocytopenia) as an indicator for the initiation of hemocyte aggregation and subsequent impairment of respiration. Lipopolysaccharide (LPS), the primary component of the gram-negative bacterial cell wall, is known to interact with crustacean hemocytes. Purified LPS was covalently bound to the surfaces of polystyrene beads resembling bacteria in size. Injection of these "LPS beads" caused a decrease in circulating hemocytes comparable to that seen with V. campbellii injection, while beads alone failed to do so. These data suggest that in general, gram-negative bacteria could stimulate hemocytopenia. To test this hypothesis, crabs were injected with different bacteria--seven gram-negative and one gram-positive species--and their effects on circulating hemocytes were assessed. With one exception, all gram-negative strains caused decreases in circulating hemocytes, suggesting an important role for LPS in the induction of this response. However, LPS is not necessary to provoke the immune response given that Bacillus coral, a gram-positive species that lacks LPS, caused a decrease in circulating hemocytes. These results suggest that a wide range of bacteria could impair metabolism in C. sapidus.

Evidence for adrenergic receptor activation in control of hemolymph flow in Procambarus clarkii

Previous studies suggest decapods have complex neural control of internal organs similar to autonomic control in vertebrates. Norepinephrine (NE) is a key controller of vertebrate cardiovascular function, yet NE regulation of vascular function in decapods remains unclear. This study investigated whether exogenous NE alters hemolymph flow to the tail muscle in freshwater crayfish Procambarus clarkii. It is hypothesized that NE decreases hemolymph flow from adrenergic constriction of the abdominal artery. P. clarkii (25–30g) were obtained from Atchafalay Biological (Raceland, LA) and allowed 2 weeks of environmental acclimation. Crayfish were sedated in an ice bath prior to study. Polyethylene tubing was inserted into the sternal sinus for injections/infusions. A laser Doppler flow probe was placed on the abdominal artery to measure hemolymph flow. NE (30mM) decreased hemolymph flow by 35.2 ± 8.4% (n=8). Co-infusion of phentolamine (α1/α2 receptor blockade; 5μg/kg/min) abolished NE mediated decreases in hemolymph flow. Stimulation of α2 receptors with clonidine (10 mM) had no effect on hemolymph flow. Thus, vasoconstriction in P. clarkii may involve α1 adrenergic mechanisms. Further pharmacological and immunhistochemical experiments will identify specific adrenergic receptor subtypes in the decapod vasculature responsible for vasoconstriction. Supported by NSF #437768 and the Ribble Foundation

Effects of crustacean cardioactive peptide on the hearts of two Orthopteran insects, and the demonstration of a Frank–Starling-like effect

Like vertebrate cardiovascular systems, the dorsal vessel of the Orthopteran insects Baculum extradentatum and Locusta migratoria is under myogenic as well as neural control, through the action of neurotransmitters, neuromodulators and neurohormones. It was previously shown that the excitatory neuropeptide, crustacean cardioactive peptide (CCAP), triggers an increase in heart rate in B. extradentatum, and CCAP-like immunoreactivity is present in the innervation to the heart in many insects. In the present study, CCAP resulted in a dose-dependent increase in heart rate and hemolymph flow velocity, or cardiac output, in B. extradentatum. In contrast, CCAP led to a significant increase in stroke volume and cardiac output in L. migratoria without modifying heart rate or aortic contraction frequency. Hemolymph flow through the excurrent ostia of L. migratoria, small openings or valves on the posterior aorta and anterior heart, was inhibited with increasing concentrations of CCAP, with complete inhibition seen at 10 −7 M CCAP. In the locust, CCAP increases the volume of hemolymph in the dorsal vessel by the synchronous closing of the excurrent ostia, resulting in more forceful heart contractions and increased stroke volume and cardiac output, without modifying heart rate through a physiological mechanism analogous to the Frank–Starling mechanism in vertebrates. Therefore, crustacean cardioactive peptide alters the contractile properties of cardiac tissue in both B. extradentatum and L. migratoria, allowing for an increase in blood flow and circulation.

Physiological homology betweenDrosophila melanogasterand vertebrate cardiovascular systems

SUMMARYThe physiology of the Drosophila melanogaster cardiovascular system remains poorly characterized compared with its vertebrate counterparts. Basic measures of physiological performance remain unknown. It also is unclear whether subtle physiological defects observed in the human cardiovascular system can be reproduced in D. melanogaster. Here we characterize the cardiovascular physiology of D. melanogaster in its pre-pupal stage by using high-speed dye angiography and optical coherence tomography. The heart has vigorous pulsatile contractions that drive intracardiac, aortic and extracellular-extravascular hemolymph flow. Several physiological measures, including weight-adjusted cardiac output, body-length-adjusted aortic velocities and intracardiac shear forces, are similar to those in the closed vertebrate cardiovascular systems, including that of humans. Extracellular-extravascular flow in the pre-pupal D. melanogaster circulation drives convection-limited fluid transport. To demonstrate homology in heart dysfunction, we showed that, at the pre-pupal stage, a troponin I mutant, held-up2 (hdp2), has impaired systolic and diastolic heart wall velocities. Impaired heart wall velocities occur in the context of a non-dilated phenotype with a mildly depressed fractional shortening. We additionally derive receiver operating characteristic curves showing that heart wall velocity is a potentially powerful discriminator of systolic heart dysfunction. Our results demonstrate physiological homology and support the use of D. melanogaster as an animal model of complex cardiovascular disease.

Contraction of the Ventral Abdomen Potentiates Extracardiac Retrograde Hemolymph Propulsion in the Mosquito Hemocoel

BackgroundHemolymph circulation in mosquitoes is primarily controlled by the contractile action of a dorsal vessel that runs underneath the dorsal midline and is subdivided into a thoracic aorta and an abdominal heart. Wave-like peristaltic contractions of the heart alternate in propelling hemolymph in anterograde and retrograde directions, where it empties into the hemocoel at the terminal ends of the insect. During our analyses of hemolymph propulsion in Anopheles gambiae, we observed periodic ventral abdominal contractions and hypothesized that they promote extracardiac hemolymph circulation in the abdominal hemocoel.Methodology/Principal FindingsWe devised methods to simultaneously analyze both heart and abdominal contractions, as well as to measure hemolymph flow in the abdominal hemocoel. Qualitative and quantitative analyses revealed that ventral abdominal contractions occur as series of bursts that propagate in the retrograde direction. Periods of ventral abdominal contraction begin only during periods of anterograde heart contraction and end immediately following a heartbeat directional reversal, suggesting that ventral abdominal contractions function to propel extracardiac hemolymph in the retrograde direction. To test this functional role, fluorescent microspheres were intrathoracically injected and their trajectory tracked throughout the hemocoel. Quantitative measurements of microsphere movement in extracardiac regions of the abdominal cavity showed that during periods of abdominal contractions hemolymph flows in dorsal and retrograde directions at a higher velocity and with greater acceleration than during periods of abdominal rest. Histochemical staining of the abdominal musculature then revealed that ventral abdominal contractions result from the contraction of intrasegmental lateral muscle fibers, intersegmental ventral muscle bands, and the ventral transverse muscles that form the ventral diaphragm.Conclusions/SignificanceThese data show that abdominal contractions potentiate extracardiac retrograde hemolymph propulsion in the abdominal hemocoel during periods of anterograde heart flow.

Heart wall velocimetry and exogenous contrast-based cardiac flow imaging in Drosophila melanogaster using Doppler optical coherence tomography

Drosophila melanogaster (fruit fly) is a central organism in biology and is becoming increasingly important in the cardiovascular sciences. Prior work in optical imaging of the D. melanogaster heart has focused on static and dynamic structural anatomy. In the study, it is demonstrated that Doppler optical coherence tomography can quantify dynamic heart wall velocity and hemolymph flow in adult D. melanogaster. Since hemolymph is optically transparent, a novel exogenous contrast technique is demonstrated to increase the backscatter-based intracardiac Doppler flow signal. The results presented here open up new possibilities for functional cardiovascular phenotyping of normal and mutant D. melanogaster.

Evidence of sternal artery vasoconstriction in Crayfish, Procambarus clarkii (Pc) but not Pacifastacus leniusculus (Pl)

Procambarus clarkii (Pc) are known to re‐distribute arterial flows when exposed to hypoxic water. In response to hypoxia, flow to the anterior aorta increases while flows to the posterior dorsal aorta and sternal artery decrease, causing an increase of hemolymph flow to the animal's head. For this study we compared isolated blood vessel function from Pc and another crayfish, Pacifastacus leniusculus (Pl). Both crayfish survive for some time out of water, but Pl is more active physically and has narrower arteries. Sternal arteries from both species were isolated. Compliance was measured in cannulated pressurized blood vessels and isometric force was measured on a wire myograph with bicarbonate‐buffered crayfish saline. After relaxation in a calcium‐free solution containing 0.1 mM EGTA, KCl depolarization produced a small transient contraction for Pc but this response has not been observed for Pl. We are now investigating compliance curves in bicarbonate‐buffered crayfish saline in the presence or absence of calcium for Pc and Pl. Vasoconstriction in arteries may have effects on blood flow resistance in Pc but not in Pl.