Carl L. Reiber

The effects of progressive hypoxia on the crustacean cardiovascular system: a comparison of the freshwater crayfish, (Procambarus clarkii ), and the lobster (Homarus americanus)

Abstract The heart rate of crayfish (Procambarus clarkii) and lobsters (Homarus americanus) decreased (bradycardia) as partial pressure of O2 (PO2) decreased, yet cardiac output (Vb) was maintained via an increased stroke volume (Sv) to PO2s of 40 mmHg and 75 mmHg for crayfish and lobsters, respectively. Vb was redistributed in both animals. Flow through the anterior aorta increased while flow dropped through the posterior aorta and sternal artery to a PO2 of 30 mmHg; below this flow was no longer maintained in crayfish. In the lobster, flow increased to the lateral arteries and the ventral thoracic artery while flow through the anterior and posterior aortas, sternal artery and ventral abdominal artery decreased to a PO2 of 75 mmHg. Anterior hemolymph flow was maintained or increased in both animals presumably to supply nervous tissue and cephalic sense organs better. Crayfish showed an increase in intracardiac and mean arterial hemolymph pressures as PO2 declined. The increased pressures combined with the net increase in cardiac filling pressure and diastolic filling time could have accounted for the increased SV. The cardiovascular response exhibited by both the crayfish and lobster was PO2 dependent; below a critical water PO2 active compensation was no longer observed.

Heart rate during development in the turtle embryo: effect of temperature

Growth and development can occur over a wide range of physical conditions in reptiles. Cardiovascular function must be critical to this ability. However, information on cardiovascular function in developing oping reptiles is lacking. Previous work indicated that in reptiles the effects of temperature on growth and metabolism are largely restricted to early development. This study examined whether the previously observed effects of temperature and different perinatal patterns of metabolism observed in amniotic vertebrates are correlated with cardiovascular function. Embryonic and hatchling carcass mass, heart mass and heart rate (HR) were compared for snapping turtle eggs (Chelydra serpentina) incubated at 24° and 29°C. Incubation time was shorter at 29 °C (56.2 days) than at 24 °C (71:1 days). Carcass and heart growth showed a sigmoidal pattern at both temperatures. However, cardiac growth showed a relative decrease as incubation proceeded. Incubation temperature significantly affected the HR pattern during development. The HR of embryos incubated at 24 °C was constant for most of incubation (51.8±4.8 min-1). A small decrease was observed just prior to and a large decrease immediately following hatching (posthatch, 22.3±4.1 min-1). At 29 °C embryonic HR was greater than at 24 °C early in development (72.3±3 min-1). The HR steadily decreased to values equivalent to those at 24 °C. The HRs of 24 °C and 29 °C hatchlings were not different. Cardiac output (estimated as the product of heart mass and HR) increased rapidly during early development and then slowed dramatically at both temperatures. These data are consistent with the suggestion that temperature exerts its effects primarily early in development. Furthermore, the changes in cardiovascular function are correlated with metabolic changes in hatching vertebrates.

Behavioral Physiology of Four Crab Species in Low Salinity

Reports focusing on the behavioral responses of crabs to exposure to low salinity have involved choice chamber experiments or quantification of changes in activity. In addition to describing changes in locomotor activity in four species of crabs of differing osmoregulatory ability, the present study describes six behaviors: increased movement of the mouthparts, cleaning of the mouthparts with the chelae, cleaning of the antennae and antennules with the maxillipeds, flicking of the antennae, retraction of the antennules, and extension of the abdomen. Callinectes sapidus and Carcinus maenas are classed as efficient osmoregulators, and in general, showed an increase in these behaviors with decreasing salinity. Cancer magister, a weak regulator, and Libinia emarginata, an osmoconformer, exhibited these behaviors to a lesser degree and became inactive in the lower salinities, tending to adopt an isolation-type response. The differences in behaviors between the species correlated closely with previously reported changes in cardiovascular function and hemolymph flow. These overt reactions are discussed in relation to the osmoregulatory physiology and ecology of each crab species.

Circulatory modification in the blue crab Callinectes sapidus, during exposure and acclimation to low salinity

Abstract A pulsed-Doppler flowmeter was used to measure heart rate and haemolymph flow rates in each arterial system of the blue crab, Callinectes sapidus, enabling calculation of stroke volume and cardiac output. During exposure to a 6–6–12 h salinity cycle of 100–25–100% seawater, there was an immediate increase in heart rate upon dilution of the medium. After this initial increase it decreased steadily, but remained elevated above levels in 100% seawater. A smaller increase in heart rate occurred when the salinity was raised, declining thereafter and reaching pre-treatment levels after 6 h in 100% SW. There was a slight decrease in stroke volume of the heart, but overall this resulted in an increase in cardiac output when the salinity was lowered. Differential haemolymph flow through each major arterial system also occurred. There was an increase in flow rates through the anterior aorta, anterolateral arteries and sternal artery during the first 2 h of low salinity exposure and smaller increases occurred again when the salinity was raised to 100% seawater. No significant changes in flow were observed in the hepatic arteries or posterior aorta. During a 72 h acclimation period in low salinity, similar increases in cardiac parameters and flow rates were observed in the first 6 h. These values declined to levels comparable to those in 100% seawater, after 40–50 h acclimation in low salinity. The changes in cardiovascular parameters are not directly related to the osmoregulatory physiology of this species, but appear to be due to specific behaviours occurring in response to low salinity. The results obtained here for this efficient osmoregulator are compared and contrasted with similar studies on Cancer magister, which is classified as a weak hyperosmoregulator.

A Review of the “Open” and “Closed” Circulatory Systems: New Terminology for Complex Invertebrate Circulatory Systems in Light of Current Findings

Invertebrate cardiovascular systems have historically been viewed as sluggish, poorly regulated, and “open”, where blood bathes the tissues directly as it moves through a system of ill-defined sinuses and/or lacunae without an endothelial boundary. When examining cardiovascular/circulatory morphology and physiology in a broader evolutionary context, one can question the very nature of the definition of a “closed” versus “open” circulatory system. Viewed in this context a number of invertebrates have evolved incomplete or even completely cell-lined vessels and or lacunae with a highly branched vasculature that allows for the production of significant driving pressures and flows to meet relatively high metabolic demands driven by active life styles. In light of our current understanding of invertebrate cardiovascular systems and their paralleled complexity to vertebrate systems, a number of long established paradigms must be questioned and new definitions presented to better align our understanding of the nature of “open” versus “closed” cardiovascular systems.

Environmental hypoxia influences hemoglobin subunit composition in the branchiopod crustacean Triops longicaudatus.

SUMMARY Hemoglobin (Hb) is a highly conserved protein that provides a vital link between environmental oxygen and its use and/or storage within an organism. While ubiquitous among vertebrates, Hb occurs frequently in invertebrate phyla as well. Many arthropod species use the copper-binding pigment hemocyanin, but unique in this phylum are the branchiopod crustaceans, which express Hb. Branchiopod Hb concentration and structure are exquisitely sensitive to environmental oxygen availability. Hemoglobin concentration and oxygen-binding affinity increase with decreasing oxygen tension in Daphnia, Artemia and Triops. The change in binding affinity is attributed to differential Hb subunit expression in Daphnia and Artemia but remains unclear for Triops. This is the first study to demonstrate developmental plasticity of Hb subunit expression in a notostracan, Triops longicaudatus, reared under conditions of varying oxygen availability. In response to variable oxygen environments, T. longicaudatus differentially express four primary Hb subunits ranging between 30 and 34 kDa, with normoxic-reared animals expressing primarily the heavier subunits, and hypoxic-reared animals expressing increased proportions of the lower molecular mass subunits. Moreover, differential Hb subunit expression is induced upon transfer of normoxic-reared adults to a hypoxic environment, such that the distribution of Hb subunits in the transferred adults becomes similar to that of hypoxic-reared animals. Two-dimensional gel electrophoresis and follow-up analyses revealed several isoforms of Hb subunits that may represent differential gene expression and/or post-translational modification. Unlike Daphnia and Artemia, the Hb hypoxic response in Triops is not reversible in that there was no significant decrease in Hb concentration or change in Hb subunit expression pattern when hypoxic-reared adults were transferred to a normoxic environment.

Assessment of the pressure-volume relationship of the single ventricle of the grass shrimp, Palaemonetes pugio.

SUMMARY The ventricular pressure–volume (PV) relationship has been used extensively to study the mechanics and energetics in multi-chambered hearts of closed circulatory system vertebrates. In the current study we applied the use of PV loops in the assessment of cardiac mechanics and energetics in the single ventricle of a decapod crustacean possessing an open circulatory system. Anatomical differences between multi-and single-chambered hearts include multiple ostia entering and valved multiple arterial systems exiting the ventricle, and the neurogenic origin of the heartbeat in decapod crustaceans. However, the microscopic architecture and excitation–contraction coupling events are similar in both systems. Ventricular pressure and area were obtained independently and integrated into pressure–area loops. Area was then converted to volume to generate PV loops. Based on the PV loops generated in this study, the ventricle of Palaemonetes pugio processes the same primary phases of the cardiac cycle as ventricles from the multi-chambered hearts of vertebrates: (1) isovolumic contraction, (2) ventricular emptying, (3) isovolumic relaxation and (4) ventricular filling. The area enclosed by the PV loop provides a measure of stroke work and when multiplied by heart rate provides an assessment of cardiac work. This initial examination of PV loops from a single-ventricle decapod crustacean demonstrates the utility of this technique to further elucidate the cardiac mechanics and energetics of this system, and in particular during times of physiological stress.

Perspectives on cardiac physiological ontogeny in crustaceans.

Crustacean embryonic and larval systems offer a unique and valuable tool for furthering our understanding of both developmental processes and physiological regulatory mechanisms. The diverse array of developmental patterns exhibited by crustaceans allows species choice to be based on the specific questions being investigated, where defined larval forms are chosen based on their developmental pattern, degree of maturation or regulatory capabilities. However, this great diversity in developmental patterns, as well as crustacean diversity, can also confound ones ability to define or identify species for investigation. These issues are addressed and suggestions put forth to clarify some of the problems. The complexity and overlapping nature of adult cardio-regulatory systems makes teasing them apart difficult. Embryonic and larval systems exhibit varying degrees of regulatory complexity depending on developmental stage and ontogenetic pattern. This can allow complex adult regulatory systems to be teased apart temporally, as the developing animal builds regulatory pathways. Equally important is the nature of crustacean larvae; many undergo dramatic metamorphoses in cases where the larvae have adaptations to environments different to those of the adult. During environmental transitions physiological adaptations to immediate change should take precedence over long-term adult adaptations. It is therefore possible to look at physiological responses as a function of developmental/environmental adaptation, independent of adult functions.

Oxygen Sensitivity in the Crayfish Procambarus clarkii: Peripheral O 2 Receptors and Their Effect on Cardiorespiratory Functions

ABSTRACT The intent of this study was to determine the location and functional influence of the branchial O2-sensitive receptors of the crayfish on cardiac and respiratory parameters. Crayfish (46.6 ± 1.7 g [±SD]) were divided into 3 groups (A) control (gills intact), (B) posterior gills removed, and (C) anterior gills removed. Animals were exposed to hypoxic water (1 h; 15 mm Hg O2) followed by left branchial injection (10 ml over 10 min) of hyperoxic water (400 mm Hg O2). The time course to changes in cardiorespiratory parameters (heart rate, cardiac output, stroke volume, arterial flows, ventilation rate, and branchial pressure) were monitored. Intact animals showed a rapid cardiovascular response to injection of hyperoxic water (2.5 s) with group C animals showing a similar response time. Group B animals exhibited a significantly longer response time (13.0 s). Ventilatory response time was similar for all 3 groups (75 s), the duration of the ventilatory response was 51.0 s in control and group C animals. Animals lacking posterior gills showed a significantly shorter response time (42.0 s). Time-course evidence may indicate the existence of 2 populations of O2 receptors, one responsible for modulation of cardiovascular functions on the posterior gills or in their cardiobranchial veins, the other responsible for ventilatory modulation located more centrally.

Changes in cardiac performance during hypoxic exposure in the grass shrimp Palaemonetes pugio.

SUMMARY In hearts of higher invertebrates as well as vertebrates, the work performed by the ventricle is a function of both rate and contractility. Decapod crustaceans experience a hypoxia-induced bradycardia that is thought to result in an overall reduction in cardiac work; however, this hypothesis has not yet been tested and is the primary purpose of this study. In the grass shrimp Palaemonetes pugio, cardiac pressure and area data were obtained simultaneously, and in vivo, under normoxic (20.2 kPa O2) and hypoxic (6.8 or 2.2 kPa O2) conditions and integrated to generate pressure–area (P–A) loops. The area enclosed by the P–A loop provides a measure of stroke work and, when multiplied by the heart rate, provides an estimate of both cardiac work and myocardial O2 consumption. Changes in intra-cardiac pressure (dp/dt) are correlated to the isovolemic contraction phase and provide an indication of stroke work. At both levels of hypoxic exposure, intra-cardiac pressure, dp/dt, stroke work and cardiac work fell significantly. The significant decrease in intra-cardiac pressure provides the primary mechanism for the decrease in stroke work, and, when coupled with the hypoxia-induced bradycardia, it contributes to an overall fall in cardiac work. Compared with normoxic P–A loops, hypoxic P–A loops (at both levels of hypoxia) become curvilinear, indicating a fall in peripheral resistance (which might account for the reduction in intra-cardiac pressure), which would reduce both stroke work and cardiac work and ultimately would serve to reduce myocardial O2 consumption. This is the most direct evidence to date indicating that the hypoxia-induced bradycardia observed in many decapod crustaceans reduces cardiac work and is therefore energetically favorable during acute exposure to conditions of low oxygen.