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Bernd Pelster

Publications and source records attributed to Bernd Pelster.

13 recordsLinked to original sources
PubMed2005

NO as a mediator during the early development of the cardiovascular system in the zebrafish.

As a general pattern innervation of the cardiovascular system appears late during development in vertebrate embryos, and cardiovascular control may be achieved by hormonal activity in early stages. However, very little is known about the onset of NO-responsiveness during development, which in adult vertebrates is known to play a key function in many physiological processes such as control of vascular tone, neurotransmission, macrophage activity, and angiogenesis. Analysis of the effect of NO on the cardiovascular system in zebrafish (Danio rerio) embryos and larvae revealed almost no effect on cardiac activity during chronic exposure to NO-producing chemicals, whereas vascular reactivity was observed in veins and arteries of the zebrafish in early developmental stages (5-6 days post fertilization). Chronic exposure also modified the development of the vascular system. The presence of an NO donor (sodium nitroprusside) did not change the patterning of the vascular bed, but it induced an earlier appearance of some blood vessels in the trunk region of the zebrafish larvae. The data reveal that NO plays an important role in the development of the cardiovascular system and in the ontogeny of the cardiovascular control system in fish.

Animals↗
PubMed2005

Importance of cytoskeletal elements in volume regulatory responses of trout hepatocytes.

The role of cytoskeletal elements in volume regulation was studied in trout hepatocytes by investigating changes in F-actin distribution during anisotonic exposure and assessing the impact of cytoskeleton disruption on volume regulatory responses. Hypotonic challenge caused a significant decrease in the ratio of cortical to cytoplasmic F-actin, whereas this ratio was unaffected in hypertonic saline. Disruption of microfilaments with cytochalasin B (CB) or cytochalasin D significantly slowed volume recovery following hypo- and hypertonic exposure in both attached and suspended cells. The decrease of net proton release and the intracellular acidification elicited by hypotonicity were unaltered by CB, whereas the increase of proton release in hypertonic saline was dramatically reduced. Because amiloride almost completely blocked the hypertonic increase of proton release and cytoskeleton disruption diminished the associated increase of intracellular pH (pH(i)), we suggest that F-actin disruption affected Na(+)/H(+) exchanger activity. In line with this, pH(i) recovery after an ammonium prepulse was significantly inhibited in CB-treated cells. The increase of cytosolic Na(+) under hypertonic conditions was not diminished but, rather, enhanced by F-actin disruption, presumably due to inhibited Na(+)-K(+)-ATPase activity and stimulated Na(+) channel activity. The elevation of cytosolic Ca(2+) in hypertonic medium was significantly reduced by CB. Altogether, our results indicate that the F-actin network is of crucial importance in the cellular responses to anisotonic conditions, possibly via interaction with the activity of ion transporters and with signalling cascades responsible for their activation. Disruption of microtubules with colchicine had no effect on any of the parameters investigated.

Actins↗
PubMed2005

Epigenetic variations in early cardiovascular performance and hematopoiesis can be explained by maternal and clutch effects in developing zebrafish (Danio rerio).

This is the first study to show maternal and clutch effects on early developmental parameters like blood cell concentration and cardiac performance (heart rate, stroke volume and cardiac output) in developing zebrafish larvae (2-8 days post fertilization, dpf). Ten individuals per parental pair and developmental stage were analyzed. A pronounced interclutch variation of heart rate has been found in younger stages (2-4 dpf), while interclutch variation of heart rate was small in later stages (8 dpf). This effect was more pronounced in offspring from parental fish nourished with living food. The opposite effect was observed in interclutch variation of blood cell concentration. Here only older stages showed significant interclutch variations. Stroke volume and cardiac output had very small interclutch variations throughout all stages. Heart rate was strongly dependent with developmental stage in all groups. Nutritional maternal effects on heart performance and also in blood cell concentration could be detected in the offspring of parent animals either fed with flake food or with living food. Red blood cell count, calculated as a product from red blood cell concentration, was not significantly different in both feeding groups. The number of spawned eggs was not different. In summary, these data indicate that "clutch effects" caused by maternal and/or genetic influences can affect the developmental pattern of cardiac performance and blood cell concentration.

Animals↗
PubMed2005

The influence of environmental P(O(2)) on hemoglobin oxygen saturation in developing zebrafish Danio rerio.

Several studies suggest that during early larval development of lower vertebrates convective blood flow is not essential to supply oxygen to the tissues, but information about the oxygenation status of larvae during the time of cutaneous respiration is still missing. If convective oxygen transport contributes to the oxygen supply to tissues, venous blood in the central circulatory system should be partly deoxygenated, and hyperoxia should increase the oxygen saturation of the hemoglobin. To analyze the changes in hemoglobin oxygen saturation induced by hyperoxic incubation, zebrafish larvae were incubated in a tiny chamber between polytetrafluoroethylene membranes (Teflon), so that the oxygen supply could be rapidly modified. Hemoglobin oxygen saturation was measured in vivo by combining video imaging techniques with a spectrophotometrical analysis of hemoglobin light absorption at specific wavelengths for maximal absorption of oxygenated and deoxygenated blood (413 nm and 431 nm, respectively) under normoxic conditions and after a 10 min period of hyperoxia (P(O(2))=100 kPa), assuming that at a P(O(2)) of 100 kPa the hemoglobin is fully saturated. The results demonstrated that red blood cell oxygenation of zebrafish larvae at 4 days post fertilization (d.p.f.), 5 d.p.f. and 12 d.p.f. could be increased by hyperoxia. The data suggest that at the time of yolk sac degradation (i.e. 4 d.p.f. and 5 d.p.f.), when the total surface area of the animal is reduced, bulk diffusion of oxygen may not be sufficient to prevent a partial deoxygenation of the hemoglobin. The decrease in hemoglobin oxygenation observed at 12 d.p.f. confirms earlier studies indicating that at 12-14 d.p.f., convective oxygen transport becomes necessary to ensure oxygen supply to the growing tissues.

Analysis of Variance↗
PubMed2005

Cardiac performance in the zebrafish breakdance mutant.

In the Tubingen screen a breakdance mutant of zebrafish (bre) was described as an arrhythmia, in which the ventricle beats only with every second atrial contraction (2:1 rhythm). Surprisingly, a careful analysis of the effect of the breakdance mutation on cardiac performance of the zebrafish during development between 3 d.p.f. and 14 d.p.f revealed that homozygous bre mutants did not always show the 2:1 rhythm. Cardiac activity was continuously recorded for a period of 20 min in each larva, and during this period we observed that heart rate randomly switched between the 2:1 rhythm and a 1:1 rhythm. Furthermore, at 28 degrees C and at 31 degrees C the expression of the 2:1 rhythm decreased with development. At 31 degrees C this was in part due to a significantly reduced survival rate of mutants beyond 4 d.p.f. Besides development, temperature had a marked effect on the expression of the 2:1 rhythm, and during the first days of development the expression of the 2:1 rhythm was significantly higher at elevated incubation temperatures. By contrast, in the 2:1 beating heart ventricular contraction rate was about 80 beats min(-1) throughout development irrespective of the temperature, and even in the 1:1 rhythm mutants showed a significant bradycardia at all three temperatures (25 degrees C, 28 degrees C or 31 degrees C). Compared to wild-type animals, cardiac output was significantly lower in bre mutants. Pressure traces recorded in the ventricle of mutants revealed a prolonged relaxation phase, indicating that the second pacemaker current could not be conveyed to the ventricle (AV-block). This phenotype is comparable to the human Long QT Syndrome, an arrhythmia caused by a modification of an ion channel involved in cardiac repolarization. The bradycardia and the modified temperature sensitivity of heart rate suggested that the activity of the pacemaker cells was also affected by this mutation.

Animals↗
PubMed2004

pH regulation and swimbladder function in fish.

Gas gland cells of the European eel (Anguilla anguilla) are specialized for the production and secretion of acidic metabolites. Although typically exposed to high oxygen partial pressures, they convert glucose mainly into lactate, but also produce CO2 in the pentose phosphate shunt. Only a very small fraction of glucose is oxidized via aerobic metabolism. Although the buffer capacity of gas gland cells appears to be high, even at low extracellular pH values intracellular pH is always kept about 0.2-0.3 pH-units more acidic. Thus, under all physiological conditions proton concentration within gas gland cells is higher than in the extracellular fluid, facilitating proton extrusion. Diffusion of CO2, Na+/H+-exchange, sodium-dependent anion exchange and a V-ATPase represent the pathways available for proton secretion. While under resting conditions the sodium-dependent pathways and diffusion of CO2 appear to be the dominating mechanisms for acid secretion, at low intracellular pH the contribution of Na+/H+-exchange and of V-ATPase appear to increase, while sodium-dependent anion exchange becomes less important. The mechanisms regulating the activity of these acid-secreting pathways and of the metabolism responsible for the production of protons are largely unknown.

Acids↗
PubMed2004

Acid-base regulation in isolated gill cells of the goldfish (Carassius auratus).

Mechanisms of acid release and intracellular pH (pH(i)) homeostasis were analysed in goldfish (Carassius auratus) gill cells in primary culture. The rate of acid secretion was measured using a cytosensor microphysiometer, and pH(i) was determined using the fluorescent probe 2',7'-bis-(3-carboxypropyl)-5-(and-6)-carboxyfluorescein (BCPCF). Amiloride, a Na(+) channel and Na(+)/H(+) exchanger (NHE) inhibitor, had no effect on pH(i), but acid secretion of the gill cells was significantly impaired. In the presence of amiloride, the intracellular acidification (achieved using the NH(4)Cl pulse technique) was more severe than in the absence of amiloride, and recovery from the acidosis was slowed down. Accordingly, acid secretion of gill cells was severely reduced in the absence of extracellular Na(+). Under steady-state conditions, 4,4'-diisothiocyanatodihydro-stilbene-2,2'-disulfonic acid (DIDS), a HCO(3)(-)-transport inhibitor, caused a slow acidification of pH(i), and acid secretion was significantly reduced. No recovery from intracellular acidification was observed in the presence of DIDS. Bafilomycin A(1), an inhibitor of V-ATPase, had no effect on steady-state pH(i) and recovery from an intracellular acidification, whereas the rate of acid secretion under steady-state conditions was slightly reduced. Immunohistochemistry clearly revealed the presence of the V-ATPase B-subunit in goldfish gill lamellae. Taken together, these results suggest that a Na(+)-dependent HCO(3)(-) transport is the dominant mechanism besides an NHE and V-ATPase to control pH(i) in goldfish gill cells.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed2004

Isoforms vatB1 and vatB2 of the vacuolar type ATPase subunit B are differentially expressed in embryos of the zebrafish (Danio rerio).

The v-type ATPase is a membrane anchored, multi-subunit proton pump, which in freshwater fish appears to play a major role in ionoregulative processes in the apical membrane of specialized gill cells. Very little is known about free-living fish embryos and larvae that are exposed to hypo-osmotic conditions with spawning but do not have their gills fully developed. By using reverse transcriptase-polymerase chain reaction and immunological methods, we could demonstrate the presence of two isoforms of the subunit B of this v-type ATPase in the early development of the zebrafish. Immunohistochemical analysis revealed the presence of one isoform (vatB1) in the apical membrane of embryonic skin cells, while vatB2 has been found ubiquitously. This differential localization of the two isoforms supports the hypothesis that vatB1 is preferentially involved in ionoregulative functions, while vatB2 may be preferentially responsible for acidification of intracellular vesicles.

Adenosine Triphosphatases↗
PubMed2004

Ontogeny of the gut motility control system in zebrafish Danio rerio embryos and larvae.

Using digital motion analysis, the ontogeny of the cholinergic, tachykinin and pituitary adenylate cyclase-activating polypeptide (PACAP) control systems was studied in zebrafish Danio rerio larvae, in vivo. For the first time we show that the regular propagating anterograde waves that occur in the zebrafish larval gut before and around the onset [at 5-6 days post fertilization (d.p.f.)] of feeding are modulated by acetylcholine or atropine, PACAP and NKA (neurokinin A). At 3 d.p.f., when no spontaneous motility has developed, application of acetylcholine did not affect the gut. However, at 4 d.p.f., acetylcholine increased and atropine reduced the frequency of propagating anterograde waves. At 5 d.p.f., NKA increased and PACAP reduced the wave frequency. This suggests that both excitatory and inhibitory pathways develop at an early stage in the gut, independent of exogenous feeding. Immunohistochemistry established the presence of gut neurons expressing PACAP and NKA in the proximal part of the developing gut from the first stage investigated (2 d.p.f.) and before regular motility was observed. 1 d.p.f. (PACAP) or 2 d.p.f. (NKA) stages later the whole gut was innervated. This supports physiological results that gut motility is under neuronal control during the period when regular motility patterns develop.

Acetylcholine↗
PubMed2003

Non-invasive imaging of blood cell concentration and blood distribution in zebrafish Danio rerio incubated in hypoxic conditions in vivo.

This is the first study to use a combination of digital imaging techniques and vital video microscopy to study hypoxia-induced changes in blood cell concentration, angiogenesis and blood redistribution in entire animals. Zebrafish Danio rerio, which are known to be independent of convective oxygen transport until about 2 weeks post-fertilization, were raised under chronic hypoxia (P(O(2))=8.7 kPa) starting at 1 day after fertilization (d.p.f.) until 15 d.p.f. In control animals, the concentration of red cells (i.e. the number of red cells per nl blood) remained constant until 7 d.p.f., and than decreased by approximately 70% until 15 d.p.f. In hypoxic animals, however, the concentration of red cells remained significantly elevated compared to control animals at 12 and 15 d.p.f. Assuming that the hemoglobin content of the red cells is similar, hypoxic animals have a higher oxygen carrying capacity in their blood. Red cell distribution within the various parts of the circulatory system, taken as an indicator for blood distribution, revealed a significant modification in the number of blood cells perfusing the organs in hypoxic animals. At 12 d.p.f., gut perfusion was reduced by almost 50% in hypoxic animals, while perfusion of the segmental muscle tissue was increased to 350% of control values. No significant changes in brain perfusion were observed under these conditions. At 15 d.p.f., the reduction in gut perfusion was abolished, although muscle perfusion was still significantly elevated. At this time, growth of hypoxic animals was less compared to control animals, revealing that hypoxia had become deleterious for further development. The vascular bed of various organs was not obviously different in hypoxic animals compared to normoxic animals.

Analysis of Variance↗
PubMed2002

Development of the swimbladder in the European eel ( Anguilla anguilla).

The swimbladder of the adult eel, Anguilla anguilla, with its bipolar countercurrent system, the rete mirabile, is a widely used model for swimbladder function, but very little is known about the development of this swimbladder. Our histological studies on the developing swimbladder revealed that during metamorphosis the swimbladder becomes present as a dorsal outgrowth of the esophagus. It is filled with surfactant, and gas was not detected in the swimbladder. In the young glass-eel, the epithelial (gas gland) cells of the swimbladder are columnar, but do not yet have the typical basolateral labyrinth established in adult animals. Few blood vessels are found in the swimbladder tissue, and the submucosa is present as a thick layer of connective tissue, giving a large diffusion distance between blood vessel and swimbladder lumen. Within the next 2 or 3 months of development, gas gland cells develop their typical basolateral labyrinth, and the thickness of the submucosa is significantly reduced, resulting in a short diffusion distance between blood vessels and the swimbladder lumen. The first filling of the swimbladder with gas is observed while the gas gland cells are still in a poorly differentiated status and it appears unlikely that these cells can accomplish their typical role in gas deposition. The presence of small gas bubbles in the swimbladder as well as in the ductus pneumaticus at the time of initial swimbladder inflation suggests that the swimbladder is filled by air gulping or possibly by taking up gas bubbles from the water.

Air Sacs↗
PubMed2002

Developmental plasticity in the cardiovascular system of fish, with special reference to the zebrafish.

During development the circulatory system of vertebrates typically starts operating earlier than any other organ. In these early stages, however, blood flow is not yet linked to metabolic requirements of tissues, as is well established for adults. While the autonomic nervous system becomes functional only quite late during development, in the early stages control of blood flow appears to be possible by blood-borne and/or local hormones. This study presents methods based on video-imaging techniques and fluorescence microscopy to visualize cardiac activity, as well as the vascular bed of developing lower vertebrates, and tests the idea that environmental factors, such as hypoxia, may modify cardiac activity, or even the early formation of blood vessels in embryos and larvae. In zebrafish larvae, adaptations of cardiovascular activity to chronic hypoxia become visible shortly after hatching, and the formation of some blood vessels is enhanced under chronic hypoxia. Exposure of early larval stages of zebrafish to a constant water current induces physiological adaptations, resulting in enhanced swimming efficiency and increased tolerance towards hypoxia. Furthermore, application of hormones such as NO can modify cardiac activity as well as peripheral resistance, and they can stimulate blood vessel formation. In consequence, even during early development of fish or amphibian larvae, the performance of cardiac muscle and of skeletal muscle can be modified by environmental influences and peripheral resistance can be adjusted. Even blood vessel formation can be stimulated by hypoxia, for example, or by the presence of specific hormones. Thus, at approximately the time of hatching the physiological performance of vertebrate larvae is already determined by the combined action of environmental influences and of genetic information.

Animals↗