Biomedical subjects

D P Hader

Publications and source records attributed to D P Hader.

At least 19 recordsLinked to original sources

PubMed2002

Life under solar UV radiation in aquatic organisms.

Aquatic photosynthetic organisms are exposed to solar ultraviolet (UV) radiation while they harvest longer wavelength radiation for energetic reasons. Solar UV-B radiation (280-315 nm) affects motility and orientation in motile organisms and impairs photosynthesis in cyanobacteria, phytoplankton and macroalgae as measured by monitoring oxygen production or pulse amplitude modulated fluorescence analysis. Upon moderate UV stress most organisms respond by photoinhibition which is an active downregulation of the photosynthetic electron transport in photosystem II by degradation of UV-damaged D1 protein. Photoinhibition is readily reversible during recovery in shaded conditions. Excessive UV stress causes photodamage which is not easily reversible. Another major target is the DNA where UV-B mainly induces thymine dimers. Cyanobacteria, phytoplankton and macroalgae produce scytonemin, mycosporine-like amino acids and other UV-absorbing substances to protect themselves from short wavelength solar radiation.

Amino Acids↗

PubMed2001

Graviperception and gravitaxis in algae.

Photosynthetic flagellates are among the most intensely studied unicellular organisms in the field of graviperception and gravitaxis. While the phenomenon of graviorientation has been known for many decades, only recently was the molecular mechanism unveiled. Earlier hypotheses tried to explain the precise orientation by a passive buoy mechanism assuming the tail end to be heavier than the front. In the photosynthetic flagellate Euglena gracilis, the whole cell body is denser than the surrounding medium, pressing onto the lower cell membrane where it seems to activate mechanosensitive ion channels specific for calcium. The calcium entering the cells during reorientation can be visualized by the fluorescence probe, Calcium Crimson. Cyclic AMP is likewise involved in the molecular pathway. Inhibitors of calcium channels and ionophores impair gravitaxis while caffeine, a blocker of the phosphodiesterase, enhances the precision of orientation.

Animals↗

PubMed2001

Physiological characterization of gravitaxis in Euglena gracilis and Astasia longa studied on sounding rocket flights.

Euglena gracilis is a photosynthetic, unicellular flagellate found in eutrophic freshwater habitats. The organisms control their vertical position in the water column using gravi- and phototaxis. Recent experiments demonstrated that negative gravitaxis cannot be explained by passive buoyancy but by an active physiological mechanism. During space experiments, the threshold of gravitaxis was determined to be between 0.08 and 0.12 x g. A strong correlation between the applied acceleration and the intracellular cAMP and Ca2+ was observed. The results support the hypothesis, that the cell body of Euglena, which is denser than the surrounding medium exerts a pressure onto the lower membrane and activates mechanosensitive Ca2+ channels. Changes in the membrane potential and the cAMP concentration are most likely subsequent elements in a signal transduction chain, which results in reorientation strokes of the flagellum.

Animals↗

PubMed2001

Calcium is involved in the gravitactic orientation in colorless flagellates.

The colorless flagellate Astasia longa shows a pronounced negative gravitaxis. The calcium fluorescence indicator Calcium Crimson was used to detect changes of the intracellular calcium concentration during gravitactical orientation. Astasia shows an increase of the fluorescence after a lag phase of about 10 s, a maximum after about 30 s and a decrease to the basic level within 60 s during gravitactic reorientation. The observed change in fluorescence corresponds to an almost doubling of the initial free calcium concentration. The influence of inhibitors, known to impair gravitaxis, on the calcium concentration of Astasia longa was tested. Addition of caffeine, an inhibitor of phosphodiesterase, increases, while addition of gadolinium, an inhibitor of mechanosensitive ion channels decreases the fluorescence signal. While gravitactic stimulation of caffeine-treated cells resulted in a kinetics of fluorescence intensity changes comparable to control cells the addition of gadolinium inhibited any calcium concentration change. Dynamic fluorescence imaging was used during a sounding rocket experiment (MAXUS 3 campaign). Different accelerations interrupted by microgravity intervals were applied to Astasia cells. The cells show an increase in the calcium signal upon acceleration and a decrease during the microgravity state. The results strongly reemphasize the working model of gravitaxis which is based on the activation of mechano-sensitive ion channels as one of the primary events in signal perception.

Acceleration↗

PubMed2001

Possible involvement of the membrane potential in the gravitactic orientation of Euglena gracilis.

Euglena gracilis, a unicellular photosynthetic flagellate, uses light and gravity as environmental hints to reach and stay in regions optimal for growth and reproduction. The current model of gravitaxis (the orientation with respect to the earth's gravitational field) is based on the specific density difference between cell body and medium. The resulting sedimentation of the cell body applies a force to the lower membrane. This force activates mechano-sensitive ion channels. The resulting ion flux changes the membrane potential, which in turn triggers reorientational movements of the trailing flagellum. One possibility for recording the predicted membrane potential changes during reorientation is the use of potential-sensitive dyes, such as Oxonol VI. The absorption changes of the dye indicating potential changes were recorded with a custom-made photometer, which allows a high precision measurement with a high temporal resolution. After a gravitactic stimulation, a short period of hyperpolarization was detected, followed by a massive depolarization of the cell. The membrane potential returned to initial values after a period of approximately 200 s. Parallel measurements of the precision of orientation and the membrane potential showed a close relationship between both phenomena. The obtained results support the current model of gravitaxis of Euglena gracilis.

Animals↗

PubMed2000

Effects of light on gravitaxis and velocity in Chlamydomonas reinhardtii.

The effects of light on gravitaxis and velocity in the bi-flagellated green alga Chlamydomonas reinhardtii were investigated using a real time automatic tracking system. Three distinct light effects on gravitaxis and velocity with parallel kinetics were found. Photosynthetically active continuous red light reversibly enhances the swimming velocity and increases or decreases the precision of gravitaxis, depending on its initial level. Blue light flashes induce fast transient increases in velocity immediately after the photophobic response, and transiently decrease or even reverse negative gravitaxis. The calcium dependence of this response, its fluence-response curve and its spectral characteristics strongly suggest the participation of chlamy-rhodopsin in this effect. The third response, a prolonged activation of velocity and gravitaxis, is also induced by blue light flashes, which can be observed even in calcium-free medium.

Animals↗

PubMed2000

Effects of solar UV-B radiation on aquatic ecosystems.

Solar UV degrades dissolved organic carbon photolytically so that they can readily be taken up by bacterioplankton. On the other hand solar UV radiation inhibits bacterioplankton activity. Bacterioplankton productivity is far greater than previously thought and is comparable to phytoplankton primary productivity. According to the "microbial loop hypothesis," bacterioplankton is seen in the center of a food web, having a similar function to phytoplankton and protists. The penetration of UV and PAR into the water column can be measured. Marine waters show large temporal and regional differences in their concentrations of dissolved and particulate absorbing substances. A network of dosimeters (ELDONET) has been installed in Europe ranging from Abisko in Northern Sweden to Gran Canaria. Cyanobacteria are capable of fixing atmospheric nitrogen which is then made available to higher plants. The agricultural potential of cyanobacteria has been recognized as a biological fertilizer for wet soils such as in rice paddies. UV-B is known to impair processes such as growth, survival, pigmentation, motility, as well as the enzymes of nitrogen metabolism and CO2 fixation. The marine phytoplankton represents the single most important ecosystem on our planet and produces about the same biomass as all terrestrial ecosystems taken together. It is the base of the aquatic food chain and any changes in the size and composition of phytoplankton communities will directly affect food production for humans from marine sources. Another important role of marine phytoplankton is to serve as a sink for atmospheric carbon dioxide. Recent investigations have shown a large sensitivity of most phytoplankton organisms toward solar short-wavelength ultraviolet radiation (UV-B); even at ambient levels of UV-B radiation many organisms seem to be under UV stress. Because of their requirement for solar energy, the phytoplankton dwell in the top layers of the water column. In this near-surface position phytoplankton will be exposed to solar ultraviolet radiation. This radiation has been shown to affect growth, photosynthesis, nitrogen incorporation and enzyme activity. Other targets of solar UV irradiation are proteins and pigments involved in photosynthesis. Whether or not screening pigments can be induced in phytoplankton to effectively shield the organisms from excessive UV irradiation needs to be determined. Macroalgae show a distinct pattern of vertical distribution in their habitat. They have developed mechanisms to regulate their photosynthetic activity to adapt to the changing light regime and protect themselves from excessive radiation. A broad survey was carried out to understand photosynthesis in aquatic ecosystems and the different adaptation strategies to solar radiation of ecologically important species of green, red and brown algae from the North Sea, Baltic Sea, Mediterranean, Atlantic, polar and tropical oceans. Photoinhibition was quantified by oxygen exchange and by PAM (pulse amplitude modulated) fluorescence measurements based on transient changes of chlorophyll fluorescence.

Animals↗

PubMed2000

On site photosynthetic performance of Atlantic green algae.

Photosynthetic performance was measured on site in four common Atlantic green algae, Asparagopsis taxiforme, Valonia utricularia, Caulerpa racemosa and Codium taylori, in Gran Canaria, Canary Islands. The photosynthetic quantum yield was determined with a portable PAM instrument and with a diving PAM in the water column. Solar radiation was measured continuously above and in the water column by means of two three-channel dosimeters, ELDONET (Real Time Computer, Möhrendorf, Germany), in the UV-A, UV-B and PAR ranges. The effective photosynthetic quantum yield was found to be affected by exposure to solar radiation in as short as 15 min but recovered in the shade in most species within several hours. Only A. taxiforme failed to recover completely, and a 30-min exposure caused severe photoinhibition from which the algae recovered only partially. While most of the effect was due to the PAR wavelength range, the UV, and especially the UV-B, range considerably enhanced photoinhibition. In all four species, a significant inhibition was found even at their growth sites in the water column, measured with the diving PAM, at high solar angles.

Chlorophyta↗

PubMed1999

Physical characterization of gravitaxis in Euglena gracilis.

Gravitaxis in unicellular microorganisms like Euglena gracilis has been known for more than 100 years. The current model explains this phenomenon on the basis of a specific density difference between cell body and surrounding medium. In order to test the feasibility of the current model in terms of physical considerations the specific density of different Euglena gracilis cultures was determined. Depending on the culture conditions the specific density was in a range between 1.046 g mL-1 and 1.054 g mL-1. Size and gravitaxis measurements were performed in parallel, which allowed to relate the force applied to the lower membrane to the kinetic properties of gravitactic reorientation. A linear relationship between force and gravitaxis kinetics was found. A comparison between estimated activation energy of the proposed stretch-sensitive ion channels and energy supplied by the displacement of the lower membrane by the sedimentation of the cell body revealed that a focusing, an amplification and/or an integration period over time must be involved in the gravitactic signal transduction chain. Analysis of stimulus-response curves revealed an integration period of about 5 seconds before a gravitactic reorientation starts. The kinetics of gravitaxis at 1 x gn, and 0.12 x gn, was found to be similar. A hypothesis is presented that explains this finding on the basis of a combination of an integration period and an all-or-none reaction during gravitactic reorientation.

Animals↗

PubMed1999

Circadian rhythm of gravitaxis in Euglena gracilis.

Euglena gracilis, a unicellular, photosynthetic flagellate is a model system for environmentally controlled behavioral reactions. One pronounced reaction is the orientation with respect to gravity. In synchronized cultures with no cell growth a distinct circadian rhythm of negative gravitactic orientation could be observed. The main maximum of sensitivity was detected 5 h after the beginning of the subjective day, the main minimum 5 h before the beginning of the subjective day. Transferring synchronized cultures to continuous light resulted in an almost instantaneous loss of rhythmicity. In contrast, after transfer to permanent darkness cells exhibited a circadian rhythm with a progressive shortening of the period for more than 5 days. These findings are in contrast to the circadian rhythm of phototaxis in Euglena, where a free-running period of 24 h was observed. Parallel measurements of negative gravitactic orientation, velocity, cell shape as well as cAMP concentration in synchronized cultures revealed a circadian rhythm of all reactions. The results are discussed with regard to the possible role of cell shape and cAMP in gravitactic orientation.

Animals↗

PubMed1999

Graviorientation in protists and plants.

Gravitaxis, gravikinesis, and gravitropism are different graviresponses found in protists and plants. The phenomena have been intensively studied under variable stimulations ranging from microgravity to hypergravity. A huge amount of information is now available, e.g. about the time course of these events, their adaptation capacity, thresholds, and interaction between gravity and other environmental stimuli. There is growing evidence that a pure physical mechanism can be excluded for orientation of protists in the gravity field. Similarly, a physiological signal transduction chain has been postulated in plants. Current investigations focus on the question whether gravity is perceived by intracellular gravireceptors (e.g. the Muller organelle of the ciliate Loxodes, barium sulfate vacuoles in Chara rhizoids or starch statoliths in higher plants) or whether the whole cell acts as a sedimenting body exerting pressure on the lower membrane. Behavioral studies in density adjusted media, effects of inhibitors of mechano-sensitive ion channels or manipulations of the proposed gravireceptor structures revealed that both mechanisms have been developed in protists and plants. The threshold values for graviresponses indicate that even 10% of the normal gravitational field can be detected, which demands a focusing and amplifying system such as the cytoskeleton and second messengers.

Animals↗

PubMed1999

Gravitaxis in unicellular microorganisms.

Orientation of organisms with respect to the gravitational field of the Earth has been studied for more than 100 years in a number of unicellular microorganisms including flagellates and ciliates. Several hypotheses have been developed how the weak stimulus is perceived. Intracellular statoliths have been found to be involved in gravitaxis of Loxodes, while no specialized organelles have been detected in other ciliates, e.g. Paramecium. Also in the slime mold Physarum no specialized gravireceptors have been identified yet. In the flagellate Euglena gracilis the whole cell body, which is denser than the surrounding medium, seems to act as a statolith pressing onto the lower membrane where it activates mechanosensitive ion channels. Similar results were obtained for the ciliate Paramecium. In contrast to the flagellate Euglena, several ciliates have been found to show gravikinesis, which is defined as a dependence of the swimming velocity on the direction of movement in the gravity field.

Animals↗

PubMed1999

Negative gravitactic behavior of Euglena gracilis can not be described by the mechanism of buoyancy-oriented upward swimming.

Gravitactic behavior of microorganisms has been known for more than a hundred years. Euglena gracilis serves as a model system for gravity-triggered behavioral responses. Two basic mechanisms are discussed for gravitaxis: one is based on a physical mechanism where an asymmetric mass distribution pulls the cell passively in the correct orientation and, in contrast, the involvement of an active sensory system. A recently developed high-resolution motion-tracking system allows the analysis of single tracks during reorientation. The results are compared to a model developed by Fukui and Asai (1985) which describes gravitaxis of Paramecium caudatum on the basis of a physical mechanism. Taking into account the different size, different density, different mass distribution as well as the different velocity, results of the adapted model description of Paramecium were applied to measured data of Euglena. General shapes as well as the time scale of the predicted reorientational movement compared to measurements were different. The analysis clearly rules out the possibility that gravitaxis of Euglena gracilis is based on a pure physical phenomenon, and gives further support to the involvement of an active reorientational system. In addition, it could be shown that cell form changes during reorientation, even in an initial period where no angular change was observed.

Animals↗

PubMed1998

Gravitaxis and graviperception in Euglena gracilis.

Gravitactic orientation in the flagellate Euglena gracilis is mediated by an active physiological receptor rather than a passive alignment of the cells. During a recent space flight on the American shuttle Columbia the cells were subjected to different accelerations between 0 and 1.5 x g and tracked by computerized real-time image analysis. The dependence of orientation on acceleration followed a sigmoidal curve with a threshold at < or = 0.16 x g and a saturation at about 0.32 x g. No adaptation of the cells to the conditions of weightlessness was observed over the duration of the space mission (12 days). Under terrestrial conditions graviorientation was eliminated when the cells were suspended in a medium the density of which (Ficoll) equaled that of the cell body (1.04 g/ml) and was reversed at higher densities indicating that the whole cytoplasm exerts a pressure on the respective lower membrane. There it probably activates stretch-sensitive calcium specific ion channels since gravitaxis can be affected by gadolinium which is a specific inhibitor of calcium transport in these structures. The sensory transduction chain could involve modulation of the membrane potential since ion channel blockers, ionophores and ATPase inhibitors impair graviperception.

Animals↗

PubMed1997

A biased random walk model for the trajectories of swimming micro-organisms.

The motion of swimming micro-organisms that have a preferred direction of travel, such as single-celled algae moving upwards (gravitaxis) or towards a light source (phototaxis), is modelled as the continuous limit of a correlated and biased random walk as the time step tends to zero. This model leads to a Fokker-Planck equation for the probability distribution function of the orientation of the cells, from which macroscopic parameters such as the mean cell swimming direction and the diffusion coefficient due to cell swimming can be calculated. The model is tested on experimental data for gravitaxis and phototaxis and used to derive values for the macroscopic parameters for future use in theories of bioconvection, for example.

Algorithms↗

PubMed1997

Graviperception and graviorientation in flagellates.

In the flagellate Euglena gracilis Klebs, gravitaxis is mediated by an active physiological receptor and is not the result of passive alignment of the cells in the water column. The threshold of this response was found at 0.08 < threshold < 0.16 g during a recent space flight on the American shuttle Columbia, where the cells were subjected to different accelerations between 0 and 1.5 g; the response saturated at 0.32 < saturation < or = 0.64 g. Over the whole duration of the mission no adaptation of the response to microgravity was observed. The whole body of the cell, rather than intracellular organelles, seems to act as statolith since suspending the cells in a density-adjusted medium (Ficoll) resulted in an inhibition of gravitaxis and even reversal of orientation at higher densities. Thus, the cytoplasm seems to exert a pressure on the respective lower membrane where it is hypothesized to activate stretch-sensitive specific ion channels, as indicated by inhibitor studies with gadolinium. One of the early steps in the sensory transduction chain seems to be a modulation of the membrane potential since ion-channel blockers, ionophores and ATPase inhibitors strongly inhibit gravitaxis in this flagellate without seriously affecting motility and phototaxis.

Adenosine Triphosphatases↗

PubMed1997

Behavioral mutants of Euglena gracilis: functional and spectroscopic characterization.

Three mutant strains of the phytoflagellate Euglena gracilis Z have been characterized in order to analyze the signal perception and signal transduction pathways involved in photo- and gravitaxis. Using the fluorescence of the chromophoric groups believed to be involved in photoperception (flavins and pterins) a method was developed for an in situ and in vivo detection of the paraxonemal body, the proposed location of the photoreceptor molecules. Two of the mutant strains, 1224-5/9f and 1224-5/1f, do not possess a stigma and also lack a paraxonemal body, as indicated by fluorescence measurements. The third strain, FB, has a small stigma, but only some cells contain a paraxonemal body. In contrast to the present hypothesis on photoorientation of Euglena, all strains were able to orient with respect to the light direction. However, the mutant strains did not show any orientation at low irradiances. At medium and high irradiances the strains 1224-5/9f and 1224-5/1f oriented perpendicular to the light direction (diaphototaxis) while cells of strains of FB showed partly negative phototaxis and partly diaphototaxis. Diaphototaxis was never observed in the wild type strain. Strains 1224-5/9f and 1224-5/1f showed normal graviresponses compared with the wild type. Astasia longa, a nonphtototactic relative of E. gracilis, as well as strain FB were both negative and positive gravitactic at all culture ages tested. This result confirmed the hypothesis that the paraxonemal body is not directly involved in graviperception.

Animals↗

PubMed1997

Effects of hypergravity on the photosynthetic flagellate, Euglena gracilis.

Euglena gracilis, a unicellular, photosynthetic flagellate, orients itself by means of gravi- and phototaxis to reach and stay in regions optimal for survival and growth. An improved version of the slow rotating centrifuge microscope, NIZEMI, was used to test wild type and mutant strains for their responses to hypergravity. Wild type cells could actively move against the acceleration vector up to 8.5 gn and were centrifuged down at higher rates. Even at 10.5 gn, the highest value tested, cells were still negative gravitactically oriented as shown by video images. In contrast, all mutant strains as well as Astasia longa, a close relative of Euglena, could move against the acceleration vector under all conditions tested. With increasing accelerations the mean orientation of the populations shifted according to a vectorial addition of gravity and acceleration. The r-value, a statistical measure of the orientation of a population, increased with moderately increased acceleration rates and decreased at higher values. While wild type Euglena and two of the three mutant strains tested were exclusively negative gravitactically, in the third strain as well as in Astasia longa half of the population reacted negative gravitactically and the other half positive gravitactically. This variation of the wild type behavior was observed at moderate acceleration rates. At high accelerations the cells became exclusively positive gravitactic. The obtained results are discussed on the basis of the current model explaining gravitaxis.

Acceleration↗