Ion channel are sensitive to gravity changes.

The effects of gravity on alamethicin doped planar lipid bilayers and on reconstituted porins of Escherichia coli outer membrane, respectively, have been investigated in this paper. The aim of the study was to find out whether and how gravity influences the highly stratified system: membrane-ion channel, in order to provide a novel approach to the explanation of gravity effects on living systems. This is necessary, as even single cells can react to gravity changes without having perceptive organelles. The mechanism of this detection is not clear yet. One possibility might be the detection of gravity by the membrane itself, or by the interaction of integral membrane proteins with gravity. Here we show for the first time that gravity directly influences the integral open state probability of native ion channels (porins) incorporated into planar lipid bilayers. Under hypergravity, especially the open state probability of porins is increased, whereas it is decreased in the microgravity case. The dependency is sigmoidal with the steepest region at 1 to 1.3 g. In the light of these experiments, a general effect of gravity on ion channels and membranes seems to be reasonable, possibly providing an explanation for several impacts of gravity on living systems.

The properties of alamethicin incorporated into planar lipid bilayers under the influence of microgravity.

During evolution, life on earth had adapted to the gravity of 1g. Due to space flight, in the last decades the question arose what happens to the brain under microgravity on the molecular level. Ion channels among others are the molecular basis of brain function. Therefore, the investigation of ion channel function under microgravity seems to be a promising approach to gather knowledge on brain function during space flight. In a first step, the ion channel forming peptide Alamethicin was used as a model channel in an artificial membrane. It is well suitable for this kind of investigation, since its properties are well described under standard gravity. For that reason, changes due to microgravity can be detected easily. All experiments were performed in the German drop tower at ZARM-FAB, Bremen. A special set-up was constructed based on the bilayer technique introduced by Mueller and Rudin. All functions of this set-up can be observed and controlled remotely. In the first set of experiments, a dramatic change of electrical properties of Alamethicin under microgravity could be observed. Mainly, the pore frequency is significantly reduced.

Distribution of gap junctions in the chicken retina.

Coupling between cells of neuronal tissue can be due to electrical or chemical synapses. The molecular basis of an electrical synapse is the gap junction channel. Gap junctions have been found between neurones and glial cells, however, in some tissue their presence in the membranes of different cell types is still under discussion. In the retina of vertebrates, which is a true part of the CNS, the presence of gap junctions in the specialised glial cells of the retina, the Müller cells is not clear for chicken. Since these cells span the whole retina vertically, for some tasks, like spatial buffering of potassium, such gap junctions would not be required, in contrast to other parts of the CNS. The spatial buffering of potassium among others plays an important role in the propagation of excitation-depression waves in neuronal tissue, especially in the chicken retina. However, gap junctions could be involved in creating an electrical syncitium of glial cells, which might also contribute to excitation-depression wave propagation. In this paper we present an about complete screening of the presence of gap junctions in the chicken retina, including the proof that the Müller cells of this retina do not have gap junctions. This finding is discussed considering the highly specialised morphological structure of the Müller cells of the chicken retina, which have an extremely extended endfeet tree.

Calcium waves in gray matter are due to voltage-sensitive glial membrane channels.

The retina is the most accessible piece of central gray matter in the vertebrate brain. Its wide dynamic operational range makes it the ideal neuronal network to study its excitability. Spreading depression waves in the retina are accompanied by strong intrinsic optical signals (IOS) and thus can be measured non-invasively with optical methods. Additionally, incubation with fluorescent dyes allows to follow calcium fluxes in parallel. The IOS can be divided into red and green scatter of light. We show that during spreading depression the red scatter signal precedes the green scatter signal and that the calcium signal matches the red scatter signal. Incubation of the retina with barium chloride leads to a reversible depression of red scatter and calcium signal whereas the green scatter signal is hardly effected. The wave propagation velocity is reduced, too. This supports the idea that the early red scatter signal is a direct visualisation of glial membrane potential and that glia cells in the chicken retina are involved in the control of extracellular calcium.

Modulation of K(+)-channels in p-neurones of the leech CNS by phosphorylation.

Two types of potassium channels of identified (p-) neurones of the leech (Hirudo medicinalis) were investigated by using the patch-clamp technique. The open-state probability of these channels in cell-attached patches can be reduced by addition of 5-hydroxytryptamine to the bath solution. After excising the patches the application of alkaline phosphatase to the cytosolic face of the patch increases the open probability. The 5-HT1A-receptor agonist buspirone mimics the effect of 5-HT. Our experiments show that the effect of 5-HT might be due to a channel phosphorylation via a 5-HT1A-receptor subtype.