Wolfgang Hanke

Two-dimensional wave patterns of spreading depolarization: Retracting, re-entrant, and stationary waves

Abstract We present spatio-temporal characteristics of spreading depolarizations (SD) in two experimental systems: retracting SD wave segments observed with intrinsic optical signals in chicken retina, and spontaneously occurring re-entrant SD waves that repeatedly spread across gyrencephalic feline cortex observed by laser speckle flowmetry. A mathematical framework of reaction–diffusion systems with augmented transmission capabilities is developed to explain the emergence and transitions between these patterns. Our prediction is that the observed patterns are reaction–diffusion patterns controlled and modulated by weak nonlocal coupling such as long-range, time-delayed, and global coupling. The described spatio-temporal characteristics of SD are of important clinical relevance under conditions of migraine and stroke. In stroke, the emergence of re-entrant SD waves is believed to worsen outcome. In migraine, retracting SD wave segments cause neurological symptoms and transitions to stationary SD wave patterns may cause persistent symptoms without evidence from noninvasive imaging of infarction.

Effects of antimigraine drugs on retinal spreading depression

It has been suggested that spreading depression may play a role in triggering classical migraine. In this study the retinal spreading depression was used as a pharmacological tool to test the neuronal effects of several common antimigraine drugs. As the chicken retina is void of any blood vessels the observed effects must be of pure neuronal origin. It is shown that propranolol, sumatriptan, methysergide, paracetamol and acetylsalicyclic acid decrease the propagation velocity of retinal spreading depression waves, accelerate the recovery of the optical and electrical signal and reduce the amplitude of the negative potential shift, concomitant with the spreading depression. Barbiturate increases the spreading velocity, and the amplitude of the potential shift. Ergotamine, clonidine, lisuride and iprazochrome have no significant influence on retinal spreading depression.

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 simplified 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.

Action potential properties are gravity dependent

The functional properties of neuronal tissue critically depend on cellular composition and intercellular comunication. A basic principle of such communication found in various types of neurons is the generation of action potentials (APs). These APs depend on the presence of voltage gated ion channels and propagate along cellular processes (e.g. axons) towards target neurons or other cells. It has already been shown that the properties of ion channels depend on gravity. To discover whether the properties of APs also depend on gravity, we examined the propagation of APs in earthworms (invertebrates) and isolated nerve fibres (i.e. bundles of axons) from earthworms under conditions of micro- and macro-gravity. In a second set of experiments we could verify our results on rat axons (vertebrates). Our experiments carried out during two parabolic flight campaigns revealed that microgravity slows AP propagation velocity and macrogravity accelerates the transmission of action potentials. The relevance for live-science related questions is considerable, taking into account that altered gravity conditions might affect AP velocity in man during space flight missions.

Pore-forming protein from Entamoeba histolytica forms voltage- and pH-controlled multi-state channels with properties similar to those of the barrel-stave aggregates

Pore-forming protein from Entamoeba histolytica forms cation-selective channels in planar bilayers. With increasing potentials, the open-state probability of these channels decreases, and channel aggregates collapse (Young, J.D.-E. and Cohn, Z.A. (1985) J. Cell. Biochem. 29, 299-308). In this communication we report the following observations: (i) incorporation of the pore in black-lipid membranes was stimulated by membrane potential, (ii) pores were rectifying, (iii) breakdown of pores resulted in a continuous spectrum of subconductance states, (iv) the open-state probability increased strongly with pH. This pattern of behaviour is similar to that of the barrel-stave aggregates (alamethicin and related toxins). We therefore conclude that the amebal pores, like those of the barrel-stave class, may consist of complexes involving variable numbers of membrane-spanning subunits.

Effects of nitric oxide on the retinal spreading depression.

The retinal spreading depression has been used as a tool to investigate the action of nitric oxide (NO) as a diffusible neurotransmitter which in many cases acts by raising the cGMP level in target cells. The role of NO as a vasodilating agents has been well-established and it has been suggested that the vasodilatation concurrent with cortical SD may be mediated by NO. In this study, we present pure neuronal effects of NO on SD as the chicken retina is void of bloodvessels. We show that NO directly decreases the velocity of retinal SD waves in a concentration-and time-dependent manner. This effect can be partially mimicked by application of membrane-permeable cGMP derivatives. Furthermore, a NO-mediated speed up of the recovery of the intrinsic optical signal after the wave-front is shown.

Excitation waves in central grey matter: the retinal spreading depression

Abstract This review deals with a phenomenon which, although it has been known for five decades, has not been recognized until recently as a suitable tool for investigations of neuronal-glial interactions and the interesting consequences of the non-linearities arising from these cooperative effects, such as the transition from quiescent states to the propagation of excitation. For neurophysiologists interested in the excitability control of neuronal populations, the retinal spreading depression wave can also be very useful, especially as a basic model for epileptic activity triggering and propagation mechanisms. In the isolated eye cup, spreading depression waves (RSDs) can be observed with ease. In fact, the RSD wave can be seen with the naked eye, which gives the observer a complete and non-invasive two-dimensional view of propagation. In the intrinsic optical signal of the RSD, several components can be separated, either by their spectral preferences or by their temporal and spatial characteristics and experimental manipulations. Glial membrane channel activity and physical changes in the composition of the extracellular medium appear to the major contributions for the optical changes during RSDs. Different components are also found in the electrophysiological concomitants of the waves, some with neuronal (synaptic terminals) and some with glial predominance. Besides the clinical and physiological aspects, the RSDs are part of a general class of self-organizing spatiotemporal structures that arise in systems far from equilibrium known as dissipative structures. In the retina, low dimensional dissipative structures self-organized, self-sustained and spatially structured wave activity, can be maintained for hours either as a soliton ‘circling wave’ or as a complex sequence of interacting spirals. Therefore, the RSD can serve as a powerful tool for cross-disciplinary research in the field of non-linear dynamics.

Correlation between the durations of refractory period and intrinsic optical signal of retinal spreading depression during temperature variations

Abstract Spreading depression (SD) is a neurophysiological phenomenon which occurs in the grey substance of the central nervous system. SD is characterised by a wave-like spread of depressed neuronal activity, by large ion shifts between intra- and extracellular space, by cellular depolarization, and by altered optical properties of the tissue giving rise to an intrinsic optical signal (IOS). In the shadow of SD further waves are difficult to trigger and such waves spread at lower velocity than usual. In this paper we examine the temperature dependence of the duration of this recovery (refractory) period and the temperature dependence of the duration of the IOS in the chicken retina. It is shown that these SD accompanying events are strongly dependent on temperature and that they are likely to depend on the metabolic rate in the tissue. The observed correlation of the duration of the IOS with the duration of the refractory period suggests that the IOS is a good indicator for the duration of the tissue recovery. Such a correlation would be of great value to the experimentalist who must know about the duration of the refractory period: while the latter is laborious to determine, recording the IOS is convenient.

Correlation of the electrical and intrinsic optical signals in the chicken spreading depression phenomenon

This paper presents some results on the correlation between the electrophysiological and intrinsic optical signals (IOS) of spreading depression waves in chicken retinae. We first show that the peak of the time derivative of the electrophysiological wave occurs precisely when the optical signal reaches the electrode tip. Second, by comparing bath applications of propranolol and glycerol it can be shown that the slow potential shift is not directly correlated to the intrinsic optical signal. Propranolol depresses the amplitude of the electrical wave, although the intrinsic optical signal continues being visible. On the other hand, we observe total absence of the IOS under glycerol, while the electrical wave is always present. Correlations of this kind are relevant for a deeper understanding of the underlying mechanisms of the spreading depression phenomenon.

Odorant-evoked electrical responses in Grueneberg ganglion neurons rely on cGMP-associated signaling proteins

The Grueneberg ganglion (GG) in the anterior nasal region of mice is considered as an olfactory compartment since its neurons were recently observed to be activated by chemical stimuli, in particular by the odorant 2,3-dimethylpyrazine (2,3-DMP). However, it is unclear whether the GG indeed serves an olfactory function since these findings are solely based on the expression of the activity-dependent gene c-Fos. Consequently, it is yet uncertain whether chemical compounds, such as given odorants, elicit electrical responses in GG neurons which are required to convey the chemosensory information to the brain. Therefore, in the present study, electrical recording experiments on tissue sections through the anterior nasal region of mice were conducted which revealed that 2,3-DMP induces electrical signals in the GG. These responses were restricted to sites harboring GG neurons, indicating that 2,3-DMP elicits an electrical signal only in these but not in other cells of the anterior nasal region. 2,3-DMP-sensitive GG neurons express signaling proteins associated with the second messenger substance cyclic guanosine monophosphate (cGMP); most notably the cyclic nucleotide-gated ion channel CNGA3 and the transmembrane guanylyl cyclase GC-G. Using mice deficient for CNGA3 or GC-G demonstrated that the 2,3-DMP-evoked electrical signals in the GG of these knockout mice were substantially lower than in the GG of wildtype conspecifics, indicating that cGMP signaling plays a crucial role for odorant-induced electrical responses in the GG.