Kerry D. Walton

Relationship between presynaptic calcium current and postsynaptic potential in squid giant synapse

The relationship between calcium current and transmitter release was studied in squid giant synapse. It was found that the voltage-dependent calcium current triggers the release of synaptic transmitter in direct proportion to its magnitude and duration. Transmitter release occurs with a delay of approximately 200 mus after the influx of calcium. A model is presented which describes these relations formally.

Postnatal changes in motoneurone electrotonic coupling studied in the in vitro rat lumbar spinal cord.

1. Electrotonic coupling between motoneurones innervating ankle flexor and extensor muscles, as well as between unidentified lumbar motoneurones, was studied using intracellular recordings in an in vitro spinal cord‐hindlimb preparation isolated from rats between birth (P0) and 13 days (P13). 2. Graded ventral root stimulation could elicit graded, short latency depolarizations (SLD) which preceded, coincided with, or followed the antidromic action potential. These SLDs were identified as electrotonic junctional potentials by their latency, relative insensitivity to changes in membrane potential and their resistance to one or more of the following: (1) high‐frequency stimulation, (2) collision with a somatofugal action potential, (3) removal of Ca2+ from the bathing solution. 3. SLDs were studied in 162 neurones and were identified in 77.2% of the cells in preparations from P0 to P3 rats (n = 57), but only in 30.8% at P8 to P13 (n = 39). 4. SLDs were largest in the youngest animals (P0 to P3), decreasing from a mean of 1.31 mV (+/‐ 0.17, n = 34) to 0.56 mV (+/‐ 0.10, n = 7) at P8 to P13. The SLDs comprised two to eight (4.3 +/‐ 0.36) all‐or‐none components as determined from twenty collision experiments. 5. Electrotonic coupling between motoneurones was specific. SLDs could be elicited in given motoneurones by stimulation of their homonymous but never of their antagonistic muscle nerves. 6. These results indicate that electrotonic coupling between lumbar motoneurones in neonatal animals exhibits a high degree of specificity and that its significance, as judged by the amplitude and frequency of occurrence of SLDs, decreases postnatally at a rate that can be correlated with the functional maturity of the motoneurones and the muscular system.

The Isolated and Perfused Brain of the Guinea‐pig In Vitro

We describe here an isolated and perfused in vitro adult guinea‐pig whole brain preparation which is an extension of the previously described in vitro brainstem–cerebellum preparation, Viability was tested by the analysis of trans‐synaptic responses along the visual pathways following the electrical stimulation of the optic nerve or the optic radiations. The evoked field potentials were recorded in the dorsal lateral geniculate, the superior colliculus and the visual cortex. The distribution of extracellular currents was studied using current source density analysis, in order to determine the amplitude, time course and spatial organization of the synaptic activity at these sites. The study indicates that field potentials were very similar to those described in vivo. These data demonstrate the survival of a complex adult sensory system in vitro and suggest that this preparation can be used for the analysis of multisynaptic circuits in the mammalian brain.

Abnormal thalamocortical activity in patients with Complex Regional Pain Syndrome (CRPS) Type I

&NA; Complex Regional Pain Syndrome (CRPS) is a neuropathic disease that presents a continuing challenge in terms of pathophysiology, diagnosis, and treatment. Recent studies of neuropathic pain, in both animals and patients, have established a direct relationship between abnormal thalamic rhythmicity related to Thalamo‐cortical Dysrhythmia (TCD) and the occurrence of central pain. Here, this relationship has been examined using magneto‐encephalographic (MEG) imaging in CRPS Type I, characterized by the absence of nerve lesions. The study addresses spontaneous MEG activity from 13 awake, adult patients (2 men, 11 women; age 15–62), with CRPS Type I of one extremity (duration range: 3 months to 10 years) and from 13 control subjects. All CRPS I patients demonstrated peaks in power spectrum in the delta (<4 Hz) and/or theta (4–9 Hz) frequency ranges resulting in a characteristically increased spectral power in those ranges when compared to control subjects. The localization of such abnormal activity, implemented using independent component analysis (ICA) of the sensor data, showed delta and/or theta range activity localized to the somatosensory cortex corresponding to the pain localization, and to orbitofrontal–temporal cortices related to the affective pain perception. Indeed, CRPS Type I patients presented abnormal brain activity typical of TCD, which has both diagnostic value indicating a central origin for this ailment and a potential treatment interest involving pharmacological and electrical stimulation therapies.

Imaging of Thalamocortical Dysrhythmia in Neuropsychiatry

Abnormal brain activity dynamics, in the sense of a thalamocortical dysrhythmia (TCD), has been proposed as the underlying mechanism for a subset of disorders that bridge the traditional delineations of neurology and neuropsychiatry. In order to test this proposal from a psychiatric perspective, a study using magnetoencephalography (MEG) was implemented in subjects with schizophrenic spectrum disorder (n = 14), obsessive–compulsive disorder (n = 10), or depressive disorder (n = 5) and in control individuals (n = 18). Detailed CNS electrophysiological analysis of these patients, using MEG, revealed the presence of abnormal theta range spectral power with typical TCD characteristics, in all cases. The use of independent component analysis and minimum-norm-based methods localized such TCD to ventromedial prefrontal and temporal cortices. The observed mode of oscillation was spectrally equivalent but spatially distinct from that of TCD observed in other related disorders, including Parkinsons disease, central tinnitus, neuropathic pain, and autism. The present results indicate that the functional basis for much of these pathologies may relate most fundamentally to the category of calcium channelopathies and serve as a model for the cellular substrate for low-frequency oscillations present in these psychiatric disorders, providing a basis for therapeutic strategies.

The effectiveness of different isomers of octanol as blockers of harmaline-induced tremor

Intracellular recording in the guinea-pig brainstem slice has demonstrated that high molecular weight alcohols block the low threshold calcium channel (LTCC) in the inferior olive (IO). These alcohols thus provide a tool for understanding the function of the pacemaking cellular networks of the olivo-cerebellar system, since the LTCC has been implicated in the oscillatory behavior of these neurons. Aspects of normal and pathological tremor are also believed to be mediated by these circuits, and thus development of effective ways of blocking the LTCC in vivo may eventually lead to novel treatments for essential tremor. The present experiments evaluated the effectiveness of the isomers of octanol in decreasing harmaline-induced tremor in vivo in the rat. Harmaline was used in this study because its tremorgenic action is mediated at the level of IO; octanol was found to be a potent antagonist of harmaline-induced tremor. Significant differences between the isomers further suggested conformational differences. This, taken in conjunction with the lack of effect of octanol in both IO lesioned rats and oxotremorine-induced tremor, implied that the action of the alcohol may be mediated at a specific binding site. These findings thus support the conclusions that the antagonism of harmaline-induced tremor by octanol occurs in the IO, and, in view of the previously reported in vitro data, that octanol may be an effective blocker of the LTCC in vivo.

Postnatal development under conditions of simulated weightlessness and space flight

The adaptability of the developing nervous system to environmental influences and the mechanisms underlying this plasticity has recently become a subject of interest in space neuroscience. Ground studies on neonatal rats using the tail suspension model of weightlessness have shown that the force of gravity clearly influences the events underlying the postnatal development of motor function. These effects depend on the age of the animal, duration of the perturbation and the motor function studied. A nine-day flight study has shown that a dam and neonates can develop under conditions of space flight. The motor function of the flight animals after landing was consistent with that seen in the tail suspension studies, being marked by limb joint extension. However, there were expected differences due to: (1) the unloading of the vestibular system in flight, which did not occur in the ground-based experiments; (2) differences between flight and suspension durations; and (3) the inability to evaluate motor function during the flight. The next step is to conduct experiments in space with the flexibility and rigor that is now limited to ground studies: an opportunity offered by the International Space Station.

Hydrogen peroxide as a source of molecular oxygen for in vitro mammalian CNS preparations

Using an isolated neonatal rat spinal cord preparation, we have studied the ability of hydrogen peroxide to act as a source of oxygen for mammalian central nervous system (CNS) maintained in vitro. We report here that hydrogen peroxide, in low concentrations (0.001-0.004%), can effectively provide the only source, or act as a supplementary source of oxygen. This is brought about via the intracellular enzyme catalase which catalyzes the conversion of H2O2 into molecular oxygen and water.

The effects of microgravity on the development of surface righting in rats.

The active interaction of neonatal animals with their environment has been shown to be a decisive factor in the postnatal development of sensory systems, which demonstrates a critical period in their maturation. The direct demonstration of such a dependence on the rearing environment has not been demonstrated for motor system function. Nor has the role of gravity in mammalian motor system development been investigated. Here we report the results of two space flight missions examining the effect of removing gravity on the development of surface righting. Since the essential stimulus that drives this synergy, gravitation, was missing, righting did not occur while the animals were in the microgravity environment. We hypothesize that this absence of contextual motor experience arrested the maturation of the motor tactics for surface righting. Such effects were permanent in rats spending 16 days (from postnatal day (P), P14 to P30), but were transient in animals spending nine days (from P15 to P24) in microgravity. Thus, active, contextual interaction with the environment during a critical period of development is necessary for the postnatal maturation of motor tactics as exemplified by surface righting, and such events must occur within a particular time period. Further, Earths gravitational field is not assumed by the developing motor system. Rather, postnatal motor system development is appropriate to the gravitational field in which the animal is reared.

Long-term effects of microgravity on the swimming behaviour of young rats

The postnatal development of sensory systems has been shown in studies over the last four decades to be influenced by experience during critical periods of development. We report here that similar experience‐dependent development can be observed in the swimming behaviour of young rats reared from postnatal day 14 (P14) to P30 in the reduced gravitational field of low earth orbit. Animals flown in space when placed in the water on the day of landing maintained their head and forelimbs in a balanced posture. However, until the animals began to swim, their hindquarters showed little lateral postural control resulting in rotation about the longitudinal axis (60°± 4 deg). Such results suggest an ‘unlinking’ of postural control of the forequarters from the hindquarters in the early hours after landing. Similar instability seen in animals age‐matched to the day of launch (97 ± 7 deg) and in ground control animals (9 ± 3 deg) was corrected within one or two rotations, even in the absence of swimming. Animals flown in space began to swim sooner after being placed in the water, and the duration of swimming strokes was shorter than in control animals. Motion analysis revealed a difference in the swimming style on landing day. In flight animals, the knee joint was more flexed throughout the stroke, there was a narrower range of movement, and the linear velocity of the tip of the foot was faster throughout most of the stroke than in age‐matched control animals. Thus, posture in the water as well as swimming speed and style were altered in the animals flown in space. Some of these characteristics persisted for as long as the animals were followed (30 days). These included the short pre‐swimming interval and short stroke duration in flight animals. These findings clearly show that an altered gravitational field influences the postnatal development of motor function. The nature of the differences between animals reared in space for 16 days and those remaining on the ground reflects an adaptation of the flight animals to the microgravity environment. The data suggest that the most fundamental of these adaptations is a resetting of the basic motor rhythm to a higher frequency.