Biomedical subjects
Publications and source records attributed to A Van Harreveld.
In addition to promoting long-lasting negative (and positive) dendritic field potentials elicited by stimulation of Schaffer collaterals in hippocampal slices, 4-aminopyridine causes a slow negative wave which may have another origin than the dendritic potentials.
Spreading depression (SD) in the chick retina is completely suppressed by 10 mM MgCl2 in the bathing solution (Mg-sensitive SD). However, after increasing the KCl concentration in the Mg solution to values between 10 and 20 mM the retina can again exhibit SDs (Mg-insensitive SD). It has been postulated that the Mg-sensitive SD is a glutamatergic phenomenon. This is supported by the effect of four gl(utamate)-antagonists--L-proline, glutamic acid diethyl ester (GDEE), D-alpha-aminoadipate (D-AA), and 2-amino-4-phosphonobutyrate (APB)--which all suppressed this type of SD. It was suggested that this effect is due to competitive binding of glutamate involved in the Mg-sensitive SD and the gl-antagonist to glutamate receptors. The suppression of SD could be reversed by washing the preparation in a physiologic salt solution. The gl-antagonists in relatively high concentrations had a cytotoxic effect which, when severe, suppressed SD and prevented the recovery of this phenomenon by washing the compound out of the tissue. The compounds examined had, in addition to their gl-antagonistic properties, a gl-agonistic effect, which was postulated to enhance the Na+ permeability of neural membranes resulting in a release of K+ into the extracellular space. In preparations bathed in 10 mM MgCl2 (which suppresses Mg-sensitive SDs) the four compounds investigated promoted Mg-insensitive SDs supposedly when the extracellular K+ concentration reached values between 10 and 20 mequiv.
The visual concomitants of spreading depression in the chick retina consist of a number of expanding concentric rings of different width and darkness around a stimulated area. The most peripheral ring consists of a narrow dark line. More central follow a light ring, then a wide dark band and most central, a light area. These rings correspond with changes in light reflected from the retina, measured with a microphotometric method (Martins-Ferreira and de Oliveira Castro, 1966). The dark outline and the dark ring correspond with minima of reflected light. It was suggested that the dark rings are caused by swelling of the Müller fibers due to a K+ release from the retinal tissue during SD. The swollen fibers would transport light to the layer of receptors where it is absorved by the choroidea, leaving less light to be reflected to the vitreal surface. The light rings would be due to an increase in reflectivity of neural tissue when invaded by SD. The dark and light bands would be caused by the local dominance of the effects of the swelling of Müller fibers, or of the increase in reflectivity of the neural tissue during SD.
A high potassium concentration (33 meq) in the solution superfusing the isolated chicken retina causes an increase in the tissue transparency. An L-glutamate (1 mM) or L-proline (10 mM) solution has the same effect. Swelling of the Müller fibers, which have a radial position in the retina, could explain the transparency increase. This possibility was investigated in electron micrographs of retinas subjected to these treatments and fixed by freeze-substitution to preserve the water distribution in the tissue. The Müller fibers in the controls had a mean diameter of 0.22 micrometer. The fibers in retinas bathed for 2 min in the high [K+] solution were more than three times as thick (0.74 micrometer); the fibers in glutamate-treated retinas were more than twice as thick (0.49 micrometer). The fibers in the proline-treated retinas had a diameter of 0.39 micrometer. The glutamate- and proline-induced swelling may be due to a K+ release from neuronal elements, acting on the Müller fibers. The fiber swelling was postulated to be the expression of different Donnan equilibriums of fibers bathed in solutions of different K+ concentrations. The observed swelling caused by the high [K+] solution was compared with the theoretical swelling of the fiber as an ideal Donnan system, postulating permeabilities for different ions of the fiber membrane. This suggested that the high [K+] solution causes an increase in Na+ permeability in addition to the permeability of the membrane for K+, Cl-, and HCO3-. Chemical fixation with glutaraldehyde and formaldehyde in an Na-phosphate buffer yielded micrographs in which the Müller fibers of retinas treated with a high [K+] or a glutamate solution had diameters similar to those of the control preparations.
Tetanic stimulation of the entorhinal area induces significant enlargement of the average dendritic spine area and perimeter in the middle and distal thirds of the dentate molecular layer 4 and 90 min following stimulation. Four minutes after stimulation, the differences between the stimulated and control animals were 20% for the dendritic spine area and 9% for the perimeter in the middle third, and in the distal third 32 and 14%, respectively. Ninety minutes after stimulation the differences were 28 and 11% for the area and perimeter in the middle third, and 33 and 18% in the distal third, respectively. Anisomycin at a dose of 25 mg/kg had no significant effect on the average spine area or perimeter in the various thirds of the dentate molecular layer in the 19 and 105 min post-application intervals. This dose of anisomycin given 15 min prior to the stimulation suppresses the stimulation-induced spine changes in the 4 min interval. In the 90 min interval when the effect of anisomycin on protein synthesis is largely terminated, spine enlargement reappears, being 21% higher than the controls in the middle and distal thirds. The differential effect of anisomycin on dendritic spines in the two post-stimulation intervals is discussed in relation to the effect of anisomycin on protein synthesis. The present experiments thus demonstrate that the stimulation-induced spine enlargement in the dentate fascia can be suppressed by a protein synthesis blocking drug.
Evidence was presented for a glutamate agonistic effect of L-proline which promotes K+-based spreading depressions (SD) in chick retinas at relatively high concentrations (5 mM), in addition to an antagonistic effect which inhibits glutamate-based SDs at lower (2 mM) concentrations. Together these effects explain the observed biphasic effect of L-proline on the incidence of SD in the retina.
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The fast as well as the slow contractions of the adductor muscle in the claw of Procambarus clarkii are inhibited by L-proline. This inhibition is dose dependent and decreases with increasing frequency of stimulation of the "slow" fiber. Contractions caused by perfusing the adductor muscle with L-glutamate solutions are also inhibited by L-proline. The inhibiting potency of L-proline is small; the effective concentration of this amino acid is 50--100 times that of the L-glutamate applied. It was postulated that the inhibitory effect of L-proline is based on competition for excitatory receptor sites of L-glutamate, which causes depolarization and contraction, and L-proline, which lacks these actions. Theoretical considerations suggested a linear relationship between the stimulating L-glutamate and the just-inhibiting L-proline concentrations. Experimental evidence supported this model.
L-Proline and some of its analogs have been shown to prevent spreading depression (SD) in the chick retina at relatively low concentrations and to impair memory processing without provoking toxic or electrophysiological disturbances. Both effects are hypothesized to be caused by inhibition of the effects of glutamate released into the extracellular space. L-Proline, its D-enantiomer, six proline analogs including two homologs (L-azetidine-2-carboxylic acid and DL-pipecolic acid), and five other compounds were examined for their effects on spreading depression and their amnestic and electrophysiological effects. L-Proline, L-baikiain, DL-3,4-dehydroproline, and L-4-hydroxyproline all reduced the incidence of SD in the chick retina and proved to be amnestic. D-Proline, L-pyroglutamic acid, L-azetidine-2-carboxylic acid, DL-pipecolic acid, L-glutamic acid diethylester, L-isoleucine and L-norleucine neither depressed SD nor caused retrograde amnesia. L-Prolyl-L-proline and L-glutamine did not depress SD at low concentrations but had significant amnestic effects. None of the listed compounds induced EEG disturbances. Implications for memory mechanisms are discussed in the light of these results.
The method used to determine the rate of fusion was based on the large difference in the dielectric constants of water and ice. A thin (50--60 micrometers) slice of a gelatin gel was used as the dielectric in a plate condenser. The slice was placed on a metal electrode built in a specimen carrier which was dropped on a silver freezing surface kept at below 70 K, forming the other plate of the condenser. Freezing of the gelatin causes a marked decrease in a 20,000 cycle current passing through the condenser. Since the thickness of the layer of frozen material was shown to be a function of the reciprocal of the current, it was possible to determine the course of fusion of the section. Freezing started at a high rate which declined during the first 5 ms but then increased again and usually became quite high at the end of fusion.
The effects of L-proline, D-proline, and L-azetidine-2-carboxylic acid (L-A.2.C., the lower homolog of L-proline) have been compared in two systems. L-Proline is more potent than either analog in causing amnesia of one-trial avoidance conditioning of the 2-day-old chick and in preventing mechanically induced spreading depression in the retina isolated from 2-3-week-old chicks. The results suggest that the L-configuration and the proper molecular size are essential for the effects of L-proline upon memory and upon spreading depression. This level of specificity is greater than that involved in protein synthesis because L-A.2.C. is incorporated into protein in place of L-proline, in several protein-synthesizing systems.
Two mechanisms have been proposed to explain spreading depression (SD): one based on a release of glutamate (Van Harreveld, 1959), and the other on a release of potassium (Grafstein 1956) from neuronal elements. Both glutamate and KCl cause transparency changes in the retina, comparable to those occurring in this tissue during SD. The glutamate effect is inhibited by MgCl2 (10mM), in contrast to the transparency change due to KCl which is not affected by Mg++. Also SD is usually inhibited by MgCl2 which suggests that such SDs are based on a glutamate release. Impairment of the tissue metabolism promotes SDs which are insensitive to MgCl2. The resulting failure of the mechanisms that transport K+ and glutamate which leak out of the intracellular compartment back into the cells and fibers, seems to be involved in the generation of Mg++ insensitive SDs. This may facilitate either K-based SDs or glutamate-based SDs since the inhibitory effect of Mg++ is counteracted by an enhanced glutamate concentration. Both proposed mechanisms for SD seem to be possible under special circumstances.
Stimulation of the perforant path induces a long-lasting increase in the area of dendritic spines, which are sites of termination of the stimulated pathway in the distal third of the dentate molecular layer. No enlarged spines were found in the proximal third of the dentate molecular layer, where the commissural afferents terminate. Following a single tetanic stimulus of 30 sec duration at 30/sec, spines became significantly larger by 15%, 38%, 35% and 23% within poststimulation intervals of 2-6 min, 10-60 min, 4-8 h, and 23 h, respectively. Axon terminals decreased their area by 15% within the 2-6 min interval and the vesicle density was decreased by 19% within the 10-60 min interval. Both changes were reversible and terminals resumed their prestimulation condition at longer intervals (greater than 4 h). The initial enlargement of spines was interpreted as being due to a glutamate-induced increase in the sodium permeability of the spine membrane, whereas for the long-lasting enlargement an increase in protein synthesis was postulated. The long-lasting enlargement of dendritic spines in the dentate molecular layer following a short train of stimuli delivered to the perforant path, supports the postulate which links such a change to the mechanism of long-lasting postactivation potentiation observed in this pathway.
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Explore the source record for details and available documents.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
Explore the source record for details and available documents.