Heide-Linde Rüthrich

Kindling and its consequences on learning in rats

To study the learning performance of pentylenetetrazol- and amygdala-kindled Wistar rats we used the following learning tests: short-term memory was tested in the response-to-change model, brightness discrimination was tested in a Y-chamber, and two-way active avoidance learning was tested in a shuttle-box. Short-term memory was not impaired by both kindling procedures. Considering two-way active avoidance learning the performance of pentylenetetrazol (PTZ)-kindled rats was significantly diminished. This effect persists over a period of 4 weeks. However, amygdala (AMY)-kindled rats acquired this task like the controls. In brightness discrimination reaction (BDR) the learning performance of PTZ-kindled animals was not influenced. Although the acquisition of BDR was nearly identical, the 24-h retention was remarkably diminished in AMY-kindled rats. It was hypothesized that the different kindling procedures interfere in different ways and extent with neuronal circuits resulting in different functional impairments.

Increased fucose incorporation into rat hippocampus during learning. A biochemical and microautoradiographic study

The incorporation of intraventricularly infected L-[1-3H]fucose into proteins of the hippocampus and visual cortex was studied during the acquisition of a shock-motivated brightness discrimination in rats. The labeling of Tris-soluble proteins from both regions was not significantly changed during learning, whereas solubilized insoluble proteins obtained from the hippocampus revealed an increased fucose incorporation in learned animals. A significant enhanced incorporation into some distinct, slow-moving, carbohydrate-rich protein bands, separated by polyacrylamide gel electrophoresis, was observed in material from CA1 and CA3 sectors as well as from area dentata of the hippocampus formation during acquisition. The corresponding gel bands from the visual cortex exhibited no differences between trained animals and controls. Jointly performed microautoradiography showed an increased fucose incorporation into most areas of the hippocampus in learned animals compared with active and passive controls. The most significant differences were found to occur mainly in substructures consisting of densely packed neuronal cells.

Time course and disposition of fucose radioactivity in rat hippocampus. A biochemical and microautoradiographic study.

Male adult rats were injected intraventricularly with L-[1-3H]fucose. At various intervals, ranging between 30 min and 11 days, one-half of the brain was prepared for microautoradiography, and the hippocampus from the other side was prepared for biochemical investigations. The TCA-precipitable proteins from the hippocampus homogenate were maximally labeled at between 8 and 24 h and remained at a high radioactivity level even 11 days after [3H]-fucose injection, the labeling being predominantly present in the solubilized insoluble proteins. Using gel electrophoretic separation, study of Tris-soluble material indicated a rapid turnover of soluble fucose-containing glycoproteins, whereas several slow-migrating bands of solubulized proteins revealed a time course suggesting the presence of fucose-containing glycoproteins with slower turnover rates. Using microautoradiography, a rapid labeling of neuronal cell bodies of the hippocampus was found, whereas the nuclei were not labeled. Perikarya were maximally labeled 4 h after [3H]fucose application. The radioactive material was continuously transported from the soma into the corresponding fiber layers, the latter being maximally labeled at a pulse interval of one day; even 10 days later a considerable amount of radioactivity could be detected in the neuropil.

Linear and cyclic β-casomorphin analogues with high analgesic activity

We investigated the antinociceptive efficacy of casomorphin (CM) derivatives using the vocalization test. Male Wistar rats received chronic microcannulae into the right lateral ventricle. One week later we examined the analgesic effect of CM derivatives 10, 30, 60, and 90 min after intraventricular injection (5 microliters). The analgesic effect was calculated as the individual percent increase in the pain threshold and was compared to controls (saline treatment). The substitution of D-lysine and D-ornithine in position 2 in connection with a cyclization through ring closure of the 2 position side chain amino group to the C-terminal glycine-COOH group resulted in high analgesic potency. The substitution of D-Pro4 was without any effect in the ineffective linear derivatives and decreased the effectiveness in the highly effective cyclic derivatives. The cyclic [D-Orn2]CM-5 and the cyclic [D-Lys2]CM-5 are the CM derivatives with the highest antinociceptive activity. The cyclic [D-Orn2]CM-5 is greater than 1000 times more effective than morphine. We conclude, on the basis of studies of receptor binding and in vitro investigations, that mu receptor activity alone is not responsible for the analgesic activity. The delta receptor and possibly also the kappa receptor could modulate the nociceptive effectiveness.

Influence of ß-casomorphins on apomorphine-induced hyperlocomotion

Abstract Derivatives of s-casomorphin Tyr-Pro-Phe-Pro-Gly and their des-Tyr 1 -derivatives were investigated on the model of apomorphine-induced hyperlocomotion (1 mg/kg = 3 μ M/kg, IP). D -Pip 4 CM 5 (5 nM) inhibited the apomorphine hypermotility completely, while D -Phe 3 CM 5 (5 nM) and D -Pro 4 CM 5 (5 nM) decreased it only to about 50%. The normal exploration was nearly completely inhibited by D -Pro 4 CM 5 (40 nM), by D -Pip 4 CM 5 (5 nM) depressed to 20%, and by D -Phe 3 CM 5 (10 nM) to 35%. The maximum inhibition of apomorphine-induced hyperlocomotion by the des-Tyr-casomorphin derivatives was about 50%. The dose-response curves were U-shaped. The exploratory activity was not significantly influenced. The mode of action and the involvement of different neurotransmitter systems in the inhibitory effect of s-casomorphin derivatives on apomorphine hyperlocomotion are discussed.

Influence of modified casomorphins on yawning behavior of rats

Apomorphine-induced yawning was completely suppressed in animals treated with 5 nmol [D-Pro4]casomorphin (CM) (ICV), 10 nmol [D-Phe3]CM (ICV) or 10 nmol [D-Pip4]CM (ICV). The apomorphine-induced yawning was also decreased, by des-Tyr analogs, but only by about 50%. Physostigmine (0.15 mg/kg, IP) induced yawning. The physostigmine-induced yawning was suppressed by 5 nmol [D-Pro4]CM and 10 nmol [D-Phe3]CM. Both [des-Tyr-D-Phe3]CM and [des-Tyr-D-Pip4]CM were without effect, whereas [des-Tyr-D-Pro4]CM increased significantly the physostigmine-induced yawning. The results suggest that dopaminergic transmission can be modulated by beta-casomorphin derivatives, thus resulting in a decrease in yawning. In the case of the des-tyrosine derivatives, we can assume a dopaminergic modulation, too. An increase in serotonergic activity might be supposed for [des-Tyr-D-Pro4]CM.

Early effects on restoration of evoked field potentials in the hippocampal CA1 region after reversible hypoxia/hypoglycemia by the radical scavenger N-tert.-butyl-α-phenylnitron

In transverse hippocampus slices a short period of hypoxia/hypoglycemia induced by perfusion with an O(2)/glucose-free medium caused early loss and incomplete restoration of evoked field potentials in the CA(1) region. In the present study a search was made for whether the formation of free oxoradicals immediately after starting the hypoxic phase could be part of the breakdown and incomplete restoration of the excitatory potentials (EPs). It was shown that preincubation and postischemic incubation with the radical scavenger PBN did not prevent the potential breakdown but significantly enhances potential restoration, even when PBN was added to the perfusion medium 40 min after hypoxia. Thus, free oxoradicals may damage membrane constituents such as receptors or channel proteins at a very early phase, before neuronal death is pronounced. The results also show that treatment with radical scavengers has a beneficial effect on early hypoxic damage.

Phe1-substituted β-casomorphin-5 analogues with analgesic activity

Abstract The antinociceptive potency of linear and cyclic β-casomorphin-5 (CM-5) analogues, modified in position 1 by substitution of the tyrosine (Tyr) by the phenylalanine (Phe) residue, was studied using the vocalization test. With the exception of the linear [Phe 1 , d -Orn 2 ]CM-5, the Phe 1 -substituted linear and cyclic casomorphin analogues exhibit remarkable analgesic potency compared to morphine, although the opioid receptor affinity and the opioid activity in vitro is diminished compared to the corresponding Tyr-containing analogues. The analgesic effect of the compounds is mediated by activation of opioid receptors, because it can be antagonized with naloxone. Furthermore, it was demonstrated that the [Phe 1 , d -Orn 2 , d -Pro 4 ]CM-5, which was about sixfold more potent than morphine, developed cross-tolerance to morphine.

Adrenalectomy attenuates the improvement of memory in rats by peripheral application of des-Tyr-d-Pro4-casomorphin

beta-Casomorphin derivatives without the N-terminal amino acid tyrosine possess memory-improving effects after central and peripheral application. We investigated the significance of adrenal glands for the memory improving effect of the systemically applied beta-casomorphin derivative des-Tyr-D-Pro4CM (Pro-Phe-D-Pro-Gly) in a learning experiment. Seven-week-old rats were adrenalectomized or sham operated. One week after surgery the rats were trained in an active avoidance task in a shuttle box. Five avoidance reactions were taken as learning criterion. After training 10 nmol/kg des-Tyr-D-Pro4CM or saline (10 ml/kg) was subcutaneously applied. There were no differences in acquisition between adrenalectomized and sham-operated rats. The memory retention of sham-operated animals was improved by des-Tyr-D-Pro4CM. In adrenalectomized rats this positive effect could not be observed. The involvement of adrenal glands in the peptide effect during learning and retention is discussed.