Elias D. Kouvelas

Changes in [3H]AMPA and [3H]kainate binding in rat caudate-putamen and nucleus accumbens after 6-hydroxydopamine lesions of the medial forebrain bundle : an autoradiographic study

The binding parameters of [3H] alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) and [3H]kainate binding were examined in caudate-putamen and nucleus accumbens of rat striatum after unilateral lesions of the right medial forebrain bundle (MFB) using in vitro receptor autoradiography. Lesioning of the dopaminergic fibres in the MFB with 6-hydroxydopamine (6-OHDA) resulted, after one or four weeks, in a significant decrease in the levels of [3H]GBR 12935 (1-[2-diphenylmethoxy)-ethyl]-4-(3-phenylpropyl) piperazine) in ipsilateral caudate-putamen and nucleus accumbens (62 and 43%, respectively). A comparison of the dissociation constants (Kd) of [3H]AMPA and [3H]kainate binding in caudate-putamen and nucleus accumbens between control and MFB-lesioned side did not indicate any significant change. However, the maximum number of [3H]AMPA and [3H]kainate binding sites (Bmax) were significantly decreased in caudate-putamen and nucleus accumbens of the MFB-lesioned side of the brain. This decrease was between 17 and 26%. Our results suggested that at least one-fourth to one-fifth of AMPA and kainate receptors in rat caudate-putamen and nucleus accumbens are localized on the presynaptic endings of dopamine fibres that follow the MFB. A role of non-NMDA glutamate receptors in the presynaptic regulation of dopamine release in rat striatum is therefore supported.

Past tense formation and comprehension of passive sentences in Parkinson’s disease: Evidence from Greek

The present study investigates the production of regular and irregular verbs in the past tense and the comprehension of passive sentences by Greek-speaking PD patients, and compares their behavior to that of normal speakers. Although the two groups manifest large scale differences at all the above constructions, the behavior of PDs is not different at regular vs. irregular past tense formation neither did we obtain strong evidence that they do not comprehend passives, most importantly, they certainly do not perform at chance. On the basis of the above, we conclude that there are no indications for a clearly linguistic deficit of the PD group, hence, their difference with the control group should be attributed to other factors, such as the computational demands of the tasks.

L-Glutamate binding sites of normal and atrophic human cerebellum

The binding kinetics, pharmacologic properties, ontogeny and localization of L-glutamate binding sites were studied in membrane preparations and sections of normal and olivopontocerebellar atrophy (OPCA) human cerebellum. One binding component was found with a Kd value in the order of 150 x 10(-9) M. No significant changes of Kd values were observed with age, whereas the highest Bmax value was observed at the age of 1 year. L-Aspartate, ibotenate, quisqualate and L-homocysteic acid were potent inhibitors of L-[3H]glutamate binding. Quantitative densitometric measurements indicated the presence of L-glutamate sites in both the molecular and granule cell layer. In OPCA cerebella a very significant decrease of L-[3H]glutamate specific binding (Bmax) was observed, whereas Kd values were found unchanged. The pharmacologic properties of L-[3H]glutamate binding sites of OPCA cerebellar tissues were similar to those of normal cerebellum. [3H]quinuclidinyl benzylate binding, expressed in fmol/mg protein, did not show significant differences between normal and OPCA cerebella.

Selective effects of neonatal handling on rat brain N-methyl-D-aspartate receptors.

Neonatal handling, an experimental model of early life experiences, is known to affect the hypothalamic-pituitary-adrenal axis function thus increasing adaptability, coping with stress, cognitive abilities and in general brain plasticity-related processes. A molecule that plays a most critical role in such processes is the N-methyl-D-aspartate (NMDA) receptor, a tetramer consisting of two obligatory, channel forming NR1 subunits and two regulatory subunits, usually a combination of NR2A and NR2B. Since the subunit composition of the NMDA receptor affects brain plasticity, in the present study we investigated the effect of neonatal handling on NR1, NR2A and NR2B mRNA levels using in situ hybridization, and on NR2B binding sites, using autoradiography of in vitro binding of [(3)H]-ifenprodil, in adult rat limbic brain areas. We found that neonatal handling specifically increased NR2B mRNA and binding sites, while it had no effect on the NR1 and NR2A subunits. More specifically, neonatally handled animals, both males and females, had higher NR2B mRNA and binding sites in the dorsal CA1 hippocampal area, as well as the prelimbic, the anterior cingulate and the somatosensory cortex, compared to the non-handled. Moreover NR2B binding sites were increased in the dorsal CA3 area of handled animals of both sexes. Furthermore, neonatal handling had a sexually dimorphic effect, increasing NR2B mRNA and binding sites in the central and medial amygdaloid nuclei only of the females. The neonatal handling-induced increase in the NR2B subunit of the NMDA receptor could underlie the higher brain plasticity, which neonatally handled animals exhibit.

Localization ofl-glutamate binding sites in chick brain by quantitative autoradiography

Quantitative autoradiography was used for the localization of L-[3H]glutamate binding sites in chick brain. The highest concentration of these sites was observed in the molecular layer of cerebellum. High concentrations were also observed in the hippocampus, area parahippocampalis, neostriatum, neostriatum intermedium and hyperstriatum ventrale. In most areas binding of L-[3H]glutamate was increased when incubation was done in chloride-containing medium. This increase was statistically significant in only few of these areas.

Sexually dimorphic long-term effects of an early life experience on AMPA receptor subunit expression in rat brain

Neonatal handling, an experimental model of early life experiences, is known to affect hypothalamic-pituitary-adrenal (HPA) axis function, thus increasing adaptability, coping with stress, cognitive abilities and in general brain plasticity-related processes. AMPA receptors (AMPARs) mediate fast synaptic transmission at excitatory glutamatergic synapses in the CNS and are crucial during neuronal development, synaptic plasticity and structural remodeling. AMPARs are composed of four types of subunits, designated as AMPA glutamate receptor subunits (GluA1, GluA2, GluA3 and GluA4), which combine to form tetramers. The present study addressed the question of whether neonatal handling (15min daily maternal separation from postnatal day 1 (PND1) to PND21) might have an effect on GluA1-4 mRNA levels in adult rat male and female brain using in situ hybridization. We have identified selective sexually dimorphic effects of neonatal handling on the mRNA expression levels of AMPAR subunits in adult rat hippocampus and nuclei of the amygdaloid complex. In the dorsal hippocampus GluA1 mRNA levels were increased in handled males, while they were decreased in handled female animals. In the ventral hippocampus and the amygdaloid complex GluA2 mRNA was lower in handled females, while no effect was observed in handled males. Furthermore, we observed that neonatal handling induced in both sexes decreases of GluA2 mRNA in the dorsal hippocampus, as well as in the somatosensory and occipital cortex, of GluA3 mRNA in most hippocampal areas, amygdaloid complex and cortical regions studied, and of GluA4 mRNA in the ventral hippocampus. These results show that glutamatergic transmission is markedly affected by an early experience. The neonatal handling-induced alterations in AMPAR subunit composition are in line with the increased brain plasticity, the more effective HPA axis function, and in general the more adaptive behavioral phenotype known to characterize the handled animals.

Pharmacological characterization, anatomical distribution and sex differences of the non-NMDA excitatory amino acid receptors in the quail brain as identified by CNQX binding

The distribution of non-N-methyl-D-aspartate binding sites was studied in coronal and sagittal sections through the brain of adult Japanese quail by quantitative autoradiography, using tritiated 6-cyano-7-nitroquinoxaline-2,3-dione as a radioligand. Saturation binding experiments were, in addition, carried out in areas showing high levels of binding (cerebellar molecular layer, nucleus anterior medialis and nucleus infundibularis) and demonstrated that the binding of tritiated ligand was specific and saturable. Competition studies with alpha-amino-3-hydroxy-methyl-4-isoxazole propionic acid and kainic acid indicated that kainic acid strongly inhibited ligand binding in all brain areas. alpha-Amino-3-hydroxy-methyl-4-isoxazole propionic acid was only a weak inhibitor in the hypothalamic nuclei whereas in the cerebellar molecular layer both high and low affinity inhibitions were detected. The highest binding levels of tritiated ligand were observed in the molecular layer of the cerebellum. Very high levels of binding were detected in various preoptic/hypothalamic sites including the nucleus suprachiasmaticus pars medialis, nucleus anterior medialis hypothalami, nucleus infundibularis, nucleus mammillaris medialis, nucleus posteromediale hypothalami and nucleus hypothalami ventromedialis. High levels of binding were also detected in the bulbus olfactorius, bed nucleus commissuralis anterior, bed nucleus commissuralis pallii, nucleus accumbens, bed nucleus striae terminalis and nucleus interpeduncularis. In the preoptic area/hypothalamus, high levels of binding were clearly present in all areas that contain gonadotropin releasing hormone cells or fibers. In the pons and mesencephalon, moderate levels of binding were associated with catecholaminergic areas such as the area ventralis tegmentalis (area ventralis of Tsai) and the locus coeruleus. Saturation analysis demonstrated the presence of a higher number of binding sites in females than in males in the cerebellar molecular layer, nucleus infundibularis and nucleus anterior medialis. This latter difference was confirmed in the one point assays that also identified higher levels of specific binding in the nucleus suprachiasmaticus pars medialis of males as compared with females. These anatomical data suggest a possible implication of non-N-methyl-D-aspartate receptors in the synthesis and/or release of both gonadotropin releasing hormone and catecholaminergic neurotransmitters that should now be tested by pharmacological experiments.

Neural Stem Cells Transplanted in a Mouse Model of Parkinson’s Disease Differentiate to Neuronal Phenotypes and Reduce Rotational Deficit

The most prominent pathological feature in Parkinsons disease (PD) is the progressive and selective loss of mesencephalic dopaminergic neurons of the nigrostriatal tract. The present study was conducted in order to investigate whether naive and or genetically modified neural stem/precursor cells (NPCs) can survive, differentiate and functionally integrate in the lesioned striatum. To this end, stereotaxic injections of 6-OHDA in the right ascending nigrostriatal dopaminergic pathway of mice and subsequent NPC transplantations were performed, followed by apomorphine-induced rotations and double-immunofluorescence experiments. Our results demonstrate that transplanted embryonic NPCs derived from the cortical ventricular zone of E14.5 transgenic mouse embryos expressing the green fluorescent protein (GFP) under control of the beta-actin promoter and cultured as neurospheres can survive in the host striatum for at least three weeks after transplantation. The percentage of surviving GFP-positive cells in the host striatum ranges from 0.2% to 0.6% of the total transplanted NPCs. Grafted cells functionally integrate in the striatum, as indicated by the statistically significant decrease of contralateral rotations after apomorphine treatment. Furthermore, we show that within the striatal environment GFP-positive cells differentiate into beta-III tubulin-expressing neurons, but not glial cells. Most importantly, GFP-positive cells further differentiate to dopaminergic (TH-positive) and medium size spiny (DARPP-32- positive) neuronal phenotypes. Over-expression of the cell cycle exit and neuronal differentiation protein Cend1 in NPCs enhances the generation of GABAergic, but not dopaminergic, neuronal phenotypes after grafting in the lesioned striatum. Our results encourage the development of strategies involving NPC transplantation for the treatment of neurodegenerative diseases.

Somatostatin, cholecystokinin and neuropeptide Y mRNAs in normal and weaver mouse brain

Summary. The distribution of mRNAs encoding for somatostatin, cholecystokinin and neuropeptide Y was determined by in situ hybridization histochemistry in the weaver (wv/wv) mouse, a model of dopamine deficiency as well as in normal (+/+) controls. Weaver mutants did not show any appreciable departure from the normal pattern of expression for mRNA encoding for neuropeptide Y. In contrast, an 82% increase in mRNA encoding for somatostatin was observed in the reticular thalamic nucleus, whereas increases in the order of 20–87% were observed in different hypothalamic nuclei of the weaver brain. In addition, a 47–103% increase of the hybridization signal encoding for choleocystokinin was observed in the cerebral cortex, hippocampus and thalamus of the weaver brain. It can be assumed that the elevated and region-specific somatostatin and choleocystokinin levels observed in the weaver brain may be due to a secondary or compensatory response under conditions of altered neurotransmitter levels.

Cajal's Brief Experimentation with Hypnotic Suggestion

Spanish histologist Santiago Ramón y Cajal, one of the most notable figures in Neuroscience, and winner, along with Camillo Golgi, of the 1906 Nobel Prize in Physiology or Medicine for his discoveries on the structure of the nervous system, did not escape experimenting with some of the psychiatric techniques available at the time, mainly hypnotic suggestion, albeit briefly. While a physician in his thirties, Cajal published a short article under the title, “Pains of labour considerably attenuated by hypnotic suggestion” in Gaceta Médica Catalana. That study may be Cajals only documented case in the field of experimental psychology. We here provide an English translation of the original Spanish text, placing it historically within Cajals involvement with some of the key scientific and philosophical issues at the time.