RESUMEN
We examined the effect of kindling on serotonergic neurotransmission in the hippocampus by measuring serotonin (5-HT) release and uptake in hippocampal synaptosomes and 5-HT1A and 5-HT4 receptor subtypes during and at different times after electrical kindling of the dentate gyrus. Using quantitative receptor autoradiography, we found that binding of 8-[3H]hydroxy-2-(di-n-propylamino)tetralin ([3H]8-OH-DPAT) to 5-HT1A receptors was selectively increased by 20% on average (p < 0.05) in the dentate gyrus of the stimulated and contralateral hippocampus 2 days after stage 2 (stereotypes and occasional retraction of a forelimb) and by 100% on average (p < 0.05) 1 week after stage 5 (tonic-clonic seizures) compared with sham-stimulated rats. A 20% increase (p < 0.05) was observed 1 month after the last generalized seizure. No changes were found after a single afterdischarge. 5-HT4 receptors, which colocalize with 5-HT1A receptors on hippocampal neurons, were not modified in kindled tissue. [3H]5-HT uptake and its release as well as the 5-HT1B autoreceptor function did not differ from shams in hippocampal synaptosomes at stages 2 and 5. Systemic administration of 100 and 1,000 microg kg(-1) 8-OH-DPAT or 1,000 microg kg(-1) WAY-100,635, 30 min before each electrical stimulation, did not significantly alter kindling progression or the occurrence of stage 5 seizures in fully kindled rats. The changes in 5-HT1A receptor density in the dentate gyrus are part of the plastic modifications occurring during kindling and may contribute to modulating tissue hyperexcitability.
Asunto(s)
Giro Dentado/fisiología , Hipocampo/fisiología , Excitación Neurológica/fisiología , Receptores de Serotonina/biosíntesis , Serotonina/metabolismo , Sinapsis/fisiología , 8-Hidroxi-2-(di-n-propilamino)tetralin/metabolismo , 8-Hidroxi-2-(di-n-propilamino)tetralin/farmacología , Animales , Corteza Cerebral/efectos de los fármacos , Corteza Cerebral/fisiología , Giro Dentado/efectos de los fármacos , Dioxanos/metabolismo , Estimulación Eléctrica , Lateralidad Funcional , Radioisótopos de Yodo , Masculino , Neuronas/efectos de los fármacos , Neuronas/fisiología , Piperazinas/farmacología , Piperidinas/metabolismo , Piridinas/farmacología , Ensayo de Unión Radioligante , Ratas , Ratas Sprague-Dawley , Receptores de Serotonina 5-HT1 , Receptores de Serotonina 5-HT4 , Antagonistas de la Serotonina/metabolismo , Antagonistas de la Serotonina/farmacología , TritioRESUMEN
Male rats were treated with 10 mg/kg D-fenfluramine (DF) i.p., twice a day for 4 days. Five days later there was a strong reduction (70-100%) in the Bmax of [3H]citalopram binding and the Vmax of [3H]5-HT uptake in cortical and hippocampal synaptosomes; 2 months after the treatment these parameters were reduced by 40-70%. The effect of treatment was also evaluated in synaptosomes preloaded with [3H]5-HT, superfused and exposed for 3 min to a releasing stimulus (15 mM K+ or 0.5 microM DF). In our experimental conditions, the stimulated [3H]5-HT release is Ca(2+)-dependent and takes place only from 5-HT nerve endings. The K(+)-stimulated release was not consistently altered by the DF treatment whereas DF-stimulated [3H]5-HT release was markedly reduced, either 5 days and 2 months after the treatment. The effect of chronic DF was different from the effect of i.c.v. 5,7-DHT, a specific 5-HT neurotoxin which completely abolished the K(+)-induced release. Since the decrease of synaptosomal [3H]5-HT uptake induced by 5,7-DHT (82%) was similar to that found after chronic DF (70-80%), these data suggest that the decrease of 5-HT uptake sites induced by chronic DF is not (only) due to neurodegeneration. That chronic DF could induce a functional down-regulation of 5-HT uptake sites (i.e. decreased density per intact nerve ending) was suggested by the decrease of DF-induced release, since the releasing activity of DF is dependent on functional 5-HT uptake sites. However, due to the characteristics of our model, our results are compatible with either the absence or the presence of a concomitant, partial neurodegeneration of 5-HT nerve endings in DF-treated rats. In summary, our data indicate that after treatment with high doses of DF, the 5-HT uptake carriers undergo a long-lasting down-regulation, thus totally or partly explaining the lower [3H]citalopram binding and the lower synaptosomal [3H]5-HT uptake.
Asunto(s)
Encéfalo/metabolismo , Regulación hacia Abajo , Fenfluramina/farmacología , Receptores de Serotonina/metabolismo , Inhibidores Selectivos de la Recaptación de Serotonina/farmacología , Animales , Relación Dosis-Respuesta a Droga , Masculino , Ratas , Ratas Sprague-Dawley , Receptores de Serotonina/efectos de los fármacos , Serotonina/metabolismo , Estereoisomerismo , Factores de TiempoRESUMEN
Chronic treatment of adult rats with DL-(E)-2-amino-4-methyl-5-phosphono-3-pentenoic carboxyethylester (CGP 39551) (30 mg/kg orally for 12 days) induced a significant increase, 72 h after the last dose, in the N-methyl-D-aspartate (NMDA)-sensitive [3H]glutamate binding in the hippocampal pyramidal layer (stratum oriens CA1, CA3: +51% on average; stratum radiatum CA1, CA3: +40% on average; stratum pyramidale CA1: +20%, CA3: +55%) and in the dentate gyrus (+43%) compared to vehicle-injected animals, as assessed by quantitative receptor autoradiography. Similar results were obtained using the NMDA receptor antagonist, [3H]DL-(E)-2-amino-4-propyl-5-phosphono-3-pentenoic acid (CGP 39653). Saturation experiments showed that the increase in [3H]CGP 39653 binding was due to the maximum number of receptors, without changes in affinity. The same regimen did not alter [3H]N-(1-[2-thienyl]-cyclohexyl)-3,4-piperidine (TCP) binding to the ion channel coupled to the receptor but prevented D-serine (5 microM)-induced enhancement of [3H]glutamate binding. NMDA (3-300 microM) enhanced [3H]noradrenaline release from hippocampal slices, and 7-Cl-kynurenic acid (5-100 microM) and (+)-5-methyl-10,11-dihydro-5H-dibenzo-[a,d]cyclo-hepten-5,10-imine maleate (MK 801) (0.03-0.3 microM), antagonists at the glycine site and ion channel respectively, antagonized this effect to the same extent in CGP 39551-treated rats and controls. Chronic CGP 39551 did not affect the neurotoxic potency of quinolinic acid, a selective agonist at the NMDA receptor, injected in the hippocampus.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
2-Amino-5-fosfonovalerato/análogos & derivados , Hipocampo/efectos de los fármacos , Receptores de N-Metil-D-Aspartato/efectos de los fármacos , 2-Amino-5-fosfonovalerato/farmacocinética , 2-Amino-5-fosfonovalerato/farmacología , Animales , Autorradiografía , Maleato de Dizocilpina/farmacología , Ácido Glutámico/metabolismo , Hipocampo/metabolismo , Ácido Quinurénico/análogos & derivados , Ácido Quinurénico/toxicidad , Masculino , Norepinefrina/metabolismo , Fenciclidina/análogos & derivados , Fenciclidina/metabolismo , Ácido Quinolínico/toxicidad , Ratas , Ratas Sprague-Dawley , Receptores de N-Metil-D-Aspartato/agonistas , Receptores de N-Metil-D-Aspartato/antagonistas & inhibidoresRESUMEN
Intracerebroventricularly (ICV) injected 5,7-dihydroxytryptamine (5,7-DHT), which reduced by 70-90% forebrain serotonin levels, significantly raised glial fibrillary acidic protein (GFAP) mRNA levels in the hippocampus and nucleus raphe dorsalis 5 days but not 15 days after the lesion. A significant increase of mitochondrial benzodiazepine receptors (MBR), measured by binding autoradiography of 3H-PK 11195, was found in the nucleus raphe dorsalis 5 and 15 days after the ICV 5,7-DHT and also in the hippocampus, ventral tegmental area, and substantia nigra at 15 days. No significant effect was observed in the striatum and cortex for either GFAP mRNA or MBR binding. Unlike the ICV route, bilateral injection of 5,7-DHT into the medial forebrain bundle, which caused a 65-90% reduction of serotonin levels in different forebrain regions, significantly raised GFAP mRNA and MBR binding only at the site of injection with no effect in hippocampus, striatum, and cortex. MBR binding slightly increased in the nucleus raphe dorsalis 15 days after the lesion. High doses of d-fenfluramine (10 mg/kg intraperitoneally twice daily for 4 days) caused 80-90% reduction of serotonin levels 5 days after the last injection but did not change the GFAP mRNA or the MBR binding in any of the brain regions considered. These findings suggest that the effect of 5,7-DHT on microglial and glial markers is probably related to a nonspecific interaction with other neuronal systems besides the serotonin or to direct interaction with glial cells; the use of these parameters for detecting selective degeneration of serotonin axons presents some obvious limitations.