RESUMO
Hippocampal long-term potentiation (LTP) is strongly associated with the acquisition of spatial memory and is attenuated by ethanol. Recent studies have shown that the inhibitory potency of ethanol against n-methyl-d-aspartate (NMDA) receptor-mediated synaptic activity is enhanced in hippocampal slices taken from juvenile rats compared to those taken from adults. In addition, ethanol has been reported to impair spatial memory acquisition at lower doses in adolescent rats compared to adults. We therefore hypothesized that the suppression of hippocampal LTP by ethanol would be more potent in hippocampal slices taken from adolescent rats compared to those taken from adults. The potency of ethanol against NMDA receptor-mediated LTP was assessed in area CA1 of hippocampal slices taken from adolescent (30 days old) and adult (90 days old) rats. In slices from adolescent rats, theta-burst stimulus trains reliably induced robust LTP in the absence of ethanol, but when the stimulus trains were presented in the presence of either 10 mM or 30 mM ethanol, LTP induction was significantly suppressed relative to controls. In contrast, there was no effect of these ethanol concentrations on the induction of LTP in hippocampal slices from adult rats. These observations indicate that ethanol suppresses LTP in the adolescent hippocampus at concentrations that do not affect than it suppresses in the adult slices, suggesting a much greater sensitivity to ethanol in adolescence.
Assuntos
Depressores do Sistema Nervoso Central/farmacologia , Etanol/farmacologia , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Hipocampo/efeitos dos fármacos , Potenciação de Longa Duração/efeitos dos fármacos , Fatores Etários , Animais , Depressores do Sistema Nervoso Central/administração & dosagem , Etanol/administração & dosagem , Hipocampo/fisiologia , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
We have developed an on-line archive of neuronal geometry to encourage the use of realistic dendritic structures in morphometry and for neuronal modeling, located at web address www.neuro.soton.ac.uk. Initially we have included full three-dimensional representations of 87 neurons from the hippocampus, obtained following intracellular staining with biocytin and reconstruction using Neurolucida. The archive system includes a structure editor for correcting any departures from valid branching geometry and which allows simple errors in the digitisation to be corrected. The editor employs a platform-independent file format which enforces the constraints that there should be no isolated branches and no closed loops. It also incorporates software for interconversion between the archive format and those used by various neuronal reconstruction and modelling packages. The raw data from digitisation software can be included in the archive as well as edited reconstructions and any further information available. Cross-referenced tables and indexes are updated automatically and are sorted according to a number of fields including the cell type, contributor, submission date and published reference. Both the archive and the structure editor should facilitate the quantitative use of full three-dimensional reconstructions of neurons from the hippocampus and other brain regions.
Assuntos
Dendritos/ultraestrutura , Hipocampo/citologia , Neurônios/citologia , Células Piramidais/citologia , Animais , Bases de Dados Factuais , Processamento de Imagem Assistida por Computador/métodos , Internet , Lisina/análogos & derivados , Modelos Anatômicos , Modelos Neurológicos , Neurônios/ultraestrutura , Células Piramidais/ultraestrutura , Reprodutibilidade dos Testes , SoftwareRESUMO
Choline supplementation during gestation in rats leads to augmentation of spatial memory in adulthood. We hypothesized that prenatal (E12-E17) choline supplementation in the rat would lead to an enhancement of hippocampal synaptic plasticity as assessed by long-term potentiation (LTP) at 3-4 mo of age. LTP was assessed blindly in area CA1 of hippocampal slices with first suprathreshold (above threshold for LTP generation in control slices) theta-burst stimulus trains. The magnitude of potentiation after these stimuli was not different between slices from control and prenatally choline supplemented animals. Next, threshold (reliably leading to LTP generation in control slices) or subthreshold theta-burst stimulus trains were applied to slices from control, prenatally choline-supplemented, and prenatally choline-deprived rats. Threshold level stimulus trains induced LTP in slices from both the control and choline-supplemented rats but not in those from the choline-deficient rats. Subthreshold stimulus trains led to LTP induction in slices from prenatally choline-supplemented rats only. These observations indicate that prenatal dietary manipulation of the amino acid, choline, leads to subsequent significant alterations of LTP induction threshold in adult animals.
Assuntos
Colina/farmacologia , Suplementos Nutricionais , Hipocampo/fisiologia , Potenciação de Longa Duração , Efeitos Tardios da Exposição Pré-Natal , Limiar Sensorial , Animais , Deficiência de Colina/fisiopatologia , Feminino , Hipocampo/embriologia , Técnicas In Vitro , Memória/fisiologia , Gravidez , Ratos , Ratos Sprague-DawleyRESUMO
Dendritic morphology and passive cable properties determine many aspects of synaptic integration in complex neurons, together with voltage-dependent membrane conductances. We investigated dendritic properties of CA1 pyramidal neurons intracellularly labeled during in vivo and in vitro physiologic recordings, by using similar intracellular staining and three-dimensional reconstruction techniques. Total dendritic length of the in vivo neurons was similar to that of the in vitro cells. After correction for shrinkage, cell extent in three-dimensional representation was not different between the two groups. Both in vivo and in vitro neurons demonstrated a variable degree of symmetry, with some neurons showing more cylindrical symmetry around the main apical axis, whereas other neurons were more elliptical, with the variation likely due to preparation and preservation conditions. Branch order analysis revealed no difference in the number of branch orders or dendritic complexity. Passive conduction of dendritic signals to the soma in these neurons shows considerable attenuation, particularly with higher frequency signals (such as synaptic potentials compared with steady-state signals), despite a relatively short electrotonic length. Essential aspects of morphometric appearance and complex dendritic integration critical to CA1 pyramidal cell functioning are preserved across neurons defined from the two different hippocampal preparations used in this study.
Assuntos
Dendritos/ultraestrutura , Células Piramidais/ultraestrutura , Animais , Técnicas In Vitro , Masculino , Potenciais da Membrana/fisiologia , Neurônios/ultraestrutura , Ratos , Ratos Endogâmicos F344 , Coloração e Rotulagem , Transmissão Sináptica/fisiologiaRESUMO
Age-related dendritic alterations were evaluated in F344 rats following a water maze assessment of spatial memory. Based on the probe trial times, 39% of the aged animals were designated impaired. CA1 pyramidal neurons were labeled intracellularly with neurobiotin in brain slices prepared from these animals. Neurons (aged: n = 15; young: n = 11) were reconstructed using a microscope-based three-dimensional system. Increased dendritic length was observed in the aged neurons both for basal dendrites (aged = 4.54 mm and young = 3.33 mm) and the entire neurons (aged = 14.8 mm and young = 10.8 mm). However, dendritic length values did not correlate with the individual animal's probe trial time. Sholl analysis revealed a diffuse increase in dendritic branch intersections in the cells from aged rats, which on branch order analysis was noted to be due to an increased number of distal branches. Mean electrotonic distance to dendritic terminals, a functional assessment of synaptic efficacy, was longer in the aged neurons (aged = 0.67 lambda and young = 0.55 lambda). These results suggest a lengthening and increased complexity of CA1 pyramidal neurons with successful aging, which may represent either an intrinsic response to aging or a reactive partial denervation response to a loss of afferent inputs.
Assuntos
Envelhecimento/fisiologia , Dendritos/fisiologia , Hipocampo/anatomia & histologia , Plasticidade Neuronal/fisiologia , Animais , Comportamento Animal/fisiologia , Masculino , Ratos , Ratos Endogâmicos F344RESUMO
CA3 pyramidal neurons were stained with biocytin during intracellular recording in rat hippocampus in vivo and reconstructed using a computer-based system. The in vivo CA3 neurons were characterized primarily according to their proximity to the hilus and secondarily with respect to the septotemporal location. Neurons measured in CA3a (n = 4), in CA3b (n = 4), and in posterior/ventral locations (n = 3) had the greatest dendritic lengths (19.8, 19.1, and 26.8 mm on average, respectively). Cells closer to the hilus showed much shorter dendritic lengths, averaging 10.4 mm for CA3c neurons (n = 4) and 11.6 mm for zone 3 neurons (n = 2). Half of the cells showed more than one major apical dendrite, and dendritic trees were highly variable even within CA3 subregions. The mean electronic length for these cell groups averaged between 0.30 lambda (CA3c) and 0.45 lambda (posterior/ventral), assuming a constant specific-membrane resistivity of 60 K omega-cm2. These CA3 neurons form a database of reconstructed neurons for further morphometric and electrical modelling studies. The large degree of variability between individual CA3 neurons indicates that both dendritic and electrical properties should be specifically calculated for each cell rather than assuming a "typical" morphology.
Assuntos
Hipocampo/citologia , Neurônios/citologia , Neurônios/fisiologia , Animais , Tamanho Celular , Células Cultivadas/citologia , Células Cultivadas/fisiologia , Dendritos/fisiologia , Condutividade Elétrica , Eletrofisiologia , Feminino , Hipocampo/fisiologia , Processamento de Imagem Assistida por Computador , Indicadores e Reagentes , Lisina/análogos & derivados , Masculino , Neurônios/ultraestrutura , Ratos , Ratos Sprague-Dawley , Ratos WistarRESUMO
Kainic acid (KA) lesions of the CA3 region of the hippocampus lead to denervation of ipsilateral CA1 neurons. To assess denervation-induced post-synaptic changes, intracellular physiological recordings were performed in the CA1 region in vitro, from both control and KA-treated tissue. The neurons were intracellularly stained with neurobiotin, reconstructed using a quantitative three-dimensional system and analyzed for morphometric and electrotonic parameters. Total dendritic length was slightly longer in the denervated CA1 cells and there was a selective and significant increase in both branches and terminals in the mid-stratum radiatum (300-550 microns from the soma using Sholl analysis) in the KA-treated rats compared to untreated controls, particularly for cells at 5 days post-lesion and later, which exhibited graded synaptically-evoked bursts. However, there was no significant difference in the basal dendritic arborization. Electrotonic modelling of the dendritic structure revealed specific membrane resistivity values of 33.4 k omega.cm2 for the normal CA1 cells and 29.8 K omega-cm2 for the KA-treated cells, assuming an internal resistivity of 200 omega.cm2, shrinkage correction of 1.57 and a spatial distribution of dendritic spines. The number of dendritic terminals of these denervated CA1 neurons at electrotonic distances between 0.5 lambda and 0.7 lambda also significantly increased in the cells from KA-treated animals. These findings indicate that there is a selective and specific increase in the number of apical terminals and dendritic branches following the unilateral kainic acid lesion. These apical branch changes may represent dendritic sprouting as a post-synaptic response to the denervation, which was particularly marked in neurons exhibiting graded synaptic bursting behavior.
Assuntos
Dendritos/fisiologia , Hipocampo/fisiologia , Ácido Caínico/toxicidade , Células Piramidais/fisiologia , Animais , Dendritos/ultraestrutura , Denervação , Estimulação Elétrica , Eletrofisiologia , Hipocampo/anatomia & histologia , Hipocampo/efeitos dos fármacos , Masculino , Modelos Biológicos , Células Piramidais/ultraestrutura , Ratos , Ratos Endogâmicos F344RESUMO
Host brain receptivity to fetal hippocampal grafts was investigated following transplantation into unilateral kainic acid (KA) lesions of adult rat hippocampus. E18-E19 hippocampal cell suspensions were labeled with rhodamine dextran amine and transplanted bilaterally into hosts at various times following the KA-lesion. After one to three and one half months survival the grafts (contained within host hippocampal slices) were analyzed using intracellular electrophysiological techniques. A nonparametric graft index was developed which assessed the overall size and distribution of the graft. Using this grading system graft development was noted to be significantly enhanced for grafts placed into hosts with KA lesions at either 2-4 days or 11-12 days following the lesion, compared to grafts placed at either 6-7 days or 27-33 days after the lesion. Also, grafts implanted at delays of either 14-16 or 28-33 days appeared to have fewer surviving cells but were more dispersed within the host brain than grafts at shorter post-lesion implant times. Synaptic responses to host stimulation were noted in most grafts. Intracellular staining of transplanted neurons showed considerable development of cell processes but atypical pyramidal cell morphology and ectopic location; numerous axons traveled into the host tissue. The time course of lesion-induced host receptivity appeared to significantly influence graft development and maturation. In this study graft survival was partially independent from cell migration. This differential effect may be due to various aspects of host brain receptivity, which in turn is influenced by the delay between the host brain lesion and grafting.
RESUMO
Effects of fetal hippocampal transplants were evaluated following a prolonged intraventricular excitotoxic lesion (1.0 mg of N-methyl-D-aspartate over two weeks infusion) in F344 rats. The septum and ipsilateral hippocampus (CA1 and dentate regions) showed extensive cell loss, decreased acquisition of spatial memory was observed and a decrease in AChE positive fiber innervation to the hippocampus was noted following the lesion. Fetal hippocampal transplants into the posterior lateral ventricle resulted in moderate graft survival and physiological analysis of graft-host interconnection in vitro demonstrated evoked field potentials. However, the transplants did not lead to significant improvement in behavior, possibly due to poor synaptic integration of the intraventricular transplants into the host hippocampus. The prolonged intraventricular NMDA lesion may be helpful to understand a mixed lesion model of both septal areas and hippocampus and also as a background lesion in which to assess the connectivity and development of various types of neural grafts.
RESUMO
Dissociated rat fetal hippocampal cells were grafted into normal adult rats. The fetal cells were incubated with one of a number of fluorescent compounds at the time of the dissociation to facilitate identification of the individual grafted cells. The fluorescent labels which were analyzed for this purpose included rhodamine latex microspheres, Cascade blue latex beads, rhodamine-dextran-amine, DiI, and carboxyfluorescein ester. The labeled cells were stereotaxically placed as a suspension into normal rat host hippocampi. The rats were sacrificed 2 to 6 weeks after the grafting for in vitro physiological recordings, and the prelabeled grafts were located using fluorescence optics. During intracellular recordings neurons within the prelabeled grafts were injected with Lucifer yellow to visualize the morphology and integration of neuronal processes within the host. Following the recordings the host slices with the grafts were fixed in 4% paraformaldehyde for anatomical analysis. The ability to prelabel cellular grafts allows subsequent anatomical and physiological analysis of the integration of grafted neurons at the resolution of a single neuron. Such an analysis will improve our understanding of the survival, differentiation, migration, and integration of the grafted neurons and their potential to replace lost function in the lesioned hippocampus.