RESUMO
Protein kinase A (PKA) are tetramers of two catalytic and two regulatory subunits, docked at precise intracellular sites to provide localized phosphorylating activity, triggered by cAMP binding to regulatory subunits and subsequent dissociation of catalytic subunits. It is unclear whether in the brain PKA dissociated subunits may also be found. PKA catalytic subunit was examined in various mouse brain areas using immunofluorescence, equilibrium binding and western blot, to reveal its location in comparison to regulatory subunits type RI and RII. In the cerebral cortex, catalytic subunits colocalized with clusters of RI, yet not all RI clusters were bound to catalytic subunits. In stria terminalis, catalytic subunits were in proximity to RI but separated from them. Catalytic subunits clusters were also present in the corpus striatum, where RII clusters were detected, whereas RI clusters were absent. Upon cAMP addition, the distribution of regulatory subunits did not change, while catalytic subunits were completely released from regulatory subunits. Unpredictably, catalytic subunits were not solubilized; instead, they re-targeted to other binding sites within the tissue, suggesting local macromolecular reorganization. Hence, the interactions between catalytic and regulatory subunits of protein kinase A consistently vary in different brain areas, supporting the idea of multiple interaction patterns.
Assuntos
Encéfalo/enzimologia , Proteína Quinase Tipo II Dependente de AMP Cíclico/metabolismo , Proteína Quinase Tipo I Dependente de AMP Cíclico/metabolismo , Animais , Córtex Cerebral/enzimologia , Corpo Estriado/enzimologia , AMP Cíclico/metabolismo , Proteína Quinase Tipo I Dependente de AMP Cíclico/genética , Proteína Quinase Tipo II Dependente de AMP Cíclico/genética , Feminino , Masculino , Camundongos , Especificidade de Órgãos , Núcleos Septais/enzimologiaRESUMO
In male quail, oestrogens produced in the brain (neuro-oestrogens) exert a dual action on male sexual behaviour: they increase sexual motivation within minutes via mechanisms activated at the membrane but facilitate sexual performance by slower, presumably nuclear-initiated, mechanisms. Recent work indicates that neuro-oestrogens are also implicated in the control of female sexual motivation despite the presence of high circulating concentrations of oestrogens of ovarian origin. Interestingly, aromatase activity (AA) in the male brain is regulated in time domains corresponding to the slow "genomic" and faster "nongenomic" modes of action of oestrogens. Furthermore, rapid changes in brain AA are observed in males after sexual interactions with a female. In the present study, we investigated whether similar rapid changes in brain AA are observed in females allowed to interact sexually with males. A significant decrease in AA was observed in the medial preoptic nucleus after interactions that lasted 2, 5 or 10 minutes, although this decrease was no longer significant after 15 minutes of interaction. In the bed nucleus of the stria terminalis, a progressive decline of average AA was observed between 2 and 15 minutes, although it never reached statistical significance. AA in this nucleus was, however, negatively correlated with the sexual receptivity of the female. These data indicate that sexual interactions affect brain AA in females as in males in an anatomically specific manner and suggest that rapid changes in brain oestrogens production could also modulate female sexual behaviour.
Assuntos
Aromatase/metabolismo , Encéfalo/enzimologia , Codorniz , Comportamento Sexual Animal , Animais , Feminino , Masculino , Área Pré-Óptica/enzimologia , Núcleos Septais/enzimologiaRESUMO
The bed nucleus of the stria terminalis (BNST) is a complex integrative centre in the forebrain, composed of multiple sub-nuclei, each with discrete populations of neurons. Progress in understanding BNST function, both in the adult and during postnatal maturation, is dependent upon a more complete characterization of neuronal phenotypes in the BNST. The aim of the current study was to define the molecular phenotype of one postnatal BNST neuronal population, in order to identify molecular factors that may underlie both (protein marker-related) immaturity, and secondly, the transience of this phenotype. This BNST population was originally identified by high, but transient expression of the EGR1 transcription factor (TF) in postnatal rat lateral intermediate BNST (BNSTLI). The current results confirm a high level of Egr1 activation in postnatal day 10 (PN10) male BNSTLI that is lost at PN40, and now demonstrate a similar pattern of transient activation in female brains. Apparent cellular immaturity in this population, as indicated by low levels of the adult neuronal marker NeuN/RBFOX3, was found to be uncorrelated with both key neuronal regulator protein expression (SOX2 and REST), and also RBFOX2 protein levels. The BNSTLI neurons have a partial catecholaminergic phenotype (tyrosine hydroxylase-positive/dopa decarboxylase-negative; TH+ve/DDC-ve) that is lost at PN40. In contrast, the co-expressed neuropeptide, somatostatin, is maintained, albeit at lower levels, at PN40. The transcriptional basis of the transient and partial catecholaminergic phenotype was investigated by analysing TFs known to maintain adult dopaminergic (TH+ve/DDC+ve) neuronal phenotypes. The BNSTLI neurons were shown to lack forkhead TFs including FOXA1, FOXA2 and FOXO1. In addition, the BNSTLI neurons had low, primarily cytoplasmic, expression of NR4A2/NURR1, an orphan nuclear receptor that is critical for adult maintenance of midbrain dopamine neurons. These results detail the molecular features of an immature neuronal phenotype, and reveal TF deficiencies that may underlie postnatal transience of the phenotype.
Assuntos
Neurônios/química , Neurônios/enzimologia , Fenótipo , Núcleos Septais/química , Núcleos Septais/enzimologia , Tirosina 3-Mono-Oxigenase/metabolismo , Animais , Animais Recém-Nascidos , Feminino , Masculino , Ratos , Ratos Transgênicos , Tirosina 3-Mono-Oxigenase/análise , Tirosina 3-Mono-Oxigenase/genéticaRESUMO
ß-amyloid precursor protein (APP) and amyloid beta peptide (Aß) are strongly implicated in Alzheimer's disease (AD) pathogenesis, although recent evidence has linked APP-ßCTF generated by BACE1 (ß-APP cleaving enzyme 1) to the development of endocytic abnormalities and cholinergic neurodegeneration in early AD. We show that partial BACE1 genetic reduction prevents these AD-related pathological features in the Ts2 mouse model of Down syndrome. Partially reducing BACE1 by deleting one BACE1 allele blocked development of age-related endosome enlargement in the medial septal nucleus, cerebral cortex, and hippocampus and loss of choline acetyltransferase (ChAT)-positive medial septal nucleus neurons. BACE1 reduction normalized APP-ßCTF elevation but did not alter Aß40 and Aß42 peptide levels in brain, supporting a critical role in vivo for APP-ßCTF in the development of these abnormalities. Although ameliorative effects of BACE1 inhibition on ß-amyloidosis and synaptic proteins levels have been previously noted in AD mouse models, our results highlight the additional potential value of BACE1 modulation in therapeutic targeting of endocytic dysfunction and cholinergic neurodegeneration in Down syndrome and AD.
Assuntos
Doença de Alzheimer/genética , Doença de Alzheimer/patologia , Secretases da Proteína Precursora do Amiloide/genética , Secretases da Proteína Precursora do Amiloide/fisiologia , Peptídeos beta-Amiloides/fisiologia , Precursor de Proteína beta-Amiloide/fisiologia , Ácido Aspártico Endopeptidases/genética , Ácido Aspártico Endopeptidases/fisiologia , Neurônios Colinérgicos/patologia , Síndrome de Down/genética , Síndrome de Down/patologia , Endossomos/patologia , Deleção de Genes , Estudos de Associação Genética , Degeneração Neural/patologia , Envelhecimento/genética , Envelhecimento/patologia , Alelos , Animais , Colina O-Acetiltransferase/metabolismo , Modelos Animais de Doenças , Endossomos/genética , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Degeneração Neural/genética , Núcleos Septais/citologia , Núcleos Septais/enzimologiaRESUMO
Over-activation of the brain renin-angiotensin system (RAS) has been implicated in the etiology of anxiety disorders. Angiotensin converting enzyme 2 (ACE2) inhibits RAS activity by converting angiotensin-II, the effector peptide of RAS, to angiotensin-(1-7), which activates the Mas receptor (MasR). Whether increasing brain ACE2 activity reduces anxiety by stimulating central MasR is unknown. To test the hypothesis that increasing brain ACE2 activity reduces anxiety-like behavior via central MasR stimulation, we generated male mice overexpressing ACE2 (ACE2 KI mice) and wild type littermate controls (WT). ACE2 KI mice explored the open arms of the elevated plus maze (EPM) significantly more than WT, suggesting increasing ACE2 activity is anxiolytic. Central delivery of diminazene aceturate, an ACE2 activator, to C57BL/6 mice also reduced anxiety-like behavior in the EPM, but centrally administering ACE2 KI mice A-779, a MasR antagonist, abolished their anxiolytic phenotype, suggesting that ACE2 reduces anxiety-like behavior by activating central MasR. To identify the brain circuits mediating these effects, we measured Fos, a marker of neuronal activation, subsequent to EPM exposure and found that ACE2 KI mice had decreased Fos in the bed nucleus of stria terminalis but had increased Fos in the basolateral amygdala (BLA). Within the BLA, we determined that â¼62% of GABAergic neurons contained MasR mRNA and expression of MasR mRNA was upregulated by ACE2 overexpression, suggesting that ACE2 may influence GABA neurotransmission within the BLA via MasR activation. Indeed, ACE2 overexpression was associated with increased frequency of spontaneous inhibitory postsynaptic currents (indicative of presynaptic release of GABA) onto BLA pyramidal neurons and central infusion of A-779 eliminated this effect. Collectively, these results suggest that ACE2 may reduce anxiety-like behavior by activating central MasR that facilitate GABA release onto pyramidal neurons within the BLA.
Assuntos
Ansiedade/enzimologia , Complexo Nuclear Basolateral da Amígdala/enzimologia , Neurônios/enzimologia , Peptidil Dipeptidase A/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Núcleos Septais/enzimologia , Angiotensina II/administração & dosagem , Angiotensina II/análogos & derivados , Enzima de Conversão de Angiotensina 2 , Animais , Complexo Nuclear Basolateral da Amígdala/efeitos dos fármacos , Potenciais Pós-Sinápticos Inibidores , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neurônios/efeitos dos fármacos , Fragmentos de Peptídeos/administração & dosagem , Proto-Oncogene Mas , Proteínas Proto-Oncogênicas/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-fos/metabolismo , RNA Mensageiro/metabolismo , Receptores Acoplados a Proteínas G/antagonistas & inibidores , Núcleos Septais/efeitos dos fármacos , Ácido gama-Aminobutírico/metabolismoRESUMO
Aromatase, which converts testosterone in estradiol, is involved in the generation of brain sex dimorphisms. Here we used the "four core genotypes" mouse model, in which the effect of gonadal sex and sex chromosome complement is dissociated, to determine if sex chromosomes influence the expression of brain aromatase. The brain of 16 days old XY mouse embryos showed higher aromatase expression in the stria terminalis and the anterior amygdaloid area than the brain of XX embryos, independent of gonadal sex. Furthermore, estradiol or dihydrotestosterone increased aromatase expression in cultures of anterior amygdala neurons derived from XX embryos, but not in those derived from XY embryos. This effect was also independent of gonadal sex. The expression of other steroidogenic molecules, estrogen receptor-α and androgen receptor was not influenced by sex chromosomes. In conclusion, sex chromosomes determine sex dimorphisms in aromatase expression and regulation in the developing mouse brain.
Assuntos
Aromatase/metabolismo , Complexo Nuclear Corticomedial/embriologia , Gônadas/enzimologia , Núcleos Septais/embriologia , Cromossomos Sexuais/metabolismo , Animais , Aromatase/genética , Células Cultivadas , Complexo Nuclear Corticomedial/citologia , Complexo Nuclear Corticomedial/enzimologia , Di-Hidrotestosterona/farmacologia , Estradiol/farmacologia , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Regulação Enzimológica da Expressão Gênica , Masculino , Camundongos , Neurônios/efeitos dos fármacos , Neurônios/enzimologia , Núcleos Septais/citologia , Núcleos Septais/enzimologia , Fatores SexuaisRESUMO
BACKGROUND: Striatal-enriched protein tyrosine phosphatase (STEP) is a brain-specific protein tyrosine phosphatase that opposes the development of synaptic strengthening and the consolidation of fear memories. In contrast, stress facilitates fear memory formation, potentially by activating corticotrophin releasing factor (CRF) neurons in the anterolateral cell group of the bed nucleus of the stria terminalis (BNSTALG). METHODS: Here, using dual-immunofluorescence, single-cell reverse transcriptase polymerase chain reaction, quantitative reverse transcriptase polymerase chain reaction, Western blot, and whole-cell patch-clamp electrophysiology, we examined the expression and role of STEP in regulating synaptic plasticity in rat BNSTALG neurons and its modulation by stress. RESULTS: Striatal-enriched protein tyrosine phosphatase was selectively expressed in CRF neurons in the oval nucleus of the BNSTALG. Following repeated restraint stress (RRS), animals displayed a significant increase in anxiety-like behavior, which was associated with a downregulation of STEP messenger RNA and protein expression in the BNSTALG, as well as selectively enhancing the magnitude of long-term potentiation (LTP) induced in Type III, putative CRF neurons. To determine if the changes in STEP expression following RRS were mechanistically related to LTP facilitation, we examined the effects of intracellular application of STEP on the induction of LTP. STEP completely blocked the RRS-induced facilitation of LTP in BNSTALG neurons. CONCLUSIONS: Hence, STEP acts to buffer CRF neurons against excessive activation, while downregulation of STEP after chronic stress may result in pathologic activation of CRF neurons in the BNSTALG and contribute to prolonged states of anxiety. Thus, targeted manipulations of STEP activity might represent a novel treatment strategy for stress-induced anxiety disorders.
Assuntos
Hormônio Liberador da Corticotropina/metabolismo , Neurônios/enzimologia , Proteínas Tirosina Fosfatases não Receptoras/metabolismo , Núcleos Septais/enzimologia , Estresse Fisiológico , Animais , Masculino , Proteínas Tirosina Fosfatases não Receptoras/fisiologia , Ratos , Ratos Sprague-Dawley , Transdução de SinaisRESUMO
Occlusal disharmony induces chronic stress, which results in learning deficits in association with the morphologic changes in the hippocampus, e.g., neuronal degeneration and increased hypertrophied glial fibrillary acidic protein-positive cells. To investigate the mechanisms underlying impaired hippocampal function resulting from occlusal disharmony, we examined the effects of the bite-raised condition on the septohippocampal cholinergic system by assessing acetylcholine release in the hippocampus and choline acetyltransferase immunoreactivity in the medial septal nucleus in aged SAMP8 mice that underwent the bite raising procedure. Aged bite-raised mice showed decreased acetylcholine release in the hippocampus and a reduced number of choline acetyltransferase-immunopositive neurons in the medial septal nucleus compared to age-matched control mice. These findings suggest that the bite-raised condition in aged SAMP8 mice enhances the age-related decline in the septohippocampal cholinergic system, leading to impaired learning.
Assuntos
Envelhecimento/metabolismo , Má Oclusão/metabolismo , Acetilcolina/metabolismo , Animais , Colina O-Acetiltransferase/metabolismo , Hipocampo/metabolismo , Imuno-Histoquímica , Masculino , Camundongos , Microdiálise , Neurônios/enzimologia , Núcleos Septais/enzimologiaRESUMO
The ventral bed nuclei of the stria terminalis (BST) and medial preoptic nucleus (MPN) of gerbils contain cells that regulate male sex behavior via a largely uncrossed pathway to the retrorubral field (RRF). Our goal was to learn more about cells at the pathway source and target. To determine if the pathway uses GABA as its transmitter, we used immunocytochemistry (ICC) to study glutamic acid decarboxlyase(67) (GAD(67)) colocalization with fluoro-gold (FG) in the ventral BST and MPN after applying FG to the RRF. To determine if the pathway is activated with mating, we studied FG-Fos colocalization in the ventral BST of recently mated males. The ventral BST expresses Fos with mating and is the major pathway source. To determine to what extent other GABAergic cells in the ventral BST are activated with mating, we studied Fos colocalization with GAD(67) mRNA visualized by in situ hybridization (ISH). We also looked for GAD(67) mRNA in RRF cells. Almost all ventral BST and MPNm cells projecting to the RRF (95-97%) and most ventral BST cells activated with mating (89%), were GABAergic. GABAergic cells were also seen in the RRF. RRF-projecting cells represented 37% of ventral BST cells activated with mating. Their activation may reflect arousal and anticipation of sexual reward. Among ventral BST cells that project to the RRF, 14% were activated with mating, consistent with how much of this pathway is needed for mating. The activated GABAergic cells that do not project to the RRF may release GABA locally and inhibit ejaculation.
Assuntos
Prosencéfalo/fisiologia , Caracteres Sexuais , Comportamento Sexual Animal/fisiologia , Tegmento Mesencefálico/fisiologia , Ácido gama-Aminobutírico/fisiologia , Animais , Ativação Enzimática/fisiologia , Feminino , Gerbillinae , Glutamato Descarboxilase/metabolismo , Glutamato Descarboxilase/fisiologia , Masculino , Vias Neurais/citologia , Vias Neurais/enzimologia , Vias Neurais/fisiologia , Área Pré-Óptica/citologia , Área Pré-Óptica/enzimologia , Área Pré-Óptica/fisiologia , Prosencéfalo/citologia , Prosencéfalo/enzimologia , Núcleos Septais/citologia , Núcleos Septais/enzimologia , Núcleos Septais/fisiologia , Tegmento Mesencefálico/citologia , Tegmento Mesencefálico/enzimologiaRESUMO
The biochemical properties, neuroanatomical location, and function of aromatase (ARO), the enzyme that converts testosterone to 17beta-estradiol, have been studied extensively in the adult quail brain. Conversely, very little is known about ARO in quail embryos. This study investigated the distribution of ARO in quail prosencephalon at embryonic days (E) 9, 11, and 15 by immunocytochemistry. ARO-immunoreactive cells were observed within the walls of the cerebral ventricles, the ventral striatum, medial preoptic nucleus (POM), medial part of the bed nucleus of the stria terminalis (BSTM), lateral part of the BST, and in the tuberal region. The BSTM and to a lesser extent the POM showed transient, female-biased sex-differences. In the BSTM, the number of the ARO-immunoreactive cells, the fractional area covered by ARO-immunoreactive structures, and the overall extension of ARO-immunoreactivity were greater in females at E9 and E11, but these differences largely disappeared at E15 and post-hatch day 1. The sex differences were confirmed at the transcriptional level by in situ hybridization. In the lateral part of the POM, females showed slightly more ARO-immunoreactivity than males at E11. Treatment of E9 male embryos with estradiol completely feminized ARO-immunoreactivity at E11. The origins and the functional significance of these sex differences remain unknown.
Assuntos
Aromatase/metabolismo , Coturnix/embriologia , Estradiol/biossíntese , Neurônios/enzimologia , Prosencéfalo/embriologia , Prosencéfalo/enzimologia , Animais , Aromatase/genética , Mapeamento Encefálico , Estradiol/farmacologia , Feminino , Imuno-Histoquímica , Masculino , Rede Nervosa/citologia , Rede Nervosa/embriologia , Rede Nervosa/enzimologia , Neurônios/citologia , Área Pré-Óptica/citologia , Área Pré-Óptica/enzimologia , Prosencéfalo/citologia , RNA Mensageiro/metabolismo , Núcleos Septais/citologia , Núcleos Septais/enzimologia , Caracteres Sexuais , Diferenciação Sexual/fisiologiaRESUMO
In a previous study, we demonstrated that androgenic-anabolic steroids increased aromatase expression in the bed nucleus of stria terminalis and preoptic area in rat brain, as evaluated using autoradiography with [11C]vorozole, a potential positron emission tomography tracer for aromatase. In this study, we explored whether the increase in aromatase binding is mediated via androgen receptors and whether this increase occurs in neurons or glial cells. Rats were given nandrolone decanoate (15 mg/kg body weight once every 3 days) and flutamide (20 mg/kg/day) alone or in combination for 20 days. Results indicated a significant increase of [11C]vorozole binding by nandrolone decanoate in the bed nucleus of the stria terminalis and preoptic area, as in our previous study. Flutamide treatment, on the other hand, decreased [11C]vorozole binding in the bed nucleus of the stria terminalis, preoptic area, and medial amygdala. Immunohistochemical examination demonstrated that upregulation of aromatase expression occurred in neurons. Our findings suggest that aromatase is regulated through an androgen receptor-mediated system. This aromatase-specific tracer and the positron emission tomography technique could be useful for exploring the role of aromatase in anabolic androgenic steroids abusers.
Assuntos
Androgênios/farmacologia , Aromatase/metabolismo , Encéfalo/efeitos dos fármacos , Flutamida/farmacologia , Neurônios/efeitos dos fármacos , Receptores Androgênicos/efeitos dos fármacos , Tonsila do Cerebelo/diagnóstico por imagem , Tonsila do Cerebelo/efeitos dos fármacos , Tonsila do Cerebelo/enzimologia , Antagonistas de Androgênios/farmacologia , Animais , Inibidores da Aromatase/metabolismo , Ligação Competitiva/efeitos dos fármacos , Ligação Competitiva/fisiologia , Encéfalo/diagnóstico por imagem , Encéfalo/enzimologia , Mapeamento Encefálico , Radioisótopos de Carbono , Masculino , Neurônios/enzimologia , Tomografia por Emissão de Pósitrons , Área Pré-Óptica/diagnóstico por imagem , Área Pré-Óptica/efeitos dos fármacos , Área Pré-Óptica/enzimologia , Ratos , Ratos Sprague-Dawley , Receptores Androgênicos/metabolismo , Núcleos Septais/diagnóstico por imagem , Núcleos Septais/efeitos dos fármacos , Núcleos Septais/enzimologia , Triazóis/metabolismo , Triazóis/farmacocinética , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/fisiologiaRESUMO
Evidence suggests that GABA might mediate the inhibitory influence of centrifugal inputs on taste-evoked responses in the parabrachial nucleus (PBN). Previous studies show that activation of the gustatory cortex (GC), bed nucleus of the stria terminalis (BNST), central nucleus of the amygdala (CeA), and lateral hypothalamus (LH) inhibits PBN taste responses, GABAergic neurons are present in these forebrain regions, and GABA reduces the input resistance of PBN neurons. The present study investigated the expression of glutamic acid decarboxylase immunoreactivity (GAD_67 ir) in GC, BNST, CeA, and LH neurons that project to the PBN in rats. After anesthesia (50 mg/kg ip Nembutal), injections of the retrograde tracer Fluorogold (FG) were made in the physiologically defined gustatory PBN. Brain tissue containing the above forebrain structures was processed and examined for FG and GAD_67 ir. Similar to previous studies, each forebrain site contained retrogradely labeled neurons. Our results suggest further that the major source of input to the PBN taste region is the CeA (608 total cells) followed by GC (257 cells), LH (106 cells), and BNST (92 cells). This suggests a differential contribution to centrifugal control of PBN taste processing. We further show that despite the presence of GAD_67 neurons in each forebrain area, colocalization was extremely rare, occurring only in 3 out of 1,063 FG-labeled cells. If we assume that the influence of centrifugal input is mediated by direct projections to the gustatory region of the PBN, then GABAergic forebrain neurons apparently are not part of this descending pathway.
Assuntos
Glutamato Descarboxilase/metabolismo , Neurônios/enzimologia , Ponte/enzimologia , Prosencéfalo/enzimologia , Tonsila do Cerebelo/enzimologia , Tonsila do Cerebelo/patologia , Animais , Eletrofisiologia , Hipotálamo/enzimologia , Hipotálamo/patologia , Masculino , Neurônios/patologia , Ponte/patologia , Prosencéfalo/patologia , Ratos , Ratos Sprague-Dawley , Núcleos Septais/enzimologia , Núcleos Septais/patologiaRESUMO
Investigations using selective lesion techniques suggest that the septohippocampal cholinergic system may not be critical for spatial orientation. These studies employ spatial tasks that provide the animal with access to both environmental and self-movement cues; therefore, intact performance may reflect spared spatial orientation or compensatory mechanisms associated with one class of spatial cues. The present study investigated the contribution of the septohippocampal cholinergic system to spatial behavior by examining performance in foraging tasks in which cue availability was manipulated. Thirteen female Long-Evans rats received selective lesions of the medial septum/vertical band with 192 IgG saporin, and 11 received sham surgeries. Rats were trained to forage for hazelnuts in an environment with access to both environmental and self-movement cues (cued condition). Manipulations include altering availability of environmental cues associated with the refuge (uncued probe), removing all visual environmental cues (dark probe), and placing environmental and self-movement cues into conflict (reversal probe). Medial septum lesions disrupted homeward segment topography only under conditions in which self-movement cues were critical for organizing food hoarding behavior (dark and reversal). These results are consistent with medial septum lesions producing a selective impairment in self-movement cue processing and suggest that these rats were able to compensate for deficits in self-movement cue processing when provided access to environmental cues.
Assuntos
Vias Aferentes/fisiologia , Fibras Colinérgicas/fisiologia , Hipocampo/fisiologia , Orientação/fisiologia , Propriocepção/fisiologia , Comportamento Espacial/fisiologia , Acetilcolinesterase/metabolismo , Vias Aferentes/citologia , Vias Aferentes/enzimologia , Animais , Fibras Colinérgicas/enzimologia , Denervação , Meio Ambiente , Comportamento Exploratório/fisiologia , Feminino , Hipocampo/citologia , Hipocampo/enzimologia , Atividade Motora/fisiologia , Distribuição Aleatória , Ratos , Ratos Long-Evans , Núcleos Septais/citologia , Núcleos Septais/enzimologia , Núcleos Septais/fisiologiaRESUMO
A 25-35% reduction of brain cytochrome oxidase (COx) activity found in Alzheimer's disease (AD) could contribute to neuronal dysfunction and cognitive impairment. The present study replicated the reduction in brain COx activity in rats by administering sodium azide (NaN(3)) for 4 weeks via Alzet minipumps at the rate of 1 mg/kg/h, and determined its effect on hippocampal cholinergic transmission, spatial and episodic memory. NaN(3) caused a selective reduction in choline acetyltransferase (ChAT) immunoreactivity in the diagonal band, a major source of cholinergic input to the hippocampus and cingulate cortex, without altering the number of cholinergic neurons. NaN(3) also induced a significant increase in vesicular acetylcholine transporter (VAChT)-immunoreactive varicosities, GAP-43 in the subgranular layer and of transferrin receptors (TfR) in the hilus of the dentate gyrus. These neurochemical changes were associated with impairment in spatial learning in the Morris water maze and in episodic memory in the object recognition test. Chronic treatment with ladostigil, a novel cholinesterase and monoamine oxidase inhibitor, prevented the decrease in ChAT in the diagonal band, the compensatory increase in synaptic plasticity and TfR and the memory deficits without restoring COx activity. Ladostigil had no significant effect on ChAT activity, synaptic plasticity or TfR in control rats. Ladostigil may have a beneficial effect on cognitive deficits in AD patients that have a reduction in cortical COx activity and cholinergic hypofunction.
Assuntos
Acetilcolina/metabolismo , Fibras Colinérgicas/efeitos dos fármacos , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Hipocampo/efeitos dos fármacos , Hipocampo/enzimologia , Indanos/farmacologia , Transtornos da Memória/enzimologia , Doença de Alzheimer/tratamento farmacológico , Doença de Alzheimer/enzimologia , Doença de Alzheimer/fisiopatologia , Animais , Fibras Colinérgicas/metabolismo , Inibidores da Colinesterase/farmacologia , Inibidores da Colinesterase/uso terapêutico , Complexo IV da Cadeia de Transporte de Elétrons/antagonistas & inibidores , Inibidores Enzimáticos/toxicidade , Proteína GAP-43/efeitos dos fármacos , Proteína GAP-43/metabolismo , Hipocampo/fisiopatologia , Indanos/uso terapêutico , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Aprendizagem em Labirinto/fisiologia , Transtornos da Memória/induzido quimicamente , Transtornos da Memória/tratamento farmacológico , Plasticidade Neuronal/efeitos dos fármacos , Plasticidade Neuronal/fisiologia , Ratos , Ratos Sprague-Dawley , Receptores da Transferrina/efeitos dos fármacos , Receptores da Transferrina/metabolismo , Núcleos Septais/efeitos dos fármacos , Núcleos Septais/enzimologia , Núcleos Septais/fisiopatologia , Azida Sódica/toxicidade , Resultado do Tratamento , Proteínas Vesiculares de Transporte de Acetilcolina/efeitos dos fármacos , Proteínas Vesiculares de Transporte de Acetilcolina/metabolismoRESUMO
In birds and mammals, aromatase activity in the preoptic-hypothalamic region (HPOA) is usually higher in males than in females. It is, however, not known whether the enzymatic sex difference reflects the differential activation of aromatase transcription or some other control mechanism. Although sex differences in aromatase activity are clearly documented in the HPOA of Japanese quail (Coturnix japonica), only minimal or even no differences at all were observed in the number of aromatase-immunoreactive (ARO-ir) cells in the medial preoptic nucleus (POM) and in the medial part of the bed nucleus striae terminalis (BSTM). We investigated by in situ hybridization the distribution and possible sex differences in aromatase mRNA expression in the brain of sexually active adult quail. The distribution of aromatase mRNA matched very closely the results of previous immunocytochemical studies with the densest signal being observed in the POM, BSTM and in the mediobasal hypothalamus (MBH). Additional weaker signals were detected in the rostral forebrain, arcopallium and mesencephalic regions. No sex difference in the optical density of the hybridization signal could be found in the POM and MBH but the area covered by mRNA was larger in males than in females, indicating a higher overall expression in males. In contrast, in the BSTM, similar areas were covered by the aromatase expression in both sexes but the density of the signal was higher in females than in males. The physiological control of aromatase is thus neuroanatomically specific and with regard to sex differences, these controls are at least partially different if one compares the level of transcription, translation and activity of the enzyme. These results also indirectly suggest that the sex difference in aromatase enzyme activity that is present in the quail HPOA largely results from differentiated controls of enzymatic activity rather than differences in enzyme concentration.
Assuntos
Aromatase/biossíntese , Coturnix/fisiologia , Área Pré-Óptica/anatomia & histologia , Área Pré-Óptica/metabolismo , RNA Mensageiro/biossíntese , Núcleos Septais/anatomia & histologia , Núcleos Septais/metabolismo , Animais , Clonagem Molecular , DNA Complementar/biossíntese , Interpretação Estatística de Dados , Feminino , Regulação Enzimológica da Expressão Gênica/fisiologia , Hibridização In Situ , Masculino , Fosforilação , Área Pré-Óptica/enzimologia , Núcleos Septais/enzimologia , Caracteres SexuaisRESUMO
In the present study, we investigated the alteration of aromatase expression in the brain by anabolic androgenic steroid treatment in male rats. The rats were given nandrolone decanoate (15 mg/kg/day) for 14 days, and the brains were used for autoradiography with [C]vorozole, which has been developed as a positron emission tomography tracer for aromatase by our group. The results indicated a significant increase of [C]vorozole binding by anabolic androgenic steroids in the bed nucleus of the stria terminalis and preoptic area. In contrast, no significant change of [C]vorozole binding was observed in the medial amygdala. Our results suggest that aromatase is significantly upregulated in the bed nucleus of the stria terminalis and preoptic area by anabolic androgenic steroids and also suggest that androgens regulate aromatase differently in these structures compared with the medial amygdala.
Assuntos
Androgênios/farmacologia , Inibidores da Aromatase/metabolismo , Hipotálamo/diagnóstico por imagem , Nandrolona/farmacologia , Tomografia por Emissão de Pósitrons/métodos , Triazóis/metabolismo , Animais , Aromatase/metabolismo , Inibidores da Aromatase/farmacologia , Autorradiografia , Radioisótopos de Carbono , Modelos Animais de Doenças , Hipotálamo/efeitos dos fármacos , Hipotálamo/enzimologia , Masculino , Ratos , Ratos Sprague-Dawley , Núcleos Septais/diagnóstico por imagem , Núcleos Septais/efeitos dos fármacos , Núcleos Septais/enzimologia , Transtornos Relacionados ao Uso de Substâncias/diagnóstico por imagem , Transtornos Relacionados ao Uso de Substâncias/metabolismo , Triazóis/farmacologiaRESUMO
The basal forebrain (BF) plays an important role in modulating cortical activity and influencing attention, learning and memory. These activities are fulfilled importantly yet not entirely by cholinergic neurons. Noncholinergic neurons also contribute and comprise GABAergic neurons and other possibly glutamatergic neurons. The aim of the present study was to estimate the total number of cells in the BF of the rat and the proportions of that total represented by cholinergic, GABAergic and glutamatergic neurons. For this purpose, cells were counted using unbiased stereological methods within the medial septum, diagonal band, magnocellular preoptic nucleus, substantia innominata and globus pallidus in sections stained for Nissl substance and/or the neurotransmitter enzymes, choline acetyltransferase (ChAT), glutamic acid decarboxylase (GAD) or phosphate-activated glutaminase (PAG). In Nissl-stained sections, the total number of neurons in the BF was estimated as approximately 355,000 and the numbers of ChAT-immuno-positive (+) as approximately 22,000, GAD+ approximately 119,000 and PAG+ approximately 316,000, corresponding to approximately 5%, approximately 35% and approximately 90% of the total. Thus, of the large population of BF neurons, only a small proportion has the capacity to synthesize acetylcholine (ACh), one third to synthesize GABA and the vast majority to synthesize glutamate (Glu). Moreover, through the presence of PAG, a proportion of ACh- and GABA-synthesizing neurons also has the capacity to synthesize Glu. In sections dual fluorescent immunostained for vesicular transporters, vesicular glutamate transporter (VGluT) 3 and not VGluT2 was present in the cell bodies of most PAG+ and ChAT+ and half the GAD+ cells. Given previous results showing that VGluT2 and not VGluT3 was present in BF axon terminals and not colocalized with VAChT or VGAT, we conclude that the BF cell population influences cortical and subcortical regions through neurons which release ACh, GABA or Glu from their terminals but which in part can also synthesize and release Glu from their soma or dendrites.
Assuntos
Colina O-Acetiltransferase/metabolismo , Glutamato Descarboxilase/metabolismo , Glutaminase/metabolismo , Neurônios/enzimologia , Substância Inominada/enzimologia , Proteínas Vesiculares de Transporte de Glutamato/metabolismo , Acetilcolina/biossíntese , Animais , Contagem de Células , Ácido Glutâmico/biossíntese , Imuno-Histoquímica , Masculino , Vias Neurais/citologia , Vias Neurais/enzimologia , Neurônios/citologia , Área Pré-Óptica/citologia , Área Pré-Óptica/enzimologia , Terminações Pré-Sinápticas/enzimologia , Terminações Pré-Sinápticas/ultraestrutura , Ratos , Ratos Wistar , Núcleos Septais/citologia , Núcleos Septais/enzimologia , Substância Inominada/citologia , Transmissão Sináptica/fisiologia , Ácido gama-Aminobutírico/biossínteseRESUMO
Prenatal exposure of mice to heroin resulted in behavioral deficits present at adulthood, and related to septohippocampal cholinergic innervation accompanied by both pre- and postsynaptic cholinergic hyperactivity; including an increase in membrane PKC activity, and a desensitization of PKC to cholinergic input, which correlated highly with the behavioral performance, and was reversed by cholinergic grafting. The effect was shown in the behaviorally relevant PKCgamma and beta whereas the less behaviorally relevant PKCalpha isoform was not affected. The present study was designed to establish the effect of heroin exposure on the expression of the PKC isoforms level and on the more functionally relevant cholinergic translocation/activation of the isoforms throughout postnatal development. The hippocampi of mice pups, exposed to heroin transplacentally, were assayed after incubation with carbachol for PKC isoforms on postnatal days (PN) 1, 7, 14, 21, 30 and 50. Prenatal heroin exposure increased basal PKCgamma, beta and alpha levels. PKCgamma and alpha levels returned to control levels on PN50. While in PKCbeta, this increase lasted until PN50. Translocation/activation of the PKC isoforms gamma and beta by cholinergic receptor stimulation was present from PN1, concurrent with the presence of the isoforms. Prenatal exposure to heroin completely abolished the translocation/activation throughout the entire postnatal development. This defect was shown from the very beginning, PN1, the day when the PKC isoforms appear. The results suggest that the PKCgamma and beta isoforms are functional concurrent with their developmental appearance. Unlike findings on some other teratogens, the prenatal heroin effect on the isoforms function is similar throughout postnatal development.
Assuntos
Fibras Colinérgicas/metabolismo , Dependência de Heroína/enzimologia , Heroína/efeitos adversos , Hipocampo/efeitos dos fármacos , Efeitos Tardios da Exposição Pré-Natal/enzimologia , Proteína Quinase C/efeitos dos fármacos , Acetilcolina/metabolismo , Analgésicos Opioides/efeitos adversos , Animais , Animais Recém-Nascidos , Agonistas Colinérgicos/farmacologia , Fibras Colinérgicas/efeitos dos fármacos , Modelos Animais de Doenças , Ativação Enzimática/efeitos dos fármacos , Ativação Enzimática/fisiologia , Feminino , Dependência de Heroína/fisiopatologia , Hipocampo/enzimologia , Hipocampo/fisiopatologia , Masculino , Camundongos , Vias Neurais/efeitos dos fármacos , Vias Neurais/enzimologia , Técnicas de Cultura de Órgãos , Gravidez , Efeitos Tardios da Exposição Pré-Natal/fisiopatologia , Isoformas de Proteínas/efeitos dos fármacos , Isoformas de Proteínas/metabolismo , Proteína Quinase C/metabolismo , Transporte Proteico/efeitos dos fármacos , Transporte Proteico/fisiologia , Receptores Colinérgicos/efeitos dos fármacos , Receptores Colinérgicos/metabolismo , Núcleos Septais/efeitos dos fármacos , Núcleos Septais/enzimologia , Núcleos Septais/fisiopatologiaRESUMO
Clinical trials show beneficial effects of acetylcholinesterase (AChE) inhibitors, including galantamine, on cognitive functions in patients with mild to moderate Alzheimer's disease. Galantamine shows a dual action profile by also acting as an allosteric modulator of nicotinic acetylcholine receptors. Nevertheless, its in vivo mechanism of action is only partly understood. Here, we first established a novel lesion model provoking significant functional impairment of the septo-hippocampal projection system without triggering massive neuronal death in the rat medial septum. Next, we studied whether galantamine, administered in doses of 1 and 3mg/kg post-lesion, promotes functional recovery of spatial navigation behaviors, and affects the output of septal cholinergic projections. Infusion of N-methyl-d-aspartate (NMDA; 30nmol/1microl) in the medial septum resulted in spatial learning deficits associated with significant shrinkage of cholinergic neurons and reduced AChE activity in the hippocampus at 7 days post-lesion. Galantamine treatment alone significantly increased the hippocampal acetylcholine concentration and attenuated the NMDA-induced spatial learning impairment. Galantamine post-treatment also affected NMDA-induced changes in AChE and choline-acetyltransferase activities. In conclusion, our data show that galantamine attenuates experimentally-induced cognitive impairments underscored by mild neuronal damage.
Assuntos
Comportamento Animal/efeitos dos fármacos , Fibras Colinérgicas/efeitos dos fármacos , Galantamina/farmacologia , Aprendizagem em Labirinto/efeitos dos fármacos , Antagonistas Nicotínicos/farmacologia , Núcleos Septais/efeitos dos fármacos , Acetilcolinesterase/efeitos dos fármacos , Acetilcolinesterase/metabolismo , Análise de Variância , Animais , Colina O-Acetiltransferase/efeitos dos fármacos , Colina O-Acetiltransferase/metabolismo , Fibras Colinérgicas/enzimologia , Inibidores da Colinesterase/farmacologia , Relação Dose-Resposta a Droga , Hipocampo/efeitos dos fármacos , Hipocampo/enzimologia , Masculino , Aprendizagem em Labirinto/fisiologia , Modelos Animais , N-Metilaspartato , Degeneração Neural/induzido quimicamente , Degeneração Neural/enzimologia , Vias Neurais/efeitos dos fármacos , Vias Neurais/enzimologia , Neurotoxinas , Ratos , Ratos Wistar , Receptores Nicotínicos/efeitos dos fármacos , Receptores Nicotínicos/metabolismo , Núcleos Septais/enzimologia , Percepção Espacial/efeitos dos fármacosRESUMO
Territorial aggression is influenced by many social and environmental factors. Since aggression is a costly behavior, individuals should account for multiple factors such as population density or reproductive status before engaging in aggression. Previous work has shown that male California mice (Peromyscus californicus) respond to winning aggressive encounters by initiating aggression more quickly in future encounters, and we investigated the physiological basis for this effect. We found that injections that produced a transient increase in testosterone (T) following an aggressive encounter caused males to behave more aggressively in an encounter the following day. Experience alone was not enough to change aggression, as males treated with saline injections showed no change in aggression. The effect of T injections on aggression was androgen-based, as the inhibition of aromatase did not block the T injections from increasing aggression. Aromatase inhibition did, however, increase aggression in the initial aggression tests (before application of T or saline injections), and aromatase activity in the bed nucleus of the stria terminalis (BNST) was negatively correlated with aggression. A previous study suggested that aromatase activity in the BNST decreases after males become fathers. Thus, distinct neuroendocrine mechanisms allow male California mice to adjust aggressive behavior in response to changes in social and reproductive status.