RESUMO
High nuclear expression of G protein-coupled receptors, including kinin B1 receptors (B1R), has been observed in several human cancers, but the clinical significance of this is unknown. We put forward the hypothesis that these "nuclearized" kinin B1R contribute to tumorigenicity and can be a new target in anticancer strategies. Our initial immunostaining and ultrastructural electron microscopy analyses demonstrated high B1R expression predominantly located at internal/nuclear compartments in the MDA-MB-231 triple-negative breast cancer (TNBC) cell line as well as in clinical samples of patients with TNBC. On the basis of these findings, in the present study, we evaluated the anticancer therapeutic potential of newly identified, cell-permeable B1R antagonists in MDA-MB-231 cells (ligand-receptor binding/activity assays and LC-MS/MS analyses). We found that these compounds (SSR240612, NG67, and N2000) were more toxic to MDA-MB-231 cells in comparison with low- or non-B1R expressing MCF-10A normal human mammary epithelial cells and COS-1 cells, respectively (clonogenic, MTT proliferative/cytocidal assays, and fluorescence-activated cell-sorting (FACS)-based apoptosis analyses). By comparison, the peptide B1R antagonist R954 unable to cross cell membrane failed to produce anticancer effects. Furthermore, the putative mechanisms underlying the anticancer activities of cell-penetrant B1R antagonists were assessed by analyzing cell cycle regulation and signaling molecules related to cell survival and apoptosis (FACS and western blot). Finally, drug combination experiments showed that cell-penetrant B1R antagonists can cooperate with suboptimal doses of chemotherapeutic agents (doxorubicin and paclitaxel) to promote TNBC death. This study provides evidence on the potential value of internally acting kinin B1R antagonists in averting growth of breast cancer.
Assuntos
Peptídeos Penetradores de Células/farmacologia , Receptor B1 da Bradicinina/genética , Receptores Acoplados a Proteínas G/genética , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Animais , Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Apoptose/efeitos dos fármacos , Antagonistas de Receptor B1 da Bradicinina/farmacologia , Células COS , Linhagem Celular Tumoral , Permeabilidade da Membrana Celular/efeitos dos fármacos , Permeabilidade da Membrana Celular/genética , Chlorocebus aethiops , Doxorrubicina/farmacologia , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Paclitaxel/farmacologia , Transdução de Sinais/efeitos dos fármacos , Neoplasias de Mama Triplo Negativas/genética , Neoplasias de Mama Triplo Negativas/patologiaRESUMO
Introduction: The aim of this manuscript is twofold: first, to investigate the relationship between rhythmic, phonological and graphomotor skills in kindergarten children; and second, to evaluate the possible impact of rhythmic training on the two other skills. Methods: To that end, we selected a sample of 78 children in Québec. Forty-two children received rhythmic training (experimental group) and 34 arts training (active control group) during the same period (10 weeks). Before and after training, children in both groups were assessed for general skills (forward and backward memory span, vocabulary, non-verbal ability), rhythmic skills (synchronization and discrimination tasks), literacy skills (phonological skills - syllable counting, syllable deletion, rhyme discrimination - and invented spelling skills) and graphomotor skills (legibility of letter writing, quality of copying of geometric shapes). Results: Results showed correlations between the child's rhythmic and literacy skills, as well as between rhythm synchronization and pen pressure. In addition, rhythmic training showed improvement in rhythmic abilities, but this did not transfer to literacy or graphomotor development (apart from a significant increase in the duration of pauses in both groups at post-test, with a larger improvement for the rhythm group). Discussion: These results are discussed in terms of duration and intensity of learning, and they highlight the possible benefits of informal rhythm practices in the classroom.
RESUMO
Opiate addiction is characterized by progressive increases in drug intake over time suggesting maladaptive changes in motivational and reward systems. These behaviors are mediated by dopaminergic neurons originating from the ventral tegmental area (VTA), and long-term changes of these dopaminergic neurons are attributed to increased postsynaptic glutamatergic activation. Indeed, chronic morphine administration is known to increase AMPA receptor glutamate receptor 1 (GluR1) subunit in the VTA. However, there is no ultrastructural evidence that morphine affects the expression or surface availability of GluR1 subunits in VTA neurons of defined distribution or transmitter phenotype. Therefore, we examined electron microscopic immunolabeling of GluR1 and tyrosine hydroxylase (TH) in two VTA regions of rats perfused 1 h after a single injection of morphine, or chronic morphine in intermittent-escalating doses for 14 d, and appropriate saline controls. Acute morphine administration produced a significant increase in GluR1 immunogold particles at the plasma membrane and postsynaptic densities in both TH- and non-TH-containing dendrites in the parabrachial VTA, a region that contains mainly prefrontal-cortical-projecting dopaminergic neurons involved in motivation and drug-seeking behavior. Chronic morphine administration maintained the increased synaptic GluR1 labeling in the parabrachial VTA, but also increased the number of GluR1-labeled synapses and TH immunoreactivity in dendrites of the paranigral VTA where substantially more dopaminergic neurons project to limbic structures implicated in locomotor activation and reward. These results demonstrate a region- and dose-dependent redistribution of GluR1-containing AMPA receptors, which is consistent with acute morphine activation of cortical-projecting VTA neurons and chronic morphine activation of limbic-projecting VTA neurons.
Assuntos
Analgésicos Opioides/administração & dosagem , Morfina/administração & dosagem , Neurônios , Receptores de AMPA/metabolismo , Área Tegmentar Ventral/citologia , Animais , Dendritos/efeitos dos fármacos , Dendritos/metabolismo , Dendritos/ultraestrutura , Relação Dose-Resposta a Droga , Esquema de Medicação , Masculino , Microscopia Imunoeletrônica/métodos , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Neurônios/ultraestrutura , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Estilbamidinas/metabolismo , Frações Subcelulares/efeitos dos fármacos , Frações Subcelulares/metabolismo , Sinapses/efeitos dos fármacos , Sinapses/metabolismo , Sinapses/ultraestrutura , Fatores de Tempo , Tirosina 3-Mono-Oxigenase/metabolismo , Área Tegmentar Ventral/efeitos dos fármacosRESUMO
Tonic activation of neurokinin-3 (NK(3)) receptors in dopamine neurons of the ventral tegmental area (VTA) has been implicated in the pathophysiology of schizophrenia. This psychiatric disorder is associated with a dysfunctional activity in VTA projection neurons that can affect cognitive function at the level of the medial prefrontal cortex (mPFC) as well as motor and motivational states controlled in part by mesolimbic output to the nucleus accumbens (Acb). To determine the relevant sites for NK(3) receptor activation within this neuronal network, we used confocal and electron microscopy to examine NK(3) receptors (Cy5; immunogold) and retrograde labeling of fluorogold (FG, FITC; immunoperoxidase) in the VTA of rats receiving either Acb or mPFC injections of FG. Comparison was made with neurokinin-1 (NK(1)) receptors, which are also present, but less abundant then NK(3) receptors, in dopaminergic and GABAergic VTA neurons. There were no observable differences between NK(3) and NK(1) receptors in their primary locations in the cytoplasm and on the plasma membrane of VTA somata and dendrites with or without FG. Dendrites labeled with FG retrogradely transported from mPFC, however, contained more NK(3) or less NK(1) immunogold particles (plasmalemmal + cytoplasmic) then those retrogradely labeled following FG injection in the Acb. Moreover, only the NK(3) receptors were detected in neuronal nuclei in the VTA and in the nuclei of human HEK-293T NK(3)-transfected cells. The enrichment of NK(3) receptors in mesocortical projection neurons and nuclear distribution of these receptors may provide insight for understanding the selective antipsychotic effectiveness of NK(3) antagonists.
Assuntos
Córtex Cerebral/metabolismo , Núcleo Accumbens/metabolismo , Receptores da Neurocinina-1/metabolismo , Receptores da Neurocinina-3/metabolismo , Área Tegmentar Ventral/metabolismo , Animais , Linhagem Celular , Membrana Celular/metabolismo , Membrana Celular/ultraestrutura , Núcleo Celular/metabolismo , Núcleo Celular/ultraestrutura , Dendritos/metabolismo , Dendritos/ultraestrutura , Vias Eferentes/citologia , Vias Eferentes/metabolismo , Humanos , Masculino , Microscopia Confocal , Microscopia Eletrônica de Transmissão , Neurônios/citologia , Neurônios/metabolismo , Ratos , Ratos Sprague-Dawley , Taquicininas/metabolismo , Área Tegmentar Ventral/citologiaRESUMO
G protein-coupled receptors (GPCRs) are integral cell-surface proteins having a central role in tumor growth and metastasis. However, several GPCRs retain an atypical intracellular/nuclear location in various types of cancer. The pathological significance of this is currently unknown. Here we extend this observation by showing that the bradykinin B2R (BK-B2R) is nuclearly expressed in the human triple-negative breast cancer (TNBC) cell line MDA-MB-231 and in human clinical specimens of TNBC. We posited that these "nuclearized" receptors could be involved in oncogenic signaling linked to aberrant growth and survival maintenance of TNBC. We used cell-penetrating BK-B2R antagonists, including FR173657 and novel transducible, cell-permeable forms of the peptide B2R antagonist HOE 140 (NG68, NG134) to demonstrate their superior efficacy over impermeable ones (HOE 140), in blocking proliferation and promoting apoptosis of MDA-MB-231 cells. Some showed an even greater antineoplastic activity over conventional chemotherapeutic drugs in vitro. The cell-permeable B2R antagonists had less to no anticancer effects on B2R shRNA-knockdown or non-B2R expressing (COS-1) cells, indicating specificity in their action. Possible mechanisms of their anticancer effects may involve activation of p38kinase/p27Kip1 pathways. Together, our data support the existence of a possible intracrine signaling pathway via internal/nuclear B2R, critical for the growth of TNBC cells, and identify new chemical entities that enable to target the corresponding intracellular GPCRs.
RESUMO
1. Systemic and regional cardiovascular changes were measured following bilateral microinjection of specific and selective opioid-receptor antagonists into the paraventricular nuclei of the hypothalamus (PVN) of awake, freely moving rats. 2. PVN microinjection of increasing doses of the specific opioid antagonist naloxone - methiodide (1 - 5.0 nmol), or a selective mu-opioid receptor antagonist, beta-funaltrexamine (0.05 - 0.5 nmol), evoked important cardiovascular changes characterized by small and transient increases in heart rate (HR) and mean arterial pressure (MAP), vasoconstriction in renal and superior mesenteric vascular beds and vasodilation in the hindquarter vascular bed. 3. No significant cardiovascular changes were observed following PVN administration of the highly selective delta-opioid-receptor antagonist, ICI 174864 (0.1 - 1 nmol), or the selective kappa-opioid-receptor antagonist, nor-binaltorphine (0.1 - 1 nmol). 4. Most of the cardiovascular responses to naloxone (3 nmol) and beta-funaltrexamine (0.5 nmol) were attenuated or abolished by an i.v. treatment with a specific vasopressin V(1) receptor antagonist. 5. These results suggest that endogenous opioid peptides and mu-type PVN opioid receptors modulate a tonically-active central depressor pathway acting on systemic and regional haemodynamic systems. Part of this influence could involve a tonic inhibition of vasopressin release.
Assuntos
Hemodinâmica/efeitos dos fármacos , Peptídeos Opioides/farmacologia , Núcleo Hipotalâmico Paraventricular/efeitos dos fármacos , Animais , Pressão Sanguínea/efeitos dos fármacos , Pressão Sanguínea/fisiologia , Frequência Cardíaca/efeitos dos fármacos , Frequência Cardíaca/fisiologia , Hemodinâmica/fisiologia , Masculino , Antagonistas de Entorpecentes/farmacologia , Inibição Neural/efeitos dos fármacos , Inibição Neural/fisiologia , Núcleo Hipotalâmico Paraventricular/fisiologia , Ratos , Ratos Wistar , Receptores Opioides/fisiologiaRESUMO
1. The role of nigral tachykinin NK(1), NK(2) and NK(3) receptors in central cardiovascular regulation was studied by measuring the effects of selective agonists and antagonists on mean arterial pressure (MAP) and heart rate (HR) after bilateral microinjection into the substantia nigra of spontaneously hypertensive rats (SHR). Quantitative in vitro autoradiography was also performed in the midbrain of SHR and Wistar-Kyoto (WKY) with the NK(3) receptor ligand [(125)I]-HPP-Senktide. 2. Tachycardia was elicited by the NK(1) ([Sar(9),Met(O(2))(11)]SP) and NK(2) ([betaAla(8)]NKA(4-10)) agonists at 25 and 100 pmol while the NK(3) agonist (senktide, 50 and 100 pmol) had no significant effect. The three agonists had no effect on behaviour, and increases in MAP were elicited by the NK(1) agonist only. 3. Whereas antagonists at NK(1) (RP 67580, 500 pmol) and NK(2) (SR 48968, 500 pmol) receptors had no significant effect on MAP and HR, the NK(3) antagonist (R-820, 500 pmol) reduced MAP for over 3 h in SHR. That anti-hypertensive effect did not occur after intracerebroventricular or intravenous injection of R-820. Also, R-820 had no cardiovascular effect in WKY. 4. The affinity (K(D): 0.7 nM) and densities of specific NK(3) receptor binding sites measured in the substantia nigra, ventral tegmental area, hippocampus and amygdala were not significantly different in SHR and WKY. 5. It is concluded that endogenous tachykinins exert a tonic activity on NK(3) receptors in the substantia nigra of SHR to maintain high blood pressure. Hence, nigral tachykinin NK(3) receptors may represent a promising therapeutic target in the treatment of arterial hypertension.
Assuntos
Hipertensão/fisiopatologia , Receptores da Neurocinina-3/fisiologia , Substância Negra/efeitos dos fármacos , Substância Negra/fisiologia , Animais , Autorradiografia/métodos , Pressão Sanguínea/efeitos dos fármacos , Pressão Sanguínea/fisiologia , Frequência Cardíaca/efeitos dos fármacos , Frequência Cardíaca/fisiologia , Hipertensão/induzido quimicamente , Masculino , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Receptores da Neurocinina-3/agonistas , Receptores da Neurocinina-3/análise , Receptores da Neurocinina-3/antagonistas & inibidores , Substância Negra/químicaRESUMO
Tachykinins are involved in the central autonomic control of blood pressure. In the present study, we examined the i.c.v. cardiovascular effects of several tachykinin receptor antagonists in awake spontaneously hypertensive rats (SHR, 15 weeks old). Results showed that two tachykinin NK(3) receptor antagonists (R-820: 3-indolylcarbonyl-Hyp-Phg-N(Me)-Bzl and SB 222200: (S)-(-)-N-(alpha-ethylbenzyl)-3-methyl-2-phenylquinoline-4-carboxamide) caused a sustained and dose-dependent reduction of blood pressure when injected i.c.v. but not i.v. The stereoselective anti-hypertensive effect of SB 222200 peaked at 3 h and faded at 6 h post-injection (if injected at 07:00 h) or had a slower onset and peaked at 8 h post-injection (if injected at 13:00 h). The effect of R-820 was maximal at 24 h and lasted up to 48 h post-injection. Both antagonists failed to alter blood pressure in normotensive Wistar-Kyoto rats (WKY) and heart rate was not affected in both strains. The anti-hypertensive effect of SB 222200 was not associated with changes in plasma levels of catecholamines and vasopressin and it remained unchanged in SHR subjected to acute bilateral nephrectomy. In contrast, blood pressure was not affected by tachykinin NK(1) (RP 67580: (+/-) 7,7-diphenyl-2[1-imino-2(2-methoxy-phenyl)-ethyl]perhydroisoindol-4-one(3aR,7aR)) and NK(2) (SR 48968: (S)-N-methyl-N[4-(4-acetylamino-4-phenylpiperidino)-2-(3,4-dichlorophenyl)butyl]benzamide) receptor antagonists. Data suggest that brain tachykinin NK(3) receptors are implicated in the maintenance of hypertension in SHR. Hence, these receptors may represent promising therapeutic target in the treatment of arterial hypertension.
Assuntos
Anti-Hipertensivos/administração & dosagem , Hipertensão/tratamento farmacológico , Receptores da Neurocinina-3/antagonistas & inibidores , Acetatos/administração & dosagem , Animais , Pressão Sanguínea/efeitos dos fármacos , Pressão Sanguínea/fisiologia , Frequência Cardíaca/efeitos dos fármacos , Frequência Cardíaca/fisiologia , Hipertensão/fisiopatologia , Injeções Intraventriculares , Masculino , Quinolinas/administração & dosagem , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Receptores da Neurocinina-3/fisiologiaRESUMO
Central neuronal circuits that relay stress information include vasopressin- (AVP) and oxytocin- (OC) containing neurons of the paraventricular nucleus of the hypothalamus (PVN). These neurons are potentially modulated by neurokinin-3 receptors (NK3Rs) of the tachykinin family of neuropeptides. NK3Rs have been localized in PVN neurons and have showed nuclear translocation following an osmotic challenge in rodents. However, their subcellular distribution in AVP or OC neurons of the PVN and plasticity following restraint stress in rats are unknown. In the present study, densities of NK3Rs in PVN AVP- or OC-labeled somatodendritic profiles were measured by quantitative immunoelectron microscopy in control or stressed rats. In resting conditions, NK3Rs were predominantly located in AVP neurons, however sparsely distributed in OC neurons of the PVN. All NK3-labeled somata of the PVN in control rats showed cytoplasmic but no nuclear immunolabeling. An acute restraint stress session of 30 min significantly increased nuclear NK3R density in AVP-labeled somata but not in OC-labeled somata. These changes were accompanied by a respective decrease and increase in plasmalemmal and cytoplamic NK3R densities in AVP-labeled but not in OC-labeled dendrites. The results of this study suggest that in the rat PVN 1) NK3R distribution is conducive to modulation of systemic and/or central AVP release through PVN inputs to the posterior pituitary and/or the amygdala and 2) acute restraint stress activates (internalizes) NK3Rs on surface and evokes nuclear NK3R translocation exclusively in AVP neurons. This trafficking might contribute to neurochemical imbalances observed in neuronal circuits involved in stress-related disorders such as anxiety.
Assuntos
Núcleo Celular/metabolismo , Células Dendríticas , Núcleo Hipotalâmico Paraventricular , Transporte Proteico , Receptores da Neurocinina-3/metabolismo , Estresse Psicológico/fisiopatologia , Vasopressinas/metabolismo , Animais , Masculino , Neurônios/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
CONTEXT: Neuronal dysfunction in cerebral cortex and other brain regions could contribute to the cognitive and behavioral defects in autism. OBJECTIVE: To characterize epigenetic signatures of autism in prefrontal cortex neurons. DESIGN: We performed fluorescence-activated sorting and separation of neuronal and nonneuronal nuclei from postmortem prefrontal cortex, digested the chromatin with micrococcal nuclease, and deeply sequenced the DNA from the mononucleosomes with trimethylated H3K4 (H3K4me3), a histone mark associated with transcriptional regulation. Approximately 15 billion base pairs of H3K4me3-enriched sequences were collected from 32 brains. SETTING: Academic medical center. PARTICIPANTS: A total of 16 subjects diagnosed as having autism and 16 control subjects ranging in age from 0.5 to 70 years. MAIN OUTCOME MEASURES: Identification of genomic loci showing autism-associated H3K4me3 changes in prefrontal cortex neurons. RESULTS: Subjects with autism showed no evidence for generalized disruption of the developmentally regulated remodeling of the H3K4me3 landscape that defines normal prefrontal cortex neurons in early infancy. However, excess spreading of H3K4me3 from the transcription start sites into downstream gene bodies and upstream promoters was observed specifically in neuronal chromatin from 4 of 16 autism cases but not in controls. Variable subsets of autism cases exhibit altered H3K4me3 peaks at numerous genes regulating neuronal connectivity, social behaviors, and cognition, often in conjunction with altered expression of the corresponding transcripts. Autism-associated H3K4me3 peaks were significantly enriched in genes and loci implicated in neurodevelopmental diseases. CONCLUSIONS: Prefrontal cortex neurons from subjects with autism show changes in chromatin structures at hundreds of loci genome-wide, revealing considerable overlap between genetic and epigenetic risk maps of developmental brain disorders.
Assuntos
Transtorno Autístico/genética , Metilação de DNA/genética , Epigênese Genética/genética , Histonas/metabolismo , Córtex Pré-Frontal/metabolismo , Adolescente , Adulto , Idoso , Transtorno Autístico/etiologia , Transtorno Autístico/metabolismo , Estudos de Casos e Controles , Criança , Pré-Escolar , Cromatina/genética , Cromatina/metabolismo , Impressões Digitais de DNA , Feminino , Citometria de Fluxo , Histonas/genética , Humanos , Masculino , Pessoa de Meia-Idade , Modelos Genéticos , Neurônios/metabolismo , Córtex Pré-Frontal/citologia , Fatores de Risco , Adulto JovemRESUMO
While genome-wide association studies are ongoing to identify sequence variation influencing susceptibility to major depressive disorder (MDD), epigenetic marks, such as DNA methylation, which can be influenced by environment, might also play a role. Here we present the first genome-wide DNA methylation (DNAm) scan in MDD. We compared 39 postmortem frontal cortex MDD samples to 26 controls. DNA was hybridized to our Comprehensive High-throughput Arrays for Relative Methylation (CHARM) platform, covering 3.5 million CpGs. CHARM identified 224 candidate regions with DNAm differences >10%. These regions are highly enriched for neuronal growth and development genes. Ten of 17 regions for which validation was attempted showed true DNAm differences; the greatest were in PRIMA1, with 12-15% increased DNAm in MDD (p = 0.0002-0.0003), and a concomitant decrease in gene expression. These results must be considered pilot data, however, as we could only test replication in a small number of additional brain samples (n = 16), which showed no significant difference in PRIMA1. Because PRIMA1 anchors acetylcholinesterase in neuronal membranes, decreased expression could result in decreased enzyme function and increased cholinergic transmission, consistent with a role in MDD. We observed decreased immunoreactivity for acetylcholinesterase in MDD brain with increased PRIMA1 DNAm, non-significant at p = 0.08.While we cannot draw firm conclusions about PRIMA1 DNAm in MDD, the involvement of neuronal development genes across the set showing differential methylation suggests a role for epigenetics in the illness. Further studies using limbic system brain regions might shed additional light on this role.
Assuntos
Metilação de DNA , Transtorno Depressivo Maior/genética , Encéfalo/metabolismo , Ilhas de CpG , Epigênese Genética , Estudos de Associação Genética , Humanos , Proteínas de Membrana/química , Proteínas de Membrana/genética , Proteínas do Tecido Nervoso/química , Proteínas do Tecido Nervoso/genéticaRESUMO
Chronic intermittent hypoxia (CIH) is a frequent concomitant of sleep apnea, which can increase sympathetic nerve activity through mechanisms involving chemoreceptor inputs to the commissural nucleus of the solitary tract (cNTS). These chemosensory inputs co-store glutamate and substance P (SP), an endogenous ligand for neurokinin-1 (NK(1)) receptors. Acute hypoxia results in internalization of NK(1) receptors, suggesting that CIH also may affect the subcellular distribution of NK(1) receptors in subpopulations of cNTS neurons, some of which may express tyrosine hydroxylase, the rate-limiting enzyme for catecholamine synthesis (TH). To test this hypothesis, we examined dual immunolabeling for the NK(1) receptor and TH in the cNTS of male mice subjected to 10days or 35days of CIH or intermittent air. Electron microscopy revealed that NK(1) receptors and TH were almost exclusively localized within separate somatodendritic profiles in cNTS of control mice. In dendrites, immunogold particles identifying NK(1) receptors were prevalent in the cytoplasm and on the plasmalemmal surface. Compared with controls, CIH produced a significant region-specific decrease in the cytoplasmic (10 and 35days, P<0.05, unpaired Student t-test) and extrasynaptic plasmalemmal (35days, P<0.01, unpaired Student t-test) density of NK(1) immunogold particles exclusively in small (<0.1microm) dendrites without TH immunoreactivity. These results suggest that CIH produces a duration-dependent reduction in the availability of NK(1) receptors preferentially in small dendrites of non-catecholaminergic neurons in the cNTS. The implications of our findings are discussed with respect to their potential involvement in the slowly developing hypertension seen in sleep apnea patients.
Assuntos
Dendritos/metabolismo , Hipóxia Encefálica/metabolismo , Receptores da Neurocinina-1/metabolismo , Síndromes da Apneia do Sono/metabolismo , Núcleo Solitário/metabolismo , Animais , Catecolaminas/metabolismo , Doença Crônica , Dendritos/patologia , Dendritos/ultraestrutura , Imunofluorescência , Hipóxia Encefálica/patologia , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microscopia Imunoeletrônica , Plasticidade Neuronal/fisiologia , Neurônios/metabolismo , Neurônios/patologia , Neurônios/ultraestrutura , Reflexo/fisiologia , Mecânica Respiratória/fisiologia , Síndromes da Apneia do Sono/patologia , Núcleo Solitário/patologia , Substância P/metabolismo , Sistema Nervoso Simpático/metabolismo , Sistema Nervoso Simpático/patologia , Núcleo Espinal do Trigêmeo/metabolismo , Núcleo Espinal do Trigêmeo/patologia , Tirosina 3-Mono-Oxigenase/metabolismoRESUMO
Estrogen receptor (ER) activation in central autonomic nuclei modulates arterial blood pressure (ABP) and counteracts the deleterious effect of hypertension. We tested the hypothesis that hypertension, in turn, influences the expression and trafficking of gonadal steroid receptors in central cardiovascular circuits. Thus, we examined whether ER- and progestin receptor (PR)-immunoreactivity (ir) are altered in medullary and hypothalamic autonomic areas of cycling rats following chronic infusion of the hypertensive agent, angiotensin II (AngII). After 1 week AngII-infusion, systolic ABP was elevated from 103+/-4 to 172+/-8 mmHg (p<0.05; N=8/group) and all rats were in diestrus (low estrogen). In AngII-infused rats the number of PR-immunoreactive nuclei was reduced (-72%) compared to saline-infused controls also in diestrus (p<0.05). Furthermore, the intensity of ERalpha-ir increased selectively in nuclei (16%) and cytoplasm (21%) of cells in the commissural nucleus of the solitary tract (cNTS; p<0.05) while neither the number nor intensity of ERbeta-labeled cells changed (p>0.05). Following chronic AngII-infusion, electron microscopy showed a higher cytoplasmic-to-nuclear ratio of ERalpha-labeling selectively in tyrosine hydroxylase (TH)-labeled neurons in the cNTS. Furthermore, AngII-infusion increased ERalpha-ir in the cytosol of TH- and non-TH neuronal perikarya and increased the amount of ERalpha-ir associated with endoplasmic reticulum only in TH-containing perikarya. The data suggest that hypertension modulates the expression and subcellular distribution of ERalpha and PR in central autonomic regions involved in blood pressure control. Considering that ERalpha counteracts the central and peripheral effects of AngII, these receptor changes may underlie adaptive responses that protect females from the deleterious effects of hypertension.