RESUMEN
n-3 Polyunsaturated fatty acids (n-3PUFAs) may be beneficial for anxiety and depression under stressful conditions. Studies however, typically utilise physical or sudden physiological stress, while gradual physiological stress is also relevant to human conditions. Using deoxycorticosterone acetate (DOCA) administration to induce gradual physiological stress, this study investigated the impact of n-3PUFAs under gradual physiological stress in rats. Animals (aged 2 months) (N=8-12/group) received daily injections of DOCA or vehicle and were concurrently fed a high n-3PUFA or control diet for eight weeks. Behavioural measures were taken throughout. Behavioural tests and physiological measures were conducted after six and eight weeks respectively. DOCA administration decreased plasma renin, plasma proteins and relative adrenal weight, and increased water intake, relative kidney weight, and anxiety in the open field. These findings demonstrate disruptions to the renin-angiotensin-aldosterone system, a result of mild physiological stress, that also impact on anxiety behaviours. No effects of n-3PUFAs were found.
Asunto(s)
Ansiedad/inducido químicamente , Depresión/inducido químicamente , Acetato de Desoxicorticosterona/administración & dosificación , Acetato de Desoxicorticosterona/efectos adversos , Ácidos Grasos Omega-3/administración & dosificación , Animales , Ansiedad/sangre , Peso Corporal/efectos de los fármacos , Depresión/sangre , Acetato de Desoxicorticosterona/farmacología , Dieta Alta en Grasa , Ingestión de Líquidos/efectos de los fármacos , Ingestión de Alimentos/efectos de los fármacos , Humanos , Masculino , Ratas , Ratas Sprague-Dawley , Renina/sangre , Estrés Fisiológico/efectos de los fármacosRESUMEN
The present study investigated the role of several 5-HT receptor subtypes in the lateral parabrachial nucleus (LPBN) in the control of sodium appetite (i.e. NaCl consumption). Male Holtzman rats had cannulas implanted bilaterally into the LPBN for the injection of 5-HT receptor agonists and antagonists in conjunction with either acute fluid depletion or 24-h sodium depletion. Following these treatments, access to 0.3 M NaCl was provided and the intakes of saline and water were measured for the next 2 h. Bilateral injections of the 5-HT2A receptor antagonist, ketanserin or the 5-HT2C receptor antagonist, mianserin into the LPBN increased 0.3 M NaCl intake without affecting water intake induced by acute fluid-depletion. Bilateral injections of the 5-HT2B receptor agonist, BW723C86 hydrochloride, had no effect on 0.3 M NaCl or water intake under these conditions. Treatment of the LPBN with the 5-HT2B/2C receptor agonist, 2-(2-methyl-4-clorophenoxy) propanoic acid (mCPP) caused dose-related reductions in 0.3 M NaCl intake after 24 h sodium depletion. The effects of mCPP were prevented by pretreating the LPBN with the 5-HT2B/2C receptor antagonist, SDZSER082. Activation of 5-HT3 receptors by the receptor agonist, 1-phenylbiguanide (PBG) caused dose-related increases in 0.3 M NaCl intake. Pretreatment of the LPBN with the 5-HT3 receptor antagonist, 1-methyl-N-[8-methyl-8-azabicyclo (3.2.1)-oct-3-yl]-1H-indazole-3-carboxamide (LY-278,584) abolished the effects of PBG, but LY-278,584 had no effects on sodium or water intake when injected by itself. PBG injected into the LPBN did not alter intake of palatable 0.06 M sucrose in fluid replete rats. The results suggest that activation of the 5-HT2A and 5-HT2C receptor subtypes inhibits sodium ingestion. In contrast, activation of the 5-HT3 receptor subtype increases sodium ingestion. Therefore, multiple serotonergic receptor subtypes in the LPBN are implicated in the control of sodium intake, sometimes by mediating opposite effects of 5-HT. The results provide new information concerning the control of sodium intake by LPBN mechanisms.
Asunto(s)
Conducta de Ingestión de Líquido/fisiología , Puente/metabolismo , Receptores de Serotonina 5-HT2/fisiología , Receptores de Serotonina 5-HT3/fisiología , Cloruro de Sodio/metabolismo , Animales , Conducta Animal , Relación Dosis-Respuesta a Droga , Ingestión de Líquidos/efectos de los fármacos , Conducta de Ingestión de Líquido/efectos de los fármacos , Interacciones Farmacológicas , Ketanserina/farmacología , Masculino , Modelos Biológicos , Puente/efectos de los fármacos , Ratas , Antagonistas de la Serotonina/farmacología , Agonistas de Receptores de Serotonina/farmacologíaRESUMEN
The mineralocorticoid (MC) receptor antagonist spironolactone (SL) improves morbidity and mortality in patients with congestive heart failure (CHF). We tested the hypothesis that the central nervous system actions of SL contribute to its beneficial effects. SL (100 ng/h for 28 days) or ethanol vehicle (VEH) was administered intracerebroventricularly or intraperitoneally to rats with CHF induced by coronary artery ligation (CL) and to SHAM-operated controls. The intracerebroventricular SL treatment prevented the increase in sodium appetite and the decreases in sodium and water excretion observed within a week of CL in VEH-treated CHF rats. Intraperitoneal SL also improved volume regulation in the CHF rats, but only after 3 wk of treatment. Four weeks of SL treatment, either intracerebroventricularly or intraperitoneally, ameliorated both the increase in sympathetic drive and the impaired baroreflex function observed in VEH-treated CHF rats. These findings suggest that activation of MC receptors in the central nervous system plays a critical role in the altered volume regulation and augmented sympathetic drive that characterize clinical heart failure.
Asunto(s)
Insuficiencia Cardíaca/tratamiento farmacológico , Insuficiencia Cardíaca/fisiopatología , Antagonistas de Receptores de Mineralocorticoides/farmacología , Espironolactona/farmacología , Sistema Nervioso Simpático/fisiología , Animales , Barorreflejo/fisiología , Presión Sanguínea/fisiología , Ingestión de Líquidos/fisiología , Corazón/inervación , Insuficiencia Cardíaca/mortalidad , Masculino , Infarto del Miocardio/tratamiento farmacológico , Infarto del Miocardio/mortalidad , Infarto del Miocardio/fisiopatología , Miocardio/patología , Tamaño de los Órganos , Ratas , Ratas Sprague-Dawley , Sistema Renina-Angiotensina/fisiología , Sodio en la Dieta/farmacología , Sodio en la Dieta/orina , Tasa de Supervivencia , Función Ventricular IzquierdaRESUMEN
This study examined whether cardiac beta-adrenoceptor (beta-AR) function was altered in conscious rats with lesions surrounding the anteroventral third ventricle (AV3V). The findings were: (1) beta(1,2)-AR-mediated tachycardia was similar in sham and AV3V-lesion rats, (2) beta(3)- and/or atypical beta-AR-mediated tachycardia elicited by isoproterenol (10 microg/kg, i.v.; ISO) was diminished in AV3V-lesion rats treated with beta(1,2)-AR antagonists, but was not in similarly-treated sham-lesion rats, and (3) the tachycardia elicited by the membrane permeable cAMP-analogue, 8-(4-chlorophenylthiol)-cAMP (10 micromol/kg, i.v.), was similar in AV3V- and sham-lesion rats. The possibility that increased plasma sodium/osmolality in AV3V-lesion rats down-regulated cardiac beta(3)- and/or atypical beta-ARs, but not beta(1,2)-ARs or intracellular cAMP signaling is discussed.
Asunto(s)
Células Quimiorreceptoras/metabolismo , AMP Cíclico/análogos & derivados , Corazón/inervación , Presorreceptores/metabolismo , Receptores Adrenérgicos beta/metabolismo , Taquicardia/metabolismo , Tercer Ventrículo/metabolismo , Equilibrio Hidroelectrolítico/fisiología , Agonistas Adrenérgicos beta/farmacología , Antagonistas Adrenérgicos beta/farmacología , Animales , Presión Sanguínea/efectos de los fármacos , Presión Sanguínea/fisiología , Células Quimiorreceptoras/efectos de los fármacos , AMP Cíclico/farmacología , Desnervación , Inhibidores Enzimáticos/farmacología , Corazón/efectos de los fármacos , Frecuencia Cardíaca/efectos de los fármacos , Frecuencia Cardíaca/fisiología , Isoproterenol/farmacología , Masculino , Donantes de Óxido Nítrico/farmacología , Presorreceptores/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Receptores Adrenérgicos beta/efectos de los fármacos , Fibras Simpáticas Posganglionares/efectos de los fármacos , Fibras Simpáticas Posganglionares/metabolismo , Taquicardia/inducido químicamente , Taquicardia/fisiopatología , Tionucleótidos/farmacología , Tercer Ventrículo/efectos de los fármacos , Equilibrio Hidroelectrolítico/efectos de los fármacosRESUMEN
In addition to the circulatory renin (REN)-angiotensin system (RAS), a tissue RAS having an important role in cardiovascular function also exists in the central nervous system. In the brain, angiotensinogen (AGT) is expressed in astrocytes and in some neurons important to cardiovascular control, but its functional role remains undefined. We generated a transgenic mouse encoding the human AGT (hAGT) gene under the control of the human glial fibrillary acidic protein (GFAP) promoter to experimentally dissect the role of brain versus systemically derived AGT. This promoter targets expression of transgene products to astrocytes, the most abundant cell type expressing AGT in brain. All transgenic lines exhibited hAGT mRNA expression in brain, with variable expression in other tissues. In one line examined in detail, transgene expression was high in brain and low in tissues outside the central nervous system, and the level of plasma hAGT was not elevated over baseline. In the brain, hAGT protein was mainly localized in astrocytes, but was present in neurons in the subfornical organ. Intracerebroventricular (ICV) injection of human REN (hREN) in conscious unrestrained mice elicited a pressor response, which was abolished by ICV preinjection of losartan. Double-transgenic mice expressing the hREN gene and the GFAP-hAGT transgene exhibited a 15-mm Hg increase in blood pressure and an increased preference for salt. Blood pressure in the hREN/GFAP-hAGT mice was lowered after ICV, but not intravenous losartan. These studies suggest that AGT synthesis in the brain has an important role in the regulation of blood pressure and electrolyte balance.
Asunto(s)
Angiotensinógeno/biosíntesis , Encéfalo/metabolismo , Proteína Ácida Fibrilar de la Glía/genética , Hipertensión/genética , Regiones Promotoras Genéticas , Angiotensinógeno/sangre , Angiotensinógeno/genética , Animales , Astrocitos/metabolismo , Presión Sanguínea/efectos de los fármacos , Presión Sanguínea/genética , Relación Dosis-Respuesta a Droga , Ingestión de Líquidos/efectos de los fármacos , Ingestión de Líquidos/genética , Expresión Génica , Humanos , Hipertensión/fisiopatología , Inyecciones Intravenosas , Inyecciones Intraventriculares , Losartán/administración & dosificación , Ratones , Ratones Transgénicos , Neuronas/metabolismo , Especificidad de Órganos/genética , ARN Mensajero/biosíntesis , Renina/administración & dosificación , Renina/antagonistas & inhibidores , Renina/genética , Órgano Subfornical/metabolismo , Transgenes , Equilibrio Hidroelectrolítico/genéticaRESUMEN
The subfornical organ (SFO) is sensitive to both ANG II and ACh, and local application of these agents produces dipsogenic responses and vasopressin release. The present study examined the effects of cholinergic drugs, ANG II, and increased extracellular osmolarity on dissociated, cultured cells of the SFO that were retrogradely labeled from the supraoptic nucleus. The effects were measured as changes in cytosolic calcium in fura 2-loaded cells by using a calcium imaging system. Both ACh and carbachol increased intracellular ionic calcium concentration ([Ca2+]i). However, in contrast to the effects of muscarinic receptor agonists on SFO neurons, manipulation of the extracellular osmolality produced no effects, and application of ANG II produced only moderate effects on [Ca2+]i in a few retrogradely labeled cells. The cholinergic effects on [Ca2+]i could be blocked with the muscarinic receptor antagonist atropine and with the more selective muscarinic receptor antagonists pirenzepine and 4-diphenylacetoxy-N-methylpiperdine methiodide (4-DAMP). In addition, the calcium in the extracellular fluid was required for the cholinergic-induced increase in [Ca2+]i. These findings indicate that ACh acts to induce a functional cellular response in SFO neurons through action on a muscarinic receptor, probably of the M1 subtype and that the increase of [Ca2+]i, at least initially, requires the entry of extracellular Ca2+. Also, consistent with a functional role of M1 receptors in the SFO are the results of immunohistochemical preparations demonstrating M1 muscarinic receptor-like protein present within this forebrain circumventricular organ.
Asunto(s)
Acetilcolina/farmacología , Angiotensina II/farmacología , Calcio/metabolismo , Neuronas/fisiología , Órgano Subfornical/fisiología , Núcleo Supraóptico/fisiología , Animales , Transporte Axonal , Carbacol/farmacología , Citosol/metabolismo , Femenino , Colorantes Fluorescentes , Fura-2/análogos & derivados , Técnicas In Vitro , Cinética , Vías Nerviosas/efectos de los fármacos , Vías Nerviosas/fisiología , Neuronas/efectos de los fármacos , Pirenzepina/farmacología , Ratas , Ratas Sprague-Dawley , Receptor Muscarínico M1 , Receptores Muscarínicos/efectos de los fármacos , Receptores Muscarínicos/fisiología , Órgano Subfornical/efectos de los fármacos , Núcleo Supraóptico/efectos de los fármacosRESUMEN
The objective of the present study was to define the optimum conditions for using replication-defective adenovirus (Ad) to transfer the gene for the green fluorescent protein (GFP) to the hypothalamic paraventricular (PVN) and supraoptic (SON) nuclei and cells of the neurohypophysis (NH). As indicated by characterizing cell survival over 15 days in culture and in electrophysiological whole cell patch-clamp studies, viral concentrations up to 2 x 10(7) pfu/coverslip did not affect viability of transfected PVN and NH cultured cells from preweanling rats. At 2 x 10(7) pfu, GFP gene expression was higher (40% of GFP-positive cells) and more sustained (up to 15 days). Using a stereotaxic approach in adult rats, we were able to directly transduce the PVN, SON, and NH and visualize gene expression in coronal brain slices and in the pituitary 4 days after injection of Ad. In animals receiving NH injections of Ad, the virus was retrogradely transported to PVN and SON neurons as indicated by the appearance of GFP-positive neurons in cultures of dissociated cells from those brain nuclei and by polymerase chain reaction and Western blot analyses of PVN and SON tissues. Adenoviral concentrations of up to 8 x 10(6) pfu injected into the NH did not affect cell viability and did not cause inflammatory responses. Adenoviral injection into the pituitary enabled the selective delivery of genes to the soma of magnocellular neurons. The experimental approaches described here provide potentially useful strategies for the treatment of disordered expression of the hormones vasopressin or oxytocin.
Asunto(s)
Adenoviridae/metabolismo , Técnicas de Transferencia de Gen , Sistema Hipotálamo-Hipofisario/metabolismo , Sistema Hipotálamo-Hipofisario/cirugía , Adenoviridae/genética , Animales , Células Cultivadas , Femenino , Expresión Génica , Proteínas Fluorescentes Verdes , Sistema Hipotálamo-Hipofisario/citología , Proteínas Luminiscentes/biosíntesis , Proteínas Luminiscentes/genética , Masculino , Núcleo Hipotalámico Paraventricular/citología , Núcleo Hipotalámico Paraventricular/metabolismo , Núcleo Hipotalámico Paraventricular/virología , Técnicas de Placa-Clamp , Neurohipófisis/citología , Neurohipófisis/metabolismo , Neurohipófisis/cirugía , Neurohipófisis/virología , Ratas , Ratas Sprague-Dawley , Núcleo Supraóptico/citología , Núcleo Supraóptico/metabolismo , Núcleo Supraóptico/virología , TransfecciónRESUMEN
Vasopressin is synthesized by magnocellular neurons in supraoptic (SON) and paraventricular (PVN) hypothalamic nuclei and released by their axon terminals in the neurohypophysis (NH). With its actions as an antidiuretic hormone and vasoactive agent, vasopressin plays a pivotal role in the control of body fluids and cardiovascular homeostasis. Because of its well-defined neurobiology and functional importance, the SON/PVN-NH system is ideal to establish methods for gene transfer of genetic material into specific pathways in the mouse central nervous system. In these studies, we compared the efficiency of transferring the gene lacZ, encoding for beta-galactosidase (beta-gal), versus a gene encoding for green fluorescent protein by using replication-deficient adenovirus (Ad) vectors in adult mice. Transfection with viral concentrations up to 2 x 10(7) plaque-forming units per coverslip of NH, PVN, and SON in dissociated, cultured cells caused efficient transfection without cytotoxicity. However, over an extended period of time, higher levels (50% to 75% of the cells) of beta-gal expression were detected in comparison with green fluorescent protein (5% to 50% of the cells). With the use of a stereotaxic approach, the pituitary glands of mice were injected with Ad (4 x 10(6) plaque-forming units). In material from these animals, we were able to visualize the expression of the beta-gal gene in the NH and in magnocellular neurons of both the PVN and SON. The results of these experiments indicate that Ad-Rous sarcoma virus promoter-beta-gal is taken up by nerve terminals at the injection site (NH) and retrogradely transported to the soma of the neurons projecting to the NH. We conclude that the application of these experimental approaches will provide powerful tools for physiological studies and potential approaches to deliver therapeutic genes to treat diseases.
Asunto(s)
Adenoviridae , Técnicas de Transferencia de Gen , Vectores Genéticos , Hipotálamo/fisiología , Animales , Femenino , Proteínas Fluorescentes Verdes , Humanos , Operón Lac , Proteínas Luminiscentes/genética , Masculino , Ratones , Ratones Endogámicos C57BL , beta-Galactosidasa/genéticaRESUMEN
We examined the role of the subfornical organ (SFO) in stimulating thirst and salt appetite using two procedures that initiate water and sodium ingestion within 1-2 h of extracellular fluid depletion. The first procedure used injections of a diuretic (furosemide, 10 mg/kg sc) and a vasodilator (minoxidil, 1-3 mg/kg ia) to produce hypotension concurrently with hypovolemia. The resulting water and sodium intakes were inhibited by intravenous administration of ANG II receptor antagonist (sarthran, 8 micrograms . kg(-1). min(-1)) or angiotensin-converting enzyme inhibitor (captopril, 2.5 mg/h). The second procedure used injections of furosemide (10 mg/kg sc) and a low dose of captopril (5 mg/kg sc) to initiate water and sodium ingestion upon formation of ANG II in the brain. Electrolytic lesions of the SFO greatly reduced the water intakes, and nearly abolished the sodium intakes, produced by these relatively acute treatments. These results contrast with earlier findings showing little effect of SFO lesions on sodium ingestion after longer-term extracellular fluid depletion.
Asunto(s)
Cloruro de Sodio Dietético , Órgano Subfornical/fisiopatología , Sed/fisiología , Angiotensina II/análogos & derivados , Angiotensina II/farmacología , Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Animales , Volumen Sanguíneo , Captopril/farmacología , Masculino , Ratas , Ratas Sprague-Dawley , Sed/efectos de los fármacos , Vasodilatación/efectos de los fármacos , Equilibrio Hidroelectrolítico/efectos de los fármacosRESUMEN
This study determined baroreceptor reflex (BR) function in conscious rats which had received sham or electrolytic lesions of the anteroventral third ventricle (AV3V) 54-56 days previously. Resting mean arterial pressure (MAP) and heart rate (HR) values of the AV3V-lesion rats were similar to those of sham-lesion rats (P>0.05 for both comparisons). The sensitivity of the BR-mediated tachycardia in AV3V-lesion was greater than in sham-lesion rats (-9. 92+/-1.00 vs. -4.54+/-0.45 bpm/mmHg, P<0.05). The sensitivity of the BR-mediated bradycardia in AV3V-lesion rats was also greater than in rats with sham lesions (-3.56+/-0.38 vs. -2.06+/-0.42 bpm/mmHg, P<0. 05). The AV3V lesions did not affect other BR parameters. These findings demonstrate that chronic lesions of the AV3V region increase the sensitivity of the baroreceptor HR reflex in conscious rats.
Asunto(s)
Ventrículos Cerebrales/fisiología , Presorreceptores/fisiología , Reflejo/fisiología , Animales , Presión Sanguínea/fisiología , Frecuencia Cardíaca/fisiología , Masculino , Ratas , Ratas Sprague-Dawley , Factores de TiempoRESUMEN
This study examined whether electrolytic ablation of the periventricular anteroventral third ventricle (AV3V) would affect the increases in mean arterial blood pressure (MAP) and heart rate (HR) in conscious rats produced by systemic injection of the centrally acting N-methyl-D-aspartate (NMDA) receptor ion-channel blocker, (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5, 10-imine (MK-801; 250 microgram/kg, i.v.). MK-801 produced a smaller increase in MAP in rats with AV3V lesions than in sham-lesion rats (+36+/-2% vs. +52+/-5%, respectively, P<0.05). In contrast, MK-801 produced similar increases in HR in the AV3V- and sham-lesion rats (+28+/-3% vs. +22+/-4%, respectively, P>0.05). These findings demonstrate that the MK-801-induced pressor response is dependent upon the integrity of the AV3V region, whereas the MK-801-induced tachycardia is not.
Asunto(s)
Presión Sanguínea/efectos de los fármacos , Ventrículos Cerebrales/fisiología , Maleato de Dizocilpina/farmacología , Antagonistas de Aminoácidos Excitadores/farmacología , Receptores de N-Metil-D-Aspartato/antagonistas & inhibidores , Animales , Inyecciones Intraarteriales , Inyecciones Intravenosas , Masculino , Ratas , Ratas Sprague-DawleyRESUMEN
Lesion of the anteroventral third-ventricle region (AV3VX) reduced saline consumption. Salt loading in AV3VX rats resulted in reduced but not completely abolished c-fos expression in the supraoptic and paraventricular nuclei. Intrinsic osmosensitivity of the magnocellular neurons, or input from other brain areas, such as the subfornical and median preoptic nuclei, may account for this residual c-fos expression. These regions showed c-fos expression following salt loading.
Asunto(s)
Ventrículos Cerebrales/fisiología , Proteínas Proto-Oncogénicas c-fos/biosíntesis , Cloruro de Sodio/farmacología , Análisis de Varianza , Animales , Ventrículos Cerebrales/citología , Masculino , Neuronas/efectos de los fármacos , Presión Osmótica , Ratas , Ratas Sprague-DawleyRESUMEN
This study examined whether a prior electrolytic lesion of the tissue surrounding the anteroventral third ventricle (AV3V) would affect the increase in mean arterial blood pressure (MAP) and the fall in heart rate (HR) produced by systemic injection of the nitric oxide synthesis (NOS) inhibitor, NG-nitro-L-arginine methyl ester (L-NAME; 25 micromol/kg, i.v.) in conscious rats. L-NAME produced a smaller increase in MAP in AV3V-lesion than in sham-lesion rats (+19+/-3 vs. +40+/-3 mmHg, respectively; P<0.05). In contrast, L-NAME produced similar falls in HR in the AV3V-lesion and sham-lesion rats (-103+/-15 vs. -97+/-8 bpm, respectively; P<0.05). These findings demonstrate that the L-NAME-induced pressor response is dependent upon the integrity of the AV3V region, whereas the L-NAME-induced bradycardia is not.
Asunto(s)
Sistema Cardiovascular/efectos de los fármacos , Ventrículos Cerebrales/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , NG-Nitroarginina Metil Éster/farmacología , Óxido Nítrico Sintasa/antagonistas & inhibidores , Animales , Inyecciones Intravenosas , Masculino , Ratas , Ratas Sprague-DawleyRESUMEN
Systemic injections of the excitatory amino acid (EAA) analogs, kainic acid (KA) and N-methyl-D-aspartate (NMDA), produce a pressor response in conscious rats that is caused by a centrally mediated activation of sympathetic drive and the release of arginine vasopressin (AVP). This study tested the hypothesis that the tissue surrounding the anteroventral part of the third ventricle (AV3V) plays a role in the expression of the pressor responses produced by systemically injected EAA analogs. Specifically, we examined whether prior electrolytic ablation of the AV3V region would affect the pressor responses to KA and NMDA (1 mg/kg iv) in conscious rats. The KA-induced pressor response was smaller in AV3V-lesioned than in sham-lesioned rats (11 +/- 2 vs. 29 +/- 2 mmHg; P < 0.05). After ganglion blockade, KA produced a pressor response in sham-lesioned but not AV3V-lesioned rats (+27 +/- 3 vs. +1 +/- 2 mmHg; P < 0.05). The KA-induced pressor response in ganglion-blocked sham-lesioned rats was abolished by a vasopressin V1-receptor antagonist. Similar results were obtained with NMDA. The pressor response to AVP (10 ng/kg iv) was slightly smaller in AV3V-lesioned than in sham-lesioned ganglion-blocked rats (45 +/- 3 vs. 57 +/- 4 mmHg; P < 0.05). This study demonstrates that the pressor responses to systemically injected EAA analogs are smaller in AV3V-lesioned rats. The EAA analogs may produce pressor responses by stimulation of EAA receptors in the AV3V region, or the AV3V region may play an important role in the expression of these responses.
Asunto(s)
Presión Sanguínea/efectos de los fármacos , Ventrículos Cerebrales/fisiología , Agonistas de Aminoácidos Excitadores/farmacología , Ácido Kaínico/farmacología , N-Metilaspartato/farmacología , Circulación Renal/fisiología , Animales , Antihipertensivos/farmacología , Arginina Vasopresina/análogos & derivados , Arginina Vasopresina/farmacología , Peso Corporal , Bradicardia/inducido químicamente , Bradicardia/fisiopatología , Clorisondamina/farmacología , Estado de Conciencia , Agonistas de Aminoácidos Excitadores/sangre , Antagonistas de Hormonas/farmacología , Ácido Kaínico/sangre , Locomoción/fisiología , Masculino , N-Metilaspartato/sangre , Ratas , Ratas Sprague-Dawley , Taquicardia/inducido químicamente , Taquicardia/fisiopatología , Vasoconstrictores/farmacologíaRESUMEN
The subfornical organ (SFO) is a forebrain structure that converts peripheral blood-borne signals reflecting the hydrational state of the body to neural signals and then through efferent fibers conveys this information to several central nervous system structures. One of the forebrain areas receiving input from the SFO is the supraoptic nucleus (SON), a source of vasopressin synthesis and control of release from the posterior pituitary. Little is known of the transduction and transmission processes by which this conversion of systemic information to brain input occurs. As a step in elucidating these mechanisms, the present study characterized the ionic currents of dissociated cells of the SFO that were identified as neurons that send efferents to the SON. A retrograde tracer was injected into the SON area in eleven-day-old rats. After three days for retrograde transport of the label, the SFOs of these animals were dissociated and plated for tissue culture. The retrograde tracer was used to identify the soma of SFO cells projecting to the SON so that voltage-dependent ionic currents using whole-cell voltage clamp methods could be studied. The three types of currents in labeled SFO neurons were characterized as a 1) rapid, transient inward current that can be blocked by tetrodotoxin (TTX) characteristic of a sodium current; 2) slow-onset sustained outward current that can be blocked by tetraethylammonium (TEA) characteristic of a delayed rectifier potassium current; and 3) remaining outward current that has a rapid-onset and transient characteristic of a potassium A-type current.
Asunto(s)
Canales Iónicos/fisiología , Neuronas/fisiología , Órgano Subfornical/fisiología , Núcleo Supraóptico/fisiología , Vías Aferentes/fisiología , Animales , Células Cultivadas , Canales Iónicos/efectos de los fármacos , Técnicas de Placa-Clamp , Bloqueadores de los Canales de Potasio , Ratas , Ratas Sprague-Dawley , Bloqueadores de los Canales de Sodio , Órgano Subfornical/citología , Tetraetilamonio/farmacología , Tetrodotoxina/farmacologíaRESUMEN
Bombesin (BN) injected to sodium depleted rats either centrally, particularly into the paraventricular nucleus (PVN) or peripherally by intraperitoneal (IP) route, exerts a potent inhibitory effect on the intake of 2% sodium chloride. To determine whether the area postrema (AP) and the lateral parabrachial nucleus (LPBN), which are known to be involved in the control of ingestive behavior, could be sites for the antinatiorexic activity of BN, we studied the effects of injections of this peptide into the LPBN or, by IP or fourth ventricular route, to surgically AP-lesioned rats. We observed that in sodium depleted rats: 1) injected into the LPBN at a dose of 50, but not of 25, ng per nucleus, BN significantly reduced the intake of 2% sodium chloride; 2) administered either intraperitoneally or into the fourth brain ventricle, BN induced a potent antinatriorexic effect in AP-sham lesioned animals; and 3) in the same experimental conditions, surgical ablation of the AP did not reduce the antinatriorexic effect of the peptide. These data indicate that the LPBN may be, with the PVN, a site for the BN-ergic inhibitory control of salt appetite and that the AP, which has been implicated in BN-induced anorexia, does not play a role in the antinatriorexic effect of this peptide.
Asunto(s)
Bombesina/farmacología , Conducta de Ingestión de Líquido/efectos de los fármacos , Natriuresis/efectos de los fármacos , Núcleo Hipotalámico Paraventricular/efectos de los fármacos , Cloruro de Sodio Dietético/antagonistas & inhibidores , Animales , Dieta Hiposódica , Inyecciones Intraventriculares , Masculino , Ratas , Ratas Wistar , Cloruro de Sodio Dietético/metabolismoRESUMEN
We have previously shown that mice transgenic for both the human renin and human angiotensinogen genes (RA+) exhibit appropriate tissue- and cell-specific expression of both transgenes, have 4-fold higher plasma angiotensin II (AII) levels, and are chronically hypertensive. However, the relative contribution of circulating and tissue-derived AII in causing hypertension in these animals is not known. We hypothesized that the brain renin-angiotensin system contributes to the elevated blood pressure in this model. To address this hypothesis, mean arterial pressure (MAP) and heart rate were measured in conscious, unrestrained mice after they were instrumented with intracerebroventricular cannulae and carotid arterial and jugular vein catheters. Intracerebroventricular administration of the selective AII type 1 (AT-1) receptor antagonist losartan (10 microgram, 1 microL) caused a significantly greater peak fall in MAP in RA+ mice than in nontransgenic RA- controls (-29+/-4 versus -4+/-2 mm Hg, P<0.01). To explore the mechanism of a central renin-angiotensin system-dependent hypertension in RA+ mice, we determined the relative depressor responses to intravenous administration of the ganglionic blocking agent hexamethonium (5 mg/kg) or an arginine vasopressin (AVP) V1 receptor antagonist (AVPX, 10 microgram/kg). Hexamethonium caused equal lowering of MAP in RA+ mice and controls (-46+/-3 versus -52+/-3, P>0.05), whereas AVPX caused a significantly greater fall in MAP in RA+ compared with RA- mice (-24+/-2 versus -6+/-1, P<0.01). Consistent with this was the observation that circulating AVP was 3-fold higher in RA+ mice than in control mice. These results suggest that increased activation of central AT-1 receptors, perhaps those located at sites involved in AVP release from the posterior pituitary gland, plays a role in the hypertension in RA+ mice. Furthermore, our finding that both human transgenes are expressed in brain regions of RA+ mice known to be involved in cardiovascular regulation raises the possibility that augmented local production of AII and increased activation of AT-1 receptors at these sites is involved.
Asunto(s)
Angiotensinas/genética , Química Encefálica/fisiología , Hipertensión/metabolismo , Sistema Renina-Angiotensina/fisiología , Renina/genética , Angiotensinas/metabolismo , Animales , Antihipertensivos/farmacología , Arginina Vasopresina/sangre , Arginina Vasopresina/farmacología , Presión Sanguínea/efectos de los fármacos , Presión Sanguínea/fisiología , Tronco Encefálico/química , Tronco Encefálico/metabolismo , Cerebelo/química , Cerebelo/metabolismo , Corteza Cerebral/química , Corteza Cerebral/metabolismo , Diencéfalo/química , Diencéfalo/metabolismo , Bloqueadores Ganglionares/farmacología , Expresión Génica/fisiología , Hexametonio/farmacología , Humanos , Inyecciones Intraventriculares , Losartán/farmacología , Ratones , Ratones Transgénicos , Flujo Pulsátil , Renina/metabolismo , Transgenes/fisiología , Vasoconstrictores/sangre , Vasoconstrictores/farmacologíaRESUMEN
Methysergide injected bilaterally into the lateral parabrachial nucleus (LPBN) increases NaCl intake in several models of renin-dependent salt appetite. The present study investigated the role of angiotensin Type 1 (AT1) receptors in the subfornical organ (SFO) on this effect. The intake of 0.3 M NaCl and water was induced by combined administration of the diuretic, furosemide (FURO), and the angiotensin-converting enzyme inhibitor, captopril (CAP). Pretreatment of the SFO with an AT1 receptor antagonist, losartan (1 microgram/200 nl), reduced water intake but not 0.3 M NaCl intake induced by subcutaneous FURO+CAP. Methysergide (4 microgram/200 nl) injected bilaterally into the LPBN increased 0.3 M NaCl intake after FURO+CAP. Losartan injected into the SFO prevented the additional 0. 3 M NaCl intake caused by LPBN methysergide injections. These results indicate that AT1 receptors located in the SFO may have a role in mediating an enhanced sodium intake produced by methysergide treatment.
Asunto(s)
Angiotensina II/fisiología , Apetito/fisiología , Metisergida/farmacología , Prosencéfalo/fisiología , Rombencéfalo/fisiología , Serotonina/fisiología , Cloruro de Sodio/metabolismo , Animales , Apetito/efectos de los fármacos , Captopril/administración & dosificación , Captopril/farmacología , Cateterismo , Furosemida/administración & dosificación , Furosemida/farmacología , Inyecciones Intraventriculares , Inyecciones Subcutáneas , Losartán/administración & dosificación , Losartán/farmacología , Masculino , Metisergida/administración & dosificación , Prosencéfalo/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Rombencéfalo/efectos de los fármacos , Órgano Subfornical/efectos de los fármacosRESUMEN
The present studies used defined cells of the subfornical organ (SFO) and supraoptic nuclei (SON) as model systems to demonstrate the efficacy of replication-deficient adenovirus (Ad) encoding green fluorescent protein (GFP) for gene transfer. The studies investigated the effects of both direct transfection of the SON and indirect transfection (i.e., via retrograde transport) of SFO neurons. The SON of rats were injected with Ad (2 x 10(6) pfu) and sacrificed 1-7 days later for cell culture of the SON and of the SFO. In the SON, GFP fluorescence was visualized in both neuronal and nonneuronal cells while only neurons in the SFO expressed GFP. Successful in vitro transfection of cultured cells from the SON and SFO was also achieved with Ad (2 x 10(6) to 2 x 10(8) pfu). The expression of GFP in in vitro transfected cells was higher in nonneuronal (approximately 28% in SON and SFO) than neuronal (approximately 4% in SON and 10% in SFO) cells. The expression of GFP was time and viral concentration related. No apparent alterations in cellular morphology of transfected cells were detected and electrophysiological characterization of transfected cells was similar between GFP-expressing and nonexpressing neurons. We conclude that (1) GFP is an effective marker for gene transfer in living SON and SFO cells, (2) Ad infects both neuronal and nonneuronal cells, (3) Ad is taken up by axonal projections from the SON and retrogradely transported to the SFO where it is expressed at detectable levels, and (4) Ad does not adversely affect neuronal viability. These results demonstrate the feasibility of using adenoviral vectors to deliver genes to the SFO-SON axis.
Asunto(s)
Adenoviridae , Técnicas de Transferencia de Gen , Indicadores y Reactivos/metabolismo , Proteínas Luminiscentes/genética , Canales de Potasio con Entrada de Voltaje , Órgano Subfornical/citología , Núcleo Supraóptico/citología , Animales , Western Blotting , Células Cultivadas , Canales de Potasio de Tipo Rectificador Tardío , Femenino , Regulación Viral de la Expresión Génica/fisiología , Genes Reporteros , Proteínas Fluorescentes Verdes , Indicadores y Reactivos/análisis , Proteínas Luminiscentes/análisis , Masculino , Potenciales de la Membrana/fisiología , Neuronas/química , Neuronas/citología , Neuronas/fisiología , Técnicas de Placa-Clamp , Canales de Potasio/fisiología , Ratas , Ratas Sprague-Dawley , Transfección , Replicación ViralRESUMEN
This study investigated the effects of bilateral injections of the local anesthetic, lidocaine, into the lateral parabrachial nucleus (LPBN) on the dipsogenic and pressor responses induced by intracerebroventricular (i.c.v.) injection of angiotensin II (ANG II). Centrally injected ANG II (50 ng/1 microliters) induced water intake (10.2 +/- 0.8 ml/h) and pressor responses (22 +/- 1 mmHg). Prior bilateral injection of 10% lidocaine (200 nl) into the LPBN increased the water intake (14.2 +/- 1.4 ml/h), but did not change the pressor response (17 +/- 1 mmHg) to i.c.v. ANG II. Lidocaine alone injected into the LPBN also induced a pressor response (23 +/- 3 mmHg). These results showing that bilateral LPBN injection of lidocaine increase water intake induced by i.c.v. ANG II are consistent with electrolytic and neurotoxic lesion studies and suggest that the LPBN is associated with inhibitory mechanisms controlling water intake induced by ANG II. These results also provide evidence that it is feasible to reversibly anesthetize this brain area to facilitate fluid-related ingestive behavior.