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1.
Am J Physiol Regul Integr Comp Physiol ; 309(4): R358-67, 2015 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-26062632

RESUMO

Both lateral hypothalamic orexinergic neurons and hindbrain catecholaminergic neurons contribute to control of feeding behavior. Orexin fibers and terminals are present in close proximity to hindbrain catecholaminergic neurons, and fourth ventricular (4V) orexin injections that increase food intake also increase c-Fos expression in hindbrain catecholamine neurons, suggesting that orexin neurons may stimulate feeding by activating catecholamine neurons. Here we examine that hypothesis in more detail. We found that 4V injection of orexin-A (0.5 nmol/rat) produced widespread activation of c-Fos in hindbrain catecholamine cell groups. In the A1 and C1 cell groups in the ventrolateral medulla, where most c-Fos-positive neurons were also dopamine ß hydroxylase (DBH) positive, direct injections of a lower dose (67 pmol/200 nl) of orexin-A also increased food intake in intact rats. Then, with the use of the retrogradely transported immunotoxin, anti-DBH conjugated to saporin (DSAP), which targets and destroys DBH-expressing catecholamine neurons, we examined the hypothesis that catecholamine neurons are required for orexin-induced feeding. Rats given paraventricular hypothalamic injections of DSAP, or unconjugated saporin (SAP) as control, were implanted with 4V or lateral ventricular (LV) cannulas and tested for feeding in response to ventricular injection of orexin-A (0.5 nmol/rat). Both LV and 4V orexin-A stimulated feeding in SAP controls, but DSAP abolished these responses. These results reveal for the first time that catecholamine neurons are required for feeding induced by injection of orexin-A into either LV or 4V.


Assuntos
Catecolaminas/metabolismo , Ventrículos Cerebrais/efeitos dos fármacos , Ingestão de Alimentos/efeitos dos fármacos , Comportamento Alimentar/efeitos dos fármacos , Peptídeos e Proteínas de Sinalização Intracelular/administração & dosagem , Fibras Nervosas/efeitos dos fármacos , Neuropeptídeos/administração & dosagem , Rombencéfalo/efeitos dos fármacos , Animais , Ventrículos Cerebrais/citologia , Ventrículos Cerebrais/imunologia , Ventrículos Cerebrais/metabolismo , Dopamina beta-Hidroxilase/imunologia , Dopamina beta-Hidroxilase/metabolismo , Imunotoxinas/administração & dosagem , Injeções Intraventriculares , Masculino , Fibras Nervosas/imunologia , Fibras Nervosas/metabolismo , Orexinas , Proteínas Proto-Oncogênicas c-fos/metabolismo , Ratos Sprague-Dawley , Rombencéfalo/citologia , Rombencéfalo/imunologia , Rombencéfalo/metabolismo , Proteínas Inativadoras de Ribossomos Tipo 1/administração & dosagem , Saporinas
2.
J Insect Physiol ; 58(4): 506-12, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22172381

RESUMO

The midgut of larval mosquitoes (Aedes aegypti) mediates a cycle of alkali secretion in the anterior segment (AMG) followed by partial reacidification in the posterior segment (PMG); both processes are serotonin-dependent. Here we report that intracellular Ca(2+)(Ca(i)(2+)) as indicated by Fura-2 fluorescence, is elevated in both tissues in response to serotonin, but the time courses differ characteristically in the two gut segments, and Ca(2+)-free solution abolishes the serotonin response in AMG, but not in PMG, whereas Thapsigargin, an inhibitor of endoplasmic Ca(2+) transport, abolished responsiveness to 5-HT in PMG. These results suggest the origins for the Ca(2+) signal differ between the two tissues. Quantitative real-time RT-PCR revealed expression of 5 putative 5-HT receptor types in AMG, including 5-HT(2)-like receptors which would be expected to initiate a Ca(2+) signal. None of these receptors were highly expressed in PMG. Cyclic AMP (cAMP) is a secretagogue for both tissues, but H89, an inhibitor of Protein Kinase A (PKA), is also a secretagogue, suggesting that the stimulatory effect of cAMP involves a non-PKA pathway. Cytochalasins B and D block the effect of 5-HT in AMG, suggesting a vesicle-fusion mechanism of activation of the basal V-ATPase in this tissue. Finally, in PMG, elevation of luminal pH increases (Ca(i)(2+)) and decreases intracellular pH as measured by BCECF fluorescence. These responses suggest that the rate of acid secretion by PMG might be responsive to local demand for luminal reacidification as well as to serosal serotonin.


Assuntos
Aedes/metabolismo , Sinalização do Cálcio , Prótons , Receptores de Serotonina/metabolismo , Serotonina/metabolismo , Animais , AMP Cíclico/metabolismo , Citocalasinas/metabolismo , Trato Gastrointestinal/metabolismo , Concentração de Íons de Hidrogênio , Larva/metabolismo
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