RESUMO
Previous studies have demonstrated that bradykinin stimulates the rapid release of inositol 1,4,5 trisphosphate (IP3) from membrane phosphatidylinositol 4,5 bisphosphate (PIP2) in Madin-Darby canine kidney (MDCK) cells. Since current evidence would suggest that the activation of phospholipase C (PLC) is mediated through a guanine nucleotide-binding protein in receptor-mediated activation of PLC, we evaluated the role of guanine nucleotide proteins in receptor-mediated (bradykinin-stimulated) activation of PLC in MDCK cells. Bradykinin at 10(-7) M produced a marked increase in IP3 formation within 10 s increasing from a basal level of 46.2 to 686.6 pmol/mg cell protein a 15-fold increase. Pretreatment of MDCK cells in culture with 200 ng/ml of pertussis toxin for 4 h reduced the bradykinin-stimulated response to 205.8 pmol/mg protein. A 41-kD protein substrate in MDCK membranes was ADP ribosylated in vitro in the presence of pertussis toxin. The ADP ribosylation in vitro was inhibited by pretreatment of the cells in culture with pertussis toxin. Membranes from MDCK cells incubated in the presence of [3H]PIP2/phosphatidyl ethanolamine liposomes demonstrated hydrolysis of [3H]PIP2 with release of [3H]IP3 when GTP 100 microM or GTP gamma S 10 microM was added. Bradykinin 10(-7) M added with GTP 100 microM markedly increased the rate of hydrolysis within 10 s, thus demonstrating a similar time course of PLC activation as intact cells. These results demonstrate that bradykinin binds to its receptor and activates a membrane-associated PLC through a pertussis toxin-sensitive, guanine nucleotide protein.
Assuntos
Bradicinina/farmacologia , Túbulos Renais/enzimologia , Fosfolipases Tipo C/metabolismo , Difosfato de Adenosina/metabolismo , Animais , Bradicinina/metabolismo , Linhagem Celular , Membrana Celular/enzimologia , Cromatografia Líquida de Alta Pressão , Cromatografia por Troca Iônica , Ativação Enzimática , Proteínas de Ligação ao GTP/metabolismo , Guanosina Difosfato/metabolismo , Guanosina Trifosfato/metabolismo , Hidrólise , Inositol 1,4,5-Trifosfato , Fosfatos de Inositol/metabolismo , Túbulos Renais/citologia , Túbulos Renais/ultraestrutura , Toxina Pertussis , Fosfatidilinositol 4,5-Difosfato , Fosfatidilinositóis/metabolismo , Fatores de Virulência de Bordetella/farmacologiaRESUMO
Rabbit renal papillary collecting tubule cells were isolated as a homogeneous population and grown in primary culture. These cells were maintained in fully defined medium to inhibit fibroblast overgrowth and to facilitate labeling of endogenous inositol phospholipids with myo-[2-3H]inositol with high specific activity. These cells demonstrated the morphology, cyclic AMP responsiveness, and prostaglandin E2 (PGE2) elaboration, consistent with previous published characterizations. When cells labeled with myo-[2-3H]inositol were stimulated by bradykinin at 10(-7) M, time-dependent and reversible changes in the distribution of inositol polyphosphates were observed. Inositol 1,4,5-triphosphate and inositol 1,4-diphosphate showed time-dependent and dose-dependent increases to maximal levels of 225 and 223% of control, respectively. These data indicate that the elaboration of inositol polyphosphates is a biochemical correlate to bradykinin stimulation and may play a role in PGE2 release in renal papillary collecting tubule cells.
Assuntos
Bradicinina/farmacologia , Medula Renal/metabolismo , Túbulos Renais Coletores/metabolismo , Túbulos Renais/metabolismo , Fosfatidilinositóis/metabolismo , Animais , Divisão Celular , Células Cultivadas , AMP Cíclico/biossíntese , Dinoprostona , Feminino , Fosfatos de Inositol/metabolismo , Medula Renal/citologia , Fosfatidilinositóis/análise , Prostaglandinas E/biossíntese , CoelhosRESUMO
Leukotriene (LT) B4 is an important pro-inflammatory autocoid. In order to investigate the potential role of this eicosanoid in renal inflammation, in this study we determined the capability of glomeruli to synthesize this mediator. Glomeruli were able to synthesize LTB4 when provided with exogenous substrate in a dose-dependent fashion in the presence of ionophore A23187. Ionophore, although by itself a weak agonist for LTB4 formation, was required for LTB4 production from exogenous arachidonate. The identity of LTB4 was confirmed by specific radioimmunoassay, high pressure liquid chromatography, and gas chromatography/mass spectrometry. The synthesis of LTB4 was inhibited by BW755C (a lipoxygenase/cyclooxygenase inhibitor) but not indomethacin. Essential fatty acid (EFA) deficiency, obtained by the deprivation of (n-6) fatty acids, is known to exert a protective effect in renal inflammatory states. This dietary manipulation markedly attenuated the ability of glomeruli to synthesize LTB4. In contrast, the synthesis of cyclooxygenase products from exogenous arachidonate was increased by EFA deficiency. Because EFA deficiency has been shown to deplete glomeruli of resident mesangial macrophages, it was hypothesized that this effect accounted for the diminished LTB4 synthesis. To test this hypothesis, glomeruli were depleted of macrophages using x-irradiation. Glomeruli from these animals exhibited a marked decrease in LTB4 synthesis. Glomerular synthesis of cyclooxygenase products was unaffected by irradiation. In sum, glomeruli have the capability to synthesize LTB4, and this capacity is correlated with the presence of glomerular macrophages. EFA deficiency attenuates the ability of glomeruli to synthesize LTB4 by depleting them of macrophages.
Assuntos
Ácidos Graxos/deficiência , Glomérulos Renais/metabolismo , Leucotrieno B4/biossíntese , 4,5-Di-Hidro-1-(3-(Trifluormetil)Fenil)-1H-Pirazol-3-Amina , Animais , Calcimicina/farmacologia , Cromatografia Líquida de Alta Pressão , Dinoprostona/farmacologia , Relação Dose-Resposta a Droga , Indometacina/farmacologia , Camundongos , Pirazóis/farmacologia , Radioimunoensaio , Ratos , Ratos Endogâmicos LewRESUMO
Interleukin 1 and nitric oxide (NO) from infiltrating macrophages and activated mesangial cells may act in concert to sustain and promote glomerular damage. To evaluate if such synergy occurs, we evaluated the effect if IL-1 beta and NO on the formation of prostaglandin (PG)E2 and cyclooxygenase (COX) expression. The NO donors, sodium nitroprusside and S-nitroso-N-acetylpenicillamine, alone did not increase basal PGE2 formation. However, these compounds amplified IL-1 beta-induced PGE2 production. Similarly, sodium nitroprusside and S-nitroso-N-acetylpenicillamine by themselves did not induce mRNA and protein for COX-2, the inducible isoform of COX; however, they both potentiated IL-1 beta-induced mRNA and protein expression of COX-2. The stimulatory effect of NO is likely to be mediated by cGMP since (a) an inhibitor of the soluble guanylate cyclase, methylene blue, reversed the stimulatory effect of NO donors on COX-2 mRNA expression; (b) the membrane-permeable cGMP analogue, 8-Br-cGMP, mimicked the stimulatory effect of NO donors on COX-2-mRNA expression; and (c) atrial natriuretic peptide, which increases cellular cGMP by activating the membrane-bound guanylate cyclase, also amplified IL-1 beta-induced COX-2 mRNA expression. These data indicate a novel interaction between NO and COX pathways.
Assuntos
Dinoprostona/biossíntese , Mesângio Glomerular/metabolismo , Interleucina-1/farmacologia , Isoenzimas/biossíntese , Óxido Nítrico/farmacologia , Prostaglandina-Endoperóxido Sintases/biossíntese , Animais , Células Cultivadas , Ciclo-Oxigenase 2 , Sinergismo Farmacológico , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Humanos , Masculino , Proteínas de Membrana , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacosRESUMO
The amygdala has emerged as an important forebrain modulator of arousal. Acetylcholine plays a role in the regulation of sleep and wakefulness, particularly rapid eye movement sleep (REM). The major cholinergic input to the amygdala comes from the basal forebrain, a region primarily linked to wakefulness. We examined sleep and the encephalogram for 8 h following bilateral microinjections into the central nucleus of the amygdala (CNA) of the cholinergic agonist, carbachol (CARB(L): 0.3 microg; CARB(H): 3.0 microg), the acetylcholinesterase inhibitor, neostigmine (NEO(L): 0.3 microg; NEO(H): 3.0 microg), the muscarinic antagonist, scopolamine (SCO(L): 0.3 microg; SCO(H): 1.0 microg), the nicotinic antagonist, mecamylamine (MEC(L): 0.3 microg; MEC(H): 1.0 microg) and saline (SAL, 0.2 microl) alone. Both doses of CARB and NEO significantly reduced REM, but did not significantly alter non-rapid eye movement sleep (NREM). Both doses of SCO significantly increased NREM, and SCO(H) also produced an initial increase in REM followed by a significant decrease. CARB(H) and NEO(H) decreased REM electroencephalogram (EEG) power in the 5.5-10 Hz band, and NEO(L) and NEO(H) decreased NREM EEG power in the 0.5-5.0 Hz band. CARB(L) decreased waking EEG power in the 0.5-5.0 Hz band, and NEO(H) decreased waking EEG power in the 5.0-10.0 Hz band. Both doses of SCO significantly increased waking EEG power in the 5.5-10.0 Hz band. Compared with SAL, MEC did not significantly alter sleep or EEG power. The reduction of REM by CARB and NEO and the alteration of sleep by SCO indicate that cholinergic regulation of the amygdala is involved in the control of arousal in rodents. In contrast, CARB microinjections into CNA increase REM in cats, though the reasons for the species difference are not known. The results are discussed in the context of anatomical inputs and species differences in the cholinergic regulation of CNA.
Assuntos
Tonsila do Cerebelo/efeitos dos fármacos , Carbacol/farmacologia , Colinérgicos/farmacologia , Neostigmina/farmacologia , Sono/efeitos dos fármacos , Vigília/efeitos dos fármacos , Animais , Eletroencefalografia/métodos , Masculino , Microinjeções/métodos , Polissonografia/métodos , Ratos , Ratos WistarRESUMO
Changes in phosphatidylinositol metabolism are suggested to be involved in the mechanism of action of many membrane active hormones. We studied the effect of vasopressin on polyphosphoinositide metabolism in rabbit papillary collecting tubule cells to assess if the hydrolysis of these phospholipids is involved in transmembrane signaling. Rabbit papillary collecting tubule cells grown in monolayers for 5 days were labeled to constant specific activity with [3H]inositol. The temporal changes in [3H]inositol-labeled phospholipids were assessed in response to vasopressin. Similarly, water-soluble inositides were monitored after separation by ion exchange chromatography. Intracellular Ca2+ was monitored by use of the fluorescent indicator dye, quin2. Vasopressin (10(-7) M) did not increase the hydrolysis of phosphoinositides over a 5 min period when compared with controls. Similarly, there was no increase in water-soluble phosphoinositols during the same interval. Pretreating the cells with LiCl (10 mM) did not produce any increase in inositol 1-phosphate when stimulated with vasopressin but did in response to bradykinin. Finally, vasopressin did not increase cytosolic Ca2+ and did not increase the release of prostaglandin E2 into the media under our experimental conditions. We conclude that vasopressin does not stimulate prostaglandin E2 in rabbit papillary collecting tubule cells, does not initiate hydrolysis of polyphosphoinositides and does not increase cytosolic Ca2+. Thus these cells lack V1 receptor coupling mechanisms.
Assuntos
Arginina Vasopressina/farmacologia , Cálcio/metabolismo , Túbulos Renais Coletores/metabolismo , Túbulos Renais/metabolismo , Fosfatidilinositóis/metabolismo , Prostaglandinas E/biossíntese , Aminoquinolinas , Animais , Cloretos/farmacologia , AMP Cíclico/fisiologia , Dinoprostona , Hidrólise , Lítio/farmacologia , Cloreto de Lítio , CoelhosRESUMO
Prostaglandin E was found to increase the formation of cyclic acdenosine 3',5'-monophosphate (cyclic AMP) by renal cortical slices. This increased release of cyclic AMP was not influenced by the absence of Ca2+ in the incubating media. The enhanced production of cyclic AMP was probably mediated by stimulation of membrane-bound adenylate cyclase activity. An increase in adenyl cyclase activity was observed with increasing concentrations of prostaglandin E. Furthermore, prostaglandin E augmented glucose production from alpha-ketoglutarate. This effect on gluconeogenesis was abolished by the removal of Ca2+ from the incubating medium. These effects are similar to those described for parathyroid hormone and suggest that the renal cortex is a prostaglandin-dependent system. Prostaglandin E decreased cyclic AMP production and glucose production (from alpha-ketoglutarate) in response to submaximal doses of parathyroid hormone, suggesting that prostaglandin may be important in modulating the intracelluar action of parathyroid hormone in the kidney cortex.
Assuntos
Adenilil Ciclases/metabolismo , AMP Cíclico/metabolismo , Córtex Renal/metabolismo , Prostaglandinas E/farmacologia , Animais , Cálcio/farmacologia , Ácidos Cetoglutáricos , Córtex Renal/efeitos dos fármacos , Hormônio Paratireóideo/farmacologia , RatosRESUMO
Rabbit renal papillary collecting tubule cells were harvested and grown in primary cultures. When labeled with myo-[2-3H]inositol and extracted under neutral conditions, a metabolite undetected under acidic extraction was observed on resolution by anion-exchange chromatography and which eluted under similar conditions with authentic DL-myo-inositol 1,2-(cyclic)phosphate; the mass spectrum of its pentakis(trimethylsilyl) derivative contains an identical ratio of selected ion fragments to the authentic standard. Bradykinin, demonstrated previously to increase labeling of free inositol polyphosphates, increases labeling of inositol 1,2 cyclic phosphate but over a time course subsequent to the formation of inositol trisphosphate. These observations are consistent with the model that bradykinin induces hydrolysis of phosphatidylinositol 4,5-bisphosphate which precedes hydrolysis of phosphatidylinositol in renal papillary collecting tubule cells.
Assuntos
Bradicinina/farmacologia , Fosfatos de Inositol/metabolismo , Túbulos Renais Coletores/metabolismo , Túbulos Renais/metabolismo , Fosfatos Açúcares/metabolismo , Animais , Cromatografia , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Fosfatidilinositóis/metabolismo , CoelhosRESUMO
The metabolic effects of imidazole were tested in rat renal cortex. Imidazole enhanced the activity of renal cortical phosphodiesterase in vitro. Imidazole inhibited glucose production in a dose-dependent fashion from a variety of substrates in the gluconeogenic pathway proximal to the triose phsophates. The stimulation in renal gluconeogenesis resulting from isoproterenol and parathyroid hormone was inhibited by imidazole. These changes correlated with an inhibition of the augmented levels of renal cortical cyclic AMP levels produced by these hormones. These studies indicate that imidazole is an effective activator of phosphodiesterase in intact renal cells and lend further support to the suggestion that the stimulation of renal gluconeogenesis produced by isoproterenol and parathyroid hormone is mediated by a release of cyclic AMP.
Assuntos
Gluconeogênese/efeitos dos fármacos , Imidazóis/farmacologia , Córtex Renal/metabolismo , 3',5'-AMP Cíclico Fosfodiesterases/metabolismo , Animais , Feminino , Concentração de Íons de Hidrogênio , Técnicas In Vitro , Isoproterenol/farmacologia , Córtex Renal/efeitos dos fármacos , Cinética , Hormônio Paratireóideo/farmacologia , RatosRESUMO
Minced rabbit pericardium actively converts [1-14C]arachidonic acid into the known prostaglandins (6-[1-14C]ketoprostaglandin F1 alpha, [1-14C]prostaglandin E2 and [1-14C]prostaglandin F2 alpha) and into several unidentified metabolites. The major metabolite was separated by C18 reverse-phase high-pressure liquid chromatography (HPLC) and identified by gas chromatography-mass spectrometry (GC-MS) to be 6,15-[1-14C]diketo-13,14-dihydroprostaglandin F1 alpha. The other nonpolar metabolites were 15-[1-14C]hydroxy-5,8,11,13-eicosa-tetraenoic acid (15-HETE), 11-[1-14C]hydroxy-5,8,12,14-eicosatetraenoic acid (11-HETE) and 12-[1-14C]hydroxy-5,8,10,14-eicosatetraenoic acid (12-HETE). Arachidonic acid metabolites actively produced by the pericardium could influence the tone of surface blood vessels on the myocardium.
Assuntos
Ácidos Araquidônicos/metabolismo , Ácidos Hidroxieicosatetraenoicos , Pericárdio/metabolismo , Ácido 12-Hidroxi-5,8,10,14-Eicosatetraenoico , 6-Cetoprostaglandina F1 alfa/análogos & derivados , 6-Cetoprostaglandina F1 alfa/metabolismo , Animais , Ácido Araquidônico , Cromatografia Líquida de Alta Pressão , Cromatografia em Camada Fina , Cromatografia Gasosa-Espectrometria de Massas , CoelhosRESUMO
Human epidermal cell cultures were examined to determine whether they were capable of histamine release. Results of these studies indicated that keratinocytes contain and release significant amounts of histamine. In the skin of some individuals, histamine content was induced after ultraviolet B light injury, and 40% of subjects demonstrated high basal histamine levels. Mass spectrometric analysis of cell supernatants showed that the histamine was released into the extracellular environment. Such release may contribute to common itching or intensify the inflammatory response in vivo.
Assuntos
Histamina/análise , Queratinócitos/química , Queratinócitos/efeitos da radiação , Raios Ultravioleta/efeitos adversos , Adulto , Células Cultivadas , Dermatite/etiologia , Liberação de Histamina/efeitos da radiação , Humanos , Queratinócitos/metabolismoRESUMO
Adrenal glomerulosa cells washed with delipidated albumin produced increased amounts of aldosterone in response to angiotensin-II (AII) or (Bu)2cAMP. Albumin treatment also increased binding of 125I-labeled AII to high affinity binding sites on adrenal cells. Lipid extracts of albumin solutions that were used to wash cells inhibited AII binding and aldosterone responses by washed glomerulosa cells. Chromatographic fractionation and mass spectroscopic analysis indicated that the inhibitors removed from cells by albumin were long chain fatty acids. Exogenous fatty acids not only inhibited AII binding, but they inhibited basal aldosterone production and increments in aldosterone caused by AII or dbcAMP, suggesting an effect on postreceptor steps in aldosteronogenesis. The most potent and most abundant fatty acids removed from adrenal cells were oleic, linoleic, and arachidonic. These fatty acids inhibited at micromolar concentrations in the absence of albumin and at somewhat higher concentrations in its presence. Cells that had been washed, then inhibited by exogenous oleic acid in vitro, were restored to their enhanced responsiveness by a second albumin wash, making it unlikely that cell damage is the mechanism of inhibition by fatty acids. Responses of fasciculata cells were not potentiated by albumin washes, and cortisol production was less sensitive than aldosterone production to exogenous fatty acids. Binding of ANP to glomerulosa cells was not affected by albumin or fatty acids. These results combined with clinical correlations make it plausible that unesterified fatty acids are naturally occurring regulators of the adrenal glomerulosa. Insulin's ability to lower plasma levels of fatty acids may be one way that it causes sodium retention.
Assuntos
Aldosterona/metabolismo , Ácidos Graxos/farmacologia , Animais , Fracionamento Químico , Ácidos Graxos/antagonistas & inibidores , Ácidos Graxos/metabolismo , Albumina Sérica/farmacologia , Irrigação Terapêutica , Zona Glomerulosa/citologia , Zona Glomerulosa/metabolismoRESUMO
Drugs that block monoamine reuptake initially suppress rapid eye movement (REM) sleep in the cat and other species. Less is known about the effects of repeated drugs administration. Desipramine (DMI) and sertraline [1S,4S-N-methyl-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydro-1 -naphthylamine] (SER), which are relatively specific in blocking norepinephrine and serotonin reuptake, respectively, were each given to cats for approximately two and a half weeks. Six-hour sleep polygraphic records were obtained under the placebo condition, after acute drug administration, and again during chronic drug administration. DMI and SER both reduced REM sleep percentage acutely and in each case. Significant tolerance then developed. These actions of DMI and SER reflected changes in mean REM sleep episode duration as well as REM sleep episode number. Such differential effects of acute and chronic monoamine reuptake blockade on REM sleep behavior in the cat may ultimately be correlated with pharmacological changes at the receptor level.
Assuntos
1-Naftilamina/farmacologia , Encéfalo/efeitos dos fármacos , Desipramina/farmacologia , Naftalenos/farmacologia , Receptores Adrenérgicos/efeitos dos fármacos , Receptores de Serotonina/efeitos dos fármacos , Antagonistas da Serotonina/farmacologia , Sono REM/efeitos dos fármacos , 1-Naftilamina/análogos & derivados , Animais , Gatos , Ritmo Circadiano/efeitos dos fármacos , Relação Dose-Resposta a Droga , Feminino , SertralinaRESUMO
The subjective sleep disturbance in posttraumatic stress disorder (PTSD), including the repetitive, stereotypical anxiety dream, suggests dysfunctional rapid eye movement (REM) sleep mechanisms. The polysomnograms of a group of physically healthy combat veterans with current PTSD were compared with those of an age-appropriate normal control group. Tonic and phasic REM sleep measures in the PTSD subjects were elevated on the second night of recorded sleep. Increased phasic REM sleep activity persisted in the PTSD group on the subsequent night. During the study, an anxiety dream occurred in a PTSD subject in REM sleep. The results are consistent with the view that a dysregulation of the REM sleep control system, particularly phasic event generation, may be involved in the pathogenesis of PTSD. The finding of a specific disturbance of sleep unique to PTSD may have significant implications for the design of effective treatments for PTSD.
Assuntos
Distúrbios de Guerra/diagnóstico , Transtornos do Sono-Vigília/diagnóstico , Sono REM , Veteranos/psicologia , Adulto , Alcoolismo/diagnóstico , Alcoolismo/psicologia , Nível de Alerta/efeitos dos fármacos , Distúrbios de Guerra/psicologia , Sonhos/efeitos dos fármacos , Humanos , Masculino , Pessoa de Meia-Idade , Polissonografia/efeitos dos fármacos , Psicotrópicos , Fases do Sono/efeitos dos fármacos , Transtornos do Sono-Vigília/psicologia , Sono REM/efeitos dos fármacos , Transtornos Relacionados ao Uso de Substâncias/diagnóstico , Transtornos Relacionados ao Uso de Substâncias/psicologiaRESUMO
BACKGROUND: Hyperarousal in posttraumatic stress disorder (PTSD) is manifested during sleep as well as waking. Elevated rapid eye movement sleep (REMS) phasic activity, likely signifying central nervous system alerting, has been identified in PTSD. The authors reasoned that PTSD compared to control subjects would show particularly increased REMS phasic activity on the first night of polysomnography, with adaptation to a novel environment. METHODS: First-night polysomnograms of 17 veterans with PTSD were compared with those of 11 control subjects. Sleep was also studied in subsets of both groups over two nights. RESULTS: On the first night, the PTSD subjects had a higher density of rapid eye movements in the first REMS period. This measure was increased on the first compared to the second night, but there was no interaction effect between night and group. CONCLUSIONS: REMS changes are again demonstrated in veterans with PTSD. Introduction to a novel environment activated a REMS phasic process, but not differentially in PTSD compared to control subjects.
Assuntos
Nível de Alerta/fisiologia , Distúrbios de Guerra/fisiopatologia , Sono REM/fisiologia , Adaptação Fisiológica , Adaptação Psicológica , Análise de Variância , Estudos de Casos e Controles , Humanos , Masculino , Pessoa de Meia-Idade , Polissonografia , Sobreviventes/psicologia , Estados Unidos , Veteranos/psicologia , VietnãRESUMO
Neurons in the medial tegmental field project directly to spinal somatic motoneurons and to cranial motoneuron pools such as the hypoglossal nucleus. The axons of these neurons may be highly collateralized, projecting to multiple levels of the spinal cord and to many diverse regions at different levels of the neuraxis. We employed a double fluorescent retrograde tracer technique to examine whether medial tegmental neurons that project to the spinal cord also project to the hypoglossal nucleus. Injections of Diamidino Yellow into the hypoglossal nucleus and Fast Blue into the spinal cord produced large numbers of double labeled neurons in the medial tegmental field, particularly in the caudal raphe nuclei and adjacent ventromedial reticular formation. In these structures the number of neurons projecting to both the hypoglossal nucleus and the spinal cord was equivalent to the number of neurons projecting to multiple levels of the spinal cord observed in control animals. Fewer neurons projecting to both the hypoglossal nucleus and the spinal cord were observed in several other nuclei and subregions of the medial tegmental field, while almost no such neurons were observed in the lateral tegmental field or other pontomedullary structures. These results demonstrate that neurons of the caudal raphe nuclei and adjacent ventromedial reticular formation project to both the spinal cord and the hypoglossal nucleus, and support the concept that the diffuse projections to motoneuron pools from the medial tegmental field globally modulate both spinal and cranial somatic motoneuron excitability.
Assuntos
Nervo Hipoglosso/anatomia & histologia , Bulbo/anatomia & histologia , Neurônios Motores/citologia , Neurônios/citologia , Ponte/anatomia & histologia , Núcleos da Rafe/anatomia & histologia , Ratos Wistar/anatomia & histologia , Medula Espinal/anatomia & histologia , Amidinas , Animais , Transporte Axonal , Corantes Fluorescentes , Masculino , RatosRESUMO
Arachidonic acid is metabolized to prostaglandin, lipoxygenase products, and products of the microsomal cytochrome P-450 enzymes of the kidney. The distribution of the metabolizing enzymes and their regulation and pharmacologic manipulation are reviewed. The mechanisms of release of arachidonic acid from membrane lipids through a surface-mediated receptor mechanism are also discussed. The localization of the various enzymes and product formation may have profound effects on glomerular filtration, renal blood flow, and electrolyte excretion. Therefore, an understanding of the potential sites of inhibition of the nonsteroidal anti-inflammatory drugs is important in assessing their effects on renal function.
Assuntos
Ácidos Araquidônicos/metabolismo , Rim/metabolismo , Animais , Ácido Araquidônico , Sítios de Ligação , Inibidores de Ciclo-Oxigenase , Sistema Enzimático do Citocromo P-450/metabolismo , Dinoprostona , Ácidos Eicosanoicos/metabolismo , Epoprostenol/metabolismo , Humanos , Rim/efeitos dos fármacos , Rim/enzimologia , Lipoxigenase/metabolismo , Oxirredução , Endoperóxidos Sintéticos de Prostaglandinas/metabolismo , Prostaglandina H2 , Prostaglandina-Endoperóxido Sintases/metabolismo , Prostaglandinas/biossíntese , Prostaglandinas E/metabolismo , Prostaglandinas H/metabolismo , Especificidade por Substrato , Tromboxanos/biossínteseRESUMO
Norepinephrine (NE) and serotonin (5HT) likely inhibit the generation of ponto-geniculo-occipital (PGO) waves. Either desipramine (DMI) or sertraline (SER:1S,4S-N-methyl-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydro-1-naphthyl amine) was administered in the cat for 2.5 weeks to probe noradrenergic and serotonergic mechanisms, respectively. Placebo days were compared with the first day of drug and with days that followed 2.5 weeks of drug (chronic). PGO rates during REM sleep and the preceding transition period were significantly decreased by either chronic DMI or SER. Cat PGO waves resemble waves that accompany alerting to intense or novel stimuli in wakefulness. Depressive disorders in humans have features of hyperarousal; PGO wave suppression by antidepressant drugs may relate to clinical antidepressant actions.
Assuntos
1-Naftilamina/análogos & derivados , Desipramina/farmacologia , Corpos Geniculados/fisiologia , Norepinefrina/fisiologia , Lobo Occipital/fisiologia , Ponte/fisiologia , Antagonistas da Serotonina/farmacologia , Serotonina/fisiologia , 1-Naftilamina/farmacologia , Animais , Transporte Biológico/efeitos dos fármacos , Gatos , Eletroencefalografia/efeitos dos fármacos , Eletromiografia , Eletroculografia , Movimentos Oculares/efeitos dos fármacos , Feminino , Corpos Geniculados/efeitos dos fármacos , Norepinefrina/metabolismo , Lobo Occipital/efeitos dos fármacos , Ponte/efeitos dos fármacos , Serotonina/metabolismo , Sertralina , Sono REM/efeitos dos fármacosRESUMO
PURPOSE: Previous work has shown that exposure of lens epithelial cells or rabbit eyes in vivo to ultraviolet B (UVB) radiation enhanced prostaglandin (PG)E2 synthesis. Such enhanced PGE2 synthesis was related to the increased DNA synthesis that followed UVB exposure. The current study examined the relationship between enhanced prostaglandin synthesis and UVB-induced cataract formation. METHODS: Seventy albino (New Zealand white) rabbit eyes were exposed to UVB radiation in vivo. Fluence of radiation at the cornea was 2.8 J/cm2, 5.6 J/cm2, or 11.2 J/cm2. Eyes were examined 24 hours after UVB exposure and for as long as 10 days by slit lamp biomicroscopy. Mass spectrometry was used to measure PGE2, PGF2 alpha, and 6-keto-PGF1 alpha content of the lens and iris-ciliary body using authentic standards. To determine the effect of inhibition of prostaglandin synthesis on UVB-induced cataract formation, animals were given indomethacin intraperitoneally. Other pharmacologic agents, such as PGE2, PGF2 alpha, and misoprostol, were applied topically to the eye. The effect of UVB on K+ pump was determined by incubating isolated lenses with [86Rb+]. RESULTS: Twenty-four hours after UVB exposure, PGE2 and PGF2 alpha concentrations in aqueous humor were increased by 100- and 30-fold, respectively. Lens PGE2 and PGF2 alpha increased by 6- and 4-fold, respectively, after UVB radiation exposure. Pretreatment of animals with indomethacin prevented the rise in lens and aqueous humor PGE2 and PGF2 alpha levels. Furthermore, indomethacin was partially protective against UVB cataract formation and lowered cataract severity from stage 3 to stage 1, but it did not prevent UVB-induced lens changes completely. Topical application of PGE2 before UVB exposure completely prevented cataract formation in the UVB-exposed eye. In contrast, topical administration of PGF2 alpha increased cataract severity. UVB-induced cataract formation preceded changes in [86Rb]+ uptake in lenses subsequently incubated in K(+)-free Tyrode's. CONCLUSIONS: Enhanced synthesis of cyclooxygenase products of arachidonic acid metabolism in the lens is associated with UVB-induced cataract formation in albino rabbit eyes, and inhibition of cyclooxygenase by indomethacin decreased the severity of cataracts. PGE2, the principal arachidonic acid metabolite, appears to have a protective role because pretreatment of the eye with topical PGE2 completely prevented UVB-induced cataract formation, whereas PGF2 alpha increased the severity of the cataract. The evidence presented for a role of PGF2 alpha in the development of cataract suggests that caution be exercised in the use of PGF2 alpha derivatives in the therapy of glaucoma.
Assuntos
Catarata/fisiopatologia , Dinoprosta/fisiologia , Dinoprostona/fisiologia , Córtex do Cristalino/fisiopatologia , Ocitócicos , Lesões Experimentais por Radiação/fisiopatologia , Raios Ultravioleta/efeitos adversos , 6-Cetoprostaglandina F1 alfa/análise , 6-Cetoprostaglandina F1 alfa/fisiologia , Animais , Catarata/etiologia , Catarata/prevenção & controle , Corpo Ciliar/química , Corpo Ciliar/efeitos da radiação , Túnica Conjuntiva/efeitos da radiação , Córnea/efeitos da radiação , Inibidores de Ciclo-Oxigenase/farmacologia , Dinoprosta/análise , Dinoprosta/farmacologia , Dinoprostona/análise , Dinoprostona/farmacologia , Indometacina/farmacologia , Iris/química , Iris/efeitos da radiação , Córtex do Cristalino/química , Córtex do Cristalino/efeitos dos fármacos , Córtex do Cristalino/efeitos da radiação , Misoprostol/farmacologia , Soluções Oftálmicas , Potássio/metabolismo , Coelhos , Lesões Experimentais por Radiação/etiologia , Lesões Experimentais por Radiação/prevenção & controle , Radioisótopos de Rubídio/metabolismo , ATPase Trocadora de Sódio-Potássio/metabolismo , ATPase Trocadora de Sódio-Potássio/efeitos da radiaçãoRESUMO
PURPOSE: The goals of this investigation were to examine the synthesis of prostaglandins after UVB exposure of lens epithelial cells and to investigate their role in cell proliferation. METHODS: Cultured rabbit lens epithelial cells (cell line N/N1003A) were exposed to low levels of UV radiation. Prostaglandins were assayed by radioimmunoassay; products of arachidonic acid metabolism were analyzed by thin-layer chromatography and mass spectroscopy. Cell proliferation was measured by [3H]thymidine incorporation and proliferative autoradiography. RESULTS: Cultured lens epithelial cells exposed to UVB radiation showed a dose-dependent increase in basal prostaglandin synthesis measured 24 hours after UV exposure. At an optimal dose (250 J/m2) of UVB, prostaglandin E2 (PGE2) synthesis was enhanced tenfold. Product identity was confirmed using thin-layer chromatography and mass spectroscopy with authentic standards. Incubation of irradiated cells with exogenous arachidonic acid followed by extraction and thin-layer chromatography revealed that the cultures produced PGE2, prostaglandin I2 (measured as 6-keto-prostaglandin F1 alpha), prostaglandin F2 alpha, and hydroxyeicosatetraenoic acid. The synthesis of all of these products was enhanced threefold in cells exposed to 250-J/m2 UVB. Indomethacin pretreatment eliminated the synthesis of prostaglandins, further confirming their identity. To discover the relationship between PGE2 synthesis and irradiation-induced cell proliferation, [3H]thymidine incorporation into DNA was determined 24 or 48 hours after exposure. These experiments revealed a fivefold increase in incorporation induced by UVB exposure. UVB-enhanced incorporation of thymidine was eliminated by pretreatment of cultures with indomethacin to eliminate PG synthesis. However, when 100 nM PGE2 was added to the indomethacin-treated irradiated cultures, incorporation of the label was restored toward the level detected in the UVB-stimulated cells. Addition of other prostaglandins to the cultures was ineffective. CONCLUSIONS: The results indicate that the synthesis of PGE2 is enhanced by exposure of lens epithelial cells to UVB radiation. PGE2 seems to play a specific role in cell proliferation after UV exposure. This increase in PGE2 synthesis may be important in posterior subcapsular cataract formation in humans and in animals exposed to UVB radiation in vivo.