RESUMEN
Dopaminergic neurons in the substantia nigra pars compacta undergo apoptosis after transection of the medial forebrain bundle. We have assessed the temporal and sequential activities of microglia in these events by examining the complement-3 (OX-42), major histocompatibility complex class II antigen presentation (OX-6) and phagocytic activity (ED1), and correlating these indicators with dopaminergic neuronal loss. Microglia in the ipsilateral substantia nigra pars reticulata evinced activation morphology at 12 h postaxotomy. Phagocytic microglia apposed dying dopaminergic neurons in the pars compacta starting at 3 days postlesion; their number increased through 14 days and slowly decreased. Nuclear chromatin condensation and significant loss of tyrosine hydroxylase-positive dopaminergic neurons occurred around 7 days postlesion. In contrast to microglial expression of interleukin-1beta and inducible nitric oxide synthase at the axotomy site, nigral microglia were interleukin-1beta and inducible nitric oxide synthase-negative. Consistently, RNase protection assays showed that interleukin-1beta and inducible nitric oxide synthase transcripts in nigra were equivocal. The present data support the idea that phagocytosis of axotomized neurons by activated microglia is not limited to dead neurons but includes dying neurons probably without cytotoxic effects of inflammatory substances, such as interleukin-1beta or nitric oxide.
Asunto(s)
Haz Prosencefálico Medial/fisiología , Microglía/metabolismo , Sustancia Negra/metabolismo , Animales , Apoptosis/fisiología , Axotomía , Citocinas/metabolismo , Masculino , Microglía/patología , Óxido Nítrico Sintasa/metabolismo , Óxido Nítrico Sintasa de Tipo II , Ratas , Ratas Wistar , Sustancia Negra/patología , Factores de Tiempo , Tirosina 3-Monooxigenasa/metabolismoRESUMEN
Electrical stimulation of the cerebellar fastigial nucleus (FN) elicits a prolonged ( approximately 10 days) and substantial (50-80%) protection against ischemic and excitotoxic injuries. The mechanism(s) of protection are unknown. We investigated whether FN stimulation directly protects brain cells against apoptotic cell death in an in vitro rat brain slice culture model. Rats were electrically stimulated in FN or, as control, the cerebellar dentate nucleus (DN). Coronal slices through the forebrain were explanted, exposed to staurosporine, harvested, and analyzed for caspase-3 activity by a fluorescence assay. FN, but not DN, stimulation significantly reduced staurosporine-induced caspase-3 activity by 39 +/- 7% at 3 h, 31 +/- 3% at 6 h and 26 +/- 4% at 10 h of incubation. Immunocytochemistry revealed FN-specific reductions in activated caspase-3 mainly in glial-like cells throughout the forebrain. FN stimulation also results in a 56.5% reduction in cytochrome c release upon staurosporine incubation. We conclude that neuroprotection elicited from FN stimulation can directly modify the sensitivity of brain cells to apoptotic stimuli and thereby suppress staurosporine induced apoptosis in adult rat brain slices. This model indicates that neuroprotection can be studied in vitro and provides new insight into the potential role of glial cells in ischemic protection of neurons induced by FN stimulation.
Asunto(s)
Apoptosis/fisiología , Cerebelo/efectos de los fármacos , Cerebelo/fisiología , Estaurosporina/farmacología , Animales , Caspasa 3 , Caspasas/metabolismo , Grupo Citocromo c/antagonistas & inhibidores , Estimulación Eléctrica , Activación Enzimática/fisiología , Técnicas In Vitro , Masculino , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
Agmatine (decarboxylated l-arginine), an endogenous ligand of imidazoline and alpha(2) adrenoreceptors, is particularly enriched in the rat hypothalamic paraventricular (PVN) and supraoptic (SON) nuclei. The present study utilized light and electron microscopic immunocytochemical methods to determine the distribution and extent of colocalization of agmatine relative to subpopulations of vasopressin- (VP) and oxytocin- (OT) producing neurons in PVN and SON nuclei. By light microscopy, agmatine-immunoreactive perikarya were found in both the magnocellular and the parvocellular neuronal subdivisions of PVN and SON. Confocal and electron microscopy revealed that agmatine-immunoreactivity (I) within neuronal perikarya was associated with the nuclear membrane as well as mitochondria, Golgi complexes, endoplasmic reticula, and plasmalemma. Additionally, agmatine-I was identified in both axons and axonal terminals, which were enriched in large dense-core vesicles. Dual and triple immunocytochemical labeling experiments also demonstrated that agmatine coexists with VP or OT in most PVN and SON magnocellular neurons. Combinations of iontophoretic injections of Fluorogold into the dorsomedullary complex with immunocytochemical labeling revealed that many retrogradely labeled neurons in the parvocellular region of the PVN contained agmatine-I and either VP or OT. These findings provide evidence that agmatine may function as a modulator of both hypothalamically mediated neuroendocrine and autonomic responses.
Asunto(s)
Agmatina/análisis , Oxitocina/análisis , Núcleo Hipotalámico Paraventricular/citología , Núcleo Supraóptico/citología , Animales , Transporte Axonal , Técnicas para Inmunoenzimas , Masculino , Neuronas/citología , Neuronas/ultraestructura , Núcleo Hipotalámico Paraventricular/ultraestructura , Ratas , Ratas Sprague-Dawley , Núcleo Supraóptico/ultraestructuraRESUMEN
We investigated whether stimulation of the functionally discrete subthalamic region, subthalamic cerebrovasodilator area (SVA), which increases cerebral blood flow (CBF) when excited, would, like stimulation of cerebellar fastigial nucleus (FN), produce central neurogenic neuroprotection. A 1-h electrical stimulation of SVA or FN reduced infarctions triggered by permanent occlusion of middle cerebral artery (MCA) by 48-55% in Sprague-Dawley rats and by 59% in Fisher rats. The salvaging effect of SVA stimulation, similar to FN, was long lasting and reduced the volume of infarctions placed 72 h or 10 days later by 58 and 26%, respectively, in Fisher rats. Bilateral lesioning of FN neurons by the microinjection of ibotenic acid 5 days before SVA stimulation did not affect SVA-evoked neuroprotection. Bilateral lesions of SVA neurons administered 5 days before FN stimulation had no effect on FN-induced neuroprotection but reversed the stimulus-locked increase in CBF accompanying FN stimulation. This study demonstrates that (1) excitation of neurons and/or fibers projecting through the SVA reduces ischemic infarctions as substantially as excitation of FN neurons; (2) the effects are long-lasting and not attributable to increases in cerebral blood flow, changes in blood gases or brain temperature, or rat strain; (3) the neuroprotective effects of SVA and FN stimulation are mutually independent and (4) FN-evoked cerebrovasodilation is mediated by SVA neurons. The SVA and FN are part of a neuronal system in CNS, which is distributed and, when excited, acts to protect the brain from ischemic injury.
Asunto(s)
Isquemia Encefálica/terapia , Núcleos Cerebelosos/fisiología , Circulación Cerebrovascular/fisiología , Terapia por Estimulación Eléctrica , Degeneración Nerviosa/prevención & control , Subtálamo/fisiología , Vasodilatación/fisiología , Animales , Isquemia Encefálica/patología , Isquemia Encefálica/fisiopatología , Núcleos Cerebelosos/citología , Desnervación , Agonistas de Aminoácidos Excitadores/farmacología , Ácido Iboténico/farmacología , Infarto de la Arteria Cerebral Media/patología , Infarto de la Arteria Cerebral Media/fisiopatología , Infarto de la Arteria Cerebral Media/terapia , Masculino , Degeneración Nerviosa/fisiopatología , Degeneración Nerviosa/terapia , Vías Nerviosas/citología , Vías Nerviosas/fisiología , Neurotoxinas/farmacología , Ratas , Ratas Endogámicas F344 , Ratas Sprague-Dawley , Subtálamo/citología , Subtálamo/efectos de los fármacos , Factores de TiempoRESUMEN
The present study examined specific effects of sodium cyanide (CN) on the membrane potential (MP), spontaneous discharge (SD) and voltage-gated ion current of the identified bulbospinal rostral ventrolateral medulla (RVLM) neuron in the rat pup brainstem slice. 125 microM CN rapidly depolarized MP in the RVLM neuron by 11.6 mV as well as enhanced the SD rate by 300%. In contrast, the same dose of CN immediately hyperpolarized unlabeled, non-RVLM neurons by 4.8 mV. 50 microM CN did not significantly affect voltage-gated Ca(++) or A-type K(+) currents. The same concentration of CN, however, rapidly and reversibly suppressed voltage-gated Na(+) currents and sustained outward K(+) currents in the RVLM neuron by 22.5% and 23%, respectively. Tetraethylammonium could mimic the effect of CN on MP, SD and sustained K(+) current in the RVLM neuron. It is concluded that: (1) like that from the adult rat, the rat pup bulbospinal RVLM neuron can be selectively and rapidly excited by CN; (2) the hypoxia-sensitive, sustained outward K(+) channel may play an important role in the acute hypoxia-induced excitation of the RVLM neurons.
Asunto(s)
Inhibidores Enzimáticos/farmacología , Activación del Canal Iónico/efectos de los fármacos , Bulbo Raquídeo/citología , Neuronas/fisiología , Cianuro de Sodio/farmacología , Animales , Canales de Calcio/metabolismo , Hipoxia de la Célula/fisiología , Bulbo Raquídeo/fisiología , Potenciales de la Membrana/efectos de los fármacos , Neuronas/efectos de los fármacos , Técnicas de Cultivo de Órganos , Oxígeno/metabolismo , Técnicas de Placa-Clamp , Canales de Potasio/metabolismo , Ratas , Canales de Sodio/metabolismoRESUMEN
Sympathoexcitatory reticulospinal neurons of the rostral ventrolateral medulla (RVLM) are oxygen detectors excited by hypoxia to globally elevate regional cerebral blood flow (rCBF). The projection, which accounts for >50% of hypoxic cerebral vasodilation, relays through the medullary vasodilator area (MCVA). However, there are no direct cortical projections from the RVLM/MCVA, suggesting a relay that diffusely innervates cortex and possibly originates in thalamic nuclei. Systematic mapping by electrical microstimulation of the thalamus and subthalamus revealed that elevations in rCBF were elicited only from a limited area, which encompassed medial pole of zona incerta, Forel's field, and prerubral zone. Stimulation (10 sec train) at an active site increased rCBF by 25 +/- 6%. Excitation of local neurons with kainic acid mimicked effects of electrical stimulation by increasing rCBF. Stimulation of the subthalamic cerebrovasodilator area (SVA) with single pulses (0.5 msec; 80 microA) triggered cortical EEG burst-CBF wave complexes with latency 24 +/- 5 msec, which were similar in shape to complexes evoked from the MCVA. Selective bilateral lesioning of the SVA neurons (ibotenic acid, 2 microg, 200 nl) blocked the vasodilation elicited from the MCVA and attenuated hypoxic cerebrovasodilation by 52 +/- 12% (p < 0.05), whereas hypercarbic vasodilation remained preserved. Lesioning of the vasodilator site in the basal forebrain failed to modify SVA-evoked rCBF increase. We conclude that (1) excitation of intrinsic neurons of functionally restricted region of subthalamus elevates rCBF, (2) these neurons relay signals from the MCVA, which elevate rCBF in response to hypoxia, and (3) the SVA is a functionally important site conveying vasodilator signal from the medulla to the telencephalon.
Asunto(s)
Corteza Cerebral/irrigación sanguínea , Hipoxia Encefálica/metabolismo , Bulbo Raquídeo/metabolismo , Neuronas/fisiología , Subtálamo/fisiología , Animales , Velocidad del Flujo Sanguíneo/efectos de los fármacos , Velocidad del Flujo Sanguíneo/fisiología , Presión Sanguínea/fisiología , Dióxido de Carbono/farmacología , Circulación Cerebrovascular/efectos de los fármacos , Circulación Cerebrovascular/fisiología , Estimulación Eléctrica , Electroencefalografía , Ácido Iboténico/administración & dosificación , Ácido Kaínico/administración & dosificación , Masculino , Bulbo Raquídeo/citología , Bulbo Raquídeo/efectos de los fármacos , Microinyecciones , Neuronas/efectos de los fármacos , Prosencéfalo/efectos de los fármacos , Prosencéfalo/fisiología , Ratas , Tiempo de Reacción/fisiología , Médula Espinal/fisiología , Subtálamo/citología , Subtálamo/efectos de los fármacos , Vasodilatación/efectos de los fármacos , Vasodilatación/fisiologíaRESUMEN
In anesthetized spinalized rat, electrical stimulation of the nucleus tractus solitarius (NTS) synchronizes the EEG by increasing the power of 4-6-Hz waves (>100%; P<0.01), and elevates cerebral blood flow (rCBF) by 18+/-5% (P<0.05). The coordinated response appears within seconds, is global, reversible, graded, evoked from the commissural sub-nucleus, and replicated by L-glutamate. The responses are markedly reduced by bilateral lesions or muscimol microinjections restricted to a region of ventral medullary reticular formation, the medullary cerebral vasodilator area (MCVA), a region from which stimulation elicits identical responses and mediates the comparable responses to hypoxic/ischemic excitation of sympathoexcitatory neurons of rostral ventrolateral medulla (RVLM). We conclude that: (a) excitation of intrinsic neurons of commissural NTS synchronizes the EEG and coordinately elevates rCBF; (b) the responses are mediated by excitation of neurons in MCVA; (c) the MCVA may be a common final pathway mediating cerebrovascular and EEG responses from multiple areas of CNS; and (d) the NTS-MCVA pathway may be a part of the anatomical substrate for behaviors, including slow-wave sleep and seizure suppression evoked by stimulation of visceral afferents terminating in NTS.
Asunto(s)
Circulación Cerebrovascular/fisiología , Electroencefalografía , Bulbo Raquídeo/fisiología , Neuronas/fisiología , Núcleo Solitario/fisiología , Animales , Tronco Encefálico/fisiología , Circulación Cerebrovascular/efectos de los fármacos , Estimulación Eléctrica , Electroencefalografía/efectos de los fármacos , Ácido Glutámico/administración & dosificación , Ácido Glutámico/farmacología , Masculino , Bulbo Raquídeo/irrigación sanguínea , Microinyecciones , Ratas , Ratas Sprague-Dawley , Núcleo Solitario/efectos de los fármacosRESUMEN
We sought to identify the medullary relay area mediating the elevations of regional cerebral blood flow (rCBF) and synchronization of the electroencephalogram (EEG) in the rat cerebral cortex elicited by hypoxic excitation of reticulospinal sympathoexcitatory neurons of the rostral ventrolateral medulla (RVLM ). In anaesthetized spinalized rats electrical stimulation of RVLM elevated rCBF (laser-Doppler flowmetry) by 31 +/- 6 %, reduced cerebrovascular resistance (CVR) by 26 +/- 8 %, and synchronized the EEG, increasing the power of the 5-6 Hz band by 98 +/- 25 %. Stimulation of a contiguous caudal region, the medullary cerebral vasodilator area (MCVA), had comparable effects which, like responses of RVLM, were replicated by microinjection of L-glutamate (5 nmol, 20 nl). Microinjection of NaCN (300 pmol in 20 nl) elevated rCBF (17 +/- 5 %) and synchronized the EEG from RVLM, but not MCVA, while nicotine (1.2 nmol in 40 nl) increased rCBF by 13 +/- 5 % and synchronized the EEG from MCVA. In intact rats nicotine lowered arterial pressure only from MCVA (101 +/- 3 to 52 +/- 9 mmHg). Bilateral electrolytic lesions of MCVA significantly reduced, by over 59 %, elevations in rCBF and, by 78 %, changes in EEG evoked from RVLM. Bilateral electrolytic lesions of RVLM did not affect responses from MCVA. Anterograde tracing with BDA demonstrated that RVLM and MCVA are interconnected. The MCVA is a nicotine-sensitive region of the medulla that relays signals elicited by excitation of oxygen-sensitive reticulospinal neurons in RVLM to reflexively elevate rCBF and slow the EEG as part of the oxygen-conserving (diving) reflex initiated in these neurons by hypoxia or ischaemia.
Asunto(s)
Tronco Encefálico/fisiología , Circulación Cerebrovascular , Electroencefalografía , Hipoxia Encefálica/fisiopatología , Bulbo Raquídeo/fisiopatología , Animales , Transporte Axonal , Presión Sanguínea , Estimulación Eléctrica , Estimulantes Ganglionares/farmacología , Masculino , Bulbo Raquídeo/irrigación sanguínea , Bulbo Raquídeo/efectos de los fármacos , Nicotina/farmacología , Ratas , Ratas Sprague-Dawley , Formación Reticular/fisiología , Cianuro de Sodio/farmacología , VasodilataciónRESUMEN
Antagonists of glutamate receptors of the N-methyl-d-aspartate subclass (NMDAR) or inhibitors of nitric oxide synthase (NOS) prevent nervous system plasticity. Inflammatory and neuropathic pain rely on plasticity, presenting a clinical opportunity for the use of NMDAR antagonists and NOS inhibitors in chronic pain. Agmatine (AG), an endogenous neuromodulator present in brain and spinal cord, has both NMDAR antagonist and NOS inhibitor activities. We report here that AG, exogenously administered to rodents, decreased hyperalgesia accompanying inflammation, normalized the mechanical hypersensitivity (allodynia/hyperalgesia) produced by chemical or mechanical nerve injury, and reduced autotomy-like behavior and lesion size after excitotoxic spinal cord injury. AG produced these effects in the absence of antinociceptive effects in acute pain tests. Endogenous AG also was detected in rodent lumbosacral spinal cord in concentrations similar to those previously detected in brain. The evidence suggests a unique antiplasticity and neuroprotective role for AG in processes underlying persistent pain and neuronal injury.
Asunto(s)
Agmatina/uso terapéutico , Analgésicos/uso terapéutico , Inflamación/complicaciones , Dolor/tratamiento farmacológico , Enfermedades del Sistema Nervioso Periférico/complicaciones , Traumatismos de la Médula Espinal/complicaciones , Animales , Inmunohistoquímica , Masculino , Ratones , N-Metilaspartato/fisiología , Dolor/etiología , Ratas , Ratas Sprague-DawleyRESUMEN
We investigated whether the medullary cerebrovasodilator area (MCVA), a region of ventral medulla mediating elevations of regional cerebral blood flow (rCBF) and electroencephalogram (EEG) synchronization elicited in cerebral cortex from stimulation of reticulospinal neurons of rostral ventrolateral medulla (RVLM), also mediates comparable responses from the cerebellar fastigial nucleus (FN). In spinalized rats, electrical stimulation of MCVA, RVLM or FN elevated rCBF and synchronized the EEG. The FN-evoked responses were significantly attenuated or blocked by bilateral lesions of MCVA. The MCVA is a novel region of medullary reticular formation mediating actions of medullary and cerebellar centers on rCBF and EEG to link visceral centers of brainstem and cerebral cortex.
Asunto(s)
Núcleos Cerebelosos/fisiología , Sincronización Cortical/métodos , Bulbo Raquídeo/fisiología , Vasodilatación/fisiología , Animales , Núcleos Cerebelosos/irrigación sanguínea , Estimulación Eléctrica/métodos , Electrólisis , Núcleos Talámicos Intralaminares/irrigación sanguínea , Masculino , Bulbo Raquídeo/irrigación sanguínea , Ratas , Ratas Sprague-Dawley , Resistencia Vascular/fisiologíaRESUMEN
The imidazoline-1 receptor (IR1) is considered a novel target for drug discovery. Toward cloning an IR1, a truncated cDNA clone was isolated from a human hippocampal lambda gt11 cDNA expression library by relying on the selectivity of two antisera directed against candidate IR proteins. Amplification reactions were performed to extend the 5' and 3' ends of this cDNA, followed by end-to-end PCR and conventional cloning. The resultant 5131-basepair molecule, designated imidazoline receptor-antisera-selected (IRAS) cDNA, was shown to encode a 1504-amino acid protein (IRAS-1). No relation exists between the amino acid sequence of IRAS-1 and proteins known to bind imidazolines (e.g., it is not an alpha2-adrenoceptor or monoamine oxidase subtype). However, certain sequences within IRAS-1 are consistent with signaling motifs found in cytokine receptors, as previously suggested for an IR1. An acidic region in IRAS-1 having an amino acid sequence nearly identical to that of ryanodine receptors led to the demonstration that ruthenium red, a dye that binds the acidic region in ryanodine receptors, also stained IRAS-1 as a 167-kD band on SDS gels and inhibited radioligand binding of native I1 sites in untransfected PC-12 cells (a source of authentic I1 binding sites). Two epitope-selective antisera were also generated against IRAS-1, and both reacted with the same 167-kD band on Western blots. In a host-cell-specific manner, transfection of IRAS cDNA into Chinese hamster ovary cells led to high-affinity I1 binding sites by criteria of nanomolar affinity for moxonidine and rilmenidine. Thus, IRAS-1 is the first protein discovered with characteristics of an IR1.
Asunto(s)
Receptores de Droga/genética , Receptores de Droga/inmunología , Receptores de Droga/metabolismo , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Animales , Western Blotting , Células CHO/metabolismo , Células COS/metabolismo , Clonidina/análogos & derivados , Clonidina/metabolismo , Clonación Molecular , Cricetinae , ADN Complementario , Epinefrina/metabolismo , Humanos , Idazoxan/metabolismo , Imidazoles/metabolismo , Receptores de Imidazolina , Sueros Inmunes , Radioisótopos de Yodo , Datos de Secuencia Molecular , Nafazolina/metabolismo , Rojo de Rutenio/química , Rojo de Rutenio/metabolismo , Lugares Marcados de Secuencia , Coloración y Etiquetado , Transfección , Yohimbina/metabolismoRESUMEN
We compared the properties of mammalian arginine decarboxylase (ADC) and ornithine decarboxylase (ODC) in rat liver and brain. Mammalian ADC is thermally unstable and associated with mitochondrial membranes. ADC decarboxylates both arginine (Km = 0.75 mM) and ornithine (Km = 0.25 mM), a reaction not inhibited by the specific ODC inhibitor, difluoromethylomithine. ADC activity is inhibited by Ca2+, Co2+, and polyamines, is present in many organs being highest in aorta and lowest in testis, and is not recognized by a specific monoclonal antibody to ODC. In contrast, ODC is thermally stable, cytosolic, and mitochondrial and is expressed at low levels in most organs except testis. Although ADC and ODC are expressed in cultured rat C6 glioma cells, the patterns of expression during growth and confluence are very different. We conclude that mammalian ADC differs from ADC isoforms expressed in plants, bacteria, or Caenorhabditis elegans and is distinct from ODC. ADC serves to synthesize agmatine in proximity to mitochondria, an organelle also harboring agmatine's degradative enzyme, agmatinase, and a class of imidazoline receptor (I2) to which agmatine binds with high affinity.
Asunto(s)
Encéfalo/enzimología , Carboxiliasas/metabolismo , Hígado/enzimología , Animales , Calcio/farmacología , Cromatografía en Capa Delgada , Estabilidad de Enzimas , Immunoblotting , Cinética , Masculino , Ornitina Descarboxilasa/metabolismo , Ratas , Ratas Sprague-Dawley , Fracciones Subcelulares/enzimología , Especificidad por Sustrato , Distribución Tisular , Células Tumorales Cultivadas/enzimologíaRESUMEN
Recent evidence suggests that agmatine, which is an intermediate in polyamine biosynthesis, might be an important neurotransmitter in mammals. Agmatine is synthesized in the brain, stored in synaptic vesicles in regionally selective neurons, accumulated by uptake, released by depolarization, and inactivated by agmatinase. Agmatine binds to alpha2-adrenoceptors and imidazoline binding sites, and blocks NMDA receptor channels and other ligand-gated cationic channels. Furthermore, agmatine inhibits nitric oxide synthase, and induces the release of some peptide hormones. As a result of its ability to inhibit both hyperalgesia and tolerance to, and withdrawal from, morphine, and its neuroprotective activity, agmatine has potential as a treatment of chronic pain, addictive states and brain injury.
Asunto(s)
Agmatina , Encéfalo/fisiología , Neurotransmisores/fisiología , Animales , Química Encefálica/fisiología , HumanosRESUMEN
The fundamental neuronal substrates of the arterial baroreceptor reflex have been elucidated by combining anatomical, neurophysiological, and pharmacological approaches. A serial pathway between neurons located in the nuclei of the solitary tract (NTS), the caudal ventrolateral medulla (CVL), and the rostral ventrolateral medulla (RVL) plays a critical role in inhibition of sympathetic outflow following stimulation of baroreceptor afferents. In this paper, we summarize our studies using tract-tracing and electron microscopic immunocytochemistry to define the potential functional sites for synaptic transmission within this circuitry. The results are discussed as they relate to the literature showing: (1) baroreceptor afferents excite second-order neurons in NTS through the release of glutamate; (2) these NTS neurons in turn send excitatory projections to neurons in the CVL; (3) GABAergic CVL neurons directly inhibit RVL sympathoexcitatory neurons; and (4) activation of this NTS-->CVL-->RVL pathway leads to disfacilitation of sympathetic preganglionic neurons by promoting withdrawal of their tonic excitatory drive, which largely arises from neurons in the RVL. Baroreceptor control may also be regulated over direct reticulospinal pathways exemplified by a newly recognized sympathoinhibitory region of the medulla, the gigantocellular depressor area. This important autonomic reflex may also be influenced by parallel, multiple, and redundant networks.
Asunto(s)
Barorreflejo/fisiología , Bulbo Raquídeo/citología , Bulbo Raquídeo/fisiología , Inhibición Neural/fisiología , Vías Nerviosas/citología , Vías Nerviosas/fisiología , Presorreceptores/citología , Presorreceptores/fisiología , Sistema Nervioso Simpático/citología , Sistema Nervioso Simpático/fisiología , Animales , Neuronas/citología , Neuronas/fisiología , RatasRESUMEN
Electrical stimulation of the cerebellar fastigial nucleus (FN) in spontaneously hypertensive (SHR), Wistar-Kyoto (WKY) and Fisher rats reduced, by approximately 50%, the infarctions produced by occlusion of the middle cerebral artery. Blockade of ATP-dependent potassium (K-ATP) channels with glibenclamide (i.c.v.) abolished salvage only in the SHR rat. While blockade of K-ATP channels failed to abolish salvage in WKY and Fisher rats, participation of potassium channels in neurogenic neuroprotection cannot be excluded.
Asunto(s)
Núcleos Cerebelosos/fisiología , Infarto Cerebral/prevención & control , Infarto Cerebral/fisiopatología , Estimulación Eléctrica , Gliburida/farmacología , Canales de Potasio/fisiología , Adenosina Trifosfato/fisiología , Animales , Núcleos Cerebelosos/fisiopatología , Ventrículos Cerebrales/efectos de los fármacos , Ventrículos Cerebrales/fisiología , Ventrículos Cerebrales/fisiopatología , Inyecciones Intraventriculares , Masculino , Bloqueadores de los Canales de Potasio , Ratas , Ratas Endogámicas F344 , Ratas Endogámicas SHR , Ratas Endogámicas WKYRESUMEN
Agmatine, an amine and organic cation, is an endogenous ligand at alpha 2-adrenergic and imidazoline (I-) receptors, to which it binds with high affinity. In addition, agmatine has properties of an endogenous neurotransmitter. Thus, agmatine (a) is locally synthesized in brain by a specific enzyme, arginine decarboxylase; (b) is stored in a large number of neurons with selective distribution in the CNS; (c) is associated with small vesicles in axon terminals that, at least in hippocampus, make synaptic asymmetric (excitatory) synapses on pyramidal cells; (d) is released from synaptosomes in a Ca(2+)-dependent manner; (e) can be enzymatically degraded by agmatinase in synaptosomes; (f) can be inactivated by selective reuptake; (g) blocks the ligand-gated NMDA receptor channel at sites distinct from ligand-binding and polyamine sites; and (h) has systemic actions when administered intraventricularly. Additionally, (i) agmatine is a precursor of brain putrescine and, hence, of higher polyamines, and (j) it competitively inhibits the activity of all isozymes of nitric oxide synthase. Agmatine meets most criteria to establish it as a novel neurotransmitter/neuromodulator in the CNS. However, agmatine differs from forms of clonidine displacing system with respect to distribution, bioactivity, and capacity to interact with antibodies raised to imidazoline-like drugs. Thus, there are multiple endogenous ligands of the imidazoline receptors, one of which is agmatine.
Asunto(s)
Agmatina/metabolismo , Encéfalo/metabolismo , Receptores de Droga/metabolismo , Agmatina/farmacología , Animales , Arginina/metabolismo , Unión Competitiva , Clonidina/metabolismo , Humanos , Receptores de Imidazolina , Cinética , Ligandos , Receptores Adrenérgicos alfa 2/metabolismoRESUMEN
We have shown that cultured vascular smooth muscle cells (VSMC) and brain astroglial cells express I-receptors of the I2 subtype. While imidazoline agents are anti-proliferative in smooth muscle cells, they increase the expression of glial fibrillary acidic protein (GFAP) in astrocytes. Because increases in GFAP suppress the induction of calcium-independent, inducible nitric oxide synthase (NOS-2), we measured whether idazoxan and related imidazolines and agmatine would also suppress the expression of NOS-2. Cultured astrocytes and macrophages, RAW 264.7 cell line, were incubated with lipopolysaccharide (LPS, 1 microgram/ml) or cytokine mixture in the presence of 1-100 microM of idazoxan, agmatine, or other imidazoline agents. Idazoxan potently (IC50 10 microM) decreased the activity of NOS-2 in astrocytes, but was less potent in RAW 264.7 cells. By contrast, agmatine was most potent in RAW 264.7 cells (IC50, 10 microM) but less potent in glial cells and VSMC. Both idazoxan and agmatine decreased the activity of NOS-2 by reducing the levels of enzyme protein as measured by immunoblot and immunocytochemistry. No specific binding of [3H]-idazoxan was observed in RAW 264.7 cell membranes. We conclude that idazoxan, agmatine, and selected imidazoline agents inhibit the expression of NOS-2 and proliferation in primary glial cells and VSMC. While the antiproliferative actions appear mediated by I-receptors of the I2 type, the anti-inflammatory response is probably not mediated by I-receptors but possibly by direct actions on signal transduction enzymes.
Asunto(s)
Astrocitos/efectos de los fármacos , Corteza Cerebral/citología , Idazoxan/farmacología , Imidazoles/farmacología , Macrófagos/citología , Receptores de Droga/fisiología , Agmatina/farmacología , Animales , Animales Recién Nacidos , Astrocitos/citología , Astrocitos/fisiología , División Celular/efectos de los fármacos , Línea Celular , Células Cultivadas , Corteza Cerebral/fisiología , Inducción Enzimática/efectos de los fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Receptores de Imidazolina , Inflamación , Macrófagos/efectos de los fármacos , Macrófagos/fisiología , Ratones , Óxido Nítrico Sintasa/biosíntesis , Óxido Nítrico Sintasa de Tipo II , Ratas , Ratas Sprague-Dawley , Receptores de Droga/genéticaRESUMEN
Electrical stimulation of the cerebellar fastigial nucleus (FN) elevates regional cerebral blood flow (rCBF) and arterial pressure (AP) and provides long-lasting protection against focal and global ischemic infarctions. We investigated which neuronal element in FN, perikarya or axons, mediates this central neurogenic neuroprotection and whether it also protects against excitotoxicity. In anesthetized rats, the FN was stimulated for 1 hr, and ibotenic acid (IBO) was microinjected unilaterally into the striatum. In unstimulated controls, the excitotoxic lesions averaged approximately 40 mm3. Stimulation of FN, but not dentate nucleus (DN), significantly reduced lesion volumes up to 80% when IBO was injected 15 min, 72 hr, or 10 d, but not 30 d, thereafter. In other rats, intrinsic neurons of FN or DN were destroyed by pretreatment with IBO. Five days later, the FN was stimulated, and 72 hr later, IBO was microinjected into the striatum. Lesions of FN, but not DN, abolished neuroprotection but not the elevations of rCBF and AP elicited from FN stimulation. Excitotoxic lesions of FN, but not DN, also abolished the 37% reduction in focal ischemic infarctions produced by middle cerebral artery occlusion. Excitation of intrinsic FN neurons provides long-lasting, substantial, and reversible protection of central neurons from excitotoxicity, as well as focal ischemia, whereas axons in the nucleus, probably collaterals of ramified brainstem neurons, mediate the elevations in rCBF, which do not contribute to neuroprotection. Long-lived protection against a range of injuries is an unrecognized function of FN neurons transmitted over pathways distinct from those regulating rCBF.
Asunto(s)
Núcleos Cerebelosos/fisiología , Ataque Isquémico Transitorio/fisiopatología , Precondicionamiento Isquémico , Neuronas/fisiología , Animales , Análisis de los Gases de la Sangre , Presión Sanguínea/fisiología , Estimulación Eléctrica , Antagonistas de Aminoácidos Excitadores/farmacología , Glucosa/metabolismo , Hematócrito , Ácido Iboténico/farmacología , Ataque Isquémico Transitorio/patología , Masculino , Vías Nerviosas/fisiología , Neuronas/patología , Ratas , Ratas Sprague-Dawley , Receptores de N-Metil-D-Aspartato/antagonistas & inhibidoresRESUMEN
Reticulospinal sympathoexcitatory neurons of rostral ventrolateral medulla (RVL) are selectively excited by hypoxia to elevate arterial pressure (AP) and cerebral blood flow (rCBF), that are elements of the oxygen-conserving (diving) reflex. We investigated whether KATP+-channels participate in this. Tolbutamide and glibenclamide, KATP+-channel blockers, microinjected into RVL in anesthetized rats, dose-dependently and site-specifically elevated AP and rCBF and potentiated responses to hypoxemia. KATP+-channels may mediate hypoxic excitation of oxygen-sensing RVL neurons.
Asunto(s)
Circulación Cerebrovascular/fisiología , Hipoxia Encefálica/fisiopatología , Bulbo Raquídeo/citología , Neuronas/fisiología , Canales de Potasio/fisiología , Adenosina Trifosfato/fisiología , Animales , Presión Sanguínea/fisiología , Química Encefálica/fisiología , Gliburida/farmacología , Hipoglucemiantes/farmacología , Hipoxia/fisiopatología , Masculino , Bulbo Raquídeo/irrigación sanguínea , Bulbo Raquídeo/química , Neuronas/química , Neuronas/efectos de los fármacos , Oxígeno/farmacología , Ratas , Ratas Sprague-Dawley , Centro Respiratorio/irrigación sanguínea , Centro Respiratorio/química , Centro Respiratorio/citología , Tolbutamida/farmacologíaRESUMEN
In rat, electrical stimulation of the cerebellar fastigial nucleus (FN) for 1 h reduces the volume of focal ischemic infarctions produced by occluding the middle cerebral artery (MCAO), even 10 days later. The mechanism by which this 'central neurogenic neuroprotection' salvages ischemic brain is not known but does not result from changes in cerebral perfusion. MCAO also triggers periodic periinfarction depolarizing waves (PIDs) in the ischemic penumbra, the territory of salvage. These may contribute to neuronal death and promote infarct expansion. Conceivably, FN stimulation, which can otherwise modify cortical excitability, may alter the development of PIDs. We investigated in anesthetized rats whether FN stimulation modifies PIDs expression and, if so, the threshold for evoking cortical spreading depression (CSD), a process sharing characteristics with PIDs and an index of cortical excitability. Stimulation of FN immediately or 72 h before MCAO decreased infarction volumes by approximately 45% (p<0.01), increased PID latency >10-fold, and decreased the number of PIDs by >50% (p<0.001). In normal rats, stimulation of FN increased the threshold current for eliciting CSD by 175% and slowed its propagation velocity by 35% (p<0.01 for each) immediately, but not 72 h, after FN stimulation. We conclude: FN stimulation elicits long-lasting suppression of PIDs in parallel with neuroprotection. However, PIDs suppression over time is unlikely to result from a major increase in cortical tolerance to depolarization and probably is not the principal mechanism of salvage.