RESUMO
Phoenixin (PNX) is a 14-amino acid amidated peptide (PNX-14) or an N-terminal extended 20-residue amidated peptide (PNX-20) recently identified in neural and non-neural tissue. Mass spectrometry analysis identified a major peak corresponding to PNX-14, with negligible PNX-20, in mouse spinal cord extracts. Using a previously characterized antiserum that recognized both PNX-14 and PNX-20, PNX-immunoreactivity (irPNX) was detected in a population of dorsal root ganglion (DRG) cells and in cell processes densely distributed to the superficial layers of the dorsal horn; irPNX cell processes were also detected in the skin. The retrograde tracer, Fluorogold, injected subcutaneously (s.c.) to the back of the cervical and thoracic spinal cord of mice, labeled a population of DRG, some of which were also irPNX. PNX-14 (2, 4 and 8 mg/kg) injected s.c.to the nape of the neck provoked dose-dependent repetitive scratching bouts directed to the back of the neck with the hindpaws. The number of scratching bouts varied from 16 to 95 in 30 min, commencing within 5 min post-injection and lasted 10-15 min. Pretreatment of mice at -20 min with nalfurafine (20 µg/kg, s.c.), the kappa opioid receptor agonist, significantly reduced the number of bouts induced by PNX-14 (4 mg/kg) compared with that of saline-pretreated mice. Our results suggest that the peptide, PNX-14, serves as one of the endogenous signal molecules transducing itch sensation in the mouse.
Assuntos
Gânglios Espinais/metabolismo , Peptídeos/metabolismo , Prurido/induzido quimicamente , Medula Espinal/metabolismo , Aminoácidos/química , Animais , Comportamento Animal/efeitos dos fármacos , Medula Cervical/metabolismo , Masculino , Camundongos , Pescoço/inervação , Peptídeos/química , Peptídeos/farmacologia , Peptídeos/fisiologia , Pele/metabolismoRESUMO
Phoenixin-14 amide, herein referred to as phoenixin, is a newly identified peptide from the rat brain. Using a previously characterized rabbit polyclonal antiserum against phoenixin, enzyme-immunoassay detected a high level (>4.5 ng/g tissue) of phoenixin-immunoreactivity (irPNX) in the rat spinal cords. Immunohistochemical studies revealed irPNX in networks of cell processes in the superficial dorsal horn, spinal trigeminal tract and nucleus of the solitary tract; and in a population of dorsal root, trigeminal and nodose ganglion cells. The pattern of distribution of irPNX in the superficial layers of the dorsal horn was similar to that of substance P immunoreactivity (irSP). Double-labeling the dorsal root ganglion sections showed that irPNX and irSP express in different populations of ganglion cells. In awake mice, intrathecal injection of phoenixin (1 or 5 µg) did not significantly affect the tail-flick latency as compared to that in animals injected with artificial cerebrospinal fluid (aCSF). Intrathecal administration of phoenixin (0.5, 1.25 or 2.5 µg) significantly reduced the number of writhes elicited by intraperitoneal injection of acetic acid (0.6%, 0.3 ml/30 g) as compared to that in mice injected with aCSF. While not affecting the tail-flick latency, phoenixin antiserum (1:100) injected intrathecally 10 min prior to the intraperitoneal injection of acetic acid significantly increased the number of writhes as compared to mice pre-treated with normal rabbit serum. Intrathecal injection of non-amidated phoenixin (2.5 µg) did not significantly alter the number of writhes evoked by acetic acid. Our result shows that phoenixin is expressed in sensory neurons of the dorsal root, nodose and trigeminal ganglia, the amidated peptide is bioactive, and exogenously administered phoenixin may preferentially suppress visceral as opposed to thermal pain.
Assuntos
Gânglios Sensitivos/fisiologia , Hormônios Hipotalâmicos/fisiologia , Hormônios Peptídicos/fisiologia , Ácido Acético , Animais , Interpretação Estatística de Dados , Gânglios Sensitivos/metabolismo , Hormônios Hipotalâmicos/metabolismo , Hormônios Hipotalâmicos/farmacologia , Técnicas Imunoenzimáticas , Imuno-Histoquímica , Masculino , Bulbo/efeitos dos fármacos , Bulbo/metabolismo , Medição da Dor/efeitos dos fármacos , Hormônios Peptídicos/metabolismo , Hormônios Peptídicos/farmacologia , Ratos , Ratos Sprague-Dawley , Tempo de Reação/efeitos dos fármacos , Tempo de Reação/fisiologia , Medula Espinal/metabolismo , Medula Espinal/fisiologiaRESUMO
Normal anterior pituitary function is essential for fertility. Release from the gland of the reproductive hormones luteinising hormone and follicle-stimulating hormone is regulated primarily by hypothalamically-derived gonadotrophin-releasing hormone (GnRH), although other releasing factors (RF) have been postulated to exist. Using a bioinformatic approach, we have identified a novel peptide, phoenixin, that regulates pituitary gonadotrophin secretion by modulating the expression of the GnRH receptor, an action with physiologically relevant consequences. Compromise of phoenixin in vivo using small interfering RNA resulted in the delayed appearance of oestrus and a reduction in GnRH receptor expression in the pituitary. Phoenixin may represent a new class of hypothalamically-derived pituitary priming factors that sensitise the pituitary to the action of other RFs, rather than directly stimulating the fusion of secretary vesicles to pituitary membranes.
Assuntos
Hormônios Hipotalâmicos/metabolismo , Hormônios Peptídicos/metabolismo , Hormônios Hipofisários/isolamento & purificação , Reprodução/genética , Sequência de Aminoácidos , Animais , Células Cultivadas , Clonagem Molecular , Relação Dose-Resposta a Droga , Feminino , Fármacos para a Fertilidade/química , Fármacos para a Fertilidade/isolamento & purificação , Fármacos para a Fertilidade/metabolismo , Fármacos para a Fertilidade/farmacologia , Hormônio Liberador de Gonadotropina/genética , Hormônio Liberador de Gonadotropina/metabolismo , Hormônios Hipotalâmicos/genética , Hormônios Hipotalâmicos/isolamento & purificação , Hormônios Hipotalâmicos/farmacologia , Hormônio Luteinizante/sangue , Masculino , Dados de Sequência Molecular , Hormônios Peptídicos/genética , Hormônios Peptídicos/isolamento & purificação , Hormônios Peptídicos/farmacologia , Hormônios Hipofisários/genética , Hormônios Hipofisários/metabolismo , Hormônios Hipofisários/farmacologia , Ratos , Ratos Sprague-Dawley , Reprodução/efeitos dos fármacos , Reprodução/fisiologia , Homologia de Sequência de AminoácidosRESUMO
Neuronostatin (NST) is a newly identified peptide of 13-amino acids encoded by the somatostatin (SST) gene. Using a rabbit polyclonal antiserum against the human NST, neuronostatin-immunoreactive (irNST) cells comparable in number and intensity to somatostatin immunoreactive (irSST) cells were detected in the hypothalamic periventricular nucleus. Fewer and/or less intensely labeled irNST cells were noted in other regions such as the hippocampus, cortex, amygdala, and cerebellum. Double-labeling hypothalamic sections with NST- and SST-antiserum revealed an extensive overlapping of irNST and irSST cells in the periventricular nucleus. Pre-absorption of the NST-antiserum with NST (1 microg/ml) but not with SST (1 microg/ml) abrogated irNST and vice versa. The activity of NST on dissociated and cultured hypothalamic neurons was assessed by the Ca(2+) imaging method. NST (10, 100, 1000 nM) concentration-dependently elevated intracellular Ca(2+) concentrations [Ca(2+)](i) in a population of hypothalamic neurons with two distinct profiles: (1) a fast and transitory increase in [Ca(2+)](i), and (2) an oscillatory response. Whereas, SST (100 nM) reduced the basal [Ca(2+)](i) in 21 of 61 hypothalamic neurons examined; an increase was not observed in any of the cells. Optical imaging with a slow-responding voltage sensitive dye DiBAC(4)(3) showed that NST (100 nM) depolarized or hyperpolarized; whereas, SST (100 nM) hyperpolarized a population of hypothalamic neurons. The result shows that NST and SST, though derived from the same precursor protein, exert different calcium mobilizing effects on cultured rat hypothalamic neurons, resulting in diverse cellular activities.
Assuntos
Cálcio/metabolismo , Hipotálamo/metabolismo , Neurônios/metabolismo , Hormônios Peptídicos/metabolismo , Somatostatina/metabolismo , Análise de Variância , Animais , Células Cultivadas , Hipotálamo/citologia , Hipotálamo/efeitos dos fármacos , Imuno-Histoquímica , Masculino , Potenciais da Membrana/efeitos dos fármacos , Neurônios/citologia , Neurônios/efeitos dos fármacos , Hormônios Peptídicos/farmacologia , Ratos , Ratos Sprague-Dawley , Somatostatina/farmacologiaRESUMO
1. The short-circuit current (Isc) technique was used to study the role of 5-hydroxytryptamine (5-HT) in the regulation of anion secretion in cultured rat cauda epididymal epithelia. 2. 5-HT, the 5-HT1B-selective agonist 5-nonyloxytryptamine (5-NOT) and the 5-HT2B-selective agonist alpha-methyl-5-hydroxytryptamine (alpha-methyl-5-HT) added basolaterally stimulated Isc in a dose-dependent manner with EC50 values of 0.4, 20 and 0.3 microM, respectively. No other agonists for 5-HT receptors had any effect. 3. The pattern of responses to 5-HT was biphasic. Pretreating the tissues with the 5-HT1B-selective antagonist isamoltane (200 microM) and the 5-HT2B-selective antagonist rauwolscine (200 microM) inhibited the rapid transient phase by 55 and 45 %, whereas the sustained phase could only be blocked by rauwolscine. 4. Removal of chloride or bicarbonate or both from the normal Krebs-Henseleit solution reduced the responses to 5-HT, 5-NOT and alpha-methyl-5-HT to varying degrees. The results suggest that 5-HT1B- and 5-HT2B-mediated responses were mainly due to chloride and bicarbonate secretion, respectively. 5. Manipulation of the cAMP and Ca2+ signal transduction pathways with chemical agents provided evidence that the responses to 5-HT were mediated through cAMP. 6. Piroxicam pretreatment abolished the Isc response to alpha-methyl-5-HT but not to 5-NOT, indicating that the 5-HT2B-mediated response, but not the 5-HT1B-mediated response, is dependent on prostaglandin synthesis. 7. Immunohistochemical studies showed that 5-HT-like immunoreactivity was detected in nerve fibres and in small granular cells surrounding the epididymal tubules. 8. It is suggested that the 5-HT released from serotonergic nerve endings and/or from mast cells regulates electrolyte and fluid secretion in the epididymis.
Assuntos
Ânions/metabolismo , Epididimo/metabolismo , Receptores de Serotonina/metabolismo , Serotonina/metabolismo , Sulfonamidas , Animais , Quelantes/farmacologia , Colforsina/farmacologia , Técnicas de Cultura , AMP Cíclico/metabolismo , Ácido Egtázico/análogos & derivados , Ácido Egtázico/farmacologia , Inibidores Enzimáticos/farmacologia , Epitélio/metabolismo , Iminas/farmacologia , Isoquinolinas/farmacologia , Masculino , Ratos , Ratos Sprague-Dawley , Receptor 5-HT1B de Serotonina , Receptor 5-HT2B de Serotonina , Receptores de Serotonina/efeitos dos fármacos , Tapsigargina/farmacologiaRESUMO
Distribution of neuronal nitric oxide synthase-immunoreactive (nNOS-IR) nerve fibers and somata in the rat epididymis and major pelvic ganglia was studied by immunohistochemical methods. In the epididymis, the supply of nNOS-IR fibers was highest in the cauda and became progressively fewer toward the caput. In the cauda and corpus, nNOS-IR fibers were distributed throughout the subepithelial tissues and around the epithelial. The pattern of distribution of vasoactive intestinal polypeptide (VIP)- and tyrosine hydroxylase (TH)-immunoreactive fibers in the epididymis was similar but the latter was generally more numerous in a given region as compared to that of nNOS-IR fibers. A population of neurons in the major pelvic ganglia were nNOS-IR-, TH- or VIP-IR. Double-labeling studies revealed that few neurons in the major pelvic ganglia contained both nNOS-IR and TH-IR. Whereas nNOS-IR and VIP-IR appeared to co-localize in the same population of the pelvic ganglion cells. Similarly, nNOS-IR fibers in the epididymis were mostly VIP-positive and TH-negative. Unilateral injection of the fluorescent tracer Fluorogold into the junction between the vas deferens and the cauda labeled a population of neurons in the right and left major pelvic ganglia, some of which were also nNOS-IR. A small number of dorsal root ganglion cells contained Fluorogold and very few expressed NOS-IR. It may be concluded that nNOS-IR nerve fibers in the rat epididymis arise mainly from neurons in the major pelvic ganglia the major of which express VIP-IR but not TH-IR. The extensive supply of nNOS-immunoreactive fibers around the epithelium and throughout the subepithelial tissues suggests that NO may be closely associated with smooth muscle contraction.
Assuntos
Epididimo/enzimologia , Fibras Nervosas/enzimologia , Neurotransmissores/análise , Óxido Nítrico Sintase/análise , Estilbamidinas , Animais , Corantes Fluorescentes , Gânglios Espinais/citologia , Gânglios Espinais/enzimologia , Plexo Hipogástrico/citologia , Plexo Hipogástrico/enzimologia , Imuno-Histoquímica , Masculino , Neurônios/enzimologia , Fenótipo , Ratos , Ratos Sprague-Dawley , Tirosina 3-Mono-Oxigenase/análise , Peptídeo Intestinal Vasoativo/análiseRESUMO
The aim of the present study was to evaluate the distribution of pituitary adenylate cyclase-activating polypeptide (PACAP)-like immunoreactivity in the mouse spinal cord using an antibody against PACAP38 and to determine the behavioral profile, particularly with respect to hyperalgesia, of PACAP38 given intrathecally (i.t.) in the mouse. Immunoreactivity to PACAP38 was detected in numerous nerve fibers in the superficial layers of the dorsal horn of cervical, thoracic, lumbar and sacral segments and a few fibers extended into the deeper layers of the spinal cord. In addition, PACAP-like immunoreactivity were seen in the intermediolateral cell column of the thoracic and sacral segments. In behavioral studies, PACAP38 (0.05-0.5 microgram) produced a dose-dependent decrease of the tail-flick latency when given i.t. in the mouse. At higher doses (1-10 micrograms), PACAP38 given i.t. elicited biting and scratching behaviors lasting 10-20 min after the injection. PACAP at high doses (1-10 micrograms) also produced licking at tail, paw and penis and intense grooming behaviors immediately after the i.t. injection. Similar to substance P, these behaviors produced by PACAP can be considered as pain-like syndrome. These findings suggest that PACAP may be a sensory neurotransmitter involved in nociceptive signalling in the mouse spinal cord.
Assuntos
Neuropeptídeos/farmacologia , Neurotransmissores/farmacologia , Medição da Dor/efeitos dos fármacos , Medula Espinal/efeitos dos fármacos , Animais , Comportamento Animal/efeitos dos fármacos , Relação Dose-Resposta a Droga , Masculino , Camundongos , Camundongos Endogâmicos ICR , Neuropeptídeos/imunologia , Neurotransmissores/imunologia , Polipeptídeo Hipofisário Ativador de Adenilato CiclaseRESUMO
Immuno-electronmicroscopic studies were performed to detect the presence and features of synaptic contacts between pituitary adenylate cyclase activating polypeptide immunoreactive (PACAP-ir) axons and cholera toxin B-horseradish peroxidase labeled preganglionic sympathetic neurons (PSNs) in the intermediolateral nucleus of the rat thoracic spinal cord. PACAP-ir axon varicosities, which contained small clear and large core synaptic vesicles, were found to form asymmetric type of synaptic contacts with dendrites and infrequently with somata of labeled preganglionic neurons. The present study provides ultrastructural evidence of PACAP-ir synaptic contacts with PSNs, raising the possibility that the peptide may function as a transmitter/modulator to these neurons.
Assuntos
Axônios/metabolismo , Neuropeptídeos/metabolismo , Medula Espinal/metabolismo , Sinapses/metabolismo , Animais , Axônios/ultraestrutura , Gânglios Simpáticos/citologia , Gânglios Simpáticos/metabolismo , Região Lombossacral , Masculino , Microscopia Imunoeletrônica , Neurônios/metabolismo , Neurônios/ultraestrutura , Polipeptídeo Hipofisário Ativador de Adenilato Ciclase , Ratos , Ratos Sprague-Dawley , Medula Espinal/citologia , Medula Espinal/ultraestrutura , Sinapses/ultraestrutura , TóraxRESUMO
Immunohistochemical studies using an antibody against pituitary adenylate cyclase activating polypeptide-38 (PACAP) were performed on spinal cords and dorsal root ganglia harvested from two human cadavers. PACAP-like immunoreactivity (PACAP-LI) was detected in nerve fibers of the superficial layers of the dorsal horn, a few of which extended into the deeper laminae and as far as the ventral horn. At the thoracic segments, additional PACAP-LI nerve fibers were seen in the lateral funiculus projecting into the intermediolateral cell column. Dorsal root ganglia contained numerous PACAP-LI cell bodies of varying intensity. As a control, immunoreactivity to calcitonin gene-related peptide (CGRP) and substance P(SP) was also studied and found to be in nerve fibers of the substantia gelatinosa of the dorsal horn and in dorsal root ganglion cells. These results show that the pattern of distribution of PACAP-LI in the human spinal cord and dorsal root ganglia is similar to that of rodents and further suggest that PACAP may participate in sensory and autonomic functions.
Assuntos
Gânglios Espinais/metabolismo , Neuropeptídeos/metabolismo , Neurotransmissores/metabolismo , Medula Espinal/metabolismo , Peptídeo Relacionado com Gene de Calcitonina/metabolismo , Humanos , Imuno-Histoquímica , Fibras Nervosas/metabolismo , Polipeptídeo Hipofisário Ativador de Adenilato Ciclase , Substância P/metabolismoRESUMO
Rat adrenal chromaffin cells were invested by a dense network of nerve fibers immunoreactive to pituitary adenylate cyclase activating polypeptide-38 (PACAP-IR). Immunohistochemical studies demonstrated the presence of PACAP-IR in nodose and dorsal root ganglion cells, but not in neurons of the intermediolateral cell column and other autonomic nuclei of the thoracic and upper lumbar spinal cord. Somata of the T7 to T12 paravertebral ganglia were PACAP-negative. A few lightly labeled neurons were occasionally noted in the dorsal motor nucleus of the vagus. Injection of the retrograde tracer Fluorogold into the left adrenal medulla 3 days prior to sacrifice resulted in the labeling of a population of neurons in the ipsilateral spinal cord intermediolateral cell column (T1 to L1), ipsilateral and contralateral nodose ganglia and ipsilateral dorsal root ganglia from T7 to T10 inclusive. A small number of lightly labeled somata was occasionally noted in the dorsal motor nucleus of the vagus. Combined retrograde tracing and PACAP immunohistochemistry showed that a population of Fluorogold-containing nodose and dorsal root ganglion cells were also PACAP-positive. Pre-treatment of the rats with capsaicin caused a marked reduction of the PACAP-IR in the adrenal gland as well as in the superficial layers of the dorsal horn and caudal spinal trigeminal nucleus. These findings, in conjunction with the apparent absence of PACAP-IR in spinal sympathetic preganglionic neurons, sympathetic postganglionic neurons, and dorsal motor nucleus of the vagus, raise the possibility that PACAP-IR fibers observed in the adrenal medulla are primarily sensory in origin. As a corollary, catecholamine secretion from chromaffin cells may be modulated by the peptidergic sensory afferents in addition to the cholinergic sympathetic preganglionic nerve fibers.
Assuntos
Medula Suprarrenal/inervação , Neurônios Aferentes/fisiologia , Neuropeptídeos/fisiologia , Neurotransmissores/fisiologia , Estilbamidinas , Medula Suprarrenal/efeitos dos fármacos , Medula Suprarrenal/metabolismo , Animais , Capsaicina/farmacologia , Sistema Cromafim/citologia , Sistema Cromafim/metabolismo , Feminino , Corantes Fluorescentes , Gânglios Espinais/citologia , Gânglios Espinais/metabolismo , Gânglios Simpáticos/citologia , Gânglios Simpáticos/metabolismo , Imuno-Histoquímica , Masculino , Fibras Nervosas/efeitos dos fármacos , Fibras Nervosas/metabolismo , Fibras Nervosas/fisiologia , Neurônios Aferentes/efeitos dos fármacos , Neurônios Aferentes/metabolismo , Neuropeptídeos/metabolismo , Neurotransmissores/metabolismo , Gânglio Nodoso/citologia , Gânglio Nodoso/metabolismo , Polipeptídeo Hipofisário Ativador de Adenilato Ciclase , Ratos , Ratos Sprague-Dawley , Medula Espinal/citologia , Medula Espinal/efeitos dos fármacos , Medula Espinal/metabolismo , Fibras Simpáticas Pós-Ganglionares/metabolismo , Fibras Simpáticas Pós-Ganglionares/fisiologiaRESUMO
Immunohistochemical detection of Fos, the protein product of the immediate-early gene c-fos, was evaluated as a functional marker of central neurons sensitive to a change of blood pressure/blood volume. Controlled hemorrhage and infusion of the hypotensive agent nitroprusside or hydralazine induced the appearance of Fos-immunoreactivity (Fos-IR) in several prominent groups of central neurons: the piriform cortex, bed nucleus of the stria terminalis, islands of Calleja, subfornical organ, central nucleus of the amygdala, parabrachial nucleus, supraoptic and paraventricular nuclei, pontine A5, locus ceruleus, ventrolateral medulla, the nucleus of the solitary tract, area postrema, and intermediolateral cell column in the spinal cord. Elevation of blood pressure by infusion of phenylephrine caused the appearance of Fos-IR in fewer groups of neurons: the bed nucleus of the stria terminalis, central nucleus of the amygdala, parabrachial nucleus, the nucleus of the solitary tract and area postrema. The differential distribution of Fos neurons in hypotensive versus hypertensive animals underscores the potential application of Fos as a metabolic marker in identifying a network of neurons responding to a specific cardiovascular challenge. Further, simultaneous characterization of the transmitter phenotype of Fos-containing neurons offers an additional advantage of this method over other conventional tract-tracing techniques.
Assuntos
Sistema Cardiovascular/inervação , Expressão Gênica , Genes fos , Neurônios/citologia , Proteínas Proto-Oncogênicas c-fos/biossíntese , Animais , Biomarcadores , Encéfalo/citologia , Encéfalo/fisiologia , Neurônios/fisiologia , Especificidade de Órgãos , RatosRESUMO
Hemorrhage caused by withdrawing 3-4 cm3 of blood from adult rats over a period of 1-2 h induced the expression of Fos proteins, which were detected by immunocytochemical methods in neurons of the hypothalamic supraoptic (SON) and paraventricular (PVN) nuclei, area postrema, nucleus of the solitary tract (NTS), ventrolateral medulla (VLM) and in intermediolateral cell column (IML) of the spinal cord. Daily intraventricular (i.c.v.) injections of c-fos antisense (50 nmol 5 microliters -1) for 2 days prior to hemorrhage markedly attenuated the extent and intensity of Fos-immunoreactivity (FOS-IR) compared with that of rats injected with c-fos sense in the areas mentioned above. The results indicate the effectiveness of i.c.v. administration of c-fos antisense in blocking Fos expression in central neurons following hemorrhage.
Assuntos
Hemorragia Cerebral/metabolismo , Neurônios/metabolismo , Oligonucleotídeos Antissenso , Proteínas Proto-Oncogênicas c-fos/genética , Animais , Sequência de Bases , Injeções Intraventriculares , Masculino , Dados de Sequência Molecular , Proteínas Proto-Oncogênicas c-fos/biossíntese , Ratos , Ratos Sprague-DawleyRESUMO
Distribution of nitric oxide synthase (NOS), somatostatin (SSN), and parvalbumin (PV) was studied in the rat hippocampus by immunohistochemical methods. The aim was to explore the interrelationship between SSN-immunoreactive (SSN-IR) neurons in the dentate hilus, which have been shown to be vulnerable to a number of pathophysiological insults, and the presence or absence of NOS and/or PV in the same subset of dentate hilar neurons. Small NOS-IR neurons were scattered in the pyramidal, oriens, and radiatum layers of the CA1-CA3 areas and in the subiculum, where larger NOS-IR neurons were occasionally noted. In the area dentata, NOS-IR neurons, which were composed of small and large polymorphic cells, appeared as a single file at the hilar border with the granule cell layer and clustered in the hilus in fairly high density. Double-labeling techniques showed that most NOS-IR neurons in the hilus were SSN-IR, whereas coexistence of NOS and PV immunoreactivity or SSN and PV immunoreactivity was low in dentate hilar neurons. In other areas of the hippocampus, colocalization of NOS and SSN in the same neurons was much less frequent. Thus, SSN-IR neurons in the dentate hilus constitute a population of neurons that contain the enzyme NOS as well. The presence of NOS coupled to the lack or low level of PV in this group of neurons may provide a neurochemical basis for their high susceptibility to certain pathophysiological insults.
Assuntos
Aminoácido Oxirredutases/metabolismo , Hipocampo/metabolismo , Somatostatina/metabolismo , Animais , Feminino , Imunofluorescência , Hipocampo/anatomia & histologia , Técnicas Imunoenzimáticas , Masculino , Óxido Nítrico Sintase , Ratos , Ratos Sprague-DawleyRESUMO
In urethane anesthetized rats one hour after lowering the systolic blood pressure to 70-75 mmHg by withdrawing 3-4 ml of blood, Fos immunoreactivity (Fos-IR), confined to the cell nucleus, was detected bilaterally in numerous cells of the nucleus of the solitary tract (NTS) and ventrolateral medulla (VLM). A few Fos-IR neurons were observed in the lateral reticular nucleus, dorsal medullary reticular nucleus, spinal trigeminal nucleus, medial inferior olive, interfasciculus hypoglossi and paramedian rostral medulla. In sham-operated animals, a much smaller number of Fos-IR neurons were scattered in the NTS, VLM and other nuclei mentioned above. Double labeling with antisera to tyrosine-hydroxylase (TH) and phenylethanolamine-N-methyltransferase (PNMT) showed that 60% of TH-positive neurons in the NTS contained Fos-IR, and 70-80% of TH-positive neurons in the caudal VLM and 50-60% of PMNT-positive neurons in the rostral VLM expressed Fos-IR. Only a few TH- or PNMT-positive neurons in the C2, C3 (paramedian rostral medulla) areas and within the medial longitudinal fasciculus were Fos-IR. About 40% of PNMT/Fos-IR neurons in the rostral VLM contained the retrograde tracer fluorogold, which was injected (< 1 microliter) into the white matter dorsolateral to the intermediolateral cell column of T2-T3 segments 2 to 3 days prior to hemorrhagic experiments. Very few TH-positive neurons in the caudal VLM contained fluorogold. Finally, clusters of Fos-IR neurons, which also labeled with antisera to choline acetyltransferase, were detected in the intermediolateral cell column of the spinal cord. The results indicate that during hemorrhage aminergic neurons in the caudal and rostral VLM and in the NTS are activated insofar as c-fos expression is concerned. As a corollary, the monoaminergic neurons in the medulla constitute an essential component in the ascending as well as descending reflex pathway involved in the adjustment of cardiovascular dynamics during hemorrhage.
Assuntos
Catecolaminas/metabolismo , Hemorragia Cerebral/metabolismo , Bulbo/metabolismo , Neurônios/metabolismo , Proteínas Proto-Oncogênicas c-fos/biossíntese , Estilbamidinas , Animais , Feminino , Corantes Fluorescentes , Expressão Gênica/fisiologia , Imuno-Histoquímica , Masculino , Feniletanolamina N-Metiltransferase/imunologia , Feniletanolamina N-Metiltransferase/metabolismo , Ratos , Ratos Sprague-Dawley , Serotonina/metabolismo , Tirosina 3-Mono-Oxigenase/imunologia , Tirosina 3-Mono-Oxigenase/metabolismoRESUMO
Immunoreactivity to Fos protein (Fos-IR) was detected in rat hypothalamic neurons within 1 h of onset of hemorrhage by withdrawing 4-5 ml of blood, which lowered the arterial blood pressure to 50-70 mm Hg. About 70% of vasopressin (AVP)-containing neurons in the supraoptic nucleus (SON) and 20% in the paraventricular nucleus (PVN) expressed Fos-IR. In contrast, 5% of oxytocin (OXY)-containing neurons in the SON and < 1% in PVN were Fos-IR. Intravenous infusion of the vasodilating agent, nitroprusside, which lowered the blood pressure to levels comparable to that attained by hemorrhage, induced Fos-IR in greater than 65% of AVP-containing neurons in the SON, while relatively few AVP neurons in the PVN were Fos positive. These results suggest that hemorrhage or hypotension preferentially induces c-fos expression in supraoptic AVP-containing neurons.
Assuntos
Genes fos , Hipotensão/metabolismo , Neurônios/metabolismo , Nitroprussiato/farmacologia , Proteínas Proto-Oncogênicas c-fos/metabolismo , Núcleo Supraóptico/metabolismo , Animais , Arginina Vasopressina/análise , Hipotensão/induzido quimicamente , Hipotensão/patologia , Masculino , Neurônios/efeitos dos fármacos , Neurônios/patologia , Ocitocina/análise , Núcleo Hipotalâmico Paraventricular/efeitos dos fármacos , Núcleo Hipotalâmico Paraventricular/metabolismo , Núcleo Hipotalâmico Paraventricular/patologia , Proteínas Proto-Oncogênicas c-fos/análise , Ratos , Ratos Sprague-Dawley , Núcleo Supraóptico/efeitos dos fármacos , Núcleo Supraóptico/patologiaRESUMO
Immunoreactivities to Fos proteins were detected in numerous neurons in the supraoptic, paraventricular and accessory neurosecretory nuclei 1 h following withdrawal of 4-5 cc of blood from the rat femoral arteries. Few or no positive cells were observed in the same nuclei in sham-operated or control animals. It is concluded that hypovolemia induces c-fos expression in hypothalamic neurons known to be associated with blood volume/pressure regulation.