RESUMO
Genetic self-incompatibility in Brassica is determined by alleles of the transmembrane serine-threonine kinase SRK, which functions in the stigma epidermis, and of the cysteine-rich peptide SCR, which functions in pollen. Using tagged versions of SRK and SCR as well as endogenous stigma and pollen proteins, we show that SCR binds the SRK ectodomain and that this binding is allele specific. Thus, SRK and SCR function as a receptor-ligand pair in the recognition of self pollen. Specificity in the self-incompatibility response derives from allele-specific formation of SRK-SCR complexes at the pollen-stigma interface.
Assuntos
Alelos , Brassica/genética , Brassica/metabolismo , Proteínas de Plantas/metabolismo , Estruturas Vegetais/metabolismo , Pólen/metabolismo , Proteínas Quinases/metabolismo , Sítios de Ligação , Fertilização/fisiologia , Ligantes , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Plantas Tóxicas , Ligação Proteica , Proteínas Quinases/química , Proteínas Quinases/genética , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Especificidade da Espécie , Especificidade por Substrato , NicotianaRESUMO
The carotid body chemoreceptor afferents have been reported to project to a discrete area located in the nucleus commissuralis of nucleus tractus solitarius [A. Vardhan et al., Am. J. Physiol., 264 (1993) R41-R50]. The afore-mentioned study was done in spontaneously breathing rats and the afferents and efferents located in the chest wall and the respiratory tract of these animals were intact. In order to exclude the role, if any, of these afferents and efferents, in the present experiments respiratory changes were monitored by recording the phrenic nerve activity instead of tracheal airflow. Experiments were carried out in pentobarbital-anesthetized, bilaterally vagotomized, paralyzed and artificially ventilated rats with a pneumothorax. The carotid body chemoreceptors were stimulated with tracheal administration of nitrogen for 7-10 s. The chemoreceptor stimulation induced an increase in the frequency and amplitude of phrenic nerve bursts. A decrease in the duration of inspiratory (T1), expiratory (TE) and total cycles (TTOT) was observed in the phrenic nerve activity. Inhibition of neuronal cell bodies by microinjections of muscimol (140 pmol/20 nl) into a discrete area in the commissural subnucleus of the nucleus tractus solitarius (coordinates in mm: 0.3 rostral to 0.5 caudal, 0 to 0.5 lateral and 0.3 to 0.5 deep with respect to the calamus scriptorius), attenuated the phrenic nerve responses to the carotid body stimulation. On the other hand, control injections of saline (0.9%) into this site did not alter the phrenic nerve response to the carotid body stimulation.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Corpo Carotídeo/fisiologia , Nervo Frênico/fisiologia , Núcleo Solitário/fisiologia , Animais , Bicuculina/farmacologia , Pressão Sanguínea/efeitos dos fármacos , Corpo Carotídeo/efeitos dos fármacos , Frequência Cardíaca/efeitos dos fármacos , Masculino , Microinjeções , Muscimol/antagonistas & inibidores , Muscimol/farmacologia , Vias Neurais/anatomia & histologia , Vias Neurais/efeitos dos fármacos , Vias Neurais/fisiologia , Nitrogênio/farmacologia , Nervo Frênico/anatomia & histologia , Nervo Frênico/efeitos dos fármacos , Ratos , Ratos Wistar , Mecânica Respiratória/efeitos dos fármacos , Núcleo Solitário/anatomia & histologia , Núcleo Solitário/efeitos dos fármacos , Estimulação QuímicaRESUMO
Microinjections of alpha 1-adrenergic receptor agonists into the intermediolateral cell column of the spinal cord (IML) elicit sympathoexcitatory responses. This observation, together with the identification of projections of epinephrine-containing cells in the rostral ventrolateral medullary pressor area (VLPA) to the IML, has prompted speculation that epinephrine may mediate pressor responses to the stimulation of the VLPA. This hypothesis was tested in pentobarbital-anesthetized, artificially ventilated, male Wistar rats. A mesenteric arterial branch was cannulated for monitoring blood pressure. Pressor responses were elicited predominantly from T8-T10 by injections (1.7 nmol/20 nl) of L-glutamate into the IML; maximum pressor responses (29.3 +/- 4 mmHg) were elicited from T9. Pressor responses were also elicited by injections of epinephrine into the IML at T9; maximum pressor effect (16.3 +/- 1.2 mmHg) was elicited by a dose of 0.05 pmol/20 nl. This effect of epinephrine at T9 was blocked by prior injections of prazosin (a selective alpha 1-adrenergic receptor blocker; 0.125 pmol/20 nl) at the same site. Stimulation of the VLPA by unilateral microinjections of glutamate elicited pressor responses (56 +/- 12 mmHg). Bilateral injections of prazosin at T8-T10, in the dose (0.125 pmol) that blocked a maximally effective dose of epinephrine, did not block the pressor responses to subsequent injections of glutamate into the VLPA. On the other hand, bilateral microinjections of AP-7 (an NMDA receptor blocker; 1 nmol/20 nl), but not DNQX (10 pmol; a non-NMDA receptor blocker), into the IML at T8-T10 blocked the pressor effects of the subsequent injections of glutamate into the VLPA.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Agonistas alfa-Adrenérgicos/administração & dosagem , Pressão Sanguínea/fisiologia , Neurônios/efeitos dos fármacos , Receptores Adrenérgicos alfa 1/fisiologia , Medula Espinal/fisiologia , Glândulas Suprarrenais/fisiologia , Aminoácidos/farmacologia , Animais , Epinefrina/farmacologia , Masculino , Artérias Mesentéricas/efeitos dos fármacos , Microinjeções , Prazosina/farmacologia , Quinoxalinas/farmacologia , Ratos , Ratos Wistar , Medula Espinal/citologia , Medula Espinal/efeitos dos fármacos , Sistema Nervoso Simpático/efeitos dos fármacos , Sistema Nervoso Simpático/fisiologiaRESUMO
Stimulation of cardio-pulmonary vagal afferent C fiber endings by right atrial injections of phenylbiguanide (PBG, 40 micrograms/kg) elicited apnea, bradycardia and hypotension. These responses were abolished by bilateral vagotomy low in the neck. Stimulation of neurons in a specific nucleus tractus solitarius site (0.5 mm rostral, 0.5 mm lateral and 0.5 mm deep with reference to the calamus scriptorius) by injections of L-glutamate produced responses similar to those following the stimulation of cardio-pulmonary vagal afferent C fiber endings by PBG. Inhibition of neurons in this NTS site by injections of muscimol abolished the responses to PBG. Specific blockade of NMDA receptors by microinjections of AP-7 (100 pmol) or non-NMDA receptors (KA and AMPA) by injections of DNQX (10-25 pmol) into this NTS site did not block the responses to PBG. Microinjections of kynurenic acid (1-4 nmol) into this NTS site blocked the responses to right atrial injections of PBG. These results indicate that: (1) blockade of either NMDA receptors or non-NMDA receptors (KA and AMPA) alone in the above-mentioned NTS site does not abolish the responses to the stimulation of cardio-pulmonary vagal afferent C fiber endings; (2) it is necessary to block NMDA as well as non-NMDA receptors in this NTS site for abolishing the responses to the stimulation of these nerve endings.
Assuntos
Terminações Nervosas/efeitos dos fármacos , Fibras Nervosas/efeitos dos fármacos , Receptores de Aminoácido/fisiologia , Núcleo Solitário/fisiologia , Nervo Vago/efeitos dos fármacos , Vias Aferentes/efeitos dos fármacos , Animais , Biguanidas/farmacologia , Coração/inervação , Pulmão/inervação , Masculino , Microinjeções , Muscimol/administração & dosagem , Quinoxalinas/farmacologia , Ratos , Ratos Wistar , Núcleo Solitário/efeitos dos fármacosRESUMO
Using yeast forward and reverse two-hybrid analysis and biochemical techniques, we present novel and definitive in vivo and in vitro evidence that both the N-terminal domain I and C-terminal domain IV of the host-encoded DnaA initiator protein of Escherichia coli interact physically with plasmid-encoded RepA initiator of pSC101. The N-terminal, but not the C-terminal, region of RepA interacted with DnaA in vitro. These protein-protein interactions are critical for two very early steps of replication initiation, namely origin unwinding and helicase loading. Neither domain I nor IV of DnaA could individually collaborate with RepA to promote pSC101 replication. However, when the two domains are co-expressed within a common cell milieu and allowed to associate non-covalently with each other via a pair of leucine zippers, replication of the plasmid was supported in vivo. Thus, the result shows that physical tethering, either non-covalent or covalent, of domain I and IV of DnaA and interaction of both domains with RepA, are critical for replication initiation. The results also provide the molecular basis for a novel, potential, replication-based bacterial two-hybrid system.
Assuntos
Proteínas de Bactérias/metabolismo , Replicação do DNA , DNA Bacteriano/biossíntese , Proteínas de Ligação a DNA/metabolismo , Proteínas/metabolismo , Transativadores , Trifosfato de Adenosina/metabolismo , Proteínas de Bactérias/genética , Sítios de Ligação , Cromatografia de Afinidade , DNA Helicases/metabolismo , Proteínas de Ligação a DNA/genética , DnaB Helicases , Ensaio de Imunoadsorção Enzimática/métodos , Escherichia coli/genética , Zíper de Leucina , Mutagênese , Plasmídeos , Dobramento de Proteína , Estrutura Terciária de Proteína , Proteínas/genética , Origem de Replicação , Saccharomyces cerevisiae , Técnicas do Sistema de Duplo-HíbridoRESUMO
Stimulation of carotid body chemoreceptors by saline saturated with 100% CO2 elicited an increase in mean arterial pressure, respiratory rate, tidal volume, and minute ventilation (VE). Microinjections of L-glutamate into a midline area 0.5-0.75 mm caudal and 0.3-0.5 mm deep with respect to the calamus scriptorius increased VE. Histological examination showed that the site was located in the commissural nucleus of the nucleus tractus solitarii (NTS). The presence of excitatory amino acid receptors [N-methyl-D-aspartic acid (NMDA); kainate, quisqualate/alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) and trans 1-amino-cyclopentane-trans-1,3-dicarboxylic acid (ACPD)] in this area was demonstrated by microinjections of appropriate agonists. Simultaneous blockade of NMDA and non-NMDA receptors by combined injections of DL-2-aminophosphonoheptanoate (AP-7; 1 nmol) and 6,7-dinitro-quinoxaline-2,3-dione (DNQX; 1 nmol) abolished the responses to stimulation of carotid body on either side. Combined injections of AP-7 and DNQX did not produce a nonspecific depression of neurons because the responses to another agonist, carbachol, remained unaltered. Inhibition of the neurons in the aforementioned area with microinjections of muscimol (which hyperpolarizes neuronal cell bodies but not fibers of passage) also abolished the responses to subsequent carotid body stimulation on either side.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Corpo Carotídeo/fisiologia , Células Quimiorreceptoras/fisiologia , Bulbo/metabolismo , Receptores de Aminoácido/fisiologia , Respiração/fisiologia , Animais , Dióxido de Carbono/farmacologia , Corpo Carotídeo/efeitos dos fármacos , Células Quimiorreceptoras/efeitos dos fármacos , Masculino , Ratos , Ratos Wistar , Receptores de Aminoácido/metabolismo , Estimulação Química , Transmissão SinápticaRESUMO
Small doses of epinephrine (0.008, 0.05, and 0.1 pmol, i.e., 20-nl volumes of 0.40, 2.5, and 5 microM solutions) produced a dose-dependent increase in heart rate when micro-injected into the right intermediolateral column (IML) at T2 spinal level. These effects were mediated via alpha 1-adrenergic receptors because prazosin blocked them. The presence of alpha 1-adrenergic receptors at this site was confirmed by microinjections of phenylephrine (a specific agonist for these receptors); phenylephrine elicited tachycardia. Larger doses of epinephrine (320, 2,000, and 3,200 pmol, i.e., 20-nl volumes of 16, 100, and 160 mM solutions) caused bradycardia when microinjected into the IML. These effects were mediated via alpha 2-adrenergic receptors because idazoxan blocked them. The presence of alpha 2-adrenergic receptors at this site was confirmed by microinjections of clonidine (a specific agonist for these receptors); clonidine elicited bradycardia. Injections of the vehicle (20 nl of normal saline containing 0.3% ascorbic acid, pH 7.4) did not evoke a response. Epinephrine, prazosin, or idazoxan did not alter the responses to L-glutamate. None of the doses of epinephrine elicited any response when injected intravenously. The aforementioned results provide pharmacological evidence for the presence of alpha 1- and alpha 2-adrenergic receptors in the IML at T2. Thus a basis is provided for investigating the role, if any, of alpha-adrenergic receptors in the IML in mediating cardiovascular responses elicited by the stimulation of different brain stem areas.
Assuntos
Epinefrina/administração & dosagem , Coração/efeitos dos fármacos , Medula Espinal/fisiologia , Animais , Bradicardia/induzido quimicamente , Clonidina/farmacologia , Relação Dose-Resposta a Droga , Frequência Cardíaca/efeitos dos fármacos , Injeções Espinhais , Masculino , Microinjeções , Fenilefrina/farmacologia , Ratos , Ratos Wistar , Taquicardia/induzido quimicamenteRESUMO
Herpes zoster is a common viral illness presenting with vesicular eruptions which are usually preceded by pain, erythema, and tenderness in a dermatomal distribution. The ophthalmic division of the trigeminal nerve is commonly involved (herpes zoster ophthalmicus). Early diagnosis before eruption of vesicles can be difficult and symptoms may be confused with other neurologic disorders. We present a patient with herpes zoster ophthalmicus who presented with face and neck pain associated with visual symptoms mimicking carotid artery dissection. Atypical presentation and benefits of early antiviral treatment are discussed.
Assuntos
Aneurisma/diagnóstico , Doenças das Artérias Carótidas/diagnóstico , Dor Facial/etiologia , Herpes Zoster Oftálmico/diagnóstico , Aciclovir/uso terapêutico , Antivirais/uso terapêutico , Diagnóstico Diferencial , Feminino , Herpes Zoster Oftálmico/complicações , Herpes Zoster Oftálmico/tratamento farmacológico , Humanos , Pessoa de Meia-IdadeRESUMO
Giant cell arteritis should not be a diagnosis of exclusion, an afterthought, or a last thought. There is urgency to establishing this diagnosis and initiating therapy. All practitioners who treat adults will be confronted with these patients. Some will have classic presentations, some will have subtle presentations. When patients complain of fever, fatigue, malaise, weight loss, or painless vision loss, GCA should be suspected. An ESR will aid in the diagnosis (although a normal ESR does not rule it out), and sometimes temporal artery biopsy will provide certainty. Giant cell arteritis is usually easy to recognize, easy to treat, and satisfying to manage.