RESUMEN
Semiconducting nanoplatelets (NPLs) have attracted great attention due to the superior photophysical properties compared to their quantum dot analogs. Understanding and tuning the optical and electronic properties of NPLs in a plasmonic environment is a new paradigm in the field of optoelectronics. Here, we report on the resonant plasmon enhancement of light emission including Raman scattering and photoluminescence from colloidal CdSe/CdS nanoplatelets deposited on arrays of Au nanodisks fabricated by electron beam lithography. The localized surface plasmon resonance (LSPR) of the Au nanodisk arrays can be tuned by varying the diameter of the disks. In the case of surface-enhanced Raman scattering (SERS), the Raman intensity profile follows a symmetric Gaussian shape matching the LSPR of the Au nanodisk arrays. The surface-enhanced photoluminescence (SEPL) profile of NPLs, however, follows an asymmetric Gaussian distribution highlighting a compromise between the excitation and emission enhancement mechanisms originating from energy transfer and Purcell effects. The SERS and SEPL enhancement factors depend on the nanodisk size and reach maximal values at 75 and 7, respectively, for the sizes, for which the LSPR energy of Au nanodisks coincides with interband transition energies in the semiconductor platelets. Finally, to explain the origin of the resonant enhancement behavior of SERS and SEPL, we apply a numerical simulation to calculate plasmon energies in Au nanodisk arrays and emission spectra from NPLs in such a plasmonic environment.
RESUMEN
Surface- and tip-enhanced resonant Raman scattering (resonant SERS and TERS) by optical phonons in a monolayer of CdSe quantum dots (QDs) is demonstrated. The SERS enhancement was achieved by employing plasmonically active substrates consisting of gold arrays with varying nanocluster diameters prepared by electron-beam lithography. The magnitude of the SERS enhancement depends on the localized surface plasmon resonance (LSPR) energy, which is determined by the structural parameters. The LSPR positions as a function of nanocluster diameter were experimentally determined from spectroscopic micro-ellipsometry, and compared to numerical simulations showing good qualitative agreement. The monolayer of CdSe QDs was deposited by the Langmuir-Blodgett-based technique on the SERS substrates. By tuning the excitation energy close to the band gap of the CdSe QDs and to the LSPR energy, resonant SERS by longitudinal optical (LO) phonons of CdSe QDs was realized. A SERS enhancement factor of 2 × 10(3) was achieved. This allowed the detection of higher order LO modes of CdSe QDs, evidencing the high crystalline quality of QDs. The dependence of LO phonon mode intensity on the size of Au nanoclusters reveals a resonant character, suggesting that the electromagnetic mechanism of the SERS enhancement is dominant. Finally, the resonant TERS spectrum from CdSe QDs was obtained using electrochemically etched gold tips providing an enhancement on the order of 10(4). This is an important step towards the detection of the phonon spectrum from a single QD.
RESUMEN
We present a new classification for the genus Conus sensu lato (family Conidae), based on molecular phylogenetic analyses of 329 species. This classification departs from both the traditional classification in only one genus and from a recently proposed shell- and radula-based classification scheme that separates members of this group into five families and 115 genera. Roughly 140 genus-group names are available for Recent cone snails. We propose to place all cone snails within a single family (Conidae) containing four genera-Conus, Conasprella, Profundiconus and Californiconus (with Conus alone encompassing about 85% of known species)-based on the clear separation of cone snails into four distinct and well-supported groups/lineages in molecular phylogenetic analyses. Within Conus and Conasprella, we recognize 57 and 11 subgenera, respectively, that represent well-supported subgroupings within these genera, which we interpret as evidence of intrageneric distinctiveness. We allocate the 803 Recent species of Conidae listed as valid in the World Register of Marine Species into these four genera and 71 subgenera, with an estimate of the confidence for placement of species in these taxonomic categories based on whether molecular or radula and/or shell data were used in these determinations. Our proposed classification effectively departs from previous schemes by (1) limiting the number of accepted genera, (2) retaining the majority of species within the genus Conus and (3) assigning members of these genera to species groups/subgenera to enable the effective communication of these groups, all of which we hope will encourage acceptance of this scheme.
RESUMEN
We present a large-scale molecular phylogeny that includes 320 of the 761 recognized valid species of the cone snails (Conus), one of the most diverse groups of marine molluscs, based on three mitochondrial genes (COI, 16S rDNA and 12S rDNA). This is the first phylogeny of the taxon to employ concatenated sequences of several genes, and it includes more than twice as many species as the last published molecular phylogeny of the entire group nearly a decade ago. Most of the numerous molecular phylogenies published during the last 15years are limited to rather small fractions of its species diversity. Bayesian and maximum likelihood analyses are mostly congruent and confirm the presence of three previously reported highly divergent lineages among cone snails, and one identified here using molecular data. About 85% of the species cluster in the single Large Major Clade; the others are divided between the Small Major Clade (â¼12%), the Conus californicus lineage (one species), and a newly defined clade (â¼3%). We also define several subclades within the Large and Small major clades, but most of their relationships remain poorly supported. To illustrate the usefulness of molecular phylogenies in addressing specific evolutionary questions, we analyse the evolution of the diet, the biogeography and the toxins of cone snails. All cone snails whose feeding biology is known inject venom into large prey animals and swallow them whole. Predation on polychaete worms is inferred as the ancestral state, and diet shifts to molluscs and fishes occurred rarely. The ancestor of cone snails probably originated from the Indo-Pacific; rather few colonisations of other biogeographic provinces have probably occurred. A new classification of the Conidae, based on the molecular phylogeny, is published in an accompanying paper.
Asunto(s)
Caracol Conus/clasificación , Filogenia , Animales , Teorema de Bayes , Caracol Conus/genética , Evolución Molecular , Genes Mitocondriales , FilogeografíaRESUMEN
Experimental observations and theoretical studies show that nonlinear internal waves occur widely in shallow water and cause acoustic propagation effects including ducting and mode coupling. Horizontal ducting results when acoustic modes travel between internal wave fronts that form waveguide boundaries. For small grazing angles between a mode trajectory and a front, an interference pattern may arise that is a horizontal Lloyd mirror pattern. An analytic description for this feature is provided along with comparisons between results from the formulated model predicting a horizontal Lloyd mirror pattern and an adiabatic mode parabolic equation. Different waveguide models are considered, including boxcar and jump sound speed profiles where change in sound speed is assumed 12 m/s. Modifications to the model are made to include multiple and moving fronts. The focus of this analysis is on different front locations relative to the source as well as on the number of fronts and their curvatures and speeds. Curvature influences mode incidence angles and thereby changes the interference patterns. For sources oriented so that the front appears concave, the areas with interference patterns shrink as curvature increases, while convexly oriented fronts cause patterns to expand.
RESUMEN
Diversity among Conus toxins mirrors the high species diversity in the Indo-Pacific region, and evolution of both is thought to stem from feeding-niche specialization derived from intra-generic competition. This study focuses on Conus californicus, a phylogenetic outlier endemic to the temperate northeast Pacific. Essentially free of congeneric competitors, it preys on a wider variety of organisms than any other cone snail. Using molecular cloning of cDNAs and mass spectrometry, we examined peptides isolated from venom ducts to elucidate the sequences and post-translational modifications of two eight-cysteine toxins (cal12a and cal12b of type 12 framework) that block voltage-gated Na(+) channels. Based on homology of leader sequence and mode of action, these toxins are related to the O-superfamily, but differ significantly from other members of that group. Six of the eight cysteine residues constitute the canonical framework of O-members, but two additional cysteine residues in the N-terminal region define an O+2 classification within the O-superfamily. Fifteen putative variants of Cal12.1 toxins have been identified by mRNAs that differ primarily in two short hypervariable regions and have been grouped into three subtypes (Cal12.1.1-3). This unique modular variation has not been described for other Conus toxins and suggests recombination as a diversity-generating mechanism. We propose that these toxin isoforms show specificity for similar molecular targets (Na(+) channels) in the many species preyed on by C. californicus and that individualistic utilization of specific toxin isoforms may involve control of gene expression.
Asunto(s)
Caracol Conus/química , Venenos de Moluscos/genética , Péptidos/genética , Bloqueadores de los Canales de Sodio/toxicidad , Animales , Secuencia de Bases , California , Clonación Molecular , Cartilla de ADN/genética , Electrofisiología , Biblioteca de Genes , Espectrometría de Masas , Datos de Secuencia Molecular , Venenos de Moluscos/análisis , Venenos de Moluscos/clasificación , Océano Pacífico , Péptidos/análisis , Péptidos/clasificación , Análisis de Secuencia de ADNRESUMEN
Acute ischemic stroke (AIS) is a significant cause of global morbidity and mortality, with functional implications for quality of life and long-term disability. The limitations of intravenous thrombolytic therapy for the treatment of AIS, especially for emergent large vessel occlusion (ELVO), have paved the way for alternative therapies and the rapidly evolving landscape of endovascular therapy (EVT). Here, we summarize the major landmark trials that have advanced the field largely due to ongoing biomedical engineering device development that have translated into significantly improved clinical outcomes. Our review describes the clinical success of EVT, and current and future trends.
Asunto(s)
Accidente Cerebrovascular Isquémico , Trombectomía , Humanos , Accidente Cerebrovascular Isquémico/cirugía , Trombectomía/métodos , Trombectomía/tendenciasRESUMEN
Tip-enhanced Raman scattering (TERS) has recently emerged as a powerful technique for studying the local properties of low dimensional materials. Being a plasmon driven system, a dramatic enhancement of the TERS sensitivity can be achieved by an appropriate choice of the plasmonic substrate in the so-called gap-mode configuration. Here, we investigate the phonon properties of CdSe nanocrystals (NCs) utilizing gap-mode TERS. Using the Langmuir-Blodgett technique, we homogeneously deposited submonolayers of colloidal CdSe NCs on two different nanostructured plasmonic substrates. Amplified by resonant gap-mode TERS, the scattering by the optical phonon modes of CdSe NCs is markedly enhanced making it possible to observe up to the third overtone of the LO mode reliably. The home-made plasmonic substrates and TERS tips allow the analysis of the TERS images of CdSe phonon modes with nanometer spatial resolution. The CdSe phonon scattering intensity is strongly correlated with the local electromagnetic field distribution across the plasmonic substrates.
RESUMEN
Understanding the evolution of ecological specialization is important for making inferences about the origins of biodiversity. Members of the predatory, marine gastropod genus Conus exhibit a variety of diets and the ability to capture prey is linked to a venom comprised of peptide neurotoxins, termed conotoxins. We identified conotoxin transcripts from Conus leopardus, a species of Conus that uniquely preys exclusively on hemichordates, and compared its venom duct transcriptome to that of four other Conus species to determine whether a shift to a specialized diet is associated with changes in the venom composition of this species. We also examined the secondary structure of predicted amino acid sequences of conotoxin transcripts of C. leopardus to identify substitutions that may be linked to specialization on hemichordates. We identified seven distinct conotoxin sequences from C. leopardus that appear to represent transcripts of seven distinct loci. Expression levels and the diversity of conotoxins expressed by C. leopardus are considerably less than those of other Conus. Moreover, gene products of two transcripts exhibited unique secondary structures that have not been previously observed from other Conus. These results suggest that transition to a specialist diet is associated with reduction in the number of components expressed in venoms of Conus and that diverse venoms of Conus are maintained in species with a broad dietary width.
Asunto(s)
Caracol Conus/genética , Venenos de Moluscos/genética , Adaptación Biológica , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Ecología , Evolución Molecular , Biblioteca de Genes , Variación Genética , Datos de Secuencia Molecular , Alineación de Secuencia , Análisis de Secuencia de ADNRESUMEN
Venoms of predatory marine gastropods of the genus Conus show amazing levels of interspecific diversity and are comprised of a cocktail of peptide neurotoxins, termed conotoxins, that are encoded by large gene families. Conotoxin gene family evolution is characterized by gene duplications and high rates of nonsynonymous substitution among paralogues; yet, what controls the differentiation of venoms among species is not clear. We compared four-loop conotoxin transcripts of six closely related Conus species to examine conotoxin expression patterns among species. The species examined appear to express different numbers of four-loop conotoxin loci and similarity in expression patterns does not seem to correspond with phylogenetic affinity. Moreover, several loci appear to have been independently silenced while others appear to have been revived from previously silenced states. Some loci also appear to exhibit coordinated expression patterns. These results suggest that the evolution of conotoxin expression patterns is incredibly dynamic and the differentiation of venoms of Conus is controlled in part by the evolution of unique conotoxin expression patterns.
Asunto(s)
Conotoxinas/genética , Caracol Conus/genética , Evolución Molecular , Perfilación de la Expresión Génica , Animales , Conotoxinas/clasificación , Variación Genética , Biología Marina , FilogeniaRESUMEN
Atrial natriuretic factor (ANF) and C-type natriuretic peptide (CNP)-activated guanylate cyclases are single-chain transmembrane-spanning proteins, containing both ligand binding and catalytic activities. In both proteins, ligand binding to the extracellular receptor domain activates the cytosolic catalytic domain, generating the second messenger cyclic GMP. Obligatory in this activation process is an ATP-dependent step. ATP directly binds to a defined ATP-regulatory module (ARM) sequence motif in the cyclases and through ARM bridges the events of ligand binding and signal transduction. These ARM sequence motifs are respectively represented by Gly503-Xa-Gly505-Xa-Xa-Xa-Gly509 and Gly499-Xa-Xa-Xa-Gly503 in the case of ANF receptor guanylate cyclase (ANF-RGC) and CNP receptor guanylate cyclase (CNP-RGC). Through genetic remodeling techniques, we now show that ARM-Gly505 in ANF-RGC and the corresponding ARM-Gly499 in CNP-RGC are critical for ANF and CNP signaling, and other ARM-Gly residues have minimal effect in the respective signaling processes.
Asunto(s)
Adenosina Trifosfato/metabolismo , Factor Natriurético Atrial/metabolismo , Guanilato Ciclasa/metabolismo , Transducción de Señal , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Sitios de Unión , Línea Celular Transformada , Chlorocebus aethiops , ADN , Datos de Secuencia MolecularRESUMEN
In the pineal gland, the membrane guanylate cyclase activity was specifically stimulated by alpha(2D/A)-adrenergic receptor (alpha(2D/A)-AR) agonists. The agonists, however, did not stimulate the cyclase activity in the cell-free membranes. It was possible to stimulate the cyclase in cell-free membranes by the addition of the pineal soluble fraction, but this stimulation was Ca2+-dependent and alpha(2D/A)-agonist-independent. It was also possible to achieve Ca2+-dependent stimulation of the cyclase by the direct addition of CD-GCAP to the isolated pineal membranes. CD-GCAP is a Ca2+-binding protein and is a specific activator of one of the two members of the ROS-GC subfamily of membrane guanylate cyclases, ROS-GC1. The soluble fraction of the pineal gland stimulated recombinant ROS-GC1 in a Ca2+-dependent fashion. The direct presence of both ROS-GC1 and CD-GCAP in the pineal was established by molecular cloning/PCR studies. The findings demonstrate the existence of a novel signal transduction mechanism--the linkage of the alpha(2D/A)-AR signaling system with ROS-GC1 transduction system, occurring through intracellular Ca2+ via CD-GCAP.
Asunto(s)
Calcio/metabolismo , Guanilato Ciclasa/fisiología , Glándula Pineal/fisiología , Receptores Adrenérgicos alfa 2/fisiología , Animales , Células COS , Bovinos , Activación Enzimática , Glándula Pineal/enzimología , Transducción de SeñalRESUMEN
Atrial natriuretic factor (ANF) and C-type natriuretic peptide (CNP)-activated guanylate cyclases are single-chain transmembrane-spanning proteins, containing both ligand binding and catalytic activities. In both proteins, ligand binding to the extracellular receptor domain activates the cytosolic catalytic domain, generating the second messenger cyclic GMP. Studies with ANF receptor guanylate cyclase (ANF-RGC) have indicated that obligatory in this activation process is an ATP-dependent step. ATP directly binds to the cyclase and bridges the events of ligand binding and signal transduction. A defined ATP-regulated module (ARM) sequence (Gly503-Arg-Gly-Ser-Asn-Tyr-Gly509) in the cyclase is critical in the ATP-mediated event. Through genetic remodeling techniques, we have now identified the core ARM sequence that is essential in both ANF and CNP signaling. This sequence is Gly-Xa-Xa-Xa-Gly, represented by Gly505-Ser-Asn-Tyr-Gly509 in the case of ANF-RGC ARM and by Gly499-Ser-Ser-Tyr-Gly503 in the CNP receptor guanylate cyclase ARM.
Asunto(s)
Adenosina Trifosfato/metabolismo , Guanilato Ciclasa/metabolismo , Receptores del Factor Natriurético Atrial/metabolismo , Receptores de Superficie Celular/metabolismo , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Membrana Celular/metabolismo , Chlorocebus aethiops , Técnicas In Vitro , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida , Oligodesoxirribonucleótidos/química , Relación Estructura-ActividadRESUMEN
Biochemical and immunological studies have established that one of the signal transducers of atrial natriuretic factor (ANF) is a 180 kDa membrane guanylate cyclase (180 kDa mGC), which is also an ANF receptor; obligatory in the transduction process is an intervening ATP-regulated step, but its mechanism is not known. GC alpha is a newly discovered member of the guanylate cyclase family whose activity is independent of the known natriuretic peptides, and the enzyme is not an ANF receptor. The genetically tailored GC alpha, GC alpha-DmutGln338Leu364, however, is not only a guanylate cyclase but also an ANF receptor and is structurally and functionally identical to the cloned wild-type ANF receptor guanylate cyclase, GC-A. We now report that the ANF-dependent guanylate cyclase activity in the particulate fractions of cells transfected with GC alpha-DmutGln338Leu364 was inhibited by the 180 kDa mGC polyclonal antibody, and with this antibody probe it was possible to purify the 130 kDa expressed receptor; the hormone-dependent cyclase activity of this receptor was exclusively dependent upon ATP; and through site-directed mutational studies with GC alpha mutants, the signaling sequence that defines ATP binding site was identified. We thus conclude that 180 kDa mGC and the mutant protein are immunologically similar, both proteins are linked to the ANF signal in the generation of cyclic GMP synthesis; and in both the ligand binding and catalytic activities are bridged through a defined ATP binding module.
Asunto(s)
Adenosina Trifosfato/metabolismo , Factor Natriurético Atrial/metabolismo , Guanilato Ciclasa/genética , Transducción de Señal , Secuencia de Aminoácidos , Animales , Secuencia de Bases , ADN , Electroforesis en Gel de Poliacrilamida , Guanilato Ciclasa/inmunología , Guanilato Ciclasa/metabolismo , Datos de Secuencia Molecular , Ratas , Receptores del Factor Natriurético Atrial , Receptores de Superficie Celular/metabolismoRESUMEN
The membrane bound guanylyl cyclase (GC) photoreceptor membrane GC1 (ROS-GCI) of photoreceptor cells synthesizes cGMP, the intracellular transmitter of vertebrate phototransduction. The activity of ROS-GCI is controlled by small Ca(2+)-binding proteins, named GC-activating proteins (GCAPs). We identified and characterized two short regulatory regions (M445-L456 and L503-1522) in the juxtamembrane domain (JMD) of ROS-GC1 by peptide competition and mutagenesis studies. Both regions are critical for the activation of ROS-GCI by GCAP-1.
Asunto(s)
Proteínas de Unión al Calcio/farmacología , Calcio/farmacología , Guanilato Ciclasa/química , Células Fotorreceptoras de Vertebrados/metabolismo , Receptores de Superficie Celular , Secuencia de Aminoácidos , Animales , Células COS , Bovinos , Activación Enzimática/efectos de los fármacos , Guanilato Ciclasa/antagonistas & inhibidores , Guanilato Ciclasa/genética , Proteínas Activadoras de la Guanilato-Ciclasa , Datos de Secuencia Molecular , Mutación , Biblioteca de Péptidos , Péptidos/química , Péptidos/farmacología , Proteínas RecombinantesRESUMEN
The effect of epidermal growth factor (EGF) on the synthesis of the components of the cholesterol side-chain cleavage enzyme complex (SCC) was studied in rat ovarian granulosa cells. The cells were cultured for 48 h in the presence or absence of EGF (15 ng/ml) and/or FSH (50 ng/ml) after which proteins were radiolabeled by incubation with [35S]methionine followed by immunoprecipitation of newly synthesized P-450scc or adrenodoxin (ISP) with polyclonal antibodies directed against the corresponding proteins from bovine adrenal cortex. In addition the action of EGF on the level of translatable RNA for P-450scc was evaluated using a cell-free translation system programmed with RNA isolated from treated and untreated cells, followed by immunoisolation of newly synthesized proteins. Immunoisolated proteins were separated by polyacrylamide-gel electrophoresis, visualized by fluorography and quantified by densitometry. EGF stimulated progesterone formation by the cells 3-fold and potentiated the FSH-induced stimulation of progesterone formation, but had no effect on cAMP accumulation. EGF also stimulated the synthesis of P-450scc and ISP, and enhanced the FSH-induced synthesis of P-450scc and ISP in a concentration-dependent fashion with a maximal stimulation attained at concentrations ranging from 1.0 to 100 ng/ml. No appreciable changes in the induction pattern were observed when EGF and dibutyryl cyclic AMP (Bt2cAMP) were added together, as compared to when Bt2cAMP was added alone. Neither treatment affected the synthesis of the constitutive mitochondrial enzyme, F1-ATPase. Immunoisolation of P-450scc from the proteins synthesized in a rabbit reticulocyte in vitro translation system programmed with RNA isolated from EGF- and/or FSH-treated cells, revealed that EGF enhanced the FSH-stimulated synthesis of the precursor form of P-450scc.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/biosíntesis , Factor de Crecimiento Epidérmico/farmacología , Células de la Granulosa/enzimología , Oxidorreductasas/biosíntesis , Animales , Células Cultivadas , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/genética , Sistema Enzimático del Citocromo P-450/biosíntesis , Sistema Enzimático del Citocromo P-450/genética , Femenino , Células de la Granulosa/citología , Progesterona/biosíntesis , Biosíntesis de Proteínas/efectos de los fármacos , ARN Mensajero/genética , Ratas , Ratas EndogámicasRESUMEN
The original concept that cyclic GMP is one of the mediators of the hormone-dependent process of steroidogenesis has been strengthened by the characterization of a 180-kDa protein from rat adrenocortical carcinoma and rat and mouse testes. This protein appears to have an unusual characteristic of containing both the atrial natriuretic factor (ANF)-binding and guanylate cyclase activities, and appears to be intimately involved in the ANF-dependent steroidogenic signal transduction. In rat adrenal glands we now demonstrate: 1) the direct presence of a 180-kDa ANF-binding protein in GTP-affinity purified membrane fraction as evidenced by affinity cross-linking technique and by the Western blot analysis of the partially purified enzyme; 2) that the enzyme is biochemically and immunologically different from the soluble guanylate cyclase as there is no antigenic cross-reactivity of 180-kDa guanylate cyclase antibody with soluble guanylate cyclase; 3) in contrast to the soluble guanylate cyclase, the particulate enzyme is not stimulated by nitrite-generating compounds and hemin; and 4) protein kinase C inhibits both the basal and ANF-dependent guanylate cyclase activity and phosphorylates the 180-kDa guanylate cyclase. These results reveal the presence of a 180-kDa protein in rat adrenal glands and support the contention that: (a) this protein contains both the guanylate cyclase and ANF receptor; (b) the 180-kDa enzyme is coupled with the ANF-dependent cyclic GMP production; (c) the 180-kDa enzyme is biochemically distinct from the nonspecific soluble guanylate cyclase; and (d) there is a protein kinase C-dependent negative regulatory loop for the operation of ANF-dependent cyclic GMP signal pathway which acts via the phosphorylation of 180-kDa guanylate cyclase.
Asunto(s)
Glándulas Suprarrenales/ultraestructura , Guanilato Ciclasa/aislamiento & purificación , Receptores de Superficie Celular/análisis , Neoplasias de la Corteza Suprarrenal/inmunología , Glándulas Suprarrenales/metabolismo , Animales , Anticuerpos/análisis , Factor Natriurético Atrial/metabolismo , Azidas/farmacología , GMP Cíclico/biosíntesis , Hemina/farmacología , Hemoglobinas/farmacología , Nitroprusiato/farmacología , Fosforilación , Proteína Quinasa C/metabolismo , Conejos , Ratas , Receptores del Factor Natriurético Atrial , Receptores de Superficie Celular/metabolismo , Azida SódicaRESUMEN
Almost three decades of research in the field of photoreceptor guanylate cyclases are discussed in this review. Primarily, it focuses on the members of membrane-bound guanylate cyclases found in the outer segments of vertebrate rods. These cyclases represent a new guanylate cyclase subfamily, termed ROS-GC, which distinguishes itself from the peptide receptor guanylate cyclase family that it is not extracellularly regulated. It is regulated, instead, by the intracellularly-generated Ca2+ signals. A remarkable feature of this regulation is that ROS-GC is a transduction switch for both the low and high Ca2+ signals. The low Ca2+ signal transduction pathway is linked to phototransduction, but the physiological relevance of the high Ca2+ signal transduction pathway is not yet clear; it may be linked to neuronal synaptic activity. The review is divided into eight sections. In Section I, the field of guanylate cyclase is introduced and the scope of the review is briefly explained; Section II covers a brief history of the investigations and ideas surrounding the discovery of rod guanylate cyclase. The first five subsections of Section III review the experimental efforts to quantify the guanylate cyclase activity of rods, including in vitro and in situ biochemistry, and also the work done since 1988 in which guanylate cyclase activity has been determined. In the remaining three subsections an analytical evaluation of the Ca2+ modulation of the rod guanylate cyclase activity related to phototransduction is presented. Section IV deals with the issues of a biochemical nature: isolation and purification, subcellular localization and functional properties of rod guanylate cyclase. Section V summarizes work on the cloning of the guanylate cyclases, analysis of their primary structures, and determination of their location with in situ hybridization. Section VI summarizes studies on the regulation of guanylate cyclases, with a focus on guanylate cyclases activating proteins. In Section VII, the evidence about the localization and functional role of guanylate cyclases in other retinal cells, especially in "on-bipolar" cells, in which guanylate cyclase most likely plays a critical role in electrical signaling, is discussed. The review concludes with Section VIII, with remarks about the future directions of research on retinal guanylate cyclases.
Asunto(s)
Guanilato Ciclasa/metabolismo , Células Fotorreceptoras/enzimología , Animales , Guanilato Ciclasa/aislamiento & purificación , Humanos , Neuronas/enzimología , Retina/enzimología , Células Fotorreceptoras Retinianas Bastones/enzimología , Células Fotorreceptoras Retinianas Bastones/fisiología , Visión OcularRESUMEN
The suspect role of the receptor-mediated cyclic GMP signaling pathway was dispelled by the discovery of a membrane guanylate cyclase that was also an atrial natriuretic factor receptor. It is now established that the membrane guanylate cyclase transduction system is linked to the signaling of natriuretic factors, guanylin/enterotoxin, and emerging evidence suggests that a new neural tissue-specific subfamily of membrane guanylate cyclases exists whose mechanism of signal transduction is different from those of the other membrane cyclases. This review will briefly discuss the fascinating, albeit turbulent, history of this signal transduction field, which will be followed by its current status and finally the direction it is heading.