RESUMEN
Animal venoms and their chemical compounds have aroused both empirical and scientific attention for ages. However, there has been a significant increase in scientific investigations in recent decades, allowing the production of various formulations that are helping in the development of many important tools for biotechnological, diagnostic, or therapeutic use, both in human and animal health, as well as in plants. Venoms are composed of biomolecules and inorganic compounds that may have physiological and pharmacological activities that are not related to their principal actions (prey immobilization, digestion, and defense). Snake venom toxins, mainly enzymatic and non-enzymatic proteins, and peptides have been identified as potential prototypes for new drugs and/or models for the development of pharmacologically active structural domains for the treatment of cancer, cardiovascular diseases, neurodegenerative and autoimmune diseases, pain, and infectious-parasitic diseases. This minireview aims to provide an overview of the biotechnological potential of animal venoms, with a focus on snakes, and to introduce the reader to the fascinating world of Applied Toxinology, where animal biodiversity can be used to develop therapeutic and diagnostic applications for humans.
Asunto(s)
Neoplasias , Venenos de Serpiente , Animales , Humanos , Venenos de Serpiente/química , Serpientes/metabolismo , Proteínas/química , Péptidos/farmacología , Neoplasias/tratamiento farmacológicoRESUMEN
We examined the behavioural responses and Fos expression pattern of rats that were exposed to snake threats from shed snakeskin and a live snake. We differentiated the behavioural responses and the pattern of Fos expression in response to the odour cues and mild threat from a live snake. Animals exposed to the snake odour alone or to the confined snake showed a great deal of risk assessment. Conversely, the intensification of odour during exposure to the live snake decreased the threat ambiguity, and the animals froze for a significantly longer period. Our Fos analysis showed that a pathway formed by the posteroventral part of the medial amygdalar nucleus to the central part of the ventromedial hypothalamic nucleus appeared to be solely responsive to odour cues. In addition, we showed increased Fos expression in a parallel circuit comprising the lateral amygdalar nucleus, ventral subiculum, lateral septum, and juxtadorsomedial region of the lateral hypothalamic area that is responsive to both the odour and mild threat from a live snake. This path is likely to process the environmental boundaries of the threat to be avoided. Both paths merge into the dorsal premammillary nucleus and periaqueductal grey sites, which all increase Fos expression in response to the snake threats and are likely to organize the defensive responses. Moreover, we found that the snake threat mobilized the Edinger-Westphal and supraoculomotor nuclei, which are involved in stress adaptation and attentional mechanisms.
Asunto(s)
Complejo Nuclear Basolateral , Conducta Animal , Animales , Complejo Nuclear Basolateral/metabolismo , Conducta Animal/fisiología , Miedo/fisiología , Sustancia Gris Periacueductal/fisiología , Proteínas Proto-Oncogénicas c-fos/metabolismo , Ratas , Serpientes/metabolismoRESUMEN
Snakes are known to express a rod visual opsin and two cone opsins, only (SWS1, LWS), a reduced palette resulting from their supposedly fossorial origins. Dipsadid snakes in the genus Helicops are highly visual predators that successfully invaded freshwater habitats from ancestral terrestrial-only habitats. Here, we report the first case of multiple SWS1 visual pigments in a vertebrate, simultaneously expressed in different photoreceptors and conferring both UV and violet sensitivity to Helicops snakes. Molecular analysis and in vitro expression confirmed the presence of two functional SWS1 opsins, likely the result of recent gene duplication. Evolutionary analyses indicate that each sws1 variant has undergone different evolutionary paths with strong purifying selection acting on the UV-sensitive copy and dN/dS â¼1 on the violet-sensitive copy. Site-directed mutagenesis points to the functional role of a single amino acid substitution, Phe86Val, in the large spectral shift between UV and violet opsins. In addition, higher densities of photoreceptors and SWS1 cones in the ventral retina suggest improved acuity in the upper visual field possibly correlated with visually guided behaviors. The expanded visual opsin repertoire and specialized retinal architecture are likely to improve photon uptake in underwater and terrestrial environments, and provide the neural substrate for a gain in chromatic discrimination, potentially conferring unique color vision in the UV-violet range. Our findings highlight the innovative solutions undertaken by a highly specialized lineage to tackle the challenges imposed by the invasion of novel photic environments and the extraordinary diversity of evolutionary trajectories taken by visual opsin-based perception in vertebrates.
Asunto(s)
Visión de Colores , Opsinas , Animales , Agua Dulce , Opsinas/genética , Opsinas/metabolismo , Filogenia , Células Fotorreceptoras Retinianas Conos/metabolismo , Opsinas de Bastones/genética , Serpientes/genética , Serpientes/metabolismoRESUMEN
MOTIVATION: Next-generation sequencing has become exceedingly common and has transformed our ability to explore nonmodel systems. In particular, transcriptomics has facilitated the study of venom and evolution of toxins in venomous lineages; however, many challenges remain. Primarily, annotation of toxins in the transcriptome is a laborious and time-consuming task. Current annotation software often fails to predict the correct coding sequence and overestimates the number of toxins present in the transcriptome. Here, we present ToxCodAn, a python script designed to perform precise annotation of snake venom gland transcriptomes. We test ToxCodAn with a set of previously curated transcriptomes and compare the results to other annotators. In addition, we provide a guide for venom gland transcriptomics to facilitate future research and use Bothrops alternatus as a case study for ToxCodAn and our guide. RESULTS: Our analysis reveals that ToxCodAn provides precise annotation of toxins present in the transcriptome of venom glands of snakes. Comparison with other annotators demonstrates that ToxCodAn has better performance with regard to run time ($>20x$ faster), coding sequence prediction ($>3x$ more accurate) and the number of toxins predicted (generating $>4x$ less false positives). In this sense, ToxCodAn is a valuable resource for toxin annotation. The ToxCodAn framework can be expanded in the future to work with other venomous lineages and detect novel toxins.
Asunto(s)
Algoritmos , Biología Computacional/métodos , Perfilación de la Expresión Génica/métodos , Venenos de Serpiente/genética , Serpientes/genética , Toxinas Biológicas/genética , Animales , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Filogenia , Venenos de Serpiente/química , Venenos de Serpiente/metabolismo , Serpientes/clasificación , Serpientes/metabolismo , Especificidad de la Especie , Toxinas Biológicas/química , Toxinas Biológicas/metabolismoRESUMEN
Novel phenotypes are commonly associated with gene duplications and neofunctionalization, less documented are the cases of phenotypic maintenance through the recruitment of novel genes. Proteolysis is the primary toxic character of many snake venoms, and ADAM metalloproteinases, named snake venom metalloproteinases (SVMPs), are largely recognized as the major effectors of this phenotype. However, by investigating original transcriptomes from 58 species of advanced snakes (Caenophidia) across their phylogeny, we discovered that a different enzyme, matrix metalloproteinase (MMP), is actually the dominant venom component in three tribes (Tachymenini, Xenodontini, and Conophiini) of rear-fanged snakes (Dipsadidae). Proteomic and functional analyses of these venoms further indicate that MMPs are likely playing an "SVMP-like" function in the proteolytic phenotype. A detailed look into the venom-specific sequences revealed a new highly expressed MMP subtype, named snake venom MMP (svMMP), which originated independently on at least three occasions from an endogenous MMP-9. We further show that by losing ancillary noncatalytic domains present in its ancestors, svMMPs followed an evolutionary path toward a simplified structure during their expansion in the genomes, thus paralleling what has been proposed for the evolution of their Viperidae counterparts, the SVMPs. Moreover, we inferred an inverse relationship between the expression of svMMPs and SVMPs along the evolutionary history of Xenodontinae, pointing out that one type of enzyme may be substituting for the other, whereas the general (metallo)proteolytic phenotype is maintained. These results provide rare evidence on how relevant phenotypic traits can be optimized via natural selection on nonhomologous genes, yielding alternate biochemical components.
Asunto(s)
Evolución Molecular , Metaloproteinasas de la Matriz/metabolismo , Venenos de Serpiente/enzimología , Serpientes/metabolismo , Animales , Metaloproteinasas de la Matriz/genética , Fenotipo , Proteolisis , Venenos de Serpiente/genética , Serpientes/genética , TranscriptomaRESUMEN
Viruses are associated with several human diseases that infect a large number of individuals, hence directly affecting global health and economy. Owing to the lack of efficient vaccines, antiviral therapy and emerging resistance strains, many viruses are considered as a potential threat to public health. Therefore, researches have been developed to identify new drug candidates for future treatments. Among them, antiviral research based on natural molecules is a promising approach. Phospholipases A2 (PLA2s) isolated from snake venom have shown significant antiviral activity against some viruses such as Dengue virus, Human Immunodeficiency virus, Hepatitis C virus and Yellow fever virus, and have emerged as an attractive alternative strategy for the development of novel antiviral therapy. Thus, this review provides an overview of remarkable findings involving PLA2s from snake venom that possess antiviral activity, and discusses the mechanisms of action mediated by PLA2s against different stages of virus replication cycle. Additionally, molecular docking simulations were performed by interacting between phospholipids from Dengue virus envelope and PLA2s from Bothrops asper snake venom. Studies on snake venom PLA2s highlight the potential use of these proteins for the development of broad-spectrum antiviral drugs.
Asunto(s)
Antivirales/farmacología , Fosfolipasas A2/farmacología , Venenos de Serpiente/enzimología , Serpientes/metabolismo , Animales , Virus del Dengue/efectos de los fármacos , Farmacorresistencia Viral/efectos de los fármacos , VIH/efectos de los fármacos , Hepacivirus/efectos de los fármacos , Simulación del Acoplamiento Molecular , Proteínas de Reptiles/farmacología , Virus de la Fiebre Amarilla/efectos de los fármacosRESUMEN
Snakes are a useful model for ecological studies because they are gape-limited predators that may undergo ontogenetic changes in diet. We analyzed carbon and nitrogen stable isotope ratios to estimate percent contributions of different prey to snake biomass, trophic positions and isotopic niche width of juveniles and adults of the snake Thamnodynastes hypoconia. We also estimated the isotopic niche overlap between the two age categories. During eight intervals over a two-year period, we collected samples of whole blood and scales at a site in southern Brazil. Isotopic ratios of carbon and nitrogen did not differ between juveniles and adults for either tissue type, nor did mean trophic positions of juveniles and adults differ. The percent contribution of prey categories to snake biomass differed to a limited extent between the two years, with Hylidae being the most important anuran group assimilated during the first year and Leptodactylidae during the second year, for both ages. The isotopic niche occupied by adult snakes was slightly larger than that of juveniles when the analysis was based on data from whole blood samples, as expected because snakes are gape-limited. We found a reverse pattern when the analysis was based on scales, which may indicate that adult snakes have a smaller niche over the long term as they become selective foragers in certain prey. Isotopic overlap between juveniles and adults occurred during the two years, but it was bigger during the second year. We infer that, despite differences in gape size, juvenile and adult snakes in the study area exploit similar prey, with the degree of trophic similarity varying interannually.
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Isótopos de Carbono , Ecosistema , Isótopos de Nitrógeno , Serpientes/metabolismo , Animales , Brasil , Dieta , Conducta AlimentariaRESUMEN
Emulsions are crucial in the treatment of snake bites to bust the antibody response of the inmunogen. The widely used Freund's emulsion typically combines 50/50 water-oil (W/O) phase. However, its use is limited because it is associated with tissue damage. We formulated and characterized a Pickering Emulsion 70/30 (W/O) that uses a chemically modified hydrophobic hydroxyapatite as surfactant. This Pickering emulsion has similar rheologic behavior to Freund's emulsion 50/50, but with lower oil and surfactant concentration. Evaluation of cell recruitment, antibody response and adhering tissue in mice immunized with B. asper of Pacific venom and treated with Freund's and Pickering 70/30 emulsions resulted in similar adjuvant activity (only 18% lower in Pickering 70/30 emulsion). However, Pickering 70/30 emulsions minimized negative side effects in the host animals and showed better ease of flow that favors injection of the host. Our results open up room for optimization and improvement of Pickering emulsion based on modified nanoparticles for medical applications.
Asunto(s)
Adyuvantes Inmunológicos/química , Anticuerpos/metabolismo , Durapatita/química , Emulsiones/química , Nanopartículas/química , Venenos de Serpiente/inmunología , Animales , Ratones , Venenos de Serpiente/química , Serpientes/metabolismo , Tensoactivos/químicaRESUMEN
Snake venom metalloproteinases (SVMPs) are abundant in the venoms of vipers and rattlesnakes, playing important roles for the snake adaptation to different environments, and are related to most of the pathological effects of these venoms in human victims. The effectiveness of SVMPs is greatly due to their functional diversity, targeting important physiological proteins or receptors in different tissues and in the coagulation system. Functional diversity is often related to the genetic diversification of the snake venom. In this review, we discuss some published evidence that posit that processing and post-translational modifications are great contributors for the generation of functional diversity and for maintaining latency or inactivation of enzymes belonging to this relevant family of venom toxins.
Asunto(s)
Metaloproteasas/química , Metaloproteasas/genética , Procesamiento Proteico-Postraduccional , Venenos de Serpiente/enzimología , Adaptación Biológica , Animales , Dominio Catalítico , Estabilidad de Enzimas , Proteolisis , Serpientes/metabolismo , Serpientes/fisiologíaRESUMEN
This study aimed at evaluating the energetic return and feeding time on Philodryas nattereri kept in captivity. Snakes were fed biweekly for 60 days (four feeding trials), in two different feeding treatments (single and multiple prey items). The energetic return revealed no significant difference between the feeding treatments; however, we found a negative relationship between snake size and prey handling time during a feed using multiple prey items. In P. nattereri, when large preys are as easy to find as small ones, there seems to be no difference in energetic return.(AU)
O objetivo deste estudo é avaliar o retorno energético e o tempo de alimentação em serpentes Philodryas nattereri mantidas em cativeiro. As serpentes foram alimentadas a cada duas semanas por 60 dias (quatro eventos alimentares) em dois tratamentos de alimentação diferentes (uma única presa e múltiplas presas). Não houve diferença significativa no retorno energético entre os tratamentos alimentares. No entanto, encontramos uma relação negativa entre o tamanho da serpente e o tempo de manipulação da presa durante uma alimentação usando várias presas. Em P. nattereri, parece não haver diferença no retorno energético quando presas grandes são tão fáceis de encontrar quanto presas pequenas.(AU)
Asunto(s)
Animales , Serpientes/crecimiento & desarrollo , Serpientes/metabolismo , Caza/análisis , Caza/métodos , Conducta Alimentaria/clasificaciónRESUMEN
Research is discussed in the areas of venomous snake taxonomy, epidemiology, species diagnosis, defining the clinical phenotype of human envenoming, pathophysiological mechanisms of envenoming, clinical testing of antivenoms and prevention of snake-bite through community education. This work was carried out over the past 40 years in many countries in four tropical continents. The help and friendship of a large number of collaborators is gratefully acknowledged.
Asunto(s)
Mordeduras de Serpientes/epidemiología , Mordeduras de Serpientes/fisiopatología , Animales , Antivenenos/uso terapéutico , Brasil/epidemiología , Educación en Salud , Humanos , Pruebas Inmunológicas/métodos , India/epidemiología , Nigeria/epidemiología , Ensayos Clínicos Controlados Aleatorios como Asunto , Mordeduras de Serpientes/tratamiento farmacológico , Mordeduras de Serpientes/prevención & control , Serpientes/clasificación , Serpientes/metabolismo , Sri Lanka/epidemiología , Tailandia/epidemiologíaRESUMEN
BACKGROUND: A long term research goal of venomics, of applied importance for improving current antivenom therapy, but also for drug discovery, is to understand the pharmacological potential of venoms. Individually or combined, proteomic and transcriptomic studies have demonstrated their feasibility to explore in depth the molecular diversity of venoms. In the absence of genome sequence, transcriptomes represent also valuable searchable databases for proteomic projects. RESULTS: The venom gland transcriptomes of 8 Costa Rican taxa from 5 genera (Crotalus, Bothrops, Atropoides, Cerrophidion, and Bothriechis) of pitvipers were investigated using high-throughput 454 pyrosequencing. 100,394 out of 330,010 masked reads produced significant hits in the available databases. 5.165,220 nucleotides (8.27%) were masked by RepeatMasker, the vast majority of which corresponding to class I (retroelements) and class II (DNA transposons) mobile elements. BLAST hits included 79,991 matches to entries of the taxonomic suborder Serpentes, of which 62,433 displayed similarity to documented venom proteins. Strong discrepancies between the transcriptome-computed and the proteome-gathered toxin compositions were obvious at first sight. Although the reasons underlaying this discrepancy are elusive, since no clear trend within or between species is apparent, the data indicate that individual mRNA species may be translationally controlled in a species-dependent manner. The minimum number of genes from each toxin family transcribed into the venom gland transcriptome of each species was calculated from multiple alignments of reads matched to a full-length reference sequence of each toxin family. Reads encoding ORF regions of Kazal-type inhibitor-like proteins were uniquely found in Bothriechis schlegelii and B. lateralis transcriptomes, suggesting a genus-specific recruitment event during the early-Middle Miocene. A transcriptome-based cladogram supports the large divergence between A. mexicanus and A. picadoi, and a closer kinship between A. mexicanus and C. godmani. CONCLUSIONS: Our comparative next-generation sequencing (NGS) analysis reveals taxon-specific trends governing the formulation of the venom arsenal. Knowledge of the venom proteome provides hints on the translation efficiency of toxin-coding transcripts, contributing thereby to a more accurate interpretation of the transcriptome. The application of NGS to the analysis of snake venom transcriptomes, may represent the tool for opening the door to systems venomics.
Asunto(s)
Perfilación de la Expresión Génica/métodos , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Glándulas Salivales/metabolismo , Análisis de Secuencia de ADN/métodos , Venenos de Serpiente/genética , Serpientes/genética , Animales , Costa Rica , Proteoma/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Serpientes/clasificación , Serpientes/metabolismoRESUMEN
We describe two geographically differentiated venom phenotypes across the wide distribution range of Bothrops atrox, from the Colombian Magdalena Medio Valley through Puerto Ayacucho and El Paují, in the Venezuelan States of Amazonas and Orinoquia, respectively, and São Bento in the Brazilian State of Maranhão. Colombian and Venezuelan venoms show an ontogenetic toxin profile phenotype whereas Brazilian venoms exhibit paedomorphic phenotypes. Venoms from each of the 16 localities sampled contain both population-specific toxins and proteins shared by neighboring B. atrox populations. Mapping the molecular similarity between conspecific populations onto a physical map of B. atrox range provides clues for tracing dispersal routes that account for the current biogeographic distribution of the species. The proteomic pattern is consistent with a model of southeast and southwest dispersal and allopatric fragmentation northern of the Amazon Basin, and trans-Amazonian expansion through the Andean Corridor and across the Amazon river between Monte Alegre and Santarém. An antivenomic approach applied to assess the efficacy towards B. atrox venoms of two antivenoms raised in Costa Rica and Brazil using Bothrops venoms different than B. atrox in the immunization mixtures showed that both antivenoms immunodepleted very efficiently the major toxins (PIII-SVMPs, serine proteinases, CRISP, LAO) of paedomorphic venoms from Puerto Ayacucho (Venezuelan Amazonia) through São Bento, but had impaired reactivity towards PLA(2) and P-I SVMP molecules abundantly present in ontogenetic venoms. The degree of immunodepletion achieved suggests that each of these antivenoms may be effective against envenomations by paedomorphic, and some ontogenetic, B. atrox venoms.
Asunto(s)
Variación Antigénica/fisiología , Antivenenos/análisis , Bothrops/metabolismo , Venenos de Crotálidos/análisis , Proteoma/análisis , Mordeduras de Serpientes/terapia , Secuencia de Aminoácidos , Animales , Antivenenos/metabolismo , Venenos de Crotálidos/metabolismo , Demografía , Femenino , Geografía , Masculino , Población , Proteoma/metabolismo , Proteómica , Ríos , Serpientes/metabolismo , Serpientes/fisiología , América del SurRESUMEN
The three distinct but related isotypes of the iodothyronine deiodinase family: D1, D2, and D3, have been amply studied in vertebrate homeotherms and to a lesser extent in ectotherms, particularly in reptiles. Here, we report the molecular and kinetic characteristics of both the native and the recombinant hepatic D3 from the pine snake Pituophis deppei (PdD3). The complete PdD3 cDNA (1680 bp) encodes a protein of 287 amino acids (aa), which is the longest type 3 deiodinase so far cloned. PdD3 shares 78% identity with chicken and 71% with its other orthologs. Interestingly, the hinge domain in D3s, including PdD3, is rich in proline. This structural feature is shared with D1s, the other inner-ring deiodinases, and deserves further study. The kinetic characteristics of both native and recombinant PdD3 were similar to those reported for D3 in other vertebrates. True K(m) values for T(3) IRD were 9 and 11 nM for native and recombinant PdD3, respectively. Both exhibited a requirement for a high concentration of cofactor (40 mM DTT), insensitivity to inhibition by PTU (>2 mM), and bisubstrate, sequential-type reaction kinetics. In summary, the present data demonstrate that the liver of the adult pine snake P. deppei expresses D3. Furthermore, this is the first report of the cloning and expression of a reptilian D3 cDNA. The finding of hepatic D3 expression in the adult pine snake P. deppei is consistent with results in adult piscine species in which the dietary T(3) content seems to regulate liver deiodinase expression. Thus, our present results support the proposal that hepatic D3 in adult vertebrates plays a sentinel role in avoiding an inappropriate overload of exogenous T(3) secondary to feeding in those species that devour the whole prey.
Asunto(s)
Yoduro Peroxidasa/metabolismo , Serpientes/metabolismo , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Yoduro Peroxidasa/química , Yoduro Peroxidasa/genética , Datos de Secuencia Molecular , Radioinmunoensayo , Proteínas de Reptiles/genética , Proteínas de Reptiles/metabolismoRESUMEN
Crotoxin (CA.CB) is a beta-neurotoxin from Crotalus durissus terrificus snake venom that is responsible for main envenomation effects upon biting by this snake. It is a heterodimer of an acidic protein (CA) devoid of any biological activity per se and a basic, enzymatically active, PLA(2) counterpart (CB). Both lethal and enzymatic activities of crotoxin have been shown to be inhibited by CNF, a protein from the blood of C. d. terrificus snakes. CNF replaces CA in the CA.CB complex, forming a stable, non-toxic complex CNF.CB. The molecular sites involved in the tight interfacial protein-protein interactions in these PLA(2)-based complexes have not been clearly determined. To help address this question, we used the peptide arrays approach to map possible interfacial interaction sites in CA.CB and CNF.CB. Amino acid stretches putatively involved in these interactions were firstly identified in the primary structure of CB. Further analysis of the interfacial availability of these stretches in the presumed biologically active structure of CB, suggested two interaction main sites, located at the amino-terminus and beta-wing regions. Peptide segments at the carboxyl-terminus of CB were also suggested to play a secondary role in the binding of both CA and CNF.
Asunto(s)
Venenos de Crotálidos/química , Crotoxina/metabolismo , Fosfolipasas A2 Grupo II/metabolismo , Serpientes/metabolismo , Animales , Crotalus , HumanosRESUMEN
The venom proteomes of Bothrops atrox from Colombia, Brazil, Ecuador, and Perú were characterized using venomic and antivenomic strategies. Our results evidence the existence of two geographically differentiated venom phenotypes. The venom from Colombia comprises at least 26 different proteins belonging to 9 different groups of toxins. PI-metalloproteinases and K49-PLA(2) molecules represent the most abundant toxins. On the other hand, the venoms from Brazilian, Ecuadorian, and Peruvian B. atrox contain predominantly PIII-metalloproteinases. These toxin profiles correlate with the venom phenotypes of adult and juvenile B. asper from Costa Rica, respectively, suggesting that paedomorphism represented a selective trend during the trans-Amazonian southward expansion of B. atrox through the Andean Corridor. The high degree of crossreactivity of a Costa Rican polyvalent (Bothrops asper, Lachesis stenophrys, Crotalus simus) antivenom against B. atrox venoms further evidenced the close evolutionary kinship between B. asper and B. atrox. This antivenom was more efficient immunodepleting proteins from the venoms of B. atrox from Brazil, Ecuador, and Perú than from Colombia. Such behaviour may be rationalized taking into account the lower content of poorly immunogenic toxins, such as PLA(2) molecules and PI-SVMPs in the paedomorphic venoms. The immunological profile of the Costa Rican antivenom strongly suggests the possibility of using this antivenom for the management of snakebites by B. atrox in Colombia and the Amazon regions of Ecuador, Perú and Brazil.
Asunto(s)
Antivenenos/análisis , Bothrops/metabolismo , Venenos de Crotálidos/análisis , Secuencia de Aminoácidos , Animales , Antivenenos/inmunología , Antivenenos/metabolismo , Brasil , Colombia , Costa Rica , Reacciones Cruzadas , Venenos de Crotálidos/inmunología , Venenos de Crotálidos/metabolismo , Ecuador , Geografía , Perú , Fenotipo , Filogenia , Proteoma/análisis , Serpientes/inmunología , Serpientes/metabolismo , Especificidad de la EspecieRESUMEN
The present study was undertaken to elucidate some aspects about the nature of the spermatozoon ultrastructure of Crotallus durissus using cytochemical methods. We also provide for the first time the ultrastructural description of this species spermatozoon. Cytochemical studies of spermatozoa have not been performed so far in the Serpentes, and species spermatozoon may prove helpful to better understand the reproductive biology of this group. Besides the synapomorphies of the Squamata and Serpentes, the C. durissus spermatozoon possess the following: circular acrosome tip; rounded perforatorium tip with a stopper-like basal modification; bilateral stratified laminar structures; central electron-dense structure within the proximal centriole; fibrous sheath extending until the level of the second mitochondrial ring; rounded mitochondria in cross-section, but with variable shape and organization in longitudinal and oblique sections, respectively; linear annulus; developed multilaminar membranes in the nuclear region and the midpiece. The formation of membrane filipin-sterol complexes occur sparsely along the head region, specially around the nucleus; the complexes were also present in the midpiece membrane and scarcely lining the flagellum. The complexes were present in the different layers of the multilaminar membranes. The ethanol-phosphotungstic acid (E-PTA) treatment relieved the presence of basic proteins in acrosome vesicle, pericentriolar material, peripheral fibers of the axoneme and fibrous sheath. The tannic acid technique revealed the microtubules of the centrioles and the axoneme; the extracellular tubules encircling the spermatozoa and those spread in the epididymal lumen were also observed. However, the immunocytochemistry assay using antibodies against alpha-tubulin and beta-tubulin, the primary microtubule monomers, does not support the existence of composition similarity between these tubular structures, since the extracellular tubules were not labeled by the antibodies. The results obtained in this work demonstrate that the utilization of electron microscopic techniques may provide relevant information to the study of ophidian reproductive biology, particularly in analyses concerning spermatozoal ultrastructure.
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Serpientes/anatomía & histología , Serpientes/metabolismo , Espermatozoides/metabolismo , Espermatozoides/ultraestructura , Animales , Histocitoquímica , Inmunohistoquímica , Masculino , Microscopía Electrónica de Transmisión , Cabeza del Espermatozoide/ultraestructuraRESUMEN
In this review, we summarize the energetic and physiological correlates of prey handling and ingestion in lizards and snakes. There were marked differences in the magnitude of aerobic metabolism during prey handling and ingestion between these two groups, although they show a similar pattern of variation as a function of relative prey mass. For lizards, the magnitude of aerobic metabolism during prey handling and ingestion also varied as a function of morphological specializations for a particular habitat, prey type, and behavior. For snakes, interspecific differences in aerobic metabolism during prey handling seem to be correlated with adaptations for prey capture (venom injection vs. constriction). During ingestion by snakes, differences in aerobic metabolism might be due to differences in cranial morphology, although allometric effects might be a potentially confounded effect. Anaerobic metabolism is used for prey handling and ingestion, but its relative contribution to total ATP production seems to be more pronounced in snakes than in lizards. The energetic costs of prey handling and ingestion are trivial for both groups and cannot be used to predict patterns of prey-size selection. For lizards, it seems that morphological and ecological factors set the constraints on prey handling and ingestion. For snakes, besides these two factors, the capacity of the cardio-respiratory system may also be an important factor constraining the capacity for prey handling and ingestion.
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Conducta Alimentaria , Lagartos/fisiología , Serpientes/fisiología , Animales , Metabolismo Energético , Lagartos/metabolismo , Conducta Predatoria , Serpientes/metabolismoRESUMEN
Snake neurotoxins (NTX) have proven to be valuable tools for the characterisation of muscular nicotinic acetylcholine receptor structure and function. It is very likely that they could also be utilised to identify subtypes of neuronal nicotinic receptors controlling specific functions within the central nervous system. In this study we examined the effects of long alpha NTX (alpha-bungarotoxin, alpha-Bgt, and alpha-cobratoxin, alpha-Cbt) and short alpha NTX (alpha-erabutoxin a, alpha-Ebt) as well as the anticholinesterase toxin fasciculin-2 (FAS), on the nicotine-evoked release of dopamine (DA) in the striatum, using the in vivo push-pull technique. The short toxins alpha-Ebt and FAS blocked the extracellular increase of DA evoked by nicotine at 4.2 microM concentrations and alpha-Ebt was more potent, as reflected by the blockade at the lower dose of 0.42 microM. In contrast, the long toxins showed a different profile of action. Alpha-Cbt did not show any blockade of the nicotine-evoked release of DA at the doses studied while alpha-Bgt did block it only at the higher dose (4.2 microM) These results indicate that short neurotoxins show a stronger interaction with striatal nicotinic receptors subtypes controlling DA release when compared to the long ones. This interaction of short neurotoxin polypeptides and presynaptic receptors may permit the further elucidation of the particular nicotinic receptor populations responsible for the modulation of striatal DA release.
Asunto(s)
Cuerpo Estriado/metabolismo , Dopamina/metabolismo , Neurotoxinas/farmacología , Nicotina/farmacología , Serpientes/metabolismo , Animales , Bungarotoxinas/farmacología , Proteínas Neurotóxicas de Elápidos/farmacología , Cuerpo Estriado/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Venenos Elapídicos/farmacología , Erabutoxinas/farmacología , Masculino , Neurotoxinas/metabolismo , Concentración Osmolar , Ratas , Ratas EndogámicasRESUMEN
The distribution of neurotensin (NT)-like immunoreactivity (LI) in the adrenal gland of the snake Waglerophis merremii has been examined immunohistochemically. Double staining, combining NT with tyrosine hydroxylase (TH) or calcitonin gene-related peptide (CGRP) antibodies and TH with CGRP antibodies, was also carried out. Results were analyzed by conventional and by confocal fluorescence microscopy. Immunostaining revealed a subpopulation of chromaffin cells containing NT-LI within the dorsal noradrenergic ribbon. In addition, there were some NT-immunoreactive (IR) fibers in this region. NT immunoreactivity was not present within adrenergic chromaffin cells or in cortical tissue. Double staining revealed CGRP-IR fibers innervating most of the chromaffin adrenergic cells. Within the dorsal noradrenergic ribbon, two groups of chromaffin TH-IR cells were present, one receiving a dense CGRP-IR innervation and another without contact with CGRP-IR terminals. The latter chromaffin cells displayed NT-LI. These results show, for the first time, the presence of a neuropeptide in chromaffin noradrenergic cells of a reptilian adrenal gland and open up the possibility that other peptides may also be present in these cells.