RESUMO
This study investigated the effects of inosine on memory acquisition and consolidation, cholinesterases activities, redox status and Na+, K+-ATPase activity in a rat model of scopolamine-induced cognitive impairment. Adult male rats were divided into four groups: control (saline), scopolamine (1 mg/kg), scopolamine plus inosine (50 mg/kg), and scopolamine plus inosine (100 mg/kg). Inosine was pre-administered for 7 days, intraperitoneally. On day 8, scopolamine was administered pre (memory acquisition protocol) or post training (memory consolidation protocol) on inhibitory avoidance tasks. The animals were subjected to the step-down inhibitory avoidance task 24 hours after the training. Scopolamine induced impairment in the acquisition and consolidation phases; however, inosine was able to prevent only the impairment in memory consolidation. Also, scopolamine increased the activity of acetylcholinesterase and reduced the activity of Na+, K+-ATPase and the treatment with inosine protected against these alterations in consolidation protocol. In the animals treated with scopolamine, inosine improved the redox status by reducing the levels of reactive oxygen species and thiobarbituric acid reactive substances and restoring the activity of the antioxidant enzymes, superoxide dismutase and catalase. Our findings suggest that inosine may offer protection against scopolamine-induced memory consolidation impairment by modulating brain redox status, cholinergic signaling and ion pump activity. This compound may provide an interesting approach in pharmacotherapy and as a prophylactic against neurodegenerative mechanisms involved in Alzheimer's disease.
Assuntos
Disfunção Cognitiva , Consolidação da Memória , Acetilcolinesterase/metabolismo , Animais , Colinérgicos/efeitos adversos , Inosina/efeitos adversos , Bombas de Íon/farmacologia , Bombas de Íon/uso terapêutico , Masculino , Aprendizagem em Labirinto , Transtornos da Memória/induzido quimicamente , Transtornos da Memória/tratamento farmacológico , Transtornos da Memória/prevenção & controle , Oxirredução , Estresse Oxidativo , Ratos , Ratos Wistar , Escopolamina/farmacologiaRESUMO
Anthocyanins (ANT) are polyphenolic flavonoids with antioxidant and neuroprotective properties. This study evaluated the effect of ANT treatment on cognitive performance and neurochemical parameters in an experimental model of sporadic dementia of Alzheimer's type (SDAT). Adult male rats were divided into four groups: control (1 ml/kg saline, once daily, by gavage), ANT (200 mg/kg, once daily, by gavage), streptozotocin (STZ, 3 mg/kg) and STZ plus ANT. STZ was administered via bilateral intracerebroventricular (ICV) injection (5 µl). ANT were administered after ICV injection for 25 days. Cognitive deficits (short-term memory and spatial memory), oxidative stress parameters, and acetylcholinesterase (AChE) and Na+-K+-ATPase activity in the cerebral cortex and hippocampus were evaluated. ANT treatment protected against the worsening of memory in STZ-induced SDAT. STZ promoted an increase in AChE and Na+-K+-ATPase total and isoform activity in both structures; ANT restored this change. STZ administration induced an increase in lipid peroxidation and decrease in the level of antioxidant enzymes, such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx), in the cerebral cortex; ANT significantly attenuated these effects. In the hippocampus, an increase in reactive oxygen species (ROS), nitrite and lipid peroxidation levels, and SOD activity and a decrease in CAT and GPx activity were seen after STZ injection. ANT protected against the changes in ROS and antioxidant enzyme levels. In conclusion, the present study showed that treatment with ANT attenuated memory deficits, protected against oxidative damage in the brain, and restored AChE and ion pump activity in an STZ-induced SDAT in rats.
Assuntos
Doença de Alzheimer/tratamento farmacológico , Antocianinas/farmacologia , Bombas de Íon/metabolismo , Transtornos da Memória/tratamento farmacológico , Estresse Oxidativo , Acetilcolinesterase/metabolismo , Doença de Alzheimer/induzido quimicamente , Animais , Antioxidantes/uso terapêutico , Encéfalo/metabolismo , Catalase/metabolismo , Córtex Cerebral/metabolismo , Cognição , Modelos Animais de Doenças , Glutationa Peroxidase/metabolismo , Hipocampo/metabolismo , Infusões Intraventriculares , Peroxidação de Lipídeos , Masculino , Aprendizagem em Labirinto , Memória/efeitos dos fármacos , Transtornos da Memória/induzido quimicamente , Fármacos Neuroprotetores/uso terapêutico , Ratos , Ratos Wistar , Estreptozocina/efeitos adversos , Compostos de Sulfidrila , Superóxido Dismutase/metabolismoRESUMO
The high-affinity K(+) transporter (HKT) family comprises a group of multifunctional cation transporters widely distributed in organisms ranging from Bacteria to Eukarya. In angiosperms, the HKT family consists primarily of nine types, whose evolutionary relationships are not fully understood. The available sequences from 31 plant species were used to perform a comprehensive evolutionary analysis, including an examination of selection pressure and estimating phylogenetic tree and gene duplication events. Our results show that a gene duplication in the HKT1;5/HKT1;4 cluster might have led to the divergence of the HKT1;5 and HKT1;4 subfamilies. Additionally, maximum likelihood analysis revealed that the HKT family has undergone a strong purifying selection. An analysis of the amino acids provided strong statistical evidence for a functional divergence between subfamilies 1 and 2. Our study was the first to provide evidence of this functional divergence between these two subfamilies. Analysis of co-evolution in HKT identified 25 co-evolved groups. These findings expanded our understanding of the evolutionary mechanisms driving functional diversification of HKT proteins.
Assuntos
Evolução Molecular , Bombas de Íon/genética , Magnoliopsida/genética , Proteínas de Plantas/genética , Potássio/metabolismo , Duplicação Gênica , Bombas de Íon/metabolismo , Magnoliopsida/classificação , Filogenia , Proteínas de Plantas/metabolismo , Seleção GenéticaRESUMO
In higher eukaryotes, the sarco-endoplasmic reticulum (ER) Ca(2+)-ATPase (SERCA) is characterized for its high sensitivity to low concentrations of thapsigargin (TG), a very specific inhibitor. In contrast, SERCA-like enzymes with different sensitivities to TG have been reported in trypanosomatids. Here, we characterized a SERCA-like enzyme from Trypanosoma evansi and evaluated its interaction with TG. Confocal fluorescence microscopy using BODIPY FL TG and specific anti-SERCA antibodies localized the T. evansi SERCA-like enzyme in the ER and confirmed its direct interaction with TG. Moreover, the use of either 1 µM TG or 25 µM 2',5'-di (tert-butyl)-1,4-benzohydroquinone prevented the reuptake of Ca(2+) and consequently produced a small increase in the parasite cytosolic calcium concentration in a calcium-free medium, which was released from the ER pool. A 3035 bp-sequence coding for a protein with an estimated molecular mass of 110.2 kDa was cloned from T. evansi. The corresponding gene product contained all the invariant residues and conserved motifs found in other P-type ATPases but lacked the calmodulin binding site. Modeling of the three-dimensional structure of the parasite enzyme revealed that the amino acid changes found in the TG-SERCA binding pocket do not compromise the interaction between the enzyme and the inhibitor. Therefore, we concluded that T. evansi possesses a SERCA-like protein that is inhibited by TG.
Assuntos
ATPases Transportadoras de Cálcio/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Bombas de Íon/efeitos dos fármacos , Tapsigargina/farmacologia , Trypanosoma/metabolismo , Sequência de Aminoácidos , Animais , Western Blotting , ATPases Transportadoras de Cálcio/antagonistas & inibidores , ATPases Transportadoras de Cálcio/genética , ATPases Transportadoras de Cálcio/imunologia , Retículo Endoplasmático/enzimologia , Doenças dos Cavalos/parasitologia , Cavalos , Bombas de Íon/metabolismo , Masculino , Microscopia Confocal , Modelos Moleculares , Dados de Sequência Molecular , Ratos , Ratos Sprague-Dawley , Alinhamento de Sequência , Trypanosoma/efeitos dos fármacos , Trypanosoma/fisiologia , Tripanossomíase/parasitologia , Tripanossomíase/veterináriaRESUMO
Intracellular pH (pHi) regulation is essential for cell function. Notably, several unique sperm ion transporters and enzymes whose elimination causes infertility are either pHi dependent or somehow related to pHi regulation. Amongst them are: CatSper, a Ca(2+) channel; Slo3, a K(+) channel; the sperm-specific Na(+)/H(+) exchanger and the soluble adenylyl cyclase. It is thus clear that pHi regulation is of the utmost importance for sperm physiology. This review briefly summarizes the key components involved in pHi regulation, their characteristics and participation in fundamental sperm functions such as motility, maturation and the acrosome reaction.
Assuntos
Espermatozoides/fisiologia , Reação Acrossômica/fisiologia , Adenilil Ciclases/metabolismo , Animais , Anidrases Carbônicas/metabolismo , Quimiotaxia/fisiologia , Humanos , Concentração de Íons de Hidrogênio , Líquido Intracelular/metabolismo , Canais Iônicos/metabolismo , Bombas de Íon/metabolismo , Masculino , Modelos Biológicos , Ouriços-do-Mar/fisiologia , Transdução de Sinais , Capacitação Espermática/fisiologia , Motilidade dos Espermatozoides/fisiologiaRESUMO
The human body is constantly exposed to many xenobiotics including environmental pollutants, food additives, therapeutic drugs, etc. The liver is considered the primary site for drug metabolism and elimination pathways, consisting in uptake, phase I and II reactions, and efflux processes, usually acting in this same order. Modulation of biotransformation and disposition of drugs of clinical application has important therapeutic and toxicological implications. We here provide a compilation and analysis of relevant, more recent literature reporting hormonal regulation of hepatic drug biotransformation and transport systems. We provide additional information on the effect of hormones that tentatively explain differences between sexes. A brief discussion on discrepancies between experimental models and species, as well as a link between gender-related differences and the hormonal mechanism explaining such differences, is also presented. Finally, we include a comment on the pathophysiological, toxicological, and pharmacological relevance of these regulations.
Assuntos
Biotransformação , Hormônios Gonadais/metabolismo , Bombas de Íon/metabolismo , Fígado/metabolismo , Animais , Feminino , Humanos , Masculino , Caracteres SexuaisRESUMO
In this study we use a theoretical approach to study the volumetric response of goldfish hepatocytes challenged by osmotic gradients and compared it with that of hepatocytes from another teleost (the trout) and a mammal (the rat). Particular focus was given to the multiple non-linear interactions of transport systems enabling hypotonically challenged cells to trigger a compensatory response known as volume regulatory decrease or RVD. For this purpose we employed a mathematical model which describes the rates of change of the intracellular concentrations of main diffusible ions, of the cell volume, and of the membrane potential. The model was fitted to experimental data on the kinetics of volume change of hepatocytes challenged by anisotonic media. In trout and rat hepatocytes, experimental results had shown that hypotonic cell swelling was followed by RVD, whereas goldfish cells swelled with no concomitant RVD (M.V. Espelt et al., 2003, J. Exp. Biol. 206, 513-522). A comparison between data predicted by the model and that obtained experimentally suggests that in trout and rat hepatocytes hypotonicity activates a sensor element and this, in turn, activates an otherwise silent efflux of KCl - whose kinetics could be successfully predicted - thereby leading to volume down-regulation. In contrast, with regard to the absence of RVD in goldfish hepatocytes the model proposed suggests that either a sensor element triggering RVD is absent or that the effector mechanism (the loss of KCl) remains inactive under the conditions employed. In line with this, we recently found that extracellular nucleotides may be required to induce RVD in these cells, indicating that our model could indeed lead to useful predictions.
Assuntos
Tamanho Celular , Hepatócitos/citologia , Modelos Biológicos , Vertebrados/metabolismo , Animais , Transporte Biológico , Peixes , Bombas de Íon/metabolismo , Soluções Isotônicas , Ligantes , Potenciais da Membrana , Osmose , Potássio/metabolismo , Ratos , Fatores de TempoRESUMO
Proteorhodopsins are light-energy-harvesting transmembrane proteins encoded by genes recently discovered in the surface waters of the world's oceans. Metagenomic data from the Global Ocean Sampling expedition (GOS) recovered 2674 proteorhodopsin-related sequences from 51 aquatic samples. Four of these samples were from non-marine environments, specifically, Lake Gatun within the Panama Canal, Delaware Bay and Chesapeake Bay and the Punta Cormorant Lagoon in Ecuador. Rhodopsins related to but phylogenetically distinct from most sequences designated proteorhodopsins were present at all four of these non-marine sites and comprised three different clades that were almost completely absent from marine samples. Phylogenomic analyses of genes adjacent to those encoding these novel rhodopsins suggest affiliation to the Actinobacteria, and hence we propose to name these divergent, non-marine rhodopsins 'actinorhodopsins'. Actinorhodopsins conserve the acidic amino acid residues critical for proton pumping and their genes lack genomic association with those encoding photo-sensory transducer proteins, thus supporting a putative ion pumping function. The ratio of recA and radA to rhodopsin genes in the different environment types sampled within the GOS indicates that rhodopsins of one type or another are abundant in microbial communities in freshwater, estuarine and lagoon ecosystems, supporting an important role for these photosystems in all aquatic environments influenced by sunlight.
Assuntos
Actinobacteria/genética , Rodopsinas Microbianas/genética , Microbiologia do Solo , Microbiologia da Água , Actinobacteria/metabolismo , Proteínas de Bactérias/genética , Proteínas de Ligação a DNA/genética , Delaware , Equador , Água Doce/microbiologia , Bombas de Íon/metabolismo , Maryland , Zona do Canal do Panamá , Filogenia , Recombinases Rec A/genéticaRESUMO
Two members of a recently discovered family of protein kinases {WNK1 and WNK4 [with no K (lysine) kinases-1 and -4]} are the cause of an inherited disease known as pseudohypoaldosteronism type II that features arterial hypertension. The family is known as WNK due to a lack of the invariant catalytic lysine in kinase subdomain II. The mechanisms by which WNKs regulate blood pressure are beginning to be understood at the physiological level from recent studies showing effects of WNK4 on several plasma membrane co-transporters and ion channels. However, little is known about the function of WNKs at the biochemical level. In this issue of the Biochemical Journal, Vitari et al. have shown that WNK1 and WNK4 interact with other kinases, SPAK (STE20/SPS1-related proline/alanine-rich kinase) and OSR1 (oxidative stress response kinase-1), which are involved in the regulation of ion transporters. WNK1 and WNK4 phosphorylate SPAK and OSR1, which in turn phosphorylate the N-terminal domain of the basolateral Na+-K+-2Cl- co-transporter, NKCCl. The phosphorylation site involved in SPAK or OSR1 activation is identified as a threonine residue within the T-loop.
Assuntos
Bombas de Íon/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Hipertensão/enzimologia , Hipertensão/genética , Família Multigênica , Fosforilação , Ligação Proteica , Proteínas Serina-Treonina Quinases/genética , Treonina/metabolismo , Fatores de Transcrição/metabolismoRESUMO
Using quantitative real-time PCR, the expression of mRNAs encoding three transport-related proteins and one putative housekeeping protein was analyzed in anterior and posterior gills of the euryhaline crab Chasmagnathus granulatus following transfer from isosmotic conditions (30 per thousand salinity) to either dilute (2 per thousand) or concentrated (45 per thousand) seawater. Modest changes were observed in the abundance of mRNAs encoding the housekeeping protein arginine kinase and the vacuolar-type H(+)-ATPase B-subunit, both of which were highly expressed under all conditions. By contrast, the expression of Na(+)/K(+)-ATPase alpha-subunit mRNA and Na(+)/K(+)/2Cl(-) cotransporter mRNA was strongly responsive to external salinity. During acclimation to dilute seawater, cotransporter mRNA increased 10-20-fold in posterior gills within the first 24 h while Na(+)/K(+)-ATPase alpha-subunit mRNA increased 35-55-fold. During acclimation to concentrated seawater, cotransporter mRNA increased 60-fold by 96 h and Na(+)/K(+)-ATPase alpha-subunit increased approximately 25-fold in posterior gills. Our results indicate a complex pattern of transcriptional regulation dependent upon the direction of salinity change and the developmental background of the gills.
Assuntos
Aclimatação/genética , Braquiúros/metabolismo , Regulação da Expressão Gênica , Bombas de Íon/metabolismo , RNA Mensageiro/metabolismo , Água do Mar/química , Análise de Variância , Animais , Primers do DNA , Brânquias/metabolismo , Bombas de Íon/genética , Reação em Cadeia da Polimerase , Fatores de TempoRESUMO
Arsenic resistance determinants from 42 environmental bacterial isolates (32 Gram negative) were analyzed by DNA: DNA hybridization using probes derived from Escherichia coli and Staphylococcus plasmid or chromosomal arsenic resistance (ars) genes. In colony hybridization assays, 11 and 1 Gram negative strains hybridized with the E. coli chromosome and plasmid probes, respectively. No hybridization was detected using a probe containing only the arsA (ATPase) gene from E. coli plasmid or with a Staphylococcus plasmid ars probe. From Southern hybridization tests of some of the positive strains it was concluded that homology to ars chromosomal genes occurred within chromosome regions, except in an E. coli isolate where hybridization occurred in both the chromosome and a 130-kb plasmid. Our results show that DNA sequences homologous to E. coli ars chromosomal genes are commonly present in the chromosomes of environmental arsenic-resistant Gram negative isolates.
Assuntos
Arsenicais/farmacologia , Bactérias/genética , Proteínas de Bactérias , DNA Bacteriano/genética , Resistência Microbiana a Medicamentos/genética , Bombas de Íon , Complexos Multienzimáticos , Fatores R/genética , Microbiologia do Solo , Adenosina Trifosfatases/genética , Arseniatos/farmacologia , ATPases Transportadoras de Arsenito , Arsenitos/farmacologia , Bactérias/efeitos dos fármacos , Cromossomos Bacterianos/genética , Escherichia coli/genética , México , Hibridização de Ácido Nucleico , Poluentes do Solo/farmacologia , Transativadores/genéticaRESUMO
Arsenic compounds, often present as environmental pollutants, are highly toxic for most microorganisms. Some microbial strains possess genetic determinants conferring resistance to arsenic derivatives. In bacteria, these determinants are usually located on plasmids, which has facilitated their analysis with molecular detail. Bacterial plasmids conferring arsenic resistance encode specific pumps that extrude arsenite (AsIII). In Gram-negative bacteria, the efflux pump consists of a complex formed by an ATPase (ArsA) associated with a membrane anion channel (ArsB). Arsenate (AsV) is converted to arsenite by a soluble reductase (ArsC). Proteins ArsB and ArsC, but not the ATPase, are also found in Gram-positive bacteria. Besides the widely spread plasmid arsenic resistance determinants, some bacteria possess the ability to enzimatically oxidize arsenite to less toxic arsenate.
Assuntos
Arsenicais/farmacologia , Bactérias/efeitos dos fármacos , Resistência Microbiana a Medicamentos , Bombas de Íon , Complexos Multienzimáticos , Adenosina Trifosfatases/fisiologia , Arseniatos/metabolismo , ATPases Transportadoras de Arsenito , Arsenitos/metabolismo , Bactérias/genética , Fenômenos Fisiológicos Bacterianos , Proteínas de Bactérias/fisiologia , Transporte Biológico Ativo , Resistência Microbiana a Medicamentos/genética , Poluentes Ambientais/farmacologia , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Negativas/fisiologia , Bactérias Gram-Positivas/efeitos dos fármacos , Bactérias Gram-Positivas/fisiologia , Canais Iônicos/fisiologia , Oxirredução , Fatores R/genéticaRESUMO
Na,K-ATPase activity of red blood cells from Chediak-Higashi syndrome (CHS) patients and relatives (gene heterozygous) was determined and compared to that of control, healthy, individuals. The enzyme activity was found to be strongly diminished in the CHS patients and slightly lower in their relatives. This reduced activity was due to a lower turnover number of the Na, K-ATPase as well as a decreased number of pumps. The reduced enzyme activity observed in these patients could be the result of an abnormal cell membrane fluidity, and the lowered number of Na, K-pumps could be explained as a consequence of an altered or deficient cell machinery caused by the CHS gene.
Assuntos
Síndrome de Chediak-Higashi/enzimologia , Membrana Eritrocítica/enzimologia , ATPase Trocadora de Sódio-Potássio/sangue , Adolescente , Adulto , Síndrome de Chediak-Higashi/sangue , Criança , Pré-Escolar , Membrana Eritrocítica/metabolismo , Eritrócitos/enzimologia , Eritrócitos/metabolismo , Heterozigoto , Humanos , Lactente , Bombas de Íon , Ouabaína/metabolismo , Ouabaína/farmacologia , Sódio/sangueRESUMO
A microsomal fraction enriched in ion pump enzymes was isolated from the gill of the carp (Cyprinus carpio Linneo). Mg(2+)-dependent (Na+ + K+), Na+, HCO3- and Ca(2+)-stimulated ATPase activities were studied following treatment with microcystin-LR-like toxin, the major toxic component isolated from Microcystis aeruginosa culture. These enzyme activities were inhibited in a dose-dependent manner. The maximum inhibition of each enzyme, induced with nM concentration of the toxin, was similar to that produced by inhibitors specific for each ATPase activity. The Mg(2+)-ATPase activity and non-specific hydrolysis of ATP were unaffected. These results strongly suggest that the massive fish death during M. aeruginosa blooms may result from the loss of ion homeostatic processes produced by the inhibitory action of microcystin on the ion pumps of gill chloride cells.
Assuntos
Toxinas Bacterianas/farmacologia , Brânquias/enzimologia , Bombas de Íon/efeitos dos fármacos , Microcystis/química , Animais , ATPases Transportadoras de Cálcio/metabolismo , Carpas , Brânquias/microbiologia , ATPase Trocadora de Sódio-Potássio/metabolismoRESUMO
1. Na(+)o-dependent Ca2+ efflux (forward Na(+)-Ca2+ exchange), and in some cases the Na(+)i-dependent Ca2+ influx (reverse Na(+)-Ca2+ exchange) were measured in internally dialysed squid axons under membrane potential control. 2. We tested the effect on the Na(+)-Ca2+ exchange of the MgATP analogue bidentate chromium adenosine-5'-triphosphate (CrATP), substrate of several kinases, and cobalt tetrammine ATP (Co(NH3)4ATP), a poor substrate of most kinases. 3. CrATP completely blocked the MgATP and MgATP-gamma-S (ATP-gamma-S) stimulation of the Na(+)o-dependent Ca2+ efflux (forward exchange) and the Na+i-dependent Ca2+ influx (reverse exchange). The analogue only blocked the nucleotide-dependent fraction of the Na(+)-Ca2+ exchange without modifying any kinetic parameters of the exchange reactions. 4. The effects of CrATP were fully reversible with a very slow time constant (t 1/2 about 30 min). 5. The MgATP stimulation of the Na(+)-Ca2+ exchange was completely saturated at 1 mM. Higher MgATP concentrations (up to 15 mM) had no additional effects. Pentalysine (internal or external), the protein kinase C inhibitor H-7 (1-(5-isoquinolinylsulphonyl)-2-methylpiperazine) and several calmodulin inhibitors did not inhibit Na(+)-Ca2+ exchange either in the absence or presence of MgATP. 6. Our results do not agree with the idea of an aminophospholipid translocase being responsible for the ATP stimulation of the Na(+)-Ca2+ exchange in squid axons; they suggest that this is due to the action of a kinase system.
Assuntos
Trifosfato de Adenosina/análogos & derivados , Trifosfato de Adenosina/farmacologia , Axônios/metabolismo , Bombas de Íon/efeitos dos fármacos , Compostos Organometálicos/farmacologia , Animais , Axônios/efeitos dos fármacos , ATPases Transportadoras de Cálcio/efeitos dos fármacos , Células Cultivadas , Decapodiformes , Diálise , Fosfotransferases/metabolismo , ATPase Trocadora de Sódio-Potássio/efeitos dos fármacosRESUMO
El transporte activo de iones se relaciona a una serie de funciones vitales para el metabolismo normal de la célula eucariótica. El estudio de los mecanismos de transporte y su regulación constituye un problema fundamental de la biología celular. La producción de anticuerpos monoclonales (Acm) permite dispones de una sola, altamente específica, que facilita el estudio de la localización, distribución y función de las proteínas transportadoras a nivel epitelial, así como también el estudio de su estructura molecular, síntesis y ensamblaje a nivel celular. La calidad del anticuerpo monoclonal dependerá de la selección acertada de cada uno de los pasos a seguir en el proceso de su producción. En este artículo, discutimos las estrategias unas comunmente utilizadas en la producción de anticuerpo monoclonales y su aplicación directa en el estudio morfológico, bioquímico y fisiológico de las bombas de iones