PubMed Health⌕ Search

Biomedical subjects

A Fernández-López

Publications and source records attributed to A Fernández-López.

At least 19 recordsLinked to original sources
PubMed2006

Pharmacological characterization and autoradiographic distribution of alpha2-adrenoceptor antagonist [3H]RX 821002 binding sites in the chicken brain.

Knowledge about the noradrenergic system in birds is very scarce even though their biological diversity and complex social behavior make them an excellent model for studying neuronal functions and developmental biology. While the role of norepinephrine has been described in depth in a large number of central and peripheral functions in mammals, reports for avian species are limited. The radioligand [(3)H]RX 821002 ([(3)H]1,4-[6,7(n)3H]-benzodioxan-2-methoxy-2-yl)-2-imidazol) has been used to map and characterize alpha(2)-adrenoceptors through the chicken brain using in vitro autoradiography and membrane homogenates binding assays. [(3)H]RX 821002 showed a saturable and high affinity binding to a site compatible with alpha(2)-adrenoceptor, and to a serotonergic component. The autoradiographic assays displayed a similar alpha(2)-adrenoceptor distribution than those previously reported in birds using other radioligands such as [(3)H]UK 14304 ([(3)H]5-bromo-N-(4,5-dihydro-1H-imidazol-2-yl)-6-quinoxalinamine) or [(3)H]clonidine. [(3)H]RX 821002 binding pharmacological characterization was carried out in different chicken brain regions using membrane homogenates for competition assays with different alpha(2)-adrenoceptor agonists and antagonists drugs (oxymetazoline, BRL 44408 [2-(2H-(1-methyl-1,3-dihydroisoindole)methyl)-4,5-dihydroimidazole] ARC 239 [2-(2-4-(O-methoxyphenyl)-piperazin-1-yl)-ethyl-4,4-dimethyl-1,3-(2H,4H)-isoquinolindione], prazosin, UK 14304 and RX 821002). The results showed alpha(2A) as the predominant alpha(2)-adrenoceptor subtype in the chicken brain while alpha(2B)- and/or alpha(2C)-adrenoceptor subtypes were detected only in the telencephalon. RX 821002, serotonin (5-HT) and 8-OH-DPAT [8-hydroxy-2-(di-n-propylamino)tetralin] competition assays, and competition binding assays performed in the presence of serotonin demonstrated that [(3)H]RX 821002 binds with higher affinity to a serotonergic component, probably 5-HT(1A) receptors, than to the alpha(2)-adrenoceptors. Similar pharmacological properties for the alpha(2)-adrenoceptor component were observed both in rat and chicken brain. The results demonstrate that the different alpha(2)-adrenoceptor subtypes are present in chicken brain and suggest that these receptors are highly conserved through evolution.

Adrenergic alpha-Agonists↗
PubMed2006

Cannabinoid system in the budgerigar brain.

Cannabinoid receptor density and cannabinoid receptor-mediated G protein stimulation were studied by autoradiographic techniques throughout the budgerigar (Melopsittacus undulatus) brain. The maximal CB(1) receptor density value (using [(3)H]CP55,940 as radioligand) was found in the molecular layer of the cerebellum (Mol), and high binding values were observed in the nucleus taeniae amygdalae (TnA), nucleus preopticus medialis, and nucleus pretectalis. The highest net-stimulated [(35)S]GTPgammaS binding values induced by the selective CB(1) receptor agonist WIN55,212-2 were observed in the nucleus paramedianus internus thalami, and high values of [(35)S]GTPgammaS binding were observed in the TnA, Mol, arcopallium dorsale and arcopallium intermedium. The distribution data suggest that in the budgerigar, as previously indicated in mammals, cannabinoid receptors may be related to the control of several brain functions in the motor system, memory, visual system, and reproductive behavior. The discrepancies between the cannabinoid receptor densities and the cannabinoid receptor-mediated stimulation found in several budgerigar brain nuclei support the hypothesis, previously described for mammals, of the existence of different G(i/o) protein populations able to associate with the cannabinoid receptors, depending on the brain structure, and could reflect the relative importance that cannabinoid transmission could exerts in each cerebral area.

Animals↗
PubMed2001

Norepinephrine, epinephrine and MHPG levels in chick brain development.

In this study we determined the norepinephrine (NE), epinephrine and methoxy-hydroxy-phenyl-glycol (MHPG) levels in dissected chick telencephalon, diencephalon/mesencephalon and cerebellum in a number of stages from the late embryonic period (E16, E17, E18 and E19) and post-hatching period (P1, P2, P3, P4, P5, P15 and P30) using HPLC coupled with a coulometric detection system. A mobile phase which permits the detection of NE, epinephrine and MHPG simultaneously is also described. During development, NE levels increase dramatically after hatching in all brain structures studied and are not correlated in the same period with an increase in the MHPG/NE ratio. The values obtained for epinephrine and MHPG were significantly lower than the NE values in all the structures and stages studied. Our results support the notion of a specific role for NE during the first days after hatching.

Animals↗
PubMed2000

Distribution of the gamma-aminobutyric acid(A) receptor complex alpha 5 subunit in chick brain. An immunocytochemical and autoradiographic study.

This work reports the distribution of the gamma-aminobutyric acid(A) (GABA(A)) receptor complex alpha5 subunit in the chick using an antibody raised against this subunit in the rat, an immunoprecipitation study and a comparative autoradiographic study using [(3)H]flunitrazepam in the presence of 1 microM zolpidem, which is considered to bind only to those areas presenting the alpha5 subunit. The specificity of the antibody for the chick GABA(A) receptor complex alpha5 subunit is supported by the similar bands obtained by Western blotting from rat and chick, the immunoprecipitation study and the general agreement in the distribution and pattern of labelling of this antibody in both species. The immunocytochemical and autoradiographic distributions in both the chick and rat are compared and some areas with disagreement between these distributions are discussed. The general conclusion is that the alpha5 subunit of the GABA(A) complex receptor seems to have been conserved along evolution.

Animals↗
PubMed2000

The subcommissural organ of the frog Rana perezi is innervated by nerve fibres containing GABA.

The innervation of the frog subcommissural organ was studied by light-microscopic and ultrastructural immunocytochemistry using antisera against serotonin, noradrenaline, dopamine, gamma-aminobutyric acid (GABA), glutamic acid decarboxylase, different GABA receptor subunits and bovine Reissner's fibre material (AFRU). In the proximity of the organ, serotonin- and noradrenaline-containing fibres were rare whereas dopamine-immunoreactive fibres were more numerous. Many GABA- and glutamic acid decarboxylase-containing nerve fibres were found at the basal portion of the ependymal cells of the subcommissural organ. Under the electron microscope, these GABA-immunolabelled nerve endings appeared to establish axoglandular synapses with secretory ependymal cells of the subcommissural organ. In addition, the secretory ependymal cells expressed high amounts of the beta2-subunit of the GABA(A) receptor. Since GABA-immunoreactive neurons were present in the frog pineal organ proper and apparently contributed axons to the pineal tract, we suggest that at least part of the GABAergic fibres innervating the frog subcommissural organ could originate from the pineal organ.

Animals↗
PubMed2000

Autoradiographic characterisation of beta-adrenoceptors in chick brain using [3H]CGP 12177.

An autoradiographic method for labelling beta(1)- and beta(2)-adrenoceptors using [3H]CGP 12177 as a radioligand is described as well as the procedure for an autoradiographic saturation kinetic study. The method afforded higher quality autoradiographs as well as an improvement in the tissue preservation when assayed in birds and chick embryos. The results confirmed the K(d) values previously reported for membrane homogenate binding. The use of different radioligands to characterise beta-adrenoceptors, the higher B(max) values found with autoradiography than those obtained by the membrane homogenate binding method and the typical errors in quantifying autoradiography are discussed. It is concluded that the method described here considerably improves autoradiographic beta-adrenergic characterisation.

Animals↗
PubMed1999

A comparative study of the beta-adrenoceptors in higher song nuclei of birds.

The present study analyses the presence of beta-adrenoceptors in the main telencephalic song nuclei of the goldfinch and parakeet, species belonging to the two most important groups of birds that reproduce learned songs: oscine songbirds and parrots, respectively. Brains of both species sectioned at appropriate levels were used to perform autoradiographic saturation studies using [3H]CGP 12177 as a radioligand. The results show similar K(D)values for both species (0.1-0.3 nM) and striking differences in Bmax. Thus, beta-adrenoceptors are abundant in the telencephalic vocal control nuclei of the parakeet but not of the goldfinch. The predominance of the beta2 subtype in the song nuclei of both species is also confirmed. We conclude that these receptors could be involved in functions unique to the parakeet and therefore may contribute to the greater flexibility of the vocal system of this species. Our findings also support the possible involvement of beta-adrenoceptors in the evolution of the avian brain.

Adrenergic beta-Agonists↗
PubMed1999

Effect of surgical stress on benzodiazepine receptors as a consequence of placebo pellet implantation in rat: an autoradiographic study.

This paper reports modifications in benzodiazepine (BZ) receptors induced by minimally invasive surgery, such as pellet implantation, a widely used surgical process for chronic drug administration. The intrinsic stress induced by this manipulation on BZ receptors was analysed in an autoradiographic saturation study determining the affinity (K(D)) and total number (Bmax) of binding sites of a number of brain areas from the mesencephalon, cerebral cortex, hippocampus, and cerebellum. The radioligand used for the study was [3H]flunitrazepam, which permitted overall characterization of BZ binding sites. Use of the specific BZ1 agonist zolpidem as an inhibitor of this radioligand permitted the direct characterization of subtype 2 (BZ2) and the indirect characterization of subtype 1 receptor (BZ1). Significant increases in Bmax and K(D) values were observed in pellet-implanted animals with respect to those not implanted. The results support the notion of an up-regulation of these receptors, mainly in BZ1 receptors, following sustained desensitization as a result of the surgical stress induced by pellet implantation.

Animals↗
PubMed1999

Seizure-refractory period after a single stimulation and inhibition of seizures after repetitive stimulation in the gerbil: effects on blood cortisol levels.

PURPOSE: To determine the epileptic response of gerbils to external shock stimulus, assessing blood cortisol levels as a parameter to determine stress conditions. METHODS: Five sets of two-month-old Mongolian gerbils were stimulated to elicit seizures by the clapping of a sheaf of papers. Stimulation was done once a week over a 10-week period to obtain a stable situation and a similar response in all the animals. Four of the sets were killed to collect blood samples: those not manipulated; those stimulated twice a day for 5 days; those stimulated once to obtain samples immediately after seizure recovery; and those stimulated once to obtain samples 30 min after seizure recovery. Blood samples from the fifth set of animals were taken in vivo from the retro-orbital plexus. RESULTS: Eliciting seizures with this stimulus, twice a day in a repetitive way, prevented further induced seizures from the second day of stimulation on. Changes in the gerbils' behavior--from exploratory to escape mode--were also observed. The blood cortisol levels found in the sets of animals killed without induced seizures were similar to the others, regardless of whether the animals had been subjected to repetitive stimulation. Additionally, significant decreases in blood cortisol levels were detected in the animals killed immediately and 30 min after recovering from an induced epileptic episode. CONCLUSIONS: The normal refractory period in gerbils can be estimated at 1 h. The lack of correlation between cortisol levels and the inhibition of seizure-elicitation through repetitive stimulation supports the environmental and exploratory hypothesis of seizure generation rather than a stress hypothesis.

Acoustic Stimulation↗
PubMed1998

Pre- and post-hatching developmental changes in beta-adrenoceptor subtypes in chick brain.

This study used [3H]CGP 12177 as a radioligand to determine the beta1 and beta2-adrenoceptor changes from the pre-hatching E17 stage, where the beta2 subtype is first detected, to the post-hatching P30 stage. While beta1-adrenoceptors were found to be present from E18 and were limited to cerebellum and hyperstriatum in all stages studied, beta2-adrenoceptors showed a wider distribution throughout the brain. In most of the structures analysed both beta1- and beta2-adrenoceptor binding values reached a maximum in the P2 stage, followed by a decrease over the following days. A second increase in both subtypes was detected again in the P15 and P30 stages. These results support the notion of a specific role for beta-adrenoceptors in neural plasticity in the first week after hatching and suggest that the beta2 subtype is the main adrenoceptor in chick brain throughout its development.

Adrenergic beta-Agonists↗
PubMed1998

A comparative study of the beta-adrenoceptors in higher visual centres of birds.

This report describes the distribution of beta-adrenergic receptors in the telencephalic visual nuclei of chick, duck, pigeon, parakeet and goldfinch using [3H]CGP 12177 (4-3-t-butylamino-2-hydroxypropoxy-[5,7(3)H] benzimidazol-2-one) as a radioligand. The results reveal a predominance of the beta2-adrenoceptor subtype in all the species studied and that this subtype fits the pharmacological profile described for mammals. It is also demonstrated that the autoradiographic interspecific differences described in previous studies are due to changes in Bmax, while KD values remain in a similar range (0.1-0.6 nM). The distribution of beta-adrenoceptors was fairly similar in the areas of the visual Wulst of all five species studied while striking differences were found in the ectostriatum, the higher centre of the tectofugal pathway. Our findings support a role for ectostriatal beta-adrenoceptors in the visual adaptation and evolution of birds.

Adrenergic beta-Agonists↗
PubMed1997

Distribution of the GABAA receptor complex beta 2/3 subunits in the brain of the frog Rana pipiens.

This report describes the distribution of labeling of the monoclonal antibody bd-17 against the beta 2/3 subunits of the mammalian GABAA receptor complex throughout the brain of the frog Rana pipiens. The distribution matches quite closely those in homologous brain regions as previously described for this antibody in fishes, birds, and mammals, indicating that this antibody also labels beta 2/3 subunits of frog. A semiquantitative analysis of the distribution of labeling throughout the brain is based upon relative optical densities with respect to the structure showing maximal optical density in each brain, using standard illumination conditions. Comparison with distributions in birds and mammals suggests that these GABAA receptor complex subunits are strongly conserved in vertebrate evolution and play an important role in the visual, auditory, olfactory and motor systems.

Animals↗
PubMed1997

The autoradiographic perspective of central benzodiazepine receptors; a short review.

1. We reviewed studies performed to characterize central benzodiazepine binding sites. 2. An overview of the different radioligands used to characterize BZ1 and BZ2 binding sites and a mapping of these central benzodiazepine sites are described. 3. Saturation studies carried out by autoradiogram quantification also are reviewed. 4. The specific use of the autoradiographic technique to carry out studies on ontogeny, development, and phylogeny is discussed, as well as studies performed using this technique on some diseases and experimental conditions, such as drug treatments or chemical and mechanical lesions.

Animals↗
PubMed1997

A comparative study of alpha2- and beta-adrenoceptor distribution in pigeon and chick brain.

The pharmacological properties and anatomical distribution of alpha2-, beta1- and beta2-adrenoceptors in pigeon and chick brains were studied by both homogenate binding and tissue section autoradiography. [3H]Bromoxidine (alpha2-adrenoceptor-), [3H]CGP 12177 (beta-adrenoceptor) and [125I]cyanopindolol (beta-adrenoceptor) were used as radioligands. In both species, [3H]bromoxidine binding to avian brain tissue showed a pharmacological profile similar to that previously reported for alpha2-adrenoceptors in mammals. Regarding the anatomical distribution, the areas with the highest densities of alpha2-adrenoceptors in the pigeon brain included the hyperstriatum, nuclei septalis, tectum opticum and some brainstem nuclei. Most beta-adrenoceptors found in tissue membranes and sections from chick and pigeon brain were of the beta2 subtype, in contrast to what has been reported in the mammalian brain, where the beta1 subtype is predominant. A striking difference was found between the two species regarding the densities of these receptors: while pigeon brain was extremely rich in [125I]cyanopindolol binding throughout the brain (mainly cerebellum) in the pigeon, the levels of labelling in the chick brain were much lower; the exception was the cerebellum, which displayed a higher density than other parts of the brain in both species. Overall, our results support the proposed anatomical equivalences between a number of structures in the avian and mammalian encephalon.

Adrenergic alpha-Agonists↗
PubMed1996

Effects of an acute dose of ethanol on dopaminergic and serotonergic systems from rat cerebral cortex and striatum.

Intraperitoneal injection 10 min before sacrifice of 1.5 g ethanol/kg weight produced an increase in rat striatal levels of homovanillic acid (HVA) (p < 0.05) but did not affect the striatal concentrations of dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA). A similar ethanol treatment led to decreases in 5-HT (p < 0.05) and 5-HIAA (p < 0.05) from cerebral cortex (prefrontal and anterior cingulate areas). The results point to several ethanol-linked alterations in central serotonergic and dopaminergic systems.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1995

An autoradiographical saturation kinetic study of the different benzodiazepine binding sites in rat brain by using [3H] flunitrazepam as a radioligand.

The present study reports on the equilibrium association constant (KD) and receptor density (Bmax) values of a number of brain areas from the mesencephalon, cerebral cortex, hippocampus, and cerebellum for the overall benzodiazepine (BZ) binding sites as well as for benzodiazepine binding site subtype 1 (BZ1) and subtype 2 (BZ2), determined by autoradiographical procedures using [3H] flunitrazepam. The differences between BZ1 and BZ2 binding sites were analyzed using the specific BZ1 agonist zolpidem as inhibitor of the radioligand. Statistically significant differences in the affinities of BZ2 with respect to BZ and BZ1 binding site were mainly found in cortical layers when pKD (negative logarithms of KD values) values were compared (ANOVA-SNK test). The distribution of Bmax, as well as the percentages of BZ1 and BZ2 and Hill coefficients which, surprisingly, are always close to 1 (> 0.9) for all the saturation kinetics analyzed, are also described. The possibility of heterogeneity related to anatomical distribution in the different subtypes is discussed.

Analysis of Variance↗
PubMed1995

Autoradiographical study of types 1 and 2 of benzodiazepine receptors in rat brain after chronic ethanol treatment and its withdrawal.

The effect of chronic ethanol treatment, and its withdrawal on benzodiazepine binding sites in rat brain is described in this autoradiographical study using the benzodiazepine agonist [3H]flunitrazepam. Several areas of the rat cerebral cortex, hippocampus, mesencephalon, cerebellum and lateral geniculate nucleus were studied in animals, chronically treated with ethanol, and 24 or 48 hr after ethanol withdrawal. The [3H]flunitrazepam binding and the relative percentages of binding to BZ1 and BZ2 sites, using zolpidem as a BZ1 site inhibitor, are described. The cerebellum and red nucleus, which only express BZ1 binding sites, appear to be areas significantly modified by ethanol and its withdrawal. In no other structure did ethanol modify [3H]flunitrazepam binding nor change the relative percentage of BZ1 and BZ2 sites.

Animals↗