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G Alonso

Publications and source records attributed to G Alonso.

At least 91 records · Page 5Linked to original sources
PubMed1994

Bromocriptine treatment increases lipolysis and steady-state levels of G proteins in adipocytes from lactating rats.

An increase in the rate of lipogenesis and a decrease in the lipolytic response to catecholamines can be observed in adipocytes after weaning or litter removal. Bromocriptine treatment of lactating rats also produces an increase in lipogenesis but fails to decrease the lipolytic response of adipocytes to catecholamines seen after weaning or litter removal. No changes in total number or affinity of beta-adrenergic receptors or adenosine A1 receptors were detected by bromocriptine treatment using radioligand binding assays. However, we observed an increase in the amount of radioactivity from [32P]NAD+ incorporated into alpha-Gs (192 +/- 26%) and alpha-Gi (178 +/- 33%) by cholera- and pertussis-toxin-catalyzed ADP-ribosylation, respectively, with the same treatment, Immunoblotting using RM/1 and AS/7 antisera, which specifically recognize alpha-Gs and alpha-Gi 1,2, respectively, confirmed the increase in the steady-state levels of these G-protein alpha-subunits. The increase in the steady-state levels of alpha-Gs may account, at least in part, for the increased lipolytic response of adipocytes to catecholamines in bromocriptine-treated rats.

Adenosine↗
PubMed1994

Immunolocalization of polysialic acid in the median eminence and neurointermediate hypophysial lobe of adult rats.

Polysialic acid (PSA) is abundant on growing axons during brain development and down regulated on maturation. However, high amounts of this carbohydrate polymer have been found to persist in some regions of the adult rat brain including the mediobasal hypothalamus. In this study, confocal laser scanning microscopy combined with double fluorescence immunostaining was used to characterize the cellular localization of PSA throughout the median eminence and neurointermediate hypophysial lobe of adult rats. In these regions, polysialic acid-immunoreactivity (PSA-IR) generally appeared associated with fiber-like structures. Double immunostaining experiments demonstrated that, in addition to large axons of the neural lobe immunoreactive to vasopressin or oxytocin, PSA was constantly associated with fibers projecting into the intermediate hypophysial lobe immunoreactive to either gamma-aminobutyric acid (GABA) or tyrosine hydroxylase. Similarly, PSA-IR was detected on most, but not all the fibers immunoreactive to GABA or tyrosine hydroxylase dispersed throughout the neural lobe and the different layers of the median eminence. On the other hand, no PSA-IR was detected on axons immunoreactive to somatostatin or to corticotropin releasing hormone projecting throughout the median eminence, or on glial cell bodies and processes immunoreactive for glial fibrillary acidic protein (GFAP) or for vimentin dispersed throughout the median eminence and the neural lobe.

Animals↗
PubMed1994

Assessment of glomerular filtration rate utilizing subcutaneously injected 51Cr-EDTA.

1. 51Cr-EDTA injected with lidocaine and epinephrine, as a subcutaneous button, is slowly absorbed, and a plasma level that is relatively stable can be maintained for a time sufficient to permit measurement of the renal clearance of EDTA, which is a measure of glomerular filtration rate (GFR). We studied this procedure in 32 normal volunteers and 24 patients with different glomerulopathies, comparing EDTA and creatinine clearances. In 20 patients these measurements were also compared with inulin clearance. 2. Creatinine clearance overestimates GFR due to tubular secretion of creatinine. This secretion is present even in patients with significantly reduced glomerular filtration rates. As a consequence, the lower the GFR the higher the overestimation will be. 3. A good correlation was obtained between the 51Cr-EDTA and inulin clearance: y(EDTA) = 4.21 + 0.88 x (inulin), r = 0.98. The procedure is simple to perform, and the radiotracer utilized is significantly less expensive than iothalamate.

Adolescent↗
PubMed1993

Neuropeptide Y-producing neurons of the arcuate nucleus regenerate axons after surgical deafferentation of the mediobasal hypothalamus.

Dorsolateral and ventomedial surgical deafferentiation of the hypothalamus were used to study the capacity of different types of neuropeptide Y-containing axons afferent to the dorsal hypothalamus to regenerate through surgical lesions. The kinetics of the postlesional responses of transected neuropeptide Y-axons was studied on 30-40 microns thick vibratome sections, either (i) by light or electron microscopy after peroxidase immunostaining for neuropeptide Y or (ii) by confocal microscopy after double fluorescence immunostaining for neuropeptide Y and for glial fibrillary acidic protein. The dorsolateral cut was found to sever 2 main pathways containing neuropeptide Y axons located, respectively, below the bed nucleus of the stria terminalis and in the perifornical region. In both regions transected fibers were found to abut onto the surgical lesion, but even 45 days after the lesion, they were very rarely observed to penetrate into the astroglial scar forming along the lesion. The ventromedial cut was found to sever numerous neuropeptide axons that originate in the underlying arcuate nucleus. Seven to 15 days after the lesion neuropeptide Y fibers located below this type of cut presented a dramatic increase in both their numerical density and their immunostaining intensity. With increasing post-surgery times, an increased number of neuropeptide Y fibers was observed to penetrate and to cross the lesional scar formed by densely packed astrocytic processes. Electron microscope observations further demonstrated that 45 days after the lesion, numerous neuropeptide Y-immunoreactive axonal profiles were included in the scar matrix, which appeared to be mainly composed of closely interdigitating astrocytic processes containing dense bundles of filaments. These data indicate that, in contrast to other neuropeptide Y neurons innervating the dorsal hypothalamus, neuropeptide Y neurons of the arcuate nucleus regenerate axons through the astroglial scar produced by a surgical lesion placed in the ventromedial hypothalamus.

Animals↗
PubMed1993

Reactive astrocytes involved in the formation of lesional scars differ in the mediobasal hypothalamus and in other forebrain regions.

The fine organization of lesional scars was studied in adult rats at the level of 2 types of surgical cuts aimed at deafferentating the dorsal hypothalamus from its neuropeptide-Y innervation. These included: (i) lesions located dorsolateral to the dorsal hypothalamus, which were shown to form a permanent obstacle to the regeneration of transected neuropeptide-Y-fibers, and (ii) lesions located in the ventromedial hypothalamus, where transected neuropeptide-Y-fibers were shown to penetrate and eventually cross the lesional area. Double labeling immunocytochemistry and conventional electron microscopy were used to identify various molecules produced by reactive astrocytes and to visualize their ultrastructural organization within the scars, respectively. In the different portions of the dorsolateral scars, the large majority of reactive astrocytes was characterized by a strong immunoreactivity to glial fibrillary acidic protein, vimentin, and embryonic (polysialylated) NCAM. Intense laminin-immunoreactivity was also observed over large patches included in the scar. Electron microscope observations further indicated that the matrix of the scar was mainly composed of tightly packed astrocytic perikarya and processes connected by extended gap junctions. All around the extracellular and perivascular spaces, these astrocyte profiles were bordered by a thick basal lamina. Only scarce axonal profiles were detected in the core of the scar, most of which exhibited degenerative features. In the ventromedial hypothalamic scars, reactive astrocytes were found to exhibit intense immunoreactivity to both glial fibrillary acidic protein and vimentin. On the other hand, only slight immunostaining to embryonic NCAM and laminin were associated with this type of lesional scar. At the ultrastructural level, the main differences with the dorsolateral scars concerned (i) the gap junctions, which were less frequent and involved shorter portions of adjacent membranes; (ii) the basal lamina, which was essentially localized to the perivascular spaces; and (iii) the axonal profiles, which were frequently observed throughout the scar matrix. These data indicate that reactive astrocytes that formed the glial scar differ in the mediobasal hypothalamus and in other forebrain regions. This provides strong support for the hypothesis that the regeneration of neuropeptide-Y axons through a mediobasal hypothalamic surgical cut depends mainly on the particular organization of the astroglial scar.

Animals↗
PubMed1993

Isolation of two pyruvate kinase activities in the parasitic protozoan Leishmania mexicana amazonensis.

Using phosphocellulose affinity chromatography we were able to separate two pyruvate kinase (EC 2.7.1.40) activities in the parasitic protozoan Leishmania mexicana amazonesis. One activity (PYK1) showed hyperbolic kinetics and was decreased by fructose 2,6-bisphosphate, whereas the second activity (PYK2) showed sigmoidal kinetics for the substrate phosphoenolpyruvate and was activated by fructose 2,6-bisphosphate. Molecular sieve chromatography (Sephacryl S-400) of PYK1 produced a single peak of apparent molecular mass around 200,000, while PYK2 eluted at a position corresponding to M(r) 55,000.

Animals↗
PubMed1993

Stimulatory effect of N-methyl-D-aspartate on somatostatin gene expression in cultured hypothalamic neurons.

The aim of the present study was to determine whether N-methyl-D-aspartate (NMDA) stimulates somatostatin gene function in primary cultures of hypothalamic neurons. Neurons were either shortly (for 3, 8, 24 and 72 h) or chronically (for 11 days) exposed to NMDA (20 microM). Medium and cellular somatostatin contents were determined by radioimmunoassay, and steady-state preprosomatostatin mRNA levels by Northern blot analysis with an oligonucleotide probe. DNA content was measured as a cellular viability control. After 8 h incubation, NMDA induced a significant 2-fold increase in somatostatin mRNA accumulation, with a maximal 4-fold increase after 24 h incubation. A significant and dose-dependent (1.7-fold and 2.5-fold at 20 and 100 microM, respectively) stimulatory effect was also observed after chronic treatment. The kinetic patterns for medium and cellular somatostatin contents were similar to those obtained for somatostatin mRNA levels. Total DNA content was not modified under any experimental condition. The augmentations in cellular somatostatin and somatostatin mRNA determined after 24 h or chronic exposure to NMDA were blocked by (+)-5-methyl-10.11-dihydro-5H-dibenzo(a,d')cyclohepten-5,10-imine hydrogen maleate (MK-801), an NMDA receptor antagonist. MK-801 alone significantly (P < 0.05) reduced somatostatin mRNA. The stimulatory effect of NMDA on somatostatin mRNA was specific since it was not accompanied by any change in glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA. After immunostaining with a specific antibody against somatostatin, no difference was observed in the number of immunostained neurons detected in control and NMDA exposed groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Additive effects of exposure to silica dust and smoking on pulmonary epithelial permeability: a radioaerosol study with technetium-99m labelled DTPA.

BACKGROUND: Increased pulmonary epithelial permeability evaluated by the rate of clearance from lung to blood of the radioaerosol solute technetium-99m labelled diethylenetriamine pentaacetate (99mTc-DTPA) has been reported in smokers and in workers exposed to silica dust. A study was carried out to determine whether there are additive effects of cigarette smoke and exposure to silica dust on clearance rates of 99mTc-DTPA in ceramic workers. METHODS: Thirty one subjects with silicosis were studied, of whom 18 smoked cigarettes and 13 were non-smokers. They had similar histories of exposure to silica dust, and radiological alterations consistent with silicosis. The results from these patients were compared with those from normal subjects and smokers previously studied by the authors. RESULTS: Pulmonary function values were normal in most patients and not significantly different among groups. The median (range) rate of clearance of 99mTc-DTPA in smokers with silicosis was 4.1 (1.9-12.7) %/minute, which was higher than the rates in non-smoking patients with silicosis of 2.2 (1.1-6.6) %/minute and in smokers without exposure to silica dust of 2.9 (1.6-4.5) %/minute. These differences were more evident and significant when the clearance rates of the lower lobes of the three groups were compared. Clearance rates higher than 3%/minute were much more frequent in smokers with silicosis (85%) than in non-smoking patients with silicosis (15%) and in smokers (40%). CONCLUSION: In ceramic workers with radiographic changes resulting from exposure to silica dust, there is an additive effect of inhalation of silica dust and cigarette smoking on clearance rates of 99mTc-DTPA.

Adult↗
PubMed1993

Effects of discrete lesions in the ventral noradrenergic ascending bundle on the corticotropic stress response depend on the site of the lesion and on the plasma levels of adrenal steroids.

Stereotaxic deletion of selected areas of the ventral noradrenergic ascending bundle (VNAB-X) by discrete bilateral injections of 6-hydroxydopamine (6-OHDA; 4 micrograms in 0.2 microliter saline) was used to explore the role of brain catecholamines (CA) and their interaction with corticosteroid feedback in stress responses of the ACTH-corticosterone (CORT) axis. The stereotaxic coordinates used for 6-OHDA lesions and the optimization of postlesion delays were determined by (a) radioautographic labeling of the VNAB axons after tracer injections into the dorsal A2/C2, or the ventral A1/C1 medullary areas, (b) histofluorescence and immunocytochemical location of interrupted CA pathways versus the postlesional scar, and (c) postlesional noradrenaline and adrenaline concentrations in whole hypothalami and paraventricular nuclei (PVN) punch samples. Two sites of 6-OHDA lesions were selected; both led to striking falls in PVN concentrations of both CA. The more dorsal lesion (dVNAB-X) was the same as that of our earlier studies and interrupted pathways originating predominantly in the A2/C2 area; the second more ventral lesion (vVNAB-X) interrupted axons stemming preferentially from the A1/C1 area. Both VNAB lesions inhibited the ether stress-induced ACTH and CORT surges in rats with intact adrenals. But the blockade (overall poststress release, amplitude and swiftness of hormonal responses) by dVNAB-X was greater than by vVNAB-X. The basal ACTH level in adrenalectomized rats (ADX) was elevated 20-fold and ether stress induced a 4-fold ACTH surge. As in sham-ADX rats, vVNAB-X in ADX rats induced only moderate inhibition of the ACTH response versus ADX + sham-vVNAB-X controls. On the other hand ADX + dVNAB-X rats showed a greatly amplified ACTH stress response over the ADX-sham dVNAB-X controls. This amplification was reversed by oral CORT supplementation. The data suggest that the CA pathways of the VNAB participating, directly or indirectly, in the poststress corticotropic activation may include subsets of CA axons of different origins, whose functional roles in stress are modulated in opposite directions by the plasma corticosteroid level.

Adrenal Cortex Hormones↗
PubMed1993

Homogeneous restriction fragment length polymorphism analysis of the ribosomal DNA repeating unit in New World Leishmania.

We have studied the Sau 3AI restriction length polymorphisms (RFLP) of the non-transcribed ribosomal spacer of Leishmania isolates from the mexicana and braziliensis complexes, using cloned sequences of Leishmania garnhami and Leishmania braziliensis. The L. garnhami probe produced very complex but conserved patterns in the homologous organisms, and these were shared by all the mexicana complex isolates at intermediate stringency conditions. The small subunit rRNA coding region within the probe also revealed a polymorphic Sau 3AI site exclusive of the braziliensis isolates. The braziliensis probe, containing only spacer sequences, yielded simple and very homogeneous patterns in all braziliensis isolates regardless of their geographical origin. Two main groups are identified in the New World isolates by the RFLP analysis in coincidence with the accepted mexicana and braziliensis complexes.

Animals↗
PubMed1992

Dipyridamole stimulates types II cAMP-dependent protein kinase in vitro.

Dipyridamole activates in vitro type II cAMP-dependent protein kinase. This agent stimulates the autophosphorylation of the regulatory subunit in the presence of cAMP but not so in the absence of the cyclic nucleotide. The activation was also observed with exogenous substrates such as casein, histone 2A and MAP2. This stimulation did not seem to be related to the cAMP binding to the R II subunit of the enzyme. Competition binding experiments showed that dipyridamole does not compete with adenosine for the A1 receptor. The results suggest that the reported regulatory properties of dipyridamole on lipid metabolism (González-Nicolás et al. Int J Biochem 21: 883-888, 1989) might be mediated through a direct action--an activation--on the catalytic subunit of a cAMP-dependent protein kinase.

Adipose Tissue↗
PubMed1992

Effects of litter removal on the lipolytic response and the regulatory components of the adenylate cyclase in adipocytes isolated from lactating rats.

The effects of litter removal on the status of different components of the hormone-sensitive adenylate cyclase system were analysed in plasma membranes of rat adipocytes. These effects were correlated with the decreased lipolytic response of adipose tissue. No changes in total number of A1 adenosine receptors or their affinity were detected in response to litter removal. In contrast, beta-adrenergic receptors showed a decrease (35%) in total number of receptors, without any significant change in their affinity. The status of alpha-GS and alpha-Gi, the alpha-subunits of G proteins which mediate stimulation and inhibition respectively of adenylate cyclase, were probed by cholera- and pertussis-toxin-catalysed ADP-ribosylation respectively and by immunoblot. Associated with litter removal, decreases of 63% and 62% in the incorporation of [alpha 32P]ADP-ribose catalysed by cholera toxin and pertussis toxin into alpha-Gs and alpha-Gi respectively were detected. Immunoblotting using RM/1 (anti-alpha-Gs) and AS/7 (anti-alpha-Gi) antisera also showed decreases in the levels of alpha-Gs (52%) and alpha-Gi (55%) in adipocyte membranes from litter-removed rats compared with lactating rats. Alterations in the status of hormone-sensitive adenylate cyclase components, such as those described herein, may be biochemical mechanism(s) by which adipose tissue shows a decreased lipolytic response during recovery from lactation.

Adenylyl Cyclases↗
PubMed1992

Identification of new world Leishmania using ribosomal gene spacer probes.

DNA probes from the nontranscribed ribosomal spacer (NTS), of Leishmania garnhami and Leishmania braziliensis were constructed and tested for sensitivity and specificity against different Leishmania isolates. The L. garnhami probes were species-specific under hybridization conditions of high stringency, but displayed specificity for the mexicana complex under conditions of intermediate stringency. The L. braziliensis probes showed 'complex' specificity. RFLP for the nontranscribed spacer within the braziliensis complex revealed very homogeneous patterns even for organisms currently accepted as different species. A PCR assay for the detection of Leishmania from the braziliensis complex is presented.

Animals↗
PubMed1992

[Decrease of pineal AMPc and TOH activity in the superior cervical ganglia after ablation of submaxillary glands in rats].

In order to investigate a possible functional relationship between the submandibular salivary gland (SSG) and the central nervous system (CNS), we have extirpated the salivary organs from thirty male rats. Twenty days after ablation both the pineal glands and the cervical superior ganglions (CSG) were dissected, homogenized and frozen until AMPc and TOH were assayed respectively. We observed a significant decrease in pineal AMPc (53.9 +/- 6.2 vs 76.1 +/- 7.6% of maximum value; p less than 0.02) which seems to be linked with a significant drop in TOH activity measured at CSG level (1.5 +/- 0.6 vs 3.7 +/- 0.9 nmoles of DOPA/h/pair GCS; p less than 0.03). Our results suggest that both findings might be due to the lack of NGF normally reaching the CSG from SSG. This data reinforces the idea of a functional link between SSG and CNS via the pineal gland.

Animals↗
PubMed1991

Microinjection of oxytocin into the dorsal vagal complex decreases pancreatic insulin secretion.

Microinjections of oxytocin and of an oxytocin antagonist into the dorsal vagal complex of the medulla oblongata were performed in order to study the possible role of the oxytocin containing axons that innervate this region in the regulation of pancreatic insulin secretion. No significant effect was produced by the intramedullary injection of the oxytocin vehicle alone or of 0.04 pM oxytocin. Injections of 4 and 20 pM oxytocin produced a reversible decrease of plasmatic insulin levels which fall to 59% of basal levels 15 min after the injection. Such an effect was abolished when 4 pM oxytocin was injected to animals which have been previously bilaterally vagotomized. In contrast to oxytocin, intramedullary injection of a specific antagonist of oxytocin to intact animals induced a marked increase of plasmatic insulin levels which raised 131% of basal levels 15 min after the injection. In animals receiving such an injection of oxytocin antagonist, a secondary injection of 4 pM oxytocin produced a slight but not significant decrease of plasmatic insulin levels. These data strongly suggest that the hypothalamic neurons producing oxytocin that densely project to the dorsal vagal complex may be involved in an inhibitory control of the vagal preganglionic neurons that innervate the pancreas.

Animals↗
PubMed1991

Differences in the immunoreactivity to phenylethanolamine-N-methyltransferase in the central adrenergic neurons of four strains of rats.

Immunocytochemistry was used to compare the immunoreactivity of adrenergic neurons to a well characterized specific immunoserum to phenylethanolamine-N-methyltransferase (PNMT) in different strains of rats commonly used in research studies. In adult animals, marked differences were found in the PNMT-immunoreactivity of neurons between Wistar rats and other strains, resulting in a lower PNMT-immunostaining intensity (i) within neuronal perikarya of the medulla oblongata, and (ii) more strikingly, within nerve fibers and terminals located in various brain regions. This low PNMT-immunoreactivity of nerve fibers was detected both in 14- and 35-day-old Wistar rats. On the other hand, the HPLC measurement of catecholamines, in particular of adrenaline in the hypothalamus and the medulla oblongata, did not show any difference between adult Wistar and Sprague-Dawley rats. These data suggest that the low PNMT-immunoreactivity observed in central adrenergic neurons of the Wistar rats is related to the poor recognition of the antigen by the PNMT-antibody used. Possibly, these nerve cells mainly display an isoform of the enzyme that is immunologically different from the PNMT contained within the adrenergic neurons of other rat strains.

Adrenergic Fibers↗