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1.
Int J Phytoremediation ; 21(11): 1112-1121, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31044606

RESUMO

Transgenic alfalfa (Medicago sativa L.) plants overexpressing the Arabidopsis ATP sulfurylase gene were generated using Agrobacterium-mediated genetic transformation to enhance their heavy metal accumulation efficiency. The ATP sulfurylase gene was cloned from Arabidopsis, following exposure to vanadium (V) and lead (Pb), and transferred into an Agrobacterium tumefaciens binary vector. This was co-cultivated with leaf explants of the alfalfa genotype Regen SY. Co-cultivated leaf explants were cultured on callus and somatic embryo induction medium, followed by regeneration medium for regenerating complete transgenic plants. The transgenic nature of the plants was confirmed using PCR and southern hybridization. The expression of Arabidopsis ATP sulfurylase gene in the transgenic plants was evaluated through RT-PCR. The selected transgenic lines showed increased tolerance to a mixture of five heavy metals and also demonstrated enhanced metal uptake ability under controlled conditions. The transgenic lines were fertile and did not exhibit any apparent morphological abnormality. The results of this study indicated an effective approach to improve the heavy metal accumulation ability of alfalfa plants which can then be used for the remediation of contaminated soil in arid regions.


Assuntos
Arabidopsis , Metais Pesados , Biodegradação Ambiental , Medicago sativa , Plantas Geneticamente Modificadas , Sulfato Adenililtransferase
2.
Avicenna J Med Biotechnol ; 10(3): 158-162, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30090209

RESUMO

BACKGROUND: The purpose of this study was to investigate the sterol profiling and predict the pharmacological potential of marine gastropod Telescopium telescopium (T. telescopium), collected from mangrove ecosystem in the South west coast of India. METHODS: Sterol fractions were separated from the crude lipids using 15% ethyl acetate. Ethyl acetate fractions were dried under ultrahigh purity N2 and analyzed using GC-MS. The biological activity was predicted using the software CLC-Pred; In silico predictions of cytotoxicity for tumor and non-tumor cell lines and PASS. RESULTS: This study proved the existence of four sterols, of which cholesterol was abundant. It was found that most of the steroids profiled from T. telescopium displayed activity against reproductive system as well as skin related diseases. CONCLUSION: The predicted anti infertility and skin related activity of the steroids identified from the marine gastropod T. telescopium is useful to attract industrial interest towards this species which will be helpful in rising new combinations with added therapeutic and nutritional worth.

3.
Mar Pollut Bull ; 118(1-2): 452-458, 2017 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-28336207

RESUMO

The present study investigates heavy metal concentrations in gastropods and bivalves collected from major fishing centers in South India. Three gastropods, Bursa spinosa, Tibia curta, and Murex trapa, and two bivalves, Perna viridis and Villoritta cyprinoids, were collected for the analysis of heavy metals. The metals in the present study followed the order Mg>Ca>Zn>Fe>Cu>Mn>Cr>Pb>Ni>Co>Cd. Trace metal concentrations in the soft tissue of the molluscs varied as follows: for Cd: 0.04-5.33, Co: 0.09-0.87, Cr: 2.18-7.59, Cu: 9.54-37.02, Mn: 1.30-8.50, Ni: 0.94-3.21, Pb: 1.16-2.64 and Zn: 68.16-113.64mgkg-1. Metal concentrations in all the species were below the limits proposed by the World Health Organization, except for Pb and Cd. This baseline study suggests that the levels of toxic metals in M. trapa, T. curta, and B. spinosa should be continuously monitored to assess the fate and effects of these metals in this fragile ecosystem.


Assuntos
Monitoramento Ambiental , Gastrópodes , Metais Pesados/análise , Moluscos , Poluentes Químicos da Água/análise , Animais , Ecossistema , Pesqueiros , Índia , Oligoelementos/análise
4.
Bull Environ Contam Toxicol ; 94(4): 496-502, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25694163

RESUMO

Surface sediment samples were collected from Chitrapuzha (Cochin) estuarine system to identify the natural and anthropogenic origin of organic matter. The distribution and sources of organic matter were assessed with the help of fatty acid and alkane biomarkers. Fatty acids ranging from C12 to C28 were identified and C16:0 was the most abundant fatty acid, which contributed between 23.5 % and 52.4 % to total fatty acids. The low levels of polyunsaturated fatty acids indicate the effective bacterial recycling of algal fatty acids during the whole settling and depositing process. Aliphatic hydrocarbons ranging from C12 to C33 were identified and the total concentration ranged from 7876 to 43,357 ng g(-1). The presence of unresolved complex mixtures and lower pristane to phytane ratios indicates the petroleum contamination in the study area.


Assuntos
Biomarcadores/análise , Monitoramento Ambiental/estatística & dados numéricos , Estuários , Sedimentos Geológicos/química , Rios , Poluentes Químicos da Água/análise , Alcanos/análise , Monitoramento Ambiental/métodos , Ácidos Graxos/análise , Hidrocarbonetos/análise , Índia , Petróleo/análise
5.
Environ Monit Assess ; 185(12): 10317-31, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23832233

RESUMO

Trace metal concentrations in the muscle of the bivalve Villorita cyprinoides from the Cochin backwaters (southwest coast of India) were investigated during the monsoon, post-monsoon and pre-monsoon periods. The seasonal average ranges of metals (µg g(-1), dry weight) in the bivalve were as follows: Fe (18,532.44-28,267.05), Co (23.25-37.58), Ni (10.56-19.28), Cu (3.58-11.35), Zn (48.45-139.15), Cd (1.06-1.50) and Pb (3.05-4.35). The marginally elevated metal concentrations in bivalve muscles are probably related to high influx of metals as a result of pollution from the industries and agricultural fields with consequent increased bioavailability of metals to the bivalve. Evaluation of the risks to human health associated with consumption of the bivalves suggested that there is no health risk for moderate shellfish consumers. A regular and continuous biomonitoring program is recommended to establish V. cyprinoides as a bioindicator for assessing the effects of trace metal pollution and to identify future changes to conserve the "health" of this fragile ecosystem.


Assuntos
Bivalves/metabolismo , Monitoramento Ambiental , Metais/metabolismo , Poluentes Químicos da Água/metabolismo , Animais , Ecossistema , Índia
6.
ScientificWorldJournal ; 2012: 972839, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22645488

RESUMO

Concentrations and distributions of trace metals (Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, and Zn) in surficial sediments of the Cochin backwaters were studied during both monsoon and pre-monsoon periods. Spatial variations were in accordance with textural charaterstics and organic matter content. A principal component analysis distinguished three zones with different metal accumulation capacity: (i) highest levels in north estuary, (ii) moderate levels in central zone, and (iii) lowest levels in southern part. Trace metal enrichments are mainly due to anthropogenic contribution of industrial, domestic, and agricultural effluents, whose effect is enhanced by settling of metals due to organic flocculation and inorganic precipitation associated with salinity changes. Enrichments factors using Fe as a normalizer showed that metal contamination was the product of anthropogenic activities. An assessment of degree of pollution-categorized sediments as moderately polluted with Cu and Pb, moderately-to-heavily polluted with Zn, and heavily-to-extremely polluted with Cd. Concentrations at many sites largely exceed NOAA ERL (e.g., Cu, Cr, and Pb) or ERM (e.g., Cd, Ni, and Zn). This means that adverse effects for benthic organisms are possible or even highly probable.


Assuntos
Sedimentos Geológicos/análise , Metais/análise , Agricultura , Animais , Ecossistema , Monitoramento Ambiental/métodos , Poluentes Ambientais/química , Geografia , Humanos , Índia , Metais/química , Compostos Orgânicos , Análise de Componente Principal , Reprodutibilidade dos Testes , Sais , Água do Mar , Poluentes Químicos da Água/análise
7.
Indian Pediatr ; 49(1): 17-9, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21719928

RESUMO

OBJECTIVE: To study the role of iron deficiency as a risk factor for simple febrile seizures. DESIGN: Case control study. SETTING: Pediatric department of a tertiary care teaching hospital. PARTICIPANTS: 154 cases and 154 controls were included in the study. Consecutive cases and concurrent controls were selected. Cases were children of age group 6 months to 3 years presenting with simple febrile seizures. Controls were children of same age group presenting with short febrile illness but without any seizures. METHODS: After informed consent, detailed history was taken and clinical examination done in both cases and controls and blood investigations were done to diagnose iron-deficiency in both cases and controls. Iron deficiency was diagnosed as per WHO criteria (hemoglobin value <11 g%, red cell distribution width of >15% and serum ferritin value <12 ng/mL). Other explanatory variables, which can be the potential confounders were also included in the study and considered for analysis. RESULTS: Highly significant association was found between iron deficiency and simple febrile seizures in both univariate and multivariate analysis. Crude odds ratio was 5.34 (CI 3.27- 8.73, P<0.001) and adjusted odds ratio in the logistic regression analysis was 4.5 (CI 2.69- 7.53, P <0.001). CONCLUSIONS: Iron deficiency is a significant risk factor for simple febrile seizures in children of age group 6 months to 3 years.


Assuntos
Anemia Ferropriva/complicações , Convulsões Febris/etiologia , Anemia Ferropriva/sangue , Estudos de Casos e Controles , Pré-Escolar , Índices de Eritrócitos , Feminino , Humanos , Lactente , Masculino , Fatores de Risco
8.
Arch Environ Contam Toxicol ; 60(4): 597-609, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-20628738

RESUMO

This article focuses on the temporal and spatial distribution of three organophosphorous pesticides-malathion, methyl parathion, and chlorpyrifos-in the sedimentary environment of a backwater ecosystem, Kuttanad backwaters, situated in Kerala, India. Based on salinity distribution, geographic characteristics, and human activities prevailing in the area, the study area was divided into three zones: zone 1 with riverine characteristics, zone 2 with freshwater characteristics during and after the monsoon season and estuarine characteristics during the premonsoon season, and zone 3 with estuarine characteristics. The organophosphorus pesticides in the study area showed the order of enrichment as chlorpyrifos >malathion >methyl parathion. While studying the variations in pesticide concentrations seasonally, higher concentrations were observed during the premonsoon monsoon season, with the concentrations being lower than the detectable level. Sediment characteristics, such as pH, texture, organic carbon, moisture content, etc., had reflective effect on the degradation rates of pesticides. The runoff water from the paddy fields made a larger contribution of pesticide pollution to the study area.


Assuntos
Agricultura , Sedimentos Geológicos/análise , Compostos Organofosforados/análise , Praguicidas/análise , Água do Mar/química , Poluentes Químicos da Água/análise , Monitoramento Ambiental , Índia , Estações do Ano , Movimentos da Água , Áreas Alagadas
9.
Indian Pediatr ; 46 Suppl: s59-62, 2009 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19279372

RESUMO

This study was conducted to find out the anthropometric, biosocial and obstetric predictors of birthweight in Kerala. The study sample consisted of 599 consecutive liveborn babies delivered at SAT Hospital, Medical College, Thiruvananthapuram and their mothers in November 2001. Details of maternal history, anthropometry, and biosocial and demographic factors were recorded. Birthweight was primary outcome variable. Multivariate analysis revealed that the biologically acceptable predictors of birthweight of a baby in our population are maternal height (P <0.001), parity (P < 0.001) gestational age (P < 0.001), pregnancy induced hypertension(P=0.05) and history of low birthweight in the previous pregnancy(P=0.05).


Assuntos
Peso ao Nascer , Peso ao Nascer/fisiologia , Estatura , Estudos Transversais , Feminino , Idade Gestacional , Humanos , Índia , Recém-Nascido , Masculino , Mães , Análise Multivariada , Paridade , Gravidez
10.
Indian Pediatr ; 46 Suppl: s83-5, 2009 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19279379

RESUMO

This study was done to examine the type and severity of disciplinary practices adopted by parents in Kerala. 1668 mothers, aged 18-49, participated in face-to-face interviews as part of a cross-sectional, population-based survey covering rural, urban and coastal areas. One child from each family was randomly selected as the index child. The interview focused primarily on disciplinary practices. 62% of the mothers reported using severe verbal discipline and 50% of mothers reported using severe physical abuse. Results suggest a high prevalence of normative and abusive practices in the community with mothers playing a prime role in disciplining the child.


Assuntos
Maus-Tratos Infantis/estatística & dados numéricos , Educação Infantil , Adolescente , Adulto , Criança , Estudos Transversais , Feminino , Humanos , Índia , Pessoa de Meia-Idade , Adulto Jovem
11.
Environ Monit Assess ; 128(1-3): 195-208, 2007 May.
Artigo em Inglês | MEDLINE | ID: mdl-17016753

RESUMO

This paper focuses on the partitioning of trace metals in five selected coral species from Lakshadweep Archipelago, which remains as one of the least studied areas in the Indian Ocean. Based on the morphological features, selected coral species are classified as massive (Porites andrewsi), ramose or branching (Lobophyllia corymbosa, Acropora formosa and Psammocora contigua) and foliaceous (Montipora digitata). Relating trace metal concentrations with morphological features in skeleton, highest concentrations of all the trace metals (except Zn) were reported for the ramose type corals. In tissue, all the metals (essential as well as non essential) showed highest concentrations within the branching type corals. Irrespective of their growth characteristics/pattern, all species except P. contigua displayed higher concentrations of Pb, Ni, Mn and Cd within their skeleton compared to tissue which may exemplify a regulatory mechanism to avoid the build up of the concentrations of these metals in their bio-part, strikingly toxic metals like Cd and Pb. The concentrations of trace metals in the skeleton and tissues of these coral species were subjected to 3 way ANOVA based on non standardized original data and the results showed significant differences between metals and between species leading to high skeleton/ tissue - species interaction as well as skeleton/tissue - metal interaction. The significant values of student's t calculated are depicted in the form of Trellis diagrams.


Assuntos
Antozoários/efeitos dos fármacos , Monitoramento Ambiental/métodos , Metais Pesados/toxicidade , Poluentes Químicos da Água/toxicidade , Animais , Oceanos e Mares
12.
Environ Geochem Health ; 28(6): 509-18, 2006 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17094013

RESUMO

The present study focuses on the determination and abundance of trace metals (viz. Cu, Ni, Zn, Cr, Co, Cd, Mn and Fe) in the surficial sediments of west coast of Arabian Sea along the Indian subcontinent. Sediment samples were collected from three transects along the western continental shelf of Arabian Sea. The enrichment of Fe and Mn in coastal oxic-sediments indicates the precipitation of these redox sensitive elements as Fe- and Mn-hydroxides and oxides, whereas the low Fe and Mn concentrations in the oxygen deficient sediments of deeper stations reflects the dissolution of their hydroxides and oxides. Concentrations of fairly redox insensitive trace metals like Cu, Ni, Zn, Cr and Cd (with the exceptions of Cr) showed higher values at nearshore sediments, then it decreased towards seaward and again showed a slight increase at oxygen minimum stations in all the three transects. This geochemical variability in their distributional characteristics is mainly associated with the extent to which the precipitation or dissolution of Fe- and Mn-oxides/hydroxides occur since the scavenging or releasing effects of Fe- and Mn-oxides/hydroxides act as significant 'sinks' or 'sources' of heavy metals. The change in wind pattern, coastal upwelling and increased productivity are also the reported factors which influence the biogeochemical cycling of trace metals in the surface sediments of west coast of India. Enrichment factor generally showed a high gradient accumulation from nearshore to shelf.


Assuntos
Sedimentos Geológicos/química , Metais Pesados/análise , Geografia , Índia , Oceanos e Mares , Oxirredução , Oxigênio/química , Cloreto de Sódio/química , Temperatura
14.
Eur J Neurosci ; 20(12): 3233-43, 2004 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-15610156

RESUMO

Removal of circulating corticosterone by adrenalectomy (ADX) leads to apoptosis after 3 days in a small population of rat dentate granule neurons, whereas most surrounding cells remain viable. Interestingly, a specific expression profile is triggered in surviving granule cells that may enhance their survival. Hippocampal slices prepared 1, 2 or 3 days after ADX or sham operation were stained ex vivo with Hoechst 33258, which serves to identify apoptotic neurons. After electrophysiological analysis, multiple gene expression in surviving individual granule cells was assessed by linear antisense RNA amplification and hybridization to slot blots containing various neuronal cDNAs. Hierarchical clustering and principal component analysis was performed on two physiological variables and 14 mRNA ratios from ADX cells from every time point. Our results indicate that surviving 3-day ADX granule cells display lower membrane capacitance, lower relative N-methyl-d-aspartate (NMDA) R1 mRNA expression and higher relative mineralocorticoid receptor (MR), alpha1A voltage-gated Ca-channel, Bcl-2 and NMDA R2C mRNA expression. Some 1- and 2-day ADX cells cluster with these 3-day survivors; therefore, one or more components of their mRNA expression profile may represent predictive markers for apoptosis resistance. The functional relevance of two candidate genes was tested by in vivo local over-expression in the same model system; of these, Bcl-2 conferred partial protection when induced shortly before ADX. Therefore, removal of corticosteroids triggers a specific gene expression profile in surviving dentate granule cells; key components of this profile may be associated with their survival.


Assuntos
Sobrevivência Celular/fisiologia , Corticosterona/deficiência , Giro Denteado/metabolismo , Perfilação da Expressão Gênica/estatística & dados numéricos , Adrenalectomia/estatística & dados numéricos , Animais , Corticosterona/biossíntese , Corticosterona/genética , Giro Denteado/citologia , Perfilação da Expressão Gênica/métodos , Técnicas In Vitro , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Ratos
15.
J Neurosci Res ; 62(2): 186-205, 2000 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-11020212

RESUMO

Developmental expression of sulfoglucuronyl carbohydrate (SGC) and its binding protein, SBP-1 was studied in the rat cerebral cortex to understand their function. Between embryonic day (ED) 14-19, SBP-1 was strongly expressed in neurons of the ventricular zone and migrating neurons throughout the cortex. SBP-1 declined at birth and by postnatal day (PD) 3 only the latest arriving neurons in the most superficial segment of the cortical plate expressed SBP-1. Between ED 14-16, SGC was expressed in a thin row of glial cells near the ventricles and on their radial processes. Between ED 16-PD 3, SGC was not in neuronal cell soma, but was in neuronal plasma membranes and processes surrounding the neuronal perikarya. The expression of SGC declined similar to SBP-1 and both of them disappeared by PD 7. The expression of SBP-1 and SGC was chronologically coordinated with neuronal migration. SBP-1 was specifically expressed in immature neuronal nuclei and plasma membranes. SBP-1 and SGC were colocalized and were available for interaction with each other on neuronal cell membranes and processes. This was confirmed with isolated neurons in culture. As in vivo, the expression of SBP-1 in neurons declined with time in culture. The dissociated cortical neurons when plated on SBP-1 as a substratum produced extensive neuritic outgrowth. HNK-1, anti-SBP-1 antibodies and sulfoglucuronyl glycolipid, SGGL specifically and severely reduced neurite outgrowth. SBP-1-SGC interactions provide a potential mechanism for guidance and cell signaling, in the processes of neuronal migration and terminal differentiation.


Assuntos
Antígenos CD57/metabolismo , Metabolismo dos Carboidratos , Movimento Celular/fisiologia , Córtex Cerebral/metabolismo , Glicolipídeos/metabolismo , Glicoproteínas/metabolismo , Neurônios/metabolismo , Animais , Animais Recém-Nascidos , Anticorpos Monoclonais , Antígenos CD57/imunologia , Adesão Celular , Células Cultivadas , Córtex Cerebral/crescimento & desenvolvimento , Embrião de Mamíferos , Epitopos , Feminino , Neuritos/metabolismo , Gravidez , Ligação Proteica , Ratos , Ratos Sprague-Dawley
16.
Brain Res Dev Brain Res ; 120(2): 165-80, 2000 Apr 14.
Artigo em Inglês | MEDLINE | ID: mdl-10775769

RESUMO

Sulfoglucuronyl carbohydrate (SGC) is expressed on several glycoproteins of the immunoglobulin superfamily of cell-adhesion molecules. Developmental expression of SGC and its binding protein, SBP-1, was studied in the rat cerebellum by immunocytochemistry to understand the function of SBP-1 and the significance of its interaction with SGC. During early postnatal development (postnatal day (PD) 3-10) SBP-1 was strongly expressed in the granule neurons of the external and internal granule cell layers (EGCL and IGCL). This expression declined by PD 15, and disappeared in the adult. Between PD 3 and 15, SGC was expressed in cellular processes surrounding the granule neurons in the IGCL, and it also declined and disappeared with development. SGC expression, however, continued in Purkinje cells and their dendrites in the molecular layer in adults. The expressions of SBP-1 and SGC were developmentally regulated and appeared to be chronologically co-ordinated with granule neuron migration from EGCL to IGCL. High magnification confocal microscopy showed that SBP-1 was primarily localized in nuclei and plasma membranes of granule neurons, whereas SGC in the IGCL was localized on neuronal plasma membranes, dendrites and glial processes, but not in cell soma. The relative localization of SBP and SGC was confirmed by cellular and subcellular markers in vivo and with dissociated cerebellar cells in culture. It is proposed that SBP-1 on plasma membranes of granule neurons interacts with SGC on the surrounding processes and membranes and this interaction could provide a potential mechanism for guidance and cell signaling, in the processes of granule neuron migration and differentiation.


Assuntos
Antígenos CD57/metabolismo , Diferenciação Celular/fisiologia , Movimento Celular/fisiologia , Cerebelo/crescimento & desenvolvimento , Cerebelo/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Proteínas de Membrana/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Neurônios/metabolismo , Envelhecimento/metabolismo , Animais , Animais Recém-Nascidos , Adesão Celular/fisiologia , Comunicação Celular/fisiologia , Compartimento Celular/fisiologia , Membrana Celular/metabolismo , Células Cultivadas , Cerebelo/citologia , Imuno-Histoquímica , Microscopia Confocal , Proteínas Associadas aos Microtúbulos/metabolismo , Moléculas de Adesão de Célula Nervosa/metabolismo , Neurônios/citologia , Ligação Proteica , Ratos , Ratos Sprague-Dawley
17.
Proc Natl Acad Sci U S A ; 96(23): 13456-61, 1999 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-10557342

RESUMO

Recent studies indicated that hyperactivity of the hypothalamo-pituitary-adrenal system is a considerable risk factor for the precipitation of affective disorders, most notably of major depression. The mechanism by which this hyperactivity eventually leads to clinical symptoms of depression is unknown. In the present animal study, we tested one possible mechanism, i.e., that long-term exposure to high corticosterone levels alters functional responses to serotonin in the hippocampus, an important area in the etiology of depression. Rats were injected daily for 3 weeks with a high dose of corticosterone; electrophysiological responses to serotonin were recorded intracellularly from CA1 pyramidal neurons in vitro. We observed that daily injections with corticosterone gradually attenuate the membrane hyperpolarization and resistance decrease mediated by serotonin-1A receptors. We next used single-cell antisense RNA amplification from identified CA1 pyramidal neurons to resolve whether the functional deficits in serotonin responsiveness are accompanied by decreased expression levels of the serotonin-1A receptor. It appeared that expression of serotonin-1A receptors in CA1 pyramidal cells is not altered; this result was supported by in situ hybridization. Expression of corticosteroid receptors in the same cells, particularly of the high-affinity mineralocorticoid receptor, was significantly reduced after long-term corticosterone treatment. The present findings indicate that prolonged elevation of the corticosteroid concentration, a possible causal factor for major depression in humans, gradually attenuates responsiveness to serotonin without necessarily decreasing serotonin-1A receptor mRNA levels in pyramidal neurons. These functional changes may occur by a posttranscriptional mechanism or by transcriptional regulation of genes other than the serotonin-1A receptor gene itself.


Assuntos
Corticosterona/farmacologia , Hipocampo/efeitos dos fármacos , Serotonina/metabolismo , Animais , Corticosterona/administração & dosagem , Hipocampo/metabolismo , Hibridização In Situ , Masculino , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Receptores de Glucocorticoides/genética , Receptores de Mineralocorticoides/genética
18.
Neuroscience ; 85(3): 759-71, 1998 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-9639270

RESUMO

Sulfoglucuronyl carbohydrate is the terminal moiety of neolacto-oligosaccharides, expressed on several glycoproteins of the immunoglobulin superfamily involved in cell-cell recognition and on two glycolipids. Sulfoglucuronyl carbohydrate is temporally and spatially regulated in the developing nervous system. It appears to be involved in neural cell recognition and in cell adhesion processes through its interaction with specific proteins on cell surfaces. Previously we have characterized a specific sulfoglucuronyl carbohydrate-binding protein in rat brain. Sulfoglucuronyl carbohydrate binding protein-1 is structurally similar to a 30,000 mol. wt adhesive and neurite outgrowth promoting protein amphoterin [Rauvala and Pihlaskari (1987) J. biol. Chem. 262, p. 16,625]. The pattern of expression of sulfoglucuronyl carbohydrate binding protein-1 in developing rat nervous system was studied to understand the significance of its interaction with sulfoglucuronyl carbohydrate-bearing molecules. Biochemical analyses showed that the expression of sulfoglucuronyl carbohydrate binding protein-1 was developmentally regulated similarly to sulfoglucuronyl carbohydrate. Immunocytochemical localization of sulfoglucuronyl carbohydrate binding protein-1 and sulfoglucuronyl carbohydrate was performed by bright-field and fluorescent confocal laser scanning microscopy. In postnatal day 7 rat cerebellum, sulfoglucuronyl carbohydrate binding protein-1 was primarily associated with neurons of the external and internal granule cell layers. The sulfoglucuronyl carbohydrate binding protein-1 immunoreactivity was absent in Purkinje cell bodies and their dendrites in the molecular layer, as well as in Bergmann glial fibres and in white matter. In contrast, sulfoglucuronyl carbohydrate (reactive with HNK-1 antibody) was localized in processes surrounding granule neurons in the internal granule cell layer. Sulfoglucuronyl carbohydrate was also expressed in Purkinje neurons and their dendrites in the molecular layer and their axonal processes in the white matter. To a lesser extent Bergmann glial fibres were also positive for sulfoglucuronyl carbohydrate. In the cerebral cortex, at embryonic day 21, sulfoglucuronyl carbohydrate binding protein-1 was mainly observed in immature neurons of the cortical plate and subplate and dividing cells near the ventricular zone. Whereas, sulfoglucuronyl carbohydrate was strongly expressed in the fibres of the subplate and marginal zone. Sulfoglucuronyl carbohydrate was also found in the processes surrounding the sulfoglucuronyl carbohydrate binding protein-1-expressing neuronal cell bodies in the cortical plate and in ventricular zone. The specific localization of sulfoglucuronyl carbohydrate binding protein- in cerebellar granule neurons and neurons of the cerebral cortex was also confirmed by immunocytochemistry of the dissociated tissue cell cultures. The complementary localization of sulfoglucuronyl carbohydrate and sulfoglucuronyl carbohydrate binding protein-1, both in cerebral cortex and cerebellum, in apposing cellular structures indicate possible interaction between the two and signalling during the process of cell migration and arrest of migration.


Assuntos
Antígenos CD57/biossíntese , Carboidratos/biossíntese , Cerebelo/química , Córtex Cerebral/química , Glicoproteínas/análise , Animais , Anticorpos , Especificidade de Anticorpos , Química Encefálica/fisiologia , Antígenos CD57/análise , Carboidratos/análise , Cerebelo/citologia , Córtex Cerebral/citologia , Imunofluorescência , Globosídeos/análise , Globosídeos/biossíntese , Glucuronatos/análise , Glucuronatos/metabolismo , Glicoproteínas/imunologia , Glicoproteínas/metabolismo , Microscopia Confocal , Neurônios/química , Neurônios/metabolismo , Coelhos , Ratos , Ratos Sprague-Dawley
19.
J Neurosci ; 18(7): 2685-96, 1998 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-9502826

RESUMO

Overexposure to corticosteroid hormones is harmful to hippocampal neuronal integrity, likely by perturbation of calcium homeostasis. To identify molecular mechanisms at the single-cell level, we characterized mRNA expression corresponding to voltage- and ligand-gated Ca channels in individual dissociated CA1 neurons in response to long-term corticosterone (CORT) exposure. Predominant mineralocorticoid receptor occupation (ADC-LO group) resulted in low levels of P/Q- and L-type Ca channel mRNAs, high levels of GluR-2 versus GluR-1, and a high ratio of NMDAR-2A to NMDAR-2B mRNA. Corresponding alterations in protein expression were consistent with the restriction of Ca influx. In contrast, additional glucocorticoid receptor occupation (ADC-HI group) altered the expression of these mRNAs in a manner consistent with enhanced Ca influx; interestingly, qualitatively similar alterations were seen in control ADX neurons. Electrophysiological data from the same neurons indicate that Ca current amplitudes also are modulated by CORT, although on a shorter time scale. Finally, principal components analysis (PCA) suggests that neuronal AMPA and NMDA receptor composition may be regulated by MR and GR activation in a complex manner. Therefore, our data implicate molecular events by which CORT may regulate Ca influx into CA1 hippocampal neurons.


Assuntos
Corticosteroides/farmacologia , Canais de Cálcio/genética , Hipocampo/citologia , Neurônios/efeitos dos fármacos , Adrenalectomia , Animais , Cálcio/metabolismo , Canais de Cálcio/metabolismo , Expressão Gênica/efeitos dos fármacos , Hipocampo/química , Ativação do Canal Iônico/fisiologia , Masculino , Neurônios/fisiologia , Técnicas de Patch-Clamp , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores de AMPA/genética , Receptores de AMPA/metabolismo , Receptores de Glucocorticoides/fisiologia , Receptores de N-Metil-D-Aspartato/genética , Receptores de N-Metil-D-Aspartato/metabolismo
20.
J Neurochem ; 68(3): 1286-97, 1997 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-9048776

RESUMO

The developmentally regulated and stage-specifically expressed HNK-1 carbohydrate found on sulfoglucuronylglycolipids (SGGLs) and certain glycoproteins has been proposed to be involved in neural cell adhesion and recognition processes through its interaction with protein "receptors." We have isolated and purified a approximately 30-kDa SGGL-binding protein (SBP-1) from neonatal rat brain. SBP-1 specifically bound to SGGLs and sulfatide both in solid-phase immunobinding and high-performance thin-layer chromatography-immunooverlay assays. N-terminal sequence analysis showed that SBP-1 is similar to an adhesive neurite outgrowth promoting protein amphoterin. Desulfation of SGGLs resulted in abolition of SBP-1 binding. However, chemical modification of glucuronic acid moiety by either esterification or reduction of the carboxyl group had no effect, suggesting requirement of the carbohydrate-linked sulfate group for SBP-1 binding. The binding of SBP-1 to SGGLs was specifically inhibited by HNK-1 antibody but not by other IgM antibodies. The binding of SBP-1 to sulfatide, however, was not inhibited by HNK-1 antibody. Heparin, fucoidan, and dextran sulfate (50K) also inhibited the binding of SBP-1 to SGGLs. During development of the rat cerebral cortex, the level of SBP-1 decreased after embryonic day 18 to an almost undetectable level by postnatal day 10, whereas in the cerebellum, the expression of SBP-1 was maximal at postnatal day 7. SBP-1 also bound specifically to the HNK-1 glycoproteins isolated from rat brain by HNK-1 immunoaffinity chromatography. Proteins without HNK-1 carbohydrate did not bind SBP-1. The binding to HNK-1 glycoproteins was inhibited by HNK-1 antibody, but not by other IgM antibodies, indicating that the binding was mediated through the HNK-1 carbohydrate moiety of the proteins. The interaction and coexpression of SBP-1 with SGGLs and HNK-1 glycoproteins, during the perinatal brain development, suggest a functional role for this protein.


Assuntos
Proteínas de Transporte/metabolismo , Glucuronatos/metabolismo , Sistema Nervoso/crescimento & desenvolvimento , Sistema Nervoso/metabolismo , Sequência de Aminoácidos , Animais , Animais Recém-Nascidos/crescimento & desenvolvimento , Anticorpos Monoclonais/farmacologia , Proteínas de Transporte/antagonistas & inibidores , Proteínas de Transporte/isolamento & purificação , Cerebelo/crescimento & desenvolvimento , Cerebelo/metabolismo , Córtex Cerebral/crescimento & desenvolvimento , Córtex Cerebral/metabolismo , Glicoproteínas/efeitos dos fármacos , Glicoproteínas/metabolismo , Concentração de Íons de Hidrogênio , Íons , Metabolismo dos Lipídeos , Concentração Osmolar , Ratos
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