[Streptococcus agalactiae meningitis in an adult with cranio-encephalic trauma and neurosurgery].
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Brain Injuries↗
Biomedical subjects
A Quesada
Publications and source records attributed to A Quesada.
Spatiotemporal gradients of differentiation of chick retina types I and II cholinergic cells: identification of a common postmitotic cell population.
The chick retina has three types of cholinergic amacrine cells. We have found that Types I and II differentiate from a common population of postmitotic cells temporarily located in the inner plexiform layer (IPL cells). Golgi staining and immunocytochemistry for choline acetyltransferase (ChAT) and gamma-aminobutyric acid (GABA) were used to trace the development and fate of IPL cells. Transformation of the shape of IPL cells into those typical of both conventional amacrine cells and those displaced to the ganglion cell layer are seen. All IPL cells are doubly immunoreactive, for ChAT and GABA, from the time they appear as a cell population within the inner plexiform layer (IPL) until their separation into the two amacrine cell populations. Polarization and early stages of shape differentiation of both types occur while they are in the IPL, starting in the dorsocentral area in the temporal retina and spreading to the rest of the retina. Three spatial gradients of differentiation are observed: from central-to-peripheral, dorsal-to-ventral, and temporal-to-nasal retina. Our findings suggest that the fate of both types of cells in the chick is determined locally, whereas their postmitotic precursors are within the IPL. The presence of GABA and acetylcholine in both types of amacrine cells at early stages of their morphogenesis, well before they have synaptic interactions, suggests a morphogenetic role for these molecules in inner retinal differentiation.
Acetylcholine↗
Quantitative and ultrastructural changes in glia and pericytes in the parietal cortex of the aging rat.
The frequency of astrocytes, microglia plus oligodendrocytes, and pericytes displaying nuclei was analyzed and quantified in 160-microm-wide strips of the parietal cortex (Par1 region) from young and aged Wistar rats. The study was performed on two groups of rats aged 3-4 and 32-36 months. Quantifications of the glial cell types and pericytes were made in 1-microm-thick sections stained with toluidine blue. Ultrathin sections were also made to analyze the ultrastructural features of these cells during aging. Astrocytes and pericytes increased in number by about 20% and 22%, respectively, with age. These increases were most significant in layers II-IV and V for both cellular types. Clusters of astrocytes were common in these layers of aging rats. The ultrastructural analysis also indicated changes in all cell types that stored inclusions and vacuoles with age, which were particularly abundant in microglial cells. End-feet astrocytes and pericytes surrounding the vascular wall also contained vacuoles and inclusions, and consequently the vascular wall increased in thickness. In conclusion, the aging process increased astrocyte and pericyte populations, but not microglia plus oligodendrocyte populations, in the rat parietal cortex. Although no significant change in nuclear size could be observed in any cell type, all glial cells as well as pericytes underwent morphological ultrastructural changes. These modifications may result from the need to correct possible homeostatic imbalances during aging.
Aging↗
Acclimation of Cyanobacterial Communities in Rice Fields and Response of Nitrogenase Activity to Light Regime
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Journal Article↗
Clustering of the nitrite reductase gene and a light-regulated gene with nitrate assimilation loci in Chlamydomonas reinhardtii.
Two new loci have been found to be clustered with five other genes for the nitrate assimilation pathway in the Chlamydomonas reinhardtii genome. One gene, located close to the 3'-end of the high-affinity nitrate transporter (HANT) gene Nrt 2; 2, corresponds to the nitrite reductase (NiR) structural gene Nii1. This is supported by a number of experimental findings: (i) NiR-deficient mutants have lost Nii1 gene expression; (ii) Nii1 mRNA accumulation is co-regulated with the expression of other structural genes of the nitrate assimilation pathway; (iii) nitrite (nitrate) utilization ability is recovered in the NiR mutants by functional complementation with a wild-type Nii1 gene; (iv) the elucidated NII1 amino acid sequence is highly similar to that of the cyanobacterial and higher-plant enzyme, and contains the predicted domains for plastidic ferredoxin-NiRs. Thus, the mutant phenotype and the mRNA sequence and expression of the Nii1 gene have been unequivocally related. Accumulation of mRNA for the second locus identified. Lde1 (light-dependent expression), was not regulated by nitrogen, but like nitrate-assimilation clustered genes, its expression was down-regulated in the dark.
Amino Acid Sequence↗
Three Nrt2 genes are differentially regulated in Chlamydomonas reinhardtii.
Two new nitrate assimilation-related genes, Nrt2;3 and Nar5, have been identified in Chlamydomonas reinhardtii. The Nrt2;3 gene is a new member of the Nrt2 family, encoding high-affinity nitrate (nitrite) transporters. Like that of the nitrate assimilation genes, expression of the Nrt2;3 gene is down-regulated by ammonium and positively controlled by Nit2, a regulatory locus specific for the pathway. The three Nrt2 genes of C. reinhardtii are differentially regulated by the nitrogen source. Expression of Nrt2;3 and of Nrt2;1, a nitrate/nitrite-bispecific transporter gene, was induced by nitrate and more efficiently by nitrite. Accumulation of mRNA of Nrt2;2, the nitrate-specific transporter gene, was only induced efficiently by nitrate. The Nar5 gene is located upstream of the Nrt2;3 genomic region and expression of its mRNA is down-regulated by ammonium. The Nrt2;3 and Nar5 genes are overexpressed in a deletion mutant that lacks nitrate assimilation loci.
Animals↗
Glutamine synthetase (GS) activity and spatial and temporal patterns of GS expression in the developing chick retina: relationship with synaptogenesis in the outer plexiform layer.
The profile of glutamine synthetase (GS) activity in the neural retina of chicken embryos and adults was studied alongside the in vivo spatio-temporal patterns of generation and morphogenesis of Müller cell and of retinal synaptogenesis. The rise of GS activity during development is not related to Müller cell differentiation but to synaptogenesis in the outer plexiform layer (opl). GS expression was investigated by immunoreaction with GS-specific antiserum. Three spatial gradients of decreasing labeling were observed between embryonic (E)15 and E18, from central to peripheral retina, dorsal to ventral, and temporal to nasal, which are in spatio-temporal relationship with synaptogenesis in the opl. GS is localized in Müller cells and apparently also in a population of astrocyte-like glial cells, located in the ganglion cell layer throughout the retina. Precocious induction by hydrocortisone, in ovo, at E10, does not show the spatial pattern of GS immunoreactivity observed in control retinas at the time of natural induction (E15). We also show that dissociated (non-cultured) Müller cells of E18-20 retinas, to which only photoreceptors or photoreceptors and neurons remain joined, maintain an immunodetectable level of GS, while those in isolated state lose GS immunoreactivity rapidly. Our results suggest that the induction of GS expression might be mediated by Müller cell-neuron interactions at the opl and also perhaps at the outer nuclear layer (onl). An analysis of our results and those of previous authors suggests that the level of GS in differentiated Müller cells could be determined by conjoint cell interactions at the onl, opl, and inner plexiform layer.
Animals↗
Sequence of the non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase from Nicotiana plumbaginifolia and phylogenetic origin of the gene family.
A cDNA-library has been constructed from Nicotiana plumbaginifolia seedlings, and the non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase (GapN, EC 1.2.1.9) was isolated by plaque hybridization using the cDNA from pea as a heterologous probe. The cDNA comprises the entire GapN coding region. A putative polyadenylation signal is identified. Phylogenetic analysis based on the deduced amino acid sequences revealed that the GapN gene family represents a separate ancient branch within the aldehyde dehydrogenase superfamily. It can be shown that the GapN gene family and other distinct branches of the superfamily have its phylogenetic origin before the separation of primary life-forms. This further demonstrates that already very early in evolution, a broad diversification of the aldehyde dehydrogenases led to the formation of the superfamily.
Aldehyde Dehydrogenase↗
Light microscopic quantification of morphological changes during aging in neurons and glia of the rat parietal cortex.
BACKGROUND: Different changes in neuronal and glial population of the aging brain have been described; however, the degree and extent of these changes are controversial. This study evaluates the quantitative and cytomorphometric effects of aging on neuronal and glial populations in the parietal cortex of the rat. METHODS: The study was performed in two groups of rats aged 4-6 and 30-32 months. Cortical volume, neuronal density, glial density, and neuronal area, and shapes of the soma and nucleus were analyzed in cortical layers I, II-IV, V, and VI using serial sections stained with cresyl-fast-violet, and quantitative morphometric techniques. RESULTS: No changes with age were found in volume of the cortex or neuronal density. Glial density increased significantly (mean for all layers 17%) in older rats. Layers II-IV, V, and VI showed an age-related decrease in the area of the neuronal soma. Neuronal shape, as revealed by the major/minor diameter ratio, also showed a decrease in old rats but only in layer II-IV. Nuclear area decreased with age only in layer VI. CONCLUSIONS: The stability of neuronal density together with the increased number of glial cells and the changes in neuronal soma size suggest that aged-related cognitive impairment could be a consequence of neuronal dysfunction rather than actual neuronal losses.
Aging↗
Environmental Factors Controlling N2 Fixation in Mediterranean Rice Fields
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Journal Article↗
PCR-identification of a Nicotiana plumbaginifolia cDNA homologous to the high-affinity nitrate transporters of the crnA family.
A family of high-affinity nitrate transporters has been identified in Aspergillus nidulans and Chlamydomonas reinhardtii, and recently homologues of this family have been cloned from a higher plant (barley). Based on six of the peptide sequences most strongly conserved between the barley and C. reinhardtii polypeptides, a set of degenerate primers was designed to permit amplification of the corresponding genes from other plant species. The utility of these primers was demonstrated by RT-PCR with cDNA made from poly(A)+ RNA from barley, C. reinhardtii and Nicotiana plumbaginifolia. A PCR fragment amplified from N. plumbaginifolia was used as probe to isolate a full-length cDNA clone which encodes a protein, NRT2;1Np, that is closely related to the previously isolated crnA homologue from barley. Genomic Southern blots indicated that there are only 1 or 2 members of the Nrt2 gene family in N. plumbaginifolia. Northern blotting showed that the Nrt2 transcripts are most strongly expressed in roots. The effects of external treatments with different N sources showed that the regulation of the Nrt2 gene(s) is very similar to that reported for nitrate reductase and nitrite reductase genes: their expression was strongly induced by nitrate but was repressed when reduced forms of N were supplied to the roots.
Amino Acid Sequence↗
Molecular identification of zeaxanthin epoxidase of Nicotiana plumbaginifolia, a gene involved in abscisic acid biosynthesis and corresponding to the ABA locus of Arabidopsis thaliana.
Abscisic acid (ABA) is a plant hormone which plays an important role in seed development and dormancy and in plant response to environmental stresses. An ABA-deficient mutant of Nicotiana plumbaginifolia, aba2, was isolated by transposon tagging using the maize Activator transposon. The aba2 mutant exhibits precocious seed germination and a severe wilty phenotype. The mutant is impaired in the first step of the ABA biosynthesis pathway, the zeaxanthin epoxidation reaction. ABA2 cDNA is able to complement N.plumbaginifolia aba2 and Arabidopsis thaliana aba mutations indicating that these mutants are homologous. ABA2 cDNA encodes a chloroplast-imported protein of 72.5 kDa, sharing similarities with different mono-oxigenases and oxidases of bacterial origin and having an ADP-binding fold and an FAD-binding domain. ABA2 protein, produced in Escherichia coli, exhibits in vitro zeaxanthin epoxidase activity. This is the first report of the isolation of a gene of the ABA biosynthetic pathway. The molecular identification of ABA2 opens the possibility to study the regulation of ABA biosynthesis and its cellular location.
Abscisic Acid↗
Nitrate and nitrate are transported by different specific transport systems and by a bispecific transporter in Chlamydomonas reinhardtii.
Nitrate transport mutants from Chlamydomonas reinhardtii and strains derived from them upon transformation with plasmids containing the C. reinhardtii nar2/Nrt2;1 or nar2/Nrt2;2 genes have been used to study nitrate and nitrite transport systems. Mutants lacking nitrate assimilation clustered genes showed a high affinity nitrite transporter activity (system 3), which was subject to ammonium inhibition and appeared to be independent of a functional nar2 gene. Transformants carrying nar2/Nrt2;2 recovered a high affinity nitrate transporter activity (system 2) and showed nitrite transport activities with properties similar to those in nonrecovered high affinity nitrate transporter activity (system 1) together with a considerably enhanced nitrite transport activity. Nitrite transport mediated by system 1 was very sensitive to inhibition by nitrate at microM concentrations. Results strongly suggest that three nitrate assimilation related high affinity transport systems operate in C. reinhardtii: one specific for nitrite, a second one encoded by nar2/Nrt2;2 specific for nitrate, and another one encoded by nar2/Nrt2;1, which is bispecific for these two anions.
Animals↗
Immunohistochemical localization of GABA in chameleon retina (Chamaleo Chamaleo).
We used a policlonal antiserum against GABA and demonstrated GABA-immunoreactivity (GABA-IR) in several populations of amacrine cells in the inner nuclear layer (INL), and other cells in the inner plexiform layer (IPL) of the central and peripheral retina of the chameleon. Horizontal cells do not contain GABA-IR and the chameleon retina is therefore an exception among non-mammals. GABA-IR was not seen in cell bodies in the position of photoreceptor, bipolar and interplexiform cells suggesting that GABA is not involved in synaptic transmission in the outer plexiform layer of chameleon retina.
Animals↗
Relationship Between Abundance of N2-fixing Cyanobacteria and Environmental Features of Spanish Rice Fields
In order to estimate the potential utilization of N2-fixing (heterocystous) cyanobacteria as natural biofertilizers in the Valencian rice fields (Spain), the distribution and seasonal variation of these microorganisms in water and sediment samples were evaluated, and the relationships among cyanobacterial abundance and physical and chemical characteristics of soil and water were investigated. N2-fixing cyanobacteria were present in all the samples analyzed (25 sampling points sampled three times per year during two years). The relative cyanobacterial abundance in soil and water followed contrasting patterns, maximum presence in soil coincided with minimum abundance in water. Correlation analysis showed that cyanobacterial abundance in the two phases (water and sediment) was influenced more by water than by soil properties. Salinity,mineralization variables, and soluble reactive phosphate (SRP) correlated positively with heterocystous cyanobacteria presence. Furthermore, dissolved inorganic nitrogen (DIN) and the ratio DIN:SRP correlated negatively with cyanobacterial abundance. However DIN:SRP ratio better described the cyanobacterial distribution, with a threshold effect: below the Redfield ratio value (7.2 in mass units) cyanobacterial abundance was clearly higher.
Journal Article↗
Early expression of a novel radial glia antigen in the chick embryo.
Monoclonal antibody 3CB2 recognizes an antigen expressed in discrete cell types derived from ectoderm and mesoderm. Biochemical and immunohistochemical studies indicate that the antigen recognized by the antibody is a 55 kDa cytoplasmic protein that may be an intermediate filament associated protein (IFAP). Developmental studies show that 3CB2 antigen is intensely expressed very early in the chick embryo, in the neural tube, myotomes, and in mesenchymal cells of visceral arches and the presumptive facial area. As development proceeds, antigen expression becomes restricted to astrocytes and radial glia cells throughout the brain. A detailed immunohistochemical study of the developing chick retina shows that the expression of 3CB2 antigen at embryonic day 8 (E8) is restricted to Müller cells, with the pattern of expression closely related to their degree of differentiation. We show, by immunocytochemistry labeling of entire Müller cells dissociated from retinas of E16-E20, that 3CB2 monoclonal is labeling the whole cell. 3CB2 selectively labels Müller cells in the rat and chameleon, but not their retinal horizontal cell axons. 3CB2 monoclonal is a very sensitive marker for early differentiating Müller cells. Our results provide evidence that 3CB2 antigen is a cytoskeletal component which may be involved in the morphogenesis of these cells, and also perhaps in the outgrowth of some axons.
Animals↗
Microglia in the avian retina: immunocytochemical demonstration in the adult quail.
Immunocytochemical techniques were used in conjunction with the QH1 antibody to study the morphological characteristics and distribution of microglia in the avascular retina of an avian species (the quail). The majority of microglial cells appeared in the outer and inner plexiform layers throughout the entire retina, whereas a few microglial cells in the nerve fiber layer were seen only in the central zone of the retina, near the optic nerve head. In the outer plexiform layer, microglial cells were star-shaped, with processes that ramified profusely in the horizontal plane. Fine process tips extended outward radially, insinuating themselves among the photoreceptors. A regular mosaic-like arrangement of microglial cells was evident in the outer plexiform layer, with no overlapping between adjacent cell territories. Microglial cells in the inner plexiform layer ramified through the entire width of this layer, showing radial and horizontal processes. Microglia in the inner plexiform layer also tended to be regularly distributed in a mosaic-like fashion, although there was slight overlapping between adjacent cell territories. Microglia density in this layer was approximately twice that in the outer plexiform layer. This pattern of microglial distribution was similar to that described in vascular retinae of several species of mammals, a finding that suggest that blood vessels are not responsible for the final locations of microglia in the adult retina, and that microglial precursors must migrate through long distances before they reach their precise destination.
Animals↗
Development of microglia in the quail optic tectum.
The development of microglia in the quail optic tectum from embryonic day 6 to adulthood was studied by using the QH1 monoclonal antibody. In youngest tecta, microglial cells were scarcely present, but their number rose in subsequent stages. A clear pattern of microglial cell distribution was observable in embryos of 9-16 days. (1) Round cells appeared close to the ventricular layer. (2) Large numbers of ameboid and round labeled cells were seen in the stratum album centrale during development. A gradient of cell density was observable in this layer, as fewer labeled cells appeared in medial regions of the tectum than in lateral regions. (3) Maturing ramified cells were found in layers external to the stratum album centrale, where they increased in number and in branching complexity during development. In adult tecta, almost all microglial cells were of the mature ramified type and were distributed homogeneously in the different tectal layers, although in some layers they had particular morphological features. The distribution of microglia in the developing tectum and in adjacent regions provided insight into the routes of microglial cell invasion of the tectum during development. Apparently, a proportion of microglial cells reached the tectal parenchyma from the meninges and from the ventricular lumen, but the majority of them migrated along nerve fiber tracts from their entry point at the pial surface of the ventromedial caudal tectum. After they reached the stratum album centrale, microglial cells continued their migration toward more external layers, where they differentiated into ramified microglia.
Aging↗