Development of cerebellar astroglia: Transitions in form and cytoskeletal content

The forms, disposition, and cytoskeletal contents of astroglia in immature mouse cerebellum were studied by immunocytochemical staining with antisera against two intermediate filament proteins, vimentin (Vim) (58,000 daltons) and glial filament protein (GF) (51,000 daltons). From embryonic (E) Day 15 to postnatal (P) Day 2, Vim is expressed in cells throughout the cerebellar anlage, including radial glia and Bergmann fibers, cells with amorphous shapes and 2–3 processes, and thick longitudinal elements oriented parallel to axons within axon tracts. GF is not expressed during the first few postnatal days, but by P7, there is a dramatic increase in GF-positive astrocyte-like cells in the putative white matter that are more densely stained and more crowded than at any other age. Between P7 and P14 all astrocytes throughout the cerebellum express both Vim and GF. From P21 on, Vim expression is progressively rarer in all astrocytes except for Bergmann fibers, and GF-positive astrocytes become less numerous. These findings raise two issues: (a) the lineage and relationships of cells expressing Vim and GF; (b) Since GF-positive cells appear as axon ingrowth ceases, axons must grow in a terrain comprised of glial cells that have a different cytoskeletal composition (vimentin), reflecting a less differentiated state, than mature astrocytes or than the GF-rich astrocytes that proliferate after injury in adult CNS.     

The detection of picomolar quantities of GD1b,GT1b and GQ1b on thin-layer chromatograms by the direct binding of125I-labeled tetanus toxin,fragment C

The¹²⁵I-labeled fragment C of tetanus toxin was found to bind specifically to the gangliosides G_D1b, G_T1b, and G_Q1b when applied to thin-layer chromatograms on which a mixture of gangliosides had been resolved. As little as 2.5 pmoles of these gangliosides could be detected by this method. In addition to factors determined by the sample, namely the amount and species of gangliosides present, optimal binding of the¹²⁵I-labeled fragment C also depended upon the iodination procedure used to generate the probe, the toxin concentration, and the concentration, buffer type, pH, and ionic strength of the binding solution. This new technique was shown to be a sensitive method for the detection and identification of specific gangliosides originating from extraneural or neural cells.Nomenclature: The gangliosides follow the nomenclature system of Svennerholm [Eur J Biochem (1977) 79:11–21] G_M3 II³NeuAc-LacCer - G_D3 II³(NeuAc)₂-LacCer - G_M1 II³NeuAc-GgOse₄Cer - G_D1a IV³NeuAc, II³NeuAc-GgOse₄Cer - G_D1b II³(NeuAc)₂-GgOse₄Cer - G_T1b IV³NeuAc, II³(NeuAc)²-GgOse₄Cer - G_Q1b IV³(Neu-Ac)₂, II³(NeuAc)₂-GgOse₄Cer - G_P1b IV³(NeuAc)₃, II³(NeuAc)₂-GgOse₄Cer     

The uptake of soil and fertilizer-nitrogen by barley growing under Scottish climatic conditions

Summary Field experiments were carried out using¹⁵N-labelled calcium nitrate, to investigate the relative uptake by barley of fertilizer-N and soil-N. On imperfectly drained till soils uptake of soil-N increased with increasing rate of fertilizer, but remained constant on a brown sand, possibly due to more efficient root exploration in the latter soil. In four out of five seasons, late uptake of soil-derived N was a major feature, and uptake from ploughed soil as compared with uptake from direct-drilled soil was correlated with seasonal rainfall patterns. Significantly higher quantities of both fertilizer- and soil-derived N were taken up by winter barley than by spring barley, reflecting the longer growth period and higher dry matter yield from the former crop.     

Purification of restriction endonuclease XcyI from Xanthomonas cyanopsidis

A new Type II restriction endonuclease XcyI, purified from Xanthomonas cyanopsidis 13D5, is an isoschizomer of SmaI and XmaI that cleaves at the nucleotide sequence 5'-C decreases CCGGG-3' of double-stranded DNA. The single restriction activity present in this strain permits rapid purification of 8000 units of cleavage activity from 10 g of freshly harvested cells. The resulting XcyI preparation is free of contaminating nuclease activities that interfere with in vitro manipulation of DNA.     

Cell culture of sixteen-day-old rat embryo cerebra and associated changes in ganglioside pattern

Abstract— Cortical slices from rat brain were incubated in Krebs-Ringer phosphate medium. Activity of the pyruvate dehydrogenase complex (PDH) was measured in homogenates of the incubated tissue. Increasing the extracellular KCI concentration from 5 to 75 mM caused a dose-dependent increase in activity of this rate-limiting mitochondrial enzyme. The increase in PDH activity, produced by high concentration of KCI. was associated with a decrease in the tissue content of ATP. Omission of calcium, or replacement of sodium by choline, reduced, and addition of ouabain prevented, the activation of the enzyme in the depolarized tissue.
The mechanism by which extracellular potassium can affect PDH activity is unknown. However, it is most likely that the alterations in enzyme activity are related to changes in properties of cell membranes during depolarization leading to intracellular events directly affecting the enzyme complex. These could include alterations in the concentrations of adenine nucleotides or free calcium ions in the cell.     

The biological half life and distribution of 125Iodide and radioiodinated protein in the imported fire ant, Solenopsis invicta

The radioisotope ¹²⁵Iodide, a gamma emittor, was used in two different forms, as ¹²⁵I mixed with egg yolk and as ¹²⁵I covalently attached to egg albumin and mixed with egg yolk, to study food flow in the imported fire ant, Solenopsis invicta Buren. The biological half life of ¹²⁵I-albumin in egg yolk powder was determined to be 96 hr in isolated workers, 108 hr in individuals held with small groups of unlabelled workers, and 1,008 hr in workers held in colonies exposed to labelled food for 48 hr. In contrast, the biological half life of free ¹²⁵I mixed with egg yolk powder was 22 hr, 20 hr, and 40 hr, respectively.The internal distribution of radioactivity was checked after 24,48, and 380 hr. There was a significant difference in distribution of ¹²⁵I in ants fed either free ¹²⁵I or ¹²⁵I-albumin. Most of the free ¹²⁵I was rapidly excreted. A high percentage of ¹²⁵I-albumin was assimilated, apparently through protein digestion pathways with eventual storage in or below the cuticle. There was no evidence of gland involvement in food flow to either larvae or queens with the radio-iodinated protein.

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Résumé L'utilisation de l'iode radio-actif (¹²⁵I) a permis d'étudier le cheminement de la nourriture chez Solenopsis invicta Buren (Myrmicinée). Deux formes différentes de l'isotope ont été étudiées. L'iode 125 a été fixé d'une manière covalente à la tyrosine dans l'albumine des oeufs en utilisant la méthode chloramine T pour ioder les protéines. L'albumine marquée a été mangée ensuite à du jaune d'oeuf en poudre.La seconde forme contenait de l'iode 125 mélangé au jaune d'oeuf en poudre en absence de tout catalyseur, ce qui empêche la fixation chimique. La demi-vie biologique (T_biol) des deux formes a été déterminée chez des ouvrières isolées, chez des individus gardés avec de petits groupes d'ouvrières non-marquées, et chez des ouvrières gardées dans des colonies exposées à la nourriture radio-active pendant 48 h. La demi-vie biologique de l'albumine marquée était de 96 h, 108 h, et 1.008 h. En contraste, la demi-vie de l'iode 125 était de 22 h, 20 h, et 40 h. L'effet de groupe créé par des échanges répétés de nourriture entre les individus était négligeable avec la nourriture protéique. L'échange répété de nourriture entre les larves et les ouvrières a beaucoup augmenté la demi-vie de l'albumine marquée à I 125. Cet effet n'était pas aussi clair avec l'iode 125 par suite de son élimination rapide.La distribution de la radio-activité a été examinée chez des ouvrières au bout de 24, 48 et 380 h après les avoir nourries avec de l'albumine marquée à I 125 et de l'iode 125. Il y avait une différence considérable de distribution, avec un haut pourcentage d'albumine assimilé, sans doute par les voies de digestion de protéines. Le radio-isotope a été ensuite conservé sous forme d'iode 125 ou d'iodotyrosine, dans (ou sous) la cuticule de l'ensemble du corps. Les fourmis ont rapidement excrété l'iode 125 libre avec 5% de la radioactivité résiduelle après 380 h, peut-être fixée aux protéines cellulaires, et ensuite transportée vers la cuticule. Les différences considérables entre les demi-vies biologiques et la distribution interne de la radioactivité chez les fourmis nourries avec de l'iode 125 ou à l'albumine marquée à I 125, soulignent le danger de croire que le cheminement de la nourriture peut être définitivement étudié en utilisant des radio-isotopes qui ne sont pas fixés chimiquement à la substance étudiée.