Multicellular antennal sensilla containing a sensory cell with a short dendrite and dense-core granules in the insect,Hypogastrura socialis (Collembola): intermolt and molting stages

Summary At the antennal tip of the collembolan insect Hypogastrura socialis two terminal-pore sensilla are located, which, in addition to normally structured and most probably chemosensitive sensory cells, also contain aberrant sensory cells. Portions of these cells resemble chemoreceptors but also shown are features that, as a rule, occur in mechanoreceptors. One cell in each sensillum is remarkable in two characteristics: (1) Its dendritic outer segment does not reach the cuticular outer structures of the sensillum; (2) it contains dense-core granules (diameter 60–110 nm) within its perikaryon, its dendritic inner segment and its axon. Additionally, these two cells do not show lengthening of their dendritic outer segment during molt as do all other sensory cells. Among the fibers of one major branch of the antennal nerve within the head capsule a single axon was observed to contain dense-core granules. This axon was traced to its termination where normal synaptical contacts were found. Based on the assumption that the axon belongs to one of the granule-containing sensory cells two alternative hypotheses are proposed: (1) an individual sensory cell of a sensillum may synthesize a transmitter that is different from that of the other sensory cells of this sensillum; (2) the aberrant cells have lost exteroceptive functions but act as neuromodulatorsSupported by the Deutsche Forschungsgemeinschaft (SFB 4/G1)     

Subcellular localization and glycoprotein nature of the invertase from the fission yeast Schizosaccharomyces pombe

The subcellular localization of the enzyme invertase in Schizosaccharomyces pombe cells, both repressed and derepressed for synthesis of the enzyme, was studied. Most of the invertase was found to be located outside the plasma membrane and only a small percentage was found to be associated to membranes. A substantial portion of the external enzyme remained firmly bound to cell-wall material.All of the invertase recovered in soluble form from cellular extracts reacted with concanavalin A and with the lectin from Bandeiraea simplicifolia seeds, indicating the presence in the enzyme of a carbohydrate moiety which probably contains terminal mannosyl (or structurally related) and galactosyl residues.The possibility of the presence of two different forms of invertase in S. pombe was considered. An intracellular, soluble form of invertase, devoid of carbohydrate, similar to the small invertase of the budding yeast Saccharomyces cerevisiae, was not found in S. pombe. However, the Michaelis constant for sucrose of the enzyme present in repressed cells was smaller than that of the invertase synthesized under derepressing conditions, although this difference could also be the result of a different pattern of glycosylation of the invertase synthesized under different growth conditions.     

Auxin action: the search for the receptor

Abstract. The molecular specificity of the substances which have auxin activity implies the existence of specific receptors. There have been many efforts to identify and isolate these receptors on the assumption that they should bind auxins with affinities coordinate to their activities in bioassays. However, the known complexity of auxin uptake and metabolism make this assumption seriously deficient. Although several such binding sites have, in fact, been identified, proof of a connection between these sites and auxin action has been lacking. Definite proof would include a requirement that the site be reconstituted, together with the appropriate macro-molecular machinery, to construct a model of an auxin response. At the moment, our ignorance of the biochemistry and molecular biology of auxin growth responses makes such a proof difficult. However, two avenues of research promise to accelerate the rate of progress. The increasingly potent tools of molecular biology should soon allow the dissection of auxin-regulated gene expression, while improved knowledge of plasma membrane proton pumps and the mechanism of cell wall biosynthesis should produce, in parallel, an understanding of the auxin regulation of acid growth.     

Insulin uptake by rat liver endothelium studied in fractionated liver cell suspensions

Summary Cellular distribution of insulin receptors was studied in fractionated rat liver cell suspensions using ¹²⁵1-insulin and a visual probe consisting of latex beads covalently linked to insulin (minibeads). Fractionation was done on metrizamide gradients which yielded two cellular fractions. The large cell fraction consisted mostly of hepatocytes and the small cell fraction consisted of 37% endothelial cells as well as Kupffer cells. The magnitude of insulin uptake by the endothelium-rich small cell fraction was at least double that of the uptake by the hepatocyte-rich fraction. The minibead technique demonstrated that in the small cell fraction only endothelial cells, and not Kupffer cells, were responsible for the insulin uptake. Our findings suggest that liver endothelium may be responsible for the uptake of circulating insulin and its transport to hepatocyte. This emphasizes the presence of a tissue-blood barrier in the liver.Abbreviations PRS phosphate-buffered saline - SEM scanning electron microscopy - TEM transmission electron microscopy     

Restriction endonuclease EcoO109 from Escherichia coli H709c with heptanucleotide recognition site 5'-PuG/GNCCPy

A new restriction endonuclease, EcoO109, has been isolated from Escherichia coli H709c by polyethyleneimine (PEI) precipitation, DEAE-cellulose chromatography and heparin agarose chromatography. The yield was high, more than 3000 units/g of wet cells. The EcoO109 endonuclease recognizes and cleaves a nucleotide sequence of (formula: see text), in the presence of 10 mM Mg2+. The enzyme will be useful for structural analysis and molecular cloning of DNA because of the stability, high yield and easy handling of the producer strain.     

Development of a high-frequency transforming vector for Aspergillus nidulans

The pyr4 gene of Neurospora crassa, which codes for orotidine-5'-phosphate decarboxylase, is capable of transforming an Aspergillus nidulans pyrG mutant by chromosomal integration, despite low homology between the transforming DNA and the recipient genome. Integration of pFB6, a plasmid carrying pyr4 and capable of replication in Escherichia coli, was not observed at the pyrG locus. The efficiency of transformation was considerably enhanced (50-100 fold) by inclusion in the transforming vector of a 3.5-kb A.nidulans chromosomal sequence, ans1. Although this sequence was isolated on the basis of replicating activity in Saccharomyces cerevisiae, there was no evidence for such activity in A.nidulans. Part of the ans1 fragment appears to be reiterated in the A.nidulans genome, though it is not yet clear whether this is directly responsible for the high transformation frequency. The efficiency of transformation of A.nidulans by plasmids bearing ans1, using an improved protocol, was approx. 5 X 10(3) stable transformants per microgram of plasmid DNA.     

Inactivation and sedimentation of mercury from organomercurials by Klebsiella pneumoniae

Abstract A susceptibility of 63 clinical isolates of Klebsiella pneumoniae to inorganic and organic mercuric compounds was determined. 18 of them were found to be resistant to fluorescein mercuric acetate (FMA) and merbromin (MB). Moreover, all the resistant strains inactivate the antibacterial effect of FMA. The changes in the amount of organic mercury at the time of inactivation of the drug and the structures of the end products were examined in detail with the plasmid-bearing strain JK9 and its transconjugants of Escherichia coli .
The results showed that FMA was inactivated by an intracellular enzyme produced inducively and was degraded to fluorescein (sodium salt, uranine), which led to the sedimentation of metallic mercury. The discovery of the genes conferring inducible organic mercury-inactivating enzymes determined by plasmids was the next step and their application in the recovery of metallic mercury from organomercurials is now imminent.