In vivo visualization of individual neurons in arthropod ganglia facilitates intracellular neuropil recording

Summary A simple method for the in vivo visualization of dye filled cells by laser illumination is used to characterize neurons in situ in the segmentai ganglia of the locust and the crayfish (Fig. 1). Neuron visualization provides the structural information necessary for identification of cells during an ongoing physiological experiment (Figs. 2, 3). Sequential penetrations of soma and neuropil as well as simultaneous double neuropil penetrations of spiking and nonspiking cells are facilitated by the visual control afforded by neuron visualization (Figs. 4, 5, 6). Furthermore, neuron visualization allows the sampling of cellular properties at multiple, predetermined sites in the dendritic and axonal arbors of identified neurons (Fig. 7) and aids in establishing synaptic connectivity through double neuropil recordings (Fig. 8).     

The effect of calmodulin and far-red light on the kinetic properties of the mitochondrial and microsomal calcium-ion transport system from corn

The kinetic properties of active Ca²⁺ transport into mitochondria and microsomal membrane vesicles prepared from coleoptiles of dark-and light-grown corn seedlings have been studied. The apparent values for K _m and V _max for Ca²⁺ of the mitochondrial transport system from dark-grown plants are about one order of magnitude higher than those from the microsomal transport system. Calmodulin has no effect on the Ca²⁺ accumulation into mitochondria whereas the apparent maximum transport velocity and affinity for Ca²⁺ of the microsomal Ca²⁺-transport system are both increased by calmodulin. When intact corn seedlings are irradiated with far-red light, the calmodulin-induced increase of the apparent maximum transport velocity and affinity for Ca²⁺ can no longer be observed. From these data it can be concluded that the low cytoplasmic Ca²⁺ concentration in the cytoplasm of coleoptile cells from dark-grown corn is maintained by a calmodulin-regulated Ca²⁺ pump. Irradiation with photomorphogenically active far-red light lowers the Ca²⁺-transport activity and thus causes an increase of the cytoplasmic, free-Ca²⁺ concentration. The physiological implications will be discussed.     

Immune response genes in inbred rats. II. Segregation studies of the GT and GA genes and their linkage to the major histocompatibility locus

Genetic analysis of the high responder-non-(or low) responder differences between WF and ACI rats to the synthetic copolymers GT and GA established singleIr-genes for both antigens and dominance for high responder status.Ir-GT andIr-GA are linked to the major histocompatibility locus. It could be demonstrated that only T cells carrying the high responderIr-GT gene undergo in vitro blast transformation to GT. The advantages of the rat systems for further studies of the regulatory role ofIr-genes on the cellular level are discussed.Abbreviations cpm counts per minute - DHR delayed hypersensitivity reaction - GA random synthetic copolymer of L-glutamic acid⁵⁰, L-alanine⁵⁰, M.W. 45,000 - GT random synthetic copolymer of L-glutamic acid⁵⁰, L-tyrosine⁵⁰, M.W. 22,000 - Ir gene immune response gene - MLC mixed lymphocyte cultures - LDH lactic dehydrogenase - (T,G)-A-L branched chain synthetic polymer of poly-L (tyrosine, glutamic acid)-poly-D, L-alanine-poly-L-lysine Rat Strains ACI ACI/MaI - WF Wistar Furth - AUG August 28807/Cr     

Enzymatic methylations: III. Cadaverine-induced conformational changes of E.coli tRNAfMet as evidenced by the availability of a specific adenosine and a specific cytidine residue for methylation+

A partially purified tRNA methylase fraction from rat liver, containing m(2)G- m(1)A- and m(5)C-methylase, was used to study the influence of Mg(++) and of the biogenic polyamine cadaverine on the enzymatic methylation of E.coli tRNA(fMet)in vitro. In presence of 1 or 10 mM Mg(++), guanosine no. 27 was methylated to m(2)G. In 1 mM Mg(++) plus 30 mM cadaverine, guanosine in position 27 and adenosine in position 59 were methylated. In presence of 30 mM cadaverine alone tRNA(fMet) accepted three methyl groups: in addition to guanosine no. 27 and adenosine no. 59 cytidine no. 49 was methylated. In order to correlate tRNA(fMet) tertiary structure changes with the methylation patterns, differentiated melting curves of tRNA(fMet) were measured under the methylation conditions. It was shown that the thermodynamic stability of tRNA(fMet) tertiary structure is different in presence of Mg(++), or Mg(++) plus cadaverine, or cadaverine alone. From the differentiated melting curves and from the methylation experiments one can conclude that at 37 degrees in the presence of Mg(++) tRNA(fMet) has a compact structure with the extra loop and the TpsiC-loop protected by tertiary structure interactions. In Mg(++) plus cadaverine, the TpsiC-loop is available, while the extra loop is yet engaged in teritary structure (G-15: C-49) interactions. In cadaverine alone, the TpsiC-loop and the extra loop are free; hence under these conditions the open tRNA(fMet) clover leaf may be the substrate for methylation. In general, cadaverine destabilizes tRNA tertiary structure in the presence of Mg(++), and stabilizes tRNA(fMet) tertiary structure in the absence of Mg(++). This may be explained by a competition of cadaverine with Mg(++) for specific binding sites on the tRNA. On the basis of these experiments a possible role of biogenic polyamines in vivo may be discussed: as essential components of procaryotic and eucaryotic ribosomes they may together with ribosomal factors facilitate tRNA-ribosome binding during protein biosynthesis by opening the tRNA tertiary structure, thus making the tRNA's TpsiC-loop available for interaction with the complementary sequence of the ribosomal 5S RNA.     

Differential staining of plant chromosomes with Giemsa

Simple Giemsa staining techniques for revealing banding patterns in somatic chromosomes of plants are described. The value of the methods in the recognition of heterochromatin was demonstrated using five monocotyledonous and two dicotyledonous species. In Trillium grandiflorum the stronger Giemsa stained chromosome segments were shown to be identical with the heterochromatic regions (H-segments) revealed by cold treatment. Preferential staining of H-segments was also observed in chromosomes from three species of Fritillaria and in Scilla sibirica. Under suitable conditions the chromosomes of Vicia faba displayed a characteristic banding pattern and the bands were identified as heterochromatin. The Giemsa techniques proved to be more sensitive than Quinacrine fluorescence in revealing a longitudinal differentiation of the chromosomes of Crepis capillaris, where plants with and without B-chromosomes were examined. Again all chromosome types had their characteristic bands but there was no difference in Giemsa staining properties between the B-chromosomes and those of the standard complement.     

Elektronenmikroskopische Untersuchungen zur Lipid-Speicherung in den Nierentubuli von Drosophila melanogaster

Zusammenfassung In den Zellen der Nierentubuli (Malpighische Gefäße) von Drosophila melanogaster werden Lipide in der gleichen Weise wie das 3-Hydroxykynurenin gespeichert. Diese Substanzen werden in Erweiterungen des endoplasmatischen Retikulums akkumuliert. Bei älteren Larven verschwinden diese Lipidtropfen. Dabei legen sich entweder Mitochondrien um die Depots oder sie werden von Membranstapeln des endoplasmatischen Retikulums umgeben und abgebaut. Die funktionelle Bedeutung dieser Befunde wird diskutiert.

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Electronmicroscopic studies on the lipid storage in the renal tubules of Drosophila melanogaster

Summary In the cells of the renal tubules (Malpighian tubules) of Drosophila melanogaster lipids are stored in the same way as 3-hydroxykynurenin. These substances are found in dilatations of the endoplasmic reticulum. In later larval stages the lipid droplets gradually disappear. In theses stages the lipid droplets are either closely associated with the mitochondria or they are removed by concentric membrane arrays of the endoplasmic reticulum. The functional significance of these findings is discussed.

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Diese Untersuchung wurde mit Unterstützung der Deutschen Forschungsgemeinschaft durchgeführt.