Union of Swiss Societies of Experimental Biology Abstracts of the 18 Th Annual Meeting
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Union of Swiss Societies of Experimental Biology Abstracts of the 18 th Annual Meeting Union des Soci&~s Suisses de Biologie Exp6rimentale Comptes rendus de la 18 e Rdunion annuelle Union Schweizerischer Gesellschaften ffir Experimentelle Biologie Berichte der 18. Jahresversammlung Basel, 20/21 March 1986 Unterstfitzt durch die Schweizerische Naturforschende Gesellschaft fiber die Schweizerische Kommission ffir Experimentelle Biologie 630 Experientia 42 (1986), Birkh/iuser Verlag, CH-4010 Basel/Switzerland Symposium I: Neuronal regeneration and transplantation Influence of adenohypophyseal tissue on the development of the rat fascia dentata in vitro Precursor cells of dorsal root ganglia in chick embryo: B. H. G~ihwiler and J. Zimmer , Preclinical Research, Sandoz Ltd, differentiation into neurons in culture CH-4002 Basel, and Institute of Anatomy B, University of Aar- hus, DK-8000 Aarhus C I. Barakat and B. Droz, Institut d'Histologie et d'Embryologie, 9, rue du Bugnon, CH-IO05 Lausanne In hippocampal slice cultures, the mossy fibers from the dentate granule cells were previously shown by Timm-stainingto retain Dissociated DRG cells from 10-day-old chick embryos were their normal connections to CA4 and CA3 pyramidal cells. cultured into two types of culture: neuron-enriched culture and During co-cultivation with an adenohypophyseal explant, the mixed DRG cell culture. In neuron-enriched cultures, the num- infrapyramidal blade of the fascia dentata largely disappeared as ber of neurons decreases with time, while in mixed DRG cell a cell layer; granule cell-like neurons would migrate toward the culture the number of neurons increases with time until 7 days of hypophyseal explant, whilst their axons still functionally inner- culture. The incorporation of 3H-thymidine into DRG ganglion vated CA3 pyramidal cells. Axon collaterals projected in the cells could not account for the increased number of cultured opposite direction and presumably terminated on dendrites, neurons. thus giving rise to the intense black labeling which was observed Microphotographs of the same area, taken every 60 rain, showed in Timm-stained preparations as a bridge connecting the two that small and flat cells can differentiate into neurons which were explants. identified by their morphological characteristic and by specific The morphological alterations induced in the fascia dentata by neuronal markers such as specific neuronal enolase and D2/N- co-cultured adenohypophysis were tissue-specific since co-cul- CAM. It is concluded that embryonic DRG in chicken contain tured neurohypophysis, pineal gland and cerebellum failed to progenitor cells which are able to differentiate into sensory neu- produce similar effects. These results suggest that cultured ade- rons in culture. nohypophyseal tissue is capable of releasing yet unidentified factors which apparently enhance neuronal migration. Concanavalin A and extracellular matrix mediate rapid outgrowth of leech neurons in saline M. Chiquet and S. Acklin, Biozentrum der Universitgit Basel, Evidence for sprouting of hippocampal cholinergic and Klingelbergstrasse 70, CH-4056 Basel aminergic fibers following partial deafferentation in rats The outgrowth of neurites in culture by single identified neurons U. Gasser and A. Dravid, Preclinical Research, Sandoz Ltd, of the leech is markedly influenced by the substrate. On cell-free CH-4002 Basel extracellular matrix, extensive sprouting occurs within a few Cholinergic enzyme activities and high affinity uptake of 3H- hours. Another substrate promoting sprouting is the plant lec- noradrenaline and 3H-serotonin were measured in septotem- tin, concanavalinA (con A). Neurons had to be attached directly poral regions of the hippocampus following partial or complete to substrate-bound con A or extracellular matrix to grow neu- fornix-fimbria transection. Subsequent to an initial reduction rites and did so in physiological saline or medium without any reflecting terminal degeneration, a significant increase in these macromolecular substances or other cells present. Sprouting on parameters was observed. Recovery of these terminal-specific con A was inhibited by the con A-specific hapten sugar ~-methyl markers most likely represents synapse replacement following mannoside. These results show that the substrate can mediate sprouting of the remaining undamaged fibers. The extent of the outgrowth of cultured leech neurons; soluble growth factors recovery seems to depend on the degree of deafferentation and are not required. However, sprouting depends not simply on was different for each transmitter system. Substantial recovery attachment to substrate but, critically, on its molecular composi- occurred within 8 weeks after partial lesions whereas after exten- tion, since other adhesive substrates did not promote sprouting. sive deafferentation the magnitude of recovery was lower and was apparent only after longer survival times. Acute facial palsy: a virological and histopathological study M. Engels, H. Felix and E. Loepfe, Institute of Virology, Univer- Evidence for the role of nerve growth factor (NGF) in sity of Ziirich, ENT-Department, University Hospital, CH-8001 central cholinergic neurons in rats Ziirieh U. Gasser, G. Weskamp, U. Otten and A. Dravid, Preclinical Virological and/or histopathological investigations have been Research, Sandoz Ltd, and Dept Pharmacology, Biocenter of the performed on 21 specimens removed from the greater petrosal University of Basel, CH-4056 Basel nerve in the course of surgical decompression of the facial nerve Cholinergie enzyme activities and NGF levels in the septum and (performed by Prof. U. Fisch) within 14 days from onset of Bell's hippocampus were measured at various times after complete Palsy. Nine specimens were examined for virus infection by a bilateral transection of the dorsal septohippocampal pathway. modified coculture method. The specimens were maintained in Within 3-7 days after transection, there was a maximal increase cell culture medium for 28 days at 37~ Vero cells were inocu- in NGF content of about 50 % in the hippocampus and 290 % in lated every day with supernatant. Infected cell supernatants un- septum followed, in both regions, by a decline to control values derwent two blind passages and, additionally, were inoculated on postlesion day 14. Cholinergic enzyme activities indicated an into the allantois and amnion of embryonated eggs. The fluids almost complete terminal degeneration in the hippocampus. In were tested for hemagglutinins. In no case any virus could be the septum, however, an initial decrease of 15 % was followed by isolated. The main morphological changes observed were nerve gradual increase to about 20% over the control levels after fiber degeneration and demyelination, and in 8 of 15 specimens longer survival time. Fornix-fimbria lesion and the resulting endoneurial fibrosis was present, In 10 cases a minimal infiltra- destruction of the cholinergic system, appears to induce en- tion of lymphocytes was observed. The etiological relationship hancement of NGF in the hippocampus and septum. These between the virological and histopathological findings in acute findings thus support the role of NGF in central cholinergic facial palsy is discussed. neurons. Experientia 42 (1986), Birkh/iuser Verlag, CH~4010 Basel/Switzerland 631 Isolation of eDNA clones from C6 rat glioma RNA cod- tube technique. The metabolism of choline and the synthesis of ing for a glia-derived neurite-promoting factor (GdNPF) acetylcholine (ACh) were investigated, in living cultures, in pres- ence of different radiolabeled precursors, and at various precur- S. Gloor, J. Giinther and D. Monard, Friedrich-Miescher-Institut, sor concentrations, and compared with the metabolism in stri- P.O. Box 2543, CH-4002 Basel atal slices of adult rat brain. Acute slice preparations of the GdNPF is a 43 kD glycoprotein which promotes neurite exten- newborn septal area and mature cultures showed different pat- sion in neuroblastoma cells and strongly inhibits serine proteases terns of choline metabolism, indicating in vitro maturation or (Giinther, J., Nick, H., and Monard, D., EMBO J. 4 (1985) regeneration of cholinergic neurons and nerve terminals. Mature 1963). RNA from C6 glioma cells was isolated, translated both cultures showed many biochemical characteristics of the tissue in in vivo and in vitro and the protein immunoprecipitated with situ, such as enhancement of ACh synthesis by depolarization polyclonal anti-GdNPF antibodies. Sucrose gradient enriched and de novo choline formation. In absence of extracellular poly(A)+ RNA was used for eDNA synthesis. The cDNA li- choline, ACh was synthesized from choline derived from the brary obtained was screened by the method of hybrid selected methylation of phosphatidylethanolamine. translation. A clone with a 800 bp insert coding for GdNPF was used for the estimation of the amount of GdNPF mRNA at different stages of rat brain development. Structural and functional aspects of glia-derived neurite promoting factor (NPF) H. Nick, J. Giinther and D. Monard, Friedrich-Miescher-Institut, Non-neuronal cells influence the cholinergic properties of P.O. Box 2543, CH-4002 Basel cultured human spinal cord neurons NPF is a protein with a single polypeptide chain of mol.wt A. C. Kato, L. Bierman and L. Erkman, Department of Pharma- 43,000 (SDS-PAGE). It promotes the formation of neurites in cology, CMU, CH-1211 Gen~ve 4 NB2a neuroblastoma cells. NPF is a strong inhibitor of serin Dissociated monolayer cultures