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Interference images of heat-shocked HeLa cells
180
Cell Biology
OF SYNAPTIC MITOCHONDRIA DURING AGING Bertoni-Freddari, Patrizia Fattoretti, Carlo Tiziana Casoli, *William Meier-Ruge, *Jurg Ulrich. Centre for Surgical Research, INRCA Research Department, Via Birarelli, 8, 60121 Ancona, Italy and *Neuropathology Division, University of Basel, Schoenbeinstrasse 40, Basel, Switzerland. Volume density (Vv), numerical density (NV). average volume (V) and shape skeleton (Sk) of synaptic mitochondria were measured by means of a computer assisted image analyser in the cerebellar granuli adult and old rats. of young, No significant difference was found in the three groups analysed with regard to Vv. A significant NV was found in adult animals when increase of compared both with young and old rats, whereas a in this parameter was significant decrease demonstrated in the old animals vs. the other V did not change between young and old groups. but was significantly smaller in the adult group, one. Sk showed a significant elongation in the shape of mitochondria during aging as demonstrated by the 55% increase of this parameter in the old animals. Since the mitochondria in the cerebellar directly glomerulus are reported to subserve synaptic transmission, the present ultrastructural findings support that age-related alterations at chemical synapses may find an explanation in and/or impaired energy mechanisms supply organelles.
P405
P407
MORPHOMRTRY
GENE E;CPRESSION
IN PEANUT MUTAHTS
RESISTANT TO LEAF SFOT DISEASE Zoventino D. Soriano, Arne C. Villanueva, Adelaida Pi. Calaguas, !?eliton T. Montejo. Natl. Acadcny of Science and Technology, TAP1 aldg., DOST Complex, Dicutan, Metro Pianila, Fhilippines The wor'x studied how to independent mutant genes are expressed to produce resistance to the leaf spot disease in peanut.
Two fungus species cause leaf spot, Cercosnora-personata which penetrates the leaf and C. arachldlcola, infects inner tissues. --The two mutant genes, Is-l and Is-2 govern inheritance of resiza?ice -to the disease. To enter the leaf. the hyphsl outgrowth of a germinating 'spore secretes enzyme cellulase which is more or less inactivated by a mutant protein in the plasma membrane of the epidermal cell. The mutant protein was found to remove the iron co-factor in cellulase polyFeptide. ;n the heterozygous condition, gene ls-2 governs synthesis of a polypeptide ?Xia? represses the action of a toxic enzyme secreted by nycelia of the fungus, n arachidicola restoring all functions ir. of chlorenchymaious cells in the leaf. The mutant polypeptide attaches to the toxic protein enzyme and displaces thiamine.
International
Reports,
P406
Vol. 14, Abstracts
MICROTUBULES (NF) IN
(MT) AGING
Supplement
1990
AND NEUROFILAMENTS RATS
J. Eyer, M. Coleman’ and J.F. Leterrier. U 298 INSERM, C.H.R.U., Angers, France and * Dpt of Immunology, St Georges Hospital Medical school. Cranmer Terrace, London, U.K. The composition and biochemical properties of MT and NF purified from young (3 months) and aged (26 months) rats ware compared on the basis of their polypeptide composition and their associated enzymatic activities. MT, purified in the presence of protease inhibitors in order to block the proteolysis of MAPS occuring in old rats (Matus and Greene, 1986), contained a lower amount of MAP2 in samples from aged animals correlated with a decrease in the amount of the MAPP-associatedcAMP dependent protein-kinase (Vallee et al, 1981) while the other MAPS were essentially unmodified. In addition, the specific volume of MT pellets was significantly decreased in samples from old animals, suggesting that the role of MAPS as spacer molecules on MT was altered by aging. NF isolated from spinal cord of the same animals were essentially undistinguishable between the young and old preparations. Both the yield of NF (mglwet weight), the polypeptidic composition and the associated protein-kinase activity were found nearly identical. The only age dependent-change in NFassociated enzymes was found with an ATPase activity which was significantly higher in NF from old rats. However, the in vitro interaction process between NF, reflected by the viscosity kinetics of filament suspensions (Leterrier and Eyer, 1987), were significantly faster in the dialysed NF samples from old than from young animals. This situation was reversed when undialysed NF were used, suggesting that dialysable factors were responsible for the distinct behaviour of both preparations. A preliminary study of dialysed molecules showed the presence of peptides reacting with a monoclonal antibody directed against the phcephorylated NF-H tail domain (from B.H. Anderton). These results argue for the lack of alteration with age of the major structural components of MT and NF, the tubulin molecule and the NF subunits, while other minor enzymes and cofactors associated with both organelles might be the privileged target of alteration induced by age, resulting in the modification of their interactions with other subcellular components (MacieiraCoehelo 1988).
NT-
P408
IMGES OF HeLa CELL!5
IWIT-SkKfXED
Silvia
Regina Pergamo de Miranda, Maria L.S.Mello. Dept. of Cell Biology, Institute of Biology, Campinas, S.P., Brazil. HeLa cells cultured for 16h at 37'C and exposed to temperatures of 40°C or 43’C (B) for Ih were studied with the aim of detecting modifications in their pattern of mass distribution promoted by the heat-shock. Cugtures that were returned to control conditions (37 C) for 4h after the thermal shock were also investigated. Control cells were maintained at 37OC. The material was observed with an interference microscope before and after a 2M NaCl pattern
treatment. In of interference changed when compared
cells,
(A)
colors to
that
of
cells,
the
was the
slightly control.These
control conditions exhibited patterns similar to heat-shocked (6; cells. After hyperthermia at 43’C the interference colors in the majority of the interphase nuclei were superior than in controls. The images for the recovered cells were similar to those of control. In preparations treated with 2M NaCl a modification in the interference colors due to removal of nuclear proteins could be assumed only for control and heat-shocked cells. Changes in when
returned
heat-shocked
to
nuclear mass distribution with heat shock were thus demonstrated for the HeLa cells and showed the to differ depending on the temperatur& of shock. A shock at the temperature of 40 C resulted
in with
a later the
response in relation to higher temperature (43’C).
observed (CAPES)
that
Cell Biology
OF SYNAPTIC MITOCHONDRIA DURING AGING Bertoni-Freddari, Patrizia Fattoretti, Carlo Tiziana Casoli, *William Meier-Ruge, *Jurg Ulrich. Centre for Surgical Research, INRCA Research Department, Via Birarelli, 8, 60121 Ancona, Italy and *Neuropathology Division, University of Basel, Schoenbeinstrasse 40, Basel, Switzerland. Volume density (Vv), numerical density (NV). average volume (V) and shape skeleton (Sk) of synaptic mitochondria were measured by means of a computer assisted image analyser in the cerebellar granuli adult and old rats. of young, No significant difference was found in the three groups analysed with regard to Vv. A significant NV was found in adult animals when increase of compared both with young and old rats, whereas a in this parameter was significant decrease demonstrated in the old animals vs. the other V did not change between young and old groups. but was significantly smaller in the adult group, one. Sk showed a significant elongation in the shape of mitochondria during aging as demonstrated by the 55% increase of this parameter in the old animals. Since the mitochondria in the cerebellar directly glomerulus are reported to subserve synaptic transmission, the present ultrastructural findings support that age-related alterations at chemical synapses may find an explanation in and/or impaired energy mechanisms supply organelles.
P405
P407
MORPHOMRTRY
GENE E;CPRESSION
IN PEANUT MUTAHTS
RESISTANT TO LEAF SFOT DISEASE Zoventino D. Soriano, Arne C. Villanueva, Adelaida Pi. Calaguas, !?eliton T. Montejo. Natl. Acadcny of Science and Technology, TAP1 aldg., DOST Complex, Dicutan, Metro Pianila, Fhilippines The wor'x studied how to independent mutant genes are expressed to produce resistance to the leaf spot disease in peanut.
Two fungus species cause leaf spot, Cercosnora-personata which penetrates the leaf and C. arachldlcola, infects inner tissues. --The two mutant genes, Is-l and Is-2 govern inheritance of resiza?ice -to the disease. To enter the leaf. the hyphsl outgrowth of a germinating 'spore secretes enzyme cellulase which is more or less inactivated by a mutant protein in the plasma membrane of the epidermal cell. The mutant protein was found to remove the iron co-factor in cellulase polyFeptide. ;n the heterozygous condition, gene ls-2 governs synthesis of a polypeptide ?Xia? represses the action of a toxic enzyme secreted by nycelia of the fungus, n arachidicola restoring all functions ir. of chlorenchymaious cells in the leaf. The mutant polypeptide attaches to the toxic protein enzyme and displaces thiamine.
International
Reports,
P406
Vol. 14, Abstracts
MICROTUBULES (NF) IN
(MT) AGING
Supplement
1990
AND NEUROFILAMENTS RATS
J. Eyer, M. Coleman’ and J.F. Leterrier. U 298 INSERM, C.H.R.U., Angers, France and * Dpt of Immunology, St Georges Hospital Medical school. Cranmer Terrace, London, U.K. The composition and biochemical properties of MT and NF purified from young (3 months) and aged (26 months) rats ware compared on the basis of their polypeptide composition and their associated enzymatic activities. MT, purified in the presence of protease inhibitors in order to block the proteolysis of MAPS occuring in old rats (Matus and Greene, 1986), contained a lower amount of MAP2 in samples from aged animals correlated with a decrease in the amount of the MAPP-associatedcAMP dependent protein-kinase (Vallee et al, 1981) while the other MAPS were essentially unmodified. In addition, the specific volume of MT pellets was significantly decreased in samples from old animals, suggesting that the role of MAPS as spacer molecules on MT was altered by aging. NF isolated from spinal cord of the same animals were essentially undistinguishable between the young and old preparations. Both the yield of NF (mglwet weight), the polypeptidic composition and the associated protein-kinase activity were found nearly identical. The only age dependent-change in NFassociated enzymes was found with an ATPase activity which was significantly higher in NF from old rats. However, the in vitro interaction process between NF, reflected by the viscosity kinetics of filament suspensions (Leterrier and Eyer, 1987), were significantly faster in the dialysed NF samples from old than from young animals. This situation was reversed when undialysed NF were used, suggesting that dialysable factors were responsible for the distinct behaviour of both preparations. A preliminary study of dialysed molecules showed the presence of peptides reacting with a monoclonal antibody directed against the phcephorylated NF-H tail domain (from B.H. Anderton). These results argue for the lack of alteration with age of the major structural components of MT and NF, the tubulin molecule and the NF subunits, while other minor enzymes and cofactors associated with both organelles might be the privileged target of alteration induced by age, resulting in the modification of their interactions with other subcellular components (MacieiraCoehelo 1988).
NT-
P408
IMGES OF HeLa CELL!5
IWIT-SkKfXED
Silvia
Regina Pergamo de Miranda, Maria L.S.Mello. Dept. of Cell Biology, Institute of Biology, Campinas, S.P., Brazil. HeLa cells cultured for 16h at 37'C and exposed to temperatures of 40°C or 43’C (B) for Ih were studied with the aim of detecting modifications in their pattern of mass distribution promoted by the heat-shock. Cugtures that were returned to control conditions (37 C) for 4h after the thermal shock were also investigated. Control cells were maintained at 37OC. The material was observed with an interference microscope before and after a 2M NaCl pattern
treatment. In of interference changed when compared
cells,
(A)
colors to
that
of
cells,
the
was the
slightly control.These
control conditions exhibited patterns similar to heat-shocked (6; cells. After hyperthermia at 43’C the interference colors in the majority of the interphase nuclei were superior than in controls. The images for the recovered cells were similar to those of control. In preparations treated with 2M NaCl a modification in the interference colors due to removal of nuclear proteins could be assumed only for control and heat-shocked cells. Changes in when
returned
heat-shocked
to
nuclear mass distribution with heat shock were thus demonstrated for the HeLa cells and showed the to differ depending on the temperatur& of shock. A shock at the temperature of 40 C resulted
in with
a later the
response in relation to higher temperature (43’C).
observed (CAPES)
that