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Morphometry of zinc-iodide-osmium (ZIO) positive synaptic vesicles during aging
178
Cell Biology
International
P398
MORPHOMETRY OF ZINC-IODIDE-OSMIUM (ZIO) POSITIVE SYNAPTIC VESICLE3 DURING AGING Tiziana Casoli, Carlo Bertoni-Freddari, Patrizia Fattoretti, *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, Base1 Switzerland. A morphometric study has been carried out on 210 positive synaptic vesicles in the cerebellar glomerulus of young, adult and old rats. Numerical density (NV), average diameter (D) and volume density (Vv) of the vesicles were measured by analyzer means of a computer-assisted image programmed by ourselves. NV showed a 25% increase in 12 month old rats as compared with the 3 month old group. In the old animals we found 49 and 862 decreases as compared with young and adult values, respectively. Vv did not show any significant change between 3 and 12 months of age, but was decreased by 54% in the 30 month old group. D was significantly decreased by 6.5 and 5.4% in the adult group as compared with young and old rats, respectively. Although the biological significance of the ZIO positive vesicles is still a matter of debate, there are some evidences supporting that they may contribute to the homeostatic control of calcium ions at synaptic terminals. We interpret our findings as an age-dependent adaptive response in controlling the intraterminal calcium ions (Bertoni-Preddari et al. Scann. Micr. Intl. Suppl. 3, 101-107, 1989)
P400
P399
Vol. 14, Abstracts
Supplement
1990
DEPRNDENTCHANGESOF POLYPHOSPMCINOSITIDE KKTRBOLISN IN RAT LIVER NUCLEI H.Previati', S.J4anziroliL M.Mazzonia, A.HatteuccilL, N.Di Giovanni', V.Bertagnolo*, Institute of Human Anatomy, University of Ferrara', CNR Institute of Cytomorphology (c/o Rizzoli) Bologna*, Italy.
MDRFHOLOGICAL AND BIOLOGICAL MODIFICATIONS IN HUMAN FIBROBLAST PRIMARY CULTURES DURING CELLULAR AGING. Pierluigi Cardelli, Susanna Scarpa, Laura Masuelli, and Andrea Modesti. Department of Medicine, University of Rome "La Sapienza" Italy. Human gengiva; fibroblast (HGF) primary culLules were obtained from biopsies of patients at different ages (20/50/65 years): each culture was passaged by trypsinization up to the total exhaustion of the cell replication activity. HGF derived from the 20 years-old donor were kept for 35 passages, the 50 years-old HGF for 19, and the 64 years-old HGF for 13 passages. Each culture was examined for growth rate and for collagen synthesis at specific pulses (every 5 starting with the second, up to passages), For growth rate determithe 35t$ passage. H-thymidine incorporation was used, nation and for pro-collagens production the proteins secreted into the culture medium were selectively ethanol precipitated and visualized by SDS-PAGE electrophoresis. The ultrastructural modifications were also analyzed by transmission electron microscoeach culture at the first and PYV comparing the last passages.
P397
Reports,
AGE
The ability of nuclei to synthesize j,~ ~&RQ phosphatidylinositol O-phosphate (PIP) phosphatidylinositol 4,5-bisphosphate (PIPa) and phosphatidie acid (PA) has been recently demonstrated. To assess the involvement of nuclear inositol lipids in cellular ageing, we have studied the lipid phosphorylation in liver nuclei isolated from rats aged between 3 and 36 months, by following the incorporation of [v-PIATP into PIP, PIPI and PA in the presence or absence of modulators of kinases Under the basal essay conditions and phosphatases. the synthesis of PIP, PIPa and PA shows a decrease in nuclei isolated from 18 and 26 month-old rats, and an increase at 12 and 30 months. In the presence of exogenous substrates and detergent it was observed that the phosphatidylinositol (PI)-kinase activity was deeply reduced in nuclei obtained from 24 month-old rats, suggesting that the drop of PIP recovery was due t.c impairment of synthesis. The activity of PIP-kinase displayed the sazm features, even though it was ma&dined to relatively higher levels. The use of inhibitors and activators of PIcycle related hydrolytic enzymes confirms that the reduction of recovery of PIP and, to a lower extent, of PIPI, was due to a decrease of kinase activity. These date indicate that an inositol lipidprotein kinase C dependent signalling system is operative in the nucleus, and that the molecular basis of aging might involve polyphosphoinositideregulated nuclear funtions.
A DECLINE M A’i’ DRIAL CONTENT
PRRCEDE DURING
A LOSS IN MITOCHON. SPLENOCYT’RS AC&MC.
Abderrhaman Maftah*, Patrick Leprat*, Marie-H&se Rstiasud**. RaymondJ&II*. * GCnius-Biorcchnologic.FruIti dcs Sciawcs. 87060 Limogcs,Frsnce.**Scrvics&Cy@m&is,Fscelt6de.M&cii87025 L.imogss,Frsaca. Numemus studies have shown that the loss of proliferative capacity of cells in vitro is due to changes in the mitochondrial compartment such as tednction of the trammembrane potential (AY) and/or membrane content A combined use of mitochondria specific probes with a potential-dependent (Rhodamine 123) or independent (Nonyl Actidine Orange) uptake may provide an appropriate method to distinguish between mitochondrial membrane potential variations and membrane content modifications during splenocytes ageing. We found that the decline in membrane potential in the mouse occuted approximately six months prior to the decrease in mitochondrial membrane content. The analysis of the Rhodamine 123/Nonyl Acridine Orange fluorescence ratio measuxed in splenocytes obtained fmm mice aged mom than 6 months showed that there was a linear loss of mitochondria efficiency. Moreovet, cells with a ratio of less than 0.85 were incapable ofproliferating and temained quiescent.
Cell Biology
International
P398
MORPHOMETRY OF ZINC-IODIDE-OSMIUM (ZIO) POSITIVE SYNAPTIC VESICLE3 DURING AGING Tiziana Casoli, Carlo Bertoni-Freddari, Patrizia Fattoretti, *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, Base1 Switzerland. A morphometric study has been carried out on 210 positive synaptic vesicles in the cerebellar glomerulus of young, adult and old rats. Numerical density (NV), average diameter (D) and volume density (Vv) of the vesicles were measured by analyzer means of a computer-assisted image programmed by ourselves. NV showed a 25% increase in 12 month old rats as compared with the 3 month old group. In the old animals we found 49 and 862 decreases as compared with young and adult values, respectively. Vv did not show any significant change between 3 and 12 months of age, but was decreased by 54% in the 30 month old group. D was significantly decreased by 6.5 and 5.4% in the adult group as compared with young and old rats, respectively. Although the biological significance of the ZIO positive vesicles is still a matter of debate, there are some evidences supporting that they may contribute to the homeostatic control of calcium ions at synaptic terminals. We interpret our findings as an age-dependent adaptive response in controlling the intraterminal calcium ions (Bertoni-Preddari et al. Scann. Micr. Intl. Suppl. 3, 101-107, 1989)
P400
P399
Vol. 14, Abstracts
Supplement
1990
DEPRNDENTCHANGESOF POLYPHOSPMCINOSITIDE KKTRBOLISN IN RAT LIVER NUCLEI H.Previati', S.J4anziroliL M.Mazzonia, A.HatteuccilL, N.Di Giovanni', V.Bertagnolo*, Institute of Human Anatomy, University of Ferrara', CNR Institute of Cytomorphology (c/o Rizzoli) Bologna*, Italy.
MDRFHOLOGICAL AND BIOLOGICAL MODIFICATIONS IN HUMAN FIBROBLAST PRIMARY CULTURES DURING CELLULAR AGING. Pierluigi Cardelli, Susanna Scarpa, Laura Masuelli, and Andrea Modesti. Department of Medicine, University of Rome "La Sapienza" Italy. Human gengiva; fibroblast (HGF) primary culLules were obtained from biopsies of patients at different ages (20/50/65 years): each culture was passaged by trypsinization up to the total exhaustion of the cell replication activity. HGF derived from the 20 years-old donor were kept for 35 passages, the 50 years-old HGF for 19, and the 64 years-old HGF for 13 passages. Each culture was examined for growth rate and for collagen synthesis at specific pulses (every 5 starting with the second, up to passages), For growth rate determithe 35t$ passage. H-thymidine incorporation was used, nation and for pro-collagens production the proteins secreted into the culture medium were selectively ethanol precipitated and visualized by SDS-PAGE electrophoresis. The ultrastructural modifications were also analyzed by transmission electron microscoeach culture at the first and PYV comparing the last passages.
P397
Reports,
AGE
The ability of nuclei to synthesize j,~ ~&RQ phosphatidylinositol O-phosphate (PIP) phosphatidylinositol 4,5-bisphosphate (PIPa) and phosphatidie acid (PA) has been recently demonstrated. To assess the involvement of nuclear inositol lipids in cellular ageing, we have studied the lipid phosphorylation in liver nuclei isolated from rats aged between 3 and 36 months, by following the incorporation of [v-PIATP into PIP, PIPI and PA in the presence or absence of modulators of kinases Under the basal essay conditions and phosphatases. the synthesis of PIP, PIPa and PA shows a decrease in nuclei isolated from 18 and 26 month-old rats, and an increase at 12 and 30 months. In the presence of exogenous substrates and detergent it was observed that the phosphatidylinositol (PI)-kinase activity was deeply reduced in nuclei obtained from 24 month-old rats, suggesting that the drop of PIP recovery was due t.c impairment of synthesis. The activity of PIP-kinase displayed the sazm features, even though it was ma&dined to relatively higher levels. The use of inhibitors and activators of PIcycle related hydrolytic enzymes confirms that the reduction of recovery of PIP and, to a lower extent, of PIPI, was due to a decrease of kinase activity. These date indicate that an inositol lipidprotein kinase C dependent signalling system is operative in the nucleus, and that the molecular basis of aging might involve polyphosphoinositideregulated nuclear funtions.
A DECLINE M A’i’ DRIAL CONTENT
PRRCEDE DURING
A LOSS IN MITOCHON. SPLENOCYT’RS AC&MC.
Abderrhaman Maftah*, Patrick Leprat*, Marie-H&se Rstiasud**. RaymondJ&II*. * GCnius-Biorcchnologic.FruIti dcs Sciawcs. 87060 Limogcs,Frsnce.**Scrvics&Cy@m&is,Fscelt6de.M&cii87025 L.imogss,Frsaca. Numemus studies have shown that the loss of proliferative capacity of cells in vitro is due to changes in the mitochondrial compartment such as tednction of the trammembrane potential (AY) and/or membrane content A combined use of mitochondria specific probes with a potential-dependent (Rhodamine 123) or independent (Nonyl Actidine Orange) uptake may provide an appropriate method to distinguish between mitochondrial membrane potential variations and membrane content modifications during splenocytes ageing. We found that the decline in membrane potential in the mouse occuted approximately six months prior to the decrease in mitochondrial membrane content. The analysis of the Rhodamine 123/Nonyl Acridine Orange fluorescence ratio measuxed in splenocytes obtained fmm mice aged mom than 6 months showed that there was a linear loss of mitochondria efficiency. Moreovet, cells with a ratio of less than 0.85 were incapable ofproliferating and temained quiescent.