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M F Counis

Publications and source records attributed to M F Counis.

30 records · Page 2Linked to original sources
PubMed1987

Collagen synthesis by long-lived mRNA in embryonic chicken lens.

Lens capsule collagen synthesis by epithelial and fiber cells was examined by immunoprecipitation and collagenase digestion in embryonic and posthatch chicken eye lens. Epithelial cells and lens fibers in the process of terminal differentiation produce alpha 1 and alpha 2 type IV collagen chains. At 6 days of embryonic development in addition to the alpha 1 (IV) and alpha 2 (IV) collagen chains, lens cells produce high molecular weight collagenase-sensitive proteins not immunologically related to type IV collagen. Lens capsule collagen components have been identified in central and outer fibers isolated from 18-day embryos and from 10-day posthatch chicken eyes. At these stages, fibers which have an increasing number of picnotic nuclei still show collagen synthesis due to long-lived mRNA. Analysis of collagen synthesis by lens cells incubated with actinomycin D suggests that stabilization of collagen mRNA occurs in lens fiber cells and to a lesser extent in epithelial cells as early as 6 days of embryonic development.

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PubMed1987

Effect of X-irradiation and vitamin C on DNA degradation and endogenous DNase in embryonic chick lens cells.

The lens is an organ in which epithelial cells become elongated fibers. During this process, nuclei are transformed and the DNA is degraded. In previous studies, we described an autodigestion of the chromatin in isolated fiber nuclei but not in epithelial nuclei, but the level of DNAase activity was found to be identical in both epithelial and fiber nuclei of lenses at 11 days of development. In this study, we have investigated the possibility that x-irradiation might stimulate the nuclear endogenous activity responsible for chromatin breakdown or epithelial cells to a level comparable to that observed in fiber cells. We have observed that x-irradiation does not increase the nuclear epithelial DNAase activity. Conversely, vitamin C, suspected to prevent cataract formation by protecting DNA against free radical formation, has a damaging effect on the DNA of the lens of chick embryo in vitro.

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PubMed1986

Nuclear endogenous Ca2+-dependent endodeoxyribonuclease in differentiating chick embryonic lens fibers.

During terminal differentiation of lens epithelial cells into fiber cells, nuclei become pycnotic and DNA degradation occurs. We investigated the putative role in this process of an endogenous DNAase. After incubation of isolated nuclei of both cell types at 37 degrees C, DNAase activity was revealed by DNA size analysis on 0.3-1% neutral and alkaline agarose, one- and two-dimensional gels. This DNAase activity is more prominent in lens fiber nuclei than in epithelial nuclei at all the embryonic stages probably because of a preexisting higher concentration of divalent cations in the former. This activity is calcium or magnesium dependent in both types of nuclei.

Animals↗
PubMed1986

Chromatin condensation and terminal differentiation process in embryonic chicken lens in vivo and in vitro.

During embryonic chick lens differentiation, the epithelial cells become transformed into elongated fibres. Concomitantly, the fibre nuclei undergo degeneration and high molecular weight (HMW) DNA breaks down due to nuclear endodeoxyribonuclease activity. An electronmicroscopic study of lens epithelial and fibre nuclei was made at different stages of chick embryonic development, both in vivo and in vitro. The in vitro conditions are conducive to the expression of endogenous endodeoxyribonuclease activity in fibres. In both conditions we observed condensation of chromatin. The organization of some nuclear material into distinct linear arrays followed by streaming of nuclear material into the cytoplasm is recorded only in vitro. Such a condition may lead to acceleration of the process of aging in lens fibres.

Animals↗
PubMed1985

Nuclear ADP-ribosylation in the chick lens during embryonic development.

Nuclear ADP-ribosyltransferase is present in cells from the chick lens throughout embryonic development. The activity does not decrease when the cells become post-mitotic and commence terminal differentiation but declines slowly in both epithelia and fibre cells. At all stages studied the enzyme retains its ability to be activated by DNA strand breaks induced either by X-irradiation or by the action of an endogenous endonuclease. There is no correlation between the enzyme activity or the levels of its substrate NAD+ and the changes in DNA repair capacity which have been observed during the development of the lens.

Animals↗
PubMed1985

DNA repeat size in chick embryonic lens epithelium, lens fiber, brain and liver cell nuclei.

The DNA repeat size is determined by micrococcal nuclease digestion kinetics and subsequent electrophoresis of the products among various chick embryonic tissues. The repeat size is found to be not significantly different from 193 to 197 bp, for brain and liver at 11 days and for lens epithelium and fiber at different embryonic stages. However, the pattern of micrococcal digestion seems to reveal an overall chromatin modification as a function of development in the lens fibers.

Animals↗
PubMed1984

Changes in the DNA breakage and crystallin synthesis of embryonic chicken lenses cultured in a tryptophan-deficient medium.

In vitro experiments were performed in order to understand the biochemistry of tryptophan-deficient cataract. Eleven-day-old embryonic chick lenses were cultured in vitro for 3 hr, one and three days in a tryptophan-deficient medium. DNA breakage was followed on sucrose gradient and water-soluble protein synthesis was analysed by SDS-PAGE coupled with fluorography. A medium lacking tryptophan delays the DNA degradation and decreases the synthesis of all soluble proteins including the crystallins.

Animals↗
PubMed1981

DNA polymerase, DNA ligase, and thymidine kinase activity in chicken lens, related to DNA X-ray lesion repair.

The activity of DNA polymerases alpha, beta, and gamma; DNA ligases I and II; and thymidine kinase in chicken lenses is determined. These enzymes are present in embryonic intact lenses freshly isolated at 6 days and 11 days of development and in lenses isolated at 11 days of development and cultured for three days. They are also found in both epithelium and fibers when separated at 10 days of embryonic development and in the epithelium of 141/2-month-old hen lenses. In the anucleate mature hen lens fibers, the only detected enzyme is thymidine kinase. Previous results showed that repair of X-ray irradiated DNA was total in 11-day-old embryonic lenses while repair was not detected at six days. On the other hand, repair was very much impaired when the 11-day-old lenses were cultured for three days. Enzymic analyses suggest that the defect in DNA repair and the spontaneous DNA degradation observed in chick lenses at certain embryonic stages, are not due to the absence of any of the above enzymic activities. Alternative explanations are discussed.

Animals↗
PubMed1980

Cycloheximide effect on DNA degradation and delta-crystallin synthesis in terminally differentiating lens cells.

Low concentrations of a protein synthesis inhibitor, cycloheximide, were added throughout the process of in vitro differentiation of 11-day old embryonic chick lens cells. We found with low concentrations of cycloheximide (0.01 to 0.03 microgram/ml, 3 days of culture), that there was an almost complete delay of DNA degradation as observed on alkaline sucrose gradient. Identical concentrations and exposure time had no blocking effect on increased delta-crystallin synthesis as detected by immunoprecipitation and electrophoresis. Higher concentrations of cycloheximide (0.1 to 1 microgram/ml) showed a marked effect on DNA size and a net inhibition on delta-crystallin synthesis. Thus a selective effect of low doses of cycloheximide was observed on terminal differentiation suggesting that there was not a relationship between DNA degradation and delta-crystallin synthesis in these short term experiments. The investigations of minor proteins could be of interest as they may have a crucial role in intact nuclei cataracts.

Animals↗
PubMed1979

DNA repair in lens cells during chick embryo development.

When chick lens epithelium is cultured in vitro, differentiation into lens fiber cells is accompanied by DNA degradation. This phenomenom of terminal differentiation was studied in the epithelium from embryos at the 6th and 11th days of development. DNA size and the ability of the cells to repair DNA damage induced by X-rays were analysed in alkaline sucrose gradients. In the 6-day epithelium a rapid degradation and complete lack of DNA repair were recorded. Similar observations have been made in previous studies on the 11-day sample, but here degradation is progressive and occurs after a lag of several days. In the younger epithelium, internal irradiation by [3H]thymidine also had a drastic effect resembling that caused by X-rays. In order to assess the process of differentiation in our experimental system the synthesis of delta- and alpha-crystallins was monitored. Stage-related modifications in the rates of synthesis were recorded. The results confirm that the DNA repair system is impaired during terminal differentiation. The differences observed between the two stages may reflect either a developmental modification in DNA repair mechanisms or a change in the relative proportions of differentiating cells. An hypothesis is proposed in support of the latter case.

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