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Metabolism of Oat Leaves during Senescence : VII. The Interaction of Carbon Dioxide and Other Atmospheric Gases with Light in Controlling Chlorophyll Loss and Senescence.

In air largely freed from CO(2), senescence of isolated oat (Avena sativa cv Victory) seedling leaves is no longer prevented by white light; instead, the leaves lose both chlorophyll and protein as rapidly as in the dark. Senescence in light is also accelerated in pure O(2), but it is greatly delayed in N(2); 100% N(2) preserves both protein and chlorophyll in light and in darkness. In light in air, most of the compounds tested that had previously been found to delay or inhibit senescence in darkness actually promote the loss of chlorophyll, but they do not promote proteolysis. Under these conditions, proteolysis can therefore be separated from chlorophyll loss. But in light minus CO(2), where chlorophyll loss is rapid in controls, two of these same reagents prevent the chlorophyll loss. Unlike the many reagents whose action in light is thus the opposite of that in darkness, abscisic acid, which promotes chlorophyll loss in the dark, also promotes it in light with or without CO(2). Kinetin, which prevents chlorophyll loss in the dark, also prevents it in light minus CO(2). In general, therefore, the responses to light minus CO(2) are similar to the responses to darkness, and (with the exception of abscisic acid and kinetin) opposite to the response to light in air.

Journal Article↗

Drop of connexin 43 in replicative senescence of human fibroblasts HEL-299 as a possible biomarker of senescence.

The expression of connexin 43 (cx43) and cell-cell communication were studied in replicative senescence of cultured HEL-299 fibroblasts. A progressive decrease in fluorescent dye transfer was detected by a scrape-loading technique in aging fibroblasts. This change was accounted for by a marked decrease in the amount of cx43 in aging cells, as detected by western blot analysis (cell extracts) and indirect fluorescence (cells in culture). However, semiquantitative RT-PCR assays of cx43 mRNA did not reveal appreciable changes, which suggests several possible explanations for the mechanism(s) underlying the decrease of cx43 in aging cells. These findings support the idea that the reduced expression of cx43 might be a biomarker of cell senescence.

Biomarkers↗

Cataract and other ophthalmic lesions in senescence accelerated mouse (SAM). Morphology and incidence of senescence associated ophthalmic changes in mice.

In a murine model of accelerated senescence (SAM), grading score and incidence in cataract, periophthalmic lesions, opacity and ulcer of the cornea were determined in mice from 4 to 24 months of age. From 4 to 6 months of age, incidence and grading score of these four categories began to increase in both the accelerated senescence prone (SAM) and resistant series with normal aging, and these increases continued with aging. As compared with the resistant series, there was a higher incidence and grading score of the four categories and a higher rate of increase in the prone series. The prone 3 series in particular showed a much higher incidence and grading score on cataract, the rate being 27.5% and 70.6% at 12 and 16 months, respectively. Histologically, the cataract was classified into two types. In one, degeneration of lens fibers, disintegration of lens cortex, and at an advanced stage, liquefaction of the lens cortex and proliferation of the anterior lens epithelial cells occurred. In the other type, lens fibers lost their distinct shapes and a homogenous mass formed at the anterior and posterior superficial cortex. The anterior lens epithelial cells had shrunk. There was an opacity and ulcer of the cornea with keratitis and the corneal epithelium was lost in case of the latter. Periophthalmic lesions included catarrhal changes of the skin of the eyelids and face and blepharitis. There were no lesions specific to each of the prone and resistant series. Thus, SAM should prove to be a suitable murine model for investigation of age-related ophthalmic lesions, including cataract in humans.

Aging↗

Senescence of aortic endothelial cells in vitro: influence of culture conditions and preliminary characterization of the senescent phenotype.

The replicative lifespan of a cloned strain of adult bovine aortic endothelial cells has been examined by serial passage in culture using conditions where the cells were either dependent on fibroblast growth factor (FGF) for rapid growth or relatively independent on FGF for growth. In FGF-dependent cultures a replicative lifespan of 130 generations was attained with a phase III period which spanned approximately 20 generations. Withdrawal of FGF at generation 55 and repeated passage of such cultures in the absence of the growth factor resulted in the loss of proliferative potential within 15 generations. The morphological changes occurring upon FGF withdrawal were different than those that occurred when cultures senesced in the presence of FGF, FGF withdrawn cells showed a homogeneous increase in cell size and after several passages overgrew one another at confluency. Endothelial cells which senesced in the presence of FGF showed a very heterogeneous distribution of enlarged cells, many of which were binucleated but continued to form a confluent monolayer at high cell densities. Under FGF-independent conditions (begun at generation 48) a replicative lifespan of 105 generations was attained in the presence of the growth factor. FGF withdrawal under these conditions only decreased the replicative lifespan to 95 generations. Under these conditions the morphological changes occurring during phase III were identical in the presence and absence of FGF. Examination of the sensitivity of endothelial cells to FGF as they entered phase III showed that their dose-response characteristics were not qualitatively altered after the onset of phase III, although the number of cells responding to FGF progressively dropped. Comparison of the patterns of proteins synthesized in phase II and phase III cultures showed that phase III cultures even when plated at sparse densities continued to synthesize proteins which were normally observed in phase II confluent cultures. The results indicate that onset of the phase III period in aortic endothelial cells cloned and maintained in the presence of fibroblast growth factor can be delayed to a greated extent if the cells are maintained under conditions where they are made dependent on FGF for rapid growth in culture.

Aging↗

A 1100-bp sequence of mitochondrial DNA is involved in senescence process in Podospora: study of senescent and mutant cultures.

In Podospora, senescence is assumed to be caused by the amplification of short sequences of the mitochondrial genome (sen-DNAs). We have characterized a 1100-bp-long mitochondrial DNA sequence which could be directly involved in the phenomenon. Indeed, by hybridization experiments, we show that this sequence is both present in all the sen-DNA molecules which originate from the beta region of the mitochondrial chromosome and rearranged in the mitochondrial genome of two mitochondrial mutants selected as resistant to senescence.

Ascomycota↗

The occurrence of vacuolation, and periodic acid-Schiff (PAS)-positive granules and plaques in the brains of C57BL/6J, AKR, senescence-prone (SAMP8) and senescence-resistant (SAMR1) mice infected with various scrapie strains.

Scrapie is a fatal, but slow, infectious disease. C57BL/6J, SAMP8 (a strain that develops early senescence), SAMR1 (a strain that is resistant to senescence) and AKR/J (a progenitor of the SAM strains) mice were infected with 22A, 139A, 22L and ME7 scrapie strains. Histopathological stains included haematoxylin and eosin (HE), and periodic acid-Schiff (PAS). Vacuolation was found in the brains of all scrapie-infected mice. The 22A strain caused more extensive vacuolation in the brains of SAMP8 and SAMR1 mice than in C57BL mice. PAS-positive plaques (PP) were found in 22A-infected mice in cortex, corpus callosum, hippocampus, subependymal zone area and thalamus. PP were significantly increased in 22A-infected SAMR1 mice compared to mice from other scrapie-infected strains. Clusters of small, round, homogeneous PAS-positive granular structures (PGS) were found in all mouse strains, especially in aging control and 22A-infected C57BL mice, predominantly in the stratum radiatum of the CA1, CA2 and CA3 areas of the hippocampus. Some of these structures were also observed in stratum oriens and piriform cortex, and in cerebellar Purkinje cell areas. Some of the PGS were associated with astrocytes and blood vessels. Each granule was 1-5 microm in diameter and there were clusters consisting of several to 40 PGS; the sizes of the clusters ranged from 10 to 80 microm in diameter. There were more PGS clusters in uninfected C57BL and AKR mice than in uninfected SAMP8 and SAMR1 mice. PGS were not increased in scrapie-infected mice. These findings suggest that PGS accumulation was more dependent on the genetic information of the mouse strain, whereas PP and vacuolation patterns depended on the scrapie strain-mouse strain combination.

Aging↗

Metabolism of Oat Leaves during Senescence : VIII. The Role of L-Serine in Modifying Senescence.

The mechanism whereby l-serine specifically promotes the dark senescence of detached oat (Avena) leaves has been examined. The fact that this promotion is strong in darkness but very weak in white light has been explained, at least in part, by the finding that added serine is partly converted to reducing sugars in light. Labeled serine gives rise to (14)C-sugars and (14)CO(2). In the absence of CO(2), serine does cause chlorophyll loss in light and undergoes a decreased conversion to sugar.As to the large promotion of protease activity which accompanies senescence in the dark, reported earlier, careful purification of the proteases shows that the l-[(14)C]serine is not incorporated into these enzymes, although it is incorporated into the total protein. Cycloheximide decreases the overall synthesis both of protease and of total protein, but again [(14)C]serine does not impart radioactivity to the purified acid proteases. Even when serine is simply added to the protease assay the proteolysis is significantly increased. It is concluded that serine promotes the protease activity by synergizing with the enzyme, or by activating an apoenzyme.

Journal Article↗

[Biological model of accelerated senescence. II. Age-related changes in the number of developing male germ cells and Sertoli cells in gonads of senescence-accelerated mice].

It was shown that in immature three- to four-week-old mice prone to accelerated senescence (SAMP1 strain), the number of spermatogonia, pachytene spermatocytes, and circular spermatids exceeded that in mice resistant to accelerated senescence (SAMR1 strain) by more than two times. Differences were found in the pattern of age-related changes in the number of meiotic and postmeiotic cells in the sexually mature SAMP1 and SAMR1 mice. In the gonads of SAMP1 and SAMR1 mice, the number of Sertoli cells was unstable.

Aging↗

Delay of leaf senescence in Medicago sativa transformed with the ipt gene controlled by the senescence-specific promoter SAG12.

We report the successfull delay of leaf senescence in Medicago sativa. A highly regenerable clone of alfalfa was transformed with the construct SAG12-IPT, an approach that has already proved efficient in other crops. Several independent transformants were obtained as determined by Southern analysis and all the transformants expressed the transgene as measured by RT-PCR. In vitro and in vivo analyses showed that SAG12-IPT plants exhibited a stay-green phenotype that has the potential to greatly improve the quantity and quality of alfalfa forage.

Alkyl and Aryl Transferases↗