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G M Martin

Publications and source records attributed to G M Martin.

At least 145 records · Page 8Linked to original sources
PubMed1985

Fidelity of DNA polymerases isolated from regenerating liver chromatin of aging Mus musculus.

DNA polymerase-alpha and -beta were fractionated from the chromatin of regenerating liver of young and old mice. The DNA polymerases were resolved from each other and partially purified by DEAE-cellulose, phosphocellulose, and DNA-cellulose column chromatography. No significant age-related difference in the kinetics of heat inactivation was observed for either DNA polymerase. No age-dependent difference was found in the fidelity with which these enzymes copied phi X174 DNA. These results suggest that the functional properties of these DNA polymerases do not change with age as is postulated in some theories of aging.

Aging↗
PubMed1985

Subpopulations of fibroblasts from mouse skeletal muscle defined by clonal variation for 5' nucleotidase expression.

A primary cloning technique has been employed for the isolation of nine spontaneously transformed cell lines from mouse skeletal muscle. Four of these lines were isolated after selection for partial resistance to the purine (adenine) analog 2'6'diaminopurine and five were isolated from non-selected control dishes. Four of the nonselected lines and three of the selected lines demonstrated a fibroblastoid morphology in vitro. The other two cell lines (one from each group) were epithelioid. Two of the three selected fibroblastoid lines were found to contain significant quantities of the enzyme 5'nucleotidase (EC3.1.3.5), whereas the four nonselected fibroblastlike lines, one selected fibroblastlike line, and the two epithelioid lines did not. In the two cell lines expressing 5'nucleotidase activity, this expression was stable in the absence of selective pressure. Histochemical staining of mouse skeletal muscle for 5'nucleotidase activity demonstrated positive staining in the cells of small blood vessels and in a subset of the connective tissue cells. The bulk of the skeletal muscle tissue, however, had no detectable 5'nucleotidase activity. We propose that the two cultivatable types of fibroblastoid cell lines represent distinct classes of fibroblastlike cells in vivo, reflecting alternative states of stable cellular differentiation involving 5'nucleotidase expression.

2-Aminopurine↗
PubMed1985

Genetics and aging; the Werner syndrome as a segmental progeroid syndrome.

The maximum lifespan potential is a constitutional feature of speciation and must be subject to polygenic controls acting both in the domain of development and in the domain of the maintenance of macromolecular integrity. The enormous genetic heterogeneity that characterizes our own species, the complexities of numerous nature-nurture interactions, and the quantitative and qualitative variations of the senescent phenotype that are observed suggest that precise patterns of aging in each of us may be unique. Patterns of aging may also differ sharply among species (for example, semelparous vs. multiparous mammals). Some potential common denominators, however, allow one to identify progeroid syndromes in man that could lead to the elucidation of important pathways of gene action. (The suffix "-oid" means "like"; it does not mean identity.) Unimodal progeroid syndromes (eg., familial dementia of the Alzheimer type, an autosomal dominant) can help us understand the pathogenesis of a particular aspect of the senescent phenotype of man. Segmental progeroid syndromes (eg. the Werner syndrome, an autosomal recessive) may be relevant to multiple aspects of the senescent phenotype. Some results of research on the Werner syndrome may be interpreted as support for "peripheral" as opposed to "central" theories of aging; they are consistent with the view that gene action in the domain of development (adolescence, in this instance) can set the stage for patterns of aging in the adult; they point to the importance of mesenchymal cell populations in the pathogenesis of age-related disorders; finally, they underscore the role of chromosomal instability, especially in the pathogenesis of neoplasia.

Aging↗
PubMed1985

Multipotent mutator strain of mouse teratocarcinoma cells.

A mutator strain [AraCr (1.5)4], isolated from mutagenized cultures of multipotent mouse teratocarcinoma cells (embryonal carcinoma stem cells), exhibited a dNTP pool imbalance, with more than a 10-fold relative increase in the intracellular concentration of dCTP. The increase in the spontaneous rate of mutation for 6-thioguanine resistance was 3.6-fold and for ouabain resistance, 7.9-fold. Normalization of the dCTP/dTTP ratio by addition of thymidine and deoxycytidine to the media was associated with normalization of the mutation rates. AraCr (1.5)4 cell retained its multipotency (including chimerization potential) when injected into blastocysts. Moreover, its differentiated progeny expressed the dNTP pool imbalance and mutator phenotype in vitro. The preliminary finding of an increased frequency of morphologically abnormal embryos derived from a series of transplanted blastocysts injected with AraC2 (1.5)4 stem cells is consistent with significant phenotypic effects in vivo.

Animals↗
PubMed1985

Formation of associations of colored and flavored food with induced sickness in five avian species.

Domestic ducks, geese, pigeons, quail, and chickens were given colored, flavored, or colored and flavored food and then injected with lithium chloride. Each species showed learning of color and taste aversions. Flavor facilitated the formation of color aversions in ducks, geese, and pigeons but not in quail or chickens. Color interfered with the formation of flavor aversions in quail and chickens but not in the other three species. These findings indicate that all birds can probably associate both colored and flavored food with induced sickness and that colored food is more easily associated with induced sickness than is colored water. Moreover, these findings suggest that the capacity to associate colored and flavored food with induced sickness and the interaction between color and flavor in food vary between species. Birds, such as quail and chickens, that eat relatively tasteless food rely more on color than on flavor cues when forming learned food aversions. Birds that can select their food on the basis of taste, such as ducks and geese, rely more on flavor than on color when forming aversions to food. Birds, such as pigeons, that are initially raised on tasty food before switching to predominantly tasteless food show tendencies that are similar to those of ducks and geese.

Animals↗
PubMed1985

Fidelity of DNA polymerase-beta in neurons from young and very aged mice.

Neurons do not divide during adult life and thus they provide a unique system to study the effects of age-accumulated damage to DNA in the absence of DNA replication. We have analyzed DNA polymerase activity in neurons isolated from young adult and very aged mice. The predominant catalytic activity is DNA polymerase-beta and it is present in similar amounts in neurons from young and old mice. This polymerase is highly error-prone in copying phi X174 DNA, the error frequency being about 1/7,000 and not significantly different when obtained from young and old animals. This high infidelity is considered with respect to DNA repair and the protein synthesis error catastrophe theory of aging.

Aging↗
PubMed1985

Induction of adenine salvage in mouse cell lines deficient in adenine phosphoribosyltransferase.

Adenine phosphoribosyltransferase (APRT) (EC 2.4.2.7) pseudorevertant cell lines were isolated under selective conditions requiring adenine salvage for survival; yet they were found to be deficient in measurable APRT activity and resistant to the purine analog 2'6'-diaminopurine (DAP) (M.S. Turker, J. A. Tischfield, P. Rabinovitch, P.J. Stambrook, J.J. Trill, A.C. Smith, C.E. Ogburn, and G.M. Martin, manuscript in preparation). Adenine salvage was examined in two APRT pseudorevertant cell lines, their two APRT homozygous deficient parental cell lines, and a genotypic APRT revertant cell line (i.e., one with measurable APRT activity and DAP sensitivity). Adenine accumulation was observed in both revertant phenotypes and was demonstrated by high-performance liquid chromatography to be linked with adenine metabolism. The ability to salvage adenine declined substantially in the pseudorevertant cell lines when they were removed from selective media containing inhibitors of de novo 5'-AMP synthesis (alanosine and azaserine); for one pseudorevertant cell line this decline was accelerated by the addition of DAP to the medium. The readdition of alanosine or azaserine to the growth medium of the pseudorevertant lines induced adenine salvage to its previous levels. An APRT-like cross-reacting material was found in the pseudorevertant cell lines, although its relationship to adenine salvage is unknown. A low level of constitutive adenine salvage was found in the parental APRT-deficient lines, and it was also possible to induce adenine salvage in these cell lines. These findings suggest a novel regulatory mechanism for adenine salvage.

2-Aminopurine↗
PubMed1984

Delayed and reduced cell replication and diminishing levels of DNA polymerase-alpha in regenerating liver of aging mice.

Kinetics of cell replication was compared in regenerating livers of Mus musculus at ages ranging between 6 and 32 months. Incorporation of [3H]-thymidine into DNA and autoradiographic analysis showed that the maximal extent of DNA replication following partial hepatectomy became delayed with age. Furthermore, the total fraction of parenchymal and nonparenchymal cells in S phase at different intervals during regeneration diminished as mice aged. The specific activity of DNA polymerase-alpha, the putative replicative enzyme, declined progressively during aging. The specific activity of DNA polymerase-beta, the purported repair enzyme, declined to an appreciably lesser extent during the lifespan of the mouse. No evidence was found for the appearance of a specific inhibitor of polymerase-alpha in senescent mouse liver. Also, the bulk of the activities of both hepatic DNA polymerase-alpha and -beta remained localized in the cell nucleus throughout the lifetime of the animal and were mainly associated with chromatin.

Aging↗
PubMed1984

A cloning assay for 6-thioguanine resistance provides evidence against certain somatic mutational theories of aging.

The frequencies of 6-thioguanine-resistant primary clones from the kidneys and skeletal muscles of aging male cohorts of two F1 hybrid strains of Mus musculus varied from 0.59 to 10.96 X 10(-5) and did not increase as a function of donor age (up to 40 months). Resistant clones were shown to be severely deficient in the activity of hypoxanthine-guanine phosphoribosyltransferase (EC 2.4.2.8). These deficiencies presumably resulted from molecular alterations at this X-linked locus, including point mutations. No alterations of the X-chromosome were observed at the level of the light microscope. These results are inconsistent with predictions of the intrinsic mutagenesis and protein synthesis error catastrophe theories of aging. They do not rule out, however, somatic mutational theories that invoke comparatively large-scale chromosomal lesions, many of which would be likely to be lethal at the cellular level.

Adenine↗
PubMed1984

High frequency "switching" at the adenine phosphoribosyltransferase locus in multipotent mouse teratocarcinoma stem cells.

Clones of multipotent mouse tetratocarcinoma stem cells, presumptively heterozygous at the adenine phosphoribosyltransferase (APRT) locus (EC 2.4.2.7), were selected for partial resistance to the purine analog 2',6'-diaminopurine (DAP). All had approximately 50% APRT activity as compared to the parental line and were found to segregate homozygous deficient cells at a high frequency (approximately 10(-2]. Homozygous deficient cells were isolated from one of the heterozygotes and were found to fall into a single class characterized by residual activity and the segregation of revertants at an equally high frequency. The revertants in turn gave rise to full mutants at comparably high frequencies. Chromosomal changes detectable with the light microscope were not associated with these transitions. Physical characterization of the APRT enzymes derived from mutant, revertant, and wild-type cells did not reveal any differences. We conclude that the reversible "switching" between heterozygosity and homozygosity is attributable to some form of gene inactivation and reactivation rather than to classical mutational events.

Adenine Phosphoribosyltransferase↗
PubMed1984

Altered differentiation, indefinite growth potential, diminished tumorigenicity, and suppressed chimerization potential of hybrids between mouse teratocarcinoma cells and thymocytes.

Hybrids between normal mouse thymocytes and a hypoxanthine-guanine phosphoribosyl transferase (HGPRT) -deficient, pseudodiploid, multipotent mouse teratocarcinoma cell line displayed predominately hypotetraploid modes with differential patterns of karyotypic variation and no apparent consistent pattern of chromosome segregation. The hybrids failed to express thymocyte antigens Thy-1.2, Lyt-1, and Lyt-2, indicating suppression of thymocyte differentiation. Like the teratocarcinoma parent, the hybrids had an embryonal carcinoma (EC) stem cell component which exhibited indefinite growth potential in vitro and produced tumors with multiple pathways of tissue differentiation when injected subcutaneously. Hybrids had decreased proportions of EC cells and individually unique patterns of differentiation. The tumorigenicity of hybrids was diminished, indicating partial suppression of the neoplastic phenotype by the genome of the thymocyte parent. The EC parent gave chimerization frequencies of 18.5-19.2%, confirming the findings of Dewey et al. that mutant teratocarcinoma cells can be used to synthesize chimeric mice. "Selfed" near-tetraploid EC cells gave a frequency of 2.3%, showing that an increase in ploidy can reduce, although not eliminate, chimerization potential. No chimeras could be detected with hybrids (frequency less than 0.65%). Thus, there was no evidence that a thymocyte could be "reprogrammed" to participate in new developmental pathways.

Animals↗
PubMed1983

Preparation of multivesicular liposomes.

A novel type of liposome, named here multivesicular liposomes, was prepared by evaporation of organic solvents from chloroform-ether spherules suspended in water. Within each spherule were numerous water droplets that contained solutes to be trapped in liposomes upon solvent evaporation. Liposome preparations of different average diameters were made, varying from 29 +/- 10 microns to 5.6 +/- 1.7 microns. The liposomes were morphologically characterized by light microscopy and transmission electron microscopy. Materials successfully trapped within the liposomes ranged in molecular size from glucose to nucleic acids. Extremely high percentages of encapsulation (up to 89%) were achieved.

Chemical Phenomena↗
PubMed1983

Mouse teratocarcinoma cells resistant to aphidicolin and arabinofuranosyl cytosine: isolation and initial characterization.

With the long-range goals of elucidating the biochemical genetics and the phenotypic expression, both in vitro and in vivo, of mutator strains of mammalian cells, we have isolated and partially characterized a series of drug-resistant mutants from a subline a feeder-dependent parental mouse teratocarcinoma line (PSA-1) known to be capable of chimerizing host blastocysts via the injection of inner cell mass. Two series of stable mutants were isolated--one (Aphr) selected on the basis of resistance to aphidicolin (Aph), a specific inhibitor of DNA polymerase-alpha, and the other (AraCr) selected by resistance to arabinofuranosyl cytosine, an analogue of cytosine. Irrespective of the method of selection, most of the 32 mutants isolated were resistant to both agents, although to different degrees, and with variations in growth characteristics, deoxynucleoside sensitivities, and sensitivities to mutagens (5-bromodeoxyuridine, N-methyl-N'-nitro-N-nitrosoguanidine and ultraviolet light). A protocol of multi-step selection provided stable mutants highly resistant to aphidicolin and only mildly resistant to AraC, even in the absence of prior mutagenesis, and is thus recommended as an approach to the isolation of candidate mutator strains of multipotent teratocarcinomas.

Animals↗
PubMed1983

Effect of pairings of pentobarbital with lithium chloride on the capacity of pentobarbital to maintain a conditioned aversion.

Rats with conditioned aversions to NaCl water were exposed to either an injection of pentobarbital (Pent) followed 30 min later by an injection of lithium chloride (LiCl) on four separate occasions or to injections of LiCl alone, Pent alone, or LiCl followed by Pent. These injections were followed by pairings of NaCl consumption with injections of Pent. The Pent leads to LiCl pairings eliminated the capacity of Pent to maintain the animals' conditioned aversion to NaCl water relative to the other groups. These findings are consistent with the idea that Pent leads to LiCl pairings cause the Pent state to elicit a compensatory response. This compensatory response seems to eliminate the properties of Pent which normally produce or maintain flavor aversions.

Animals↗
PubMed1983

Mammalian mutator mutant with an aphidicolin-resistant DNA polymerase alpha.

The Chinese hamster V79 cell mutant aphr-4-2, selected for its resistance to aphidicolin, a specific inhibitor of DNA polymerase alpha (DNA nucleotidyltransferase, EC 2.7.7.7), is characterized by slow growth, UV sensitivity, and hypersensitivity to UV-induced mutation. DNA polymerase alpha has been purified from mitochondria-free crude extracts of the mutant and its parental wild-type cells by sequential column chromatography on DEAE-cellulose and phosphocellulose. The major DNA polymerase activity from both cell lines was found to have characteristics of the alpha-type polymerase: sensitivity to 0.2 M KCl, resistance to heat denaturation (45 degrees C for 15 min), an apparent Km of 5 microM for dATP, and an ability to copy poly(dT)X(rA)10 but not poly(rA)X(dT)12. The crude extracts and purified DNA polymerase alpha from the mutant cells are not inhibited by aphidicolin (greater than 0.6 microM). The apparent Km for dCTP with DNA polymerase alpha is 1.0 +/- 0.4 microM (mean +/- SD) for the mutant enzyme. The polymerase from the parental cells, similarly purified, is sensitive to aphidicolin and has an apparent Km for dCTP of 10 +/- 4 microM. The spontaneous mutation rate (per cell per division), determined by fluctuation analysis at the Na+/K+-ATPase (EC 3.6.1.8) locus, is higher for mutant cells (42-73 x 10(-8)) than for parental cells (3-16 x 10(-8)). These data suggest a mechanism for aphidicolin resistance of the mutant--i.e., a decrease in the Km for dCTP. The results also indicate that an altered DNA polymerase alpha may be intrinsically mutagenic during normal semiconservative replicative as well as during UV-induced repair syntheses.

Animals↗