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PubMed1983

Genetic selection, general characterization, and histology of atherosclerosis-susceptible and -resistant Japanese quail.

A new animal model of atherosclerosis has been developed through genetic selection of Japanese quail (Coturnix coturnix japonica) into susceptible (SUS) and resistant (RES) lines. Characterization of the selected quail has shown that the RES birds were resistant to the disease and developed little atherosclerosis on a diet containing 1% cholesterol. The SUS birds were sensitive and developed severe atherosclerosis in 8-9 wks on a diet containing only 0.5% cholesterol. The histology of the progression of atherosclerosis in the SUS quail was studied. It bore a morphological similarity to that of human arterial atherosclerosis. The atherosclerotic plaque was characterized by intimal thickening, the presence of foam cells, the proliferation of smooth muscle cells and/or fibroblasts, and the formation of scar with collagen deposition. We believe that these two lines of quail may serve as a valid animal model for the study of the genetic and biochemical basis of cholesterol-induced atherosclerosis.

Animals↗
PubMed1993

A genetic selection elucidates structural determinants of arginine versus lysine specificity in trypsin.

A genetic selection has been used to isolate variants of the serine protease, trypsin (Tsn), altered in specificity toward lysine- and arginine-containing substrates. Growth of a lysine auxotroph of Escherichia coli was coupled to activation by Tsn of a non-nutritive source of lysine present in selective media. Nine Tsn variants possessing partial activities were isolated from a random library encompassing amino acids 189 and 190 at the base of the primary specificity pocket. Functional analysis of these isolates indicates that preservation of activity toward lysine-containing substrates is more tolerant to mutation than is activity toward equivalent arginine-containing substrates. Both the position, as well as the accessibility to substrate, of a negatively charged group in the binding pocket appear critical to maintenance of high-level catalytic potency by Tsn.

Arginine↗
PubMed1988

Maize glutamine synthetase cDNAs: isolation by direct genetic selection in Escherichia coli.

Maize glutamine synthetase cDNA clones were isolated by genetic selection for functional rescue of an Escherichia coli delta glnA mutant growing on medium lacking glutamine. The Black Mexican Sweet cDNA library used in this study was constructed in pUC13 such that cDNA sense strands were transcribed under the control of the lac promoter. E. coli delta glnA cells were transformed with cDNA library plasmid DNA, grown briefly in rich medium to allow phenotypic expression of the cDNAs and the pUC13 ampr gene, and challenged to grow on agar medium lacking glutamine. Large numbers of glutamine synthetase cDNA clones have been identified in individual 150-mm Petri dishes; all characterized cDNA clones carry complete coding sequences. Two cDNAs identical except for different 5' and 3' termini have been sequenced. The major open reading frame predicts a protein with an amino acid sequence that exhibits striking similarity to the amino acid sequences of the predicted products of previously sequenced eukaryotic glutamine synthetase cDNAs and genes. In addition, the maize glutamine synthetase cDNAs were shown to contain a 5' mini-ORF of 29 codons separated by 37 nucleotide pairs from the major ORF. This mini-ORF was shown not to be essential for the functional rescue of the E. coli delta glnA mutant. Expression of the cDNAs in E. coli is presumed to be due to the function of a polycistronic hybrid lac messenger RNA or translational fusions encoded by the pUC plasmids. Proteins of the expected sizes encoded by two different pUC clones were shown to react with antibodies to tobacco glutamine synthetase.

Amino Acid Sequence↗
PubMed1985

Regulatory function of Thy-1-negative cells: V. A lymphokine of B cell origin (BEF) induces in vitro high antibody response in genetically selected low responder mice.

Studies were conducted on two lines of mice genetically selected, respectively, for high (AB/H) and low (AB/L) antibody production, in order to identify the mechanism by which genes involved in the selection express their functions. It was found that B cell-derived enhancing factor (BEF), a lymphokine of B cell origin which acts on T cells by preventing the activation of suppressor cells, is effective in inducing high responses in low responder mice, whereas it is ineffective in modulating antibody production in high responder mice. As a result, no difference was found between the responses of AB/H and AB/L mice when spleen cells were stimulated in vitro in the presence of BEF. AB/H and AB/L mice do not seem to differ in their B cell functions since no difference was found in the capacity of B cells of either line to synthesize antibodies in the presence of T cell-replacing factor (TRF), or to produce endogenous BEF. These data indicate that, at least in vitro, the character of general responsiveness of these two mouse lines mainly reflects differential reactivity of T suppressor cells. Since the two lines represent, respectively, the maximal and minimal responsiveness of an outbred population of animals, it is suggested that the individual difference in antibody response is related to individual differences of T suppressor cells in responding to the immunogen.

Animals↗
PubMed1996

Behavioral stress response of genetically selected aggressive and nonaggressive wild house mice in the shock-probe/defensive burying test.

Genetically selected aggressive and nonaggressive male wild house mice were tested in the shock-probe/defensive burying test: Five distinct behaviors (burying, immobility, rearing, grooming, and exploration) were recorded in two environmental situations: fresh and home cage sawdust. Nonaggressive animals, characterized by a Long Attack Latency (LAL), showed more immobility in both test situations than animals having Short Attack Latencies (SAL), whereas SAL males displayed more defensive burying than LAL ones when tested with fresh sawdust. Testing with home cage sawdust, however, resulted in the same duration of defensive burying in SAL and LAL. These results support earlier findings about the existence of two heritable, fundamentally different strategies to cope with aversive situations. Aggressive (SAL) animal react actively to environmental challenges, whereas nonaggressive animals react actively or passively, depending on the characteristics of the stressful environment. These mouse lines, selected for attack latency, i.e., aggression, may, therefore, be important tools to unravel the genetic architecture underlying the physiological and neuronal mechanisms of behavioral strategies towards stressful events.

Aggression↗
PubMed1997

Diet-induced thermogenesis in cockerels is modulated by genetic selection for high or low residual feed intake.

Energy balance of adult cockerels genetically selected for high (R+) or low (R-) residual feed consumption was investigated by using indirect calorimetry. Although no between-line difference was found in digestion of ingested energy, the true metabolizable energy (ME) intake was 40% greater in R+ than in R- birds. Basal heat production (HP) did not significantly differ between genotypes. Diet-induced thermogenesis (DIT) was significantly enhanced in R+ compared with R- birds, i.e., +84% when expressed as the difference in kilojoules of heat production determined in feed-deprived and fed cockerels, +31% when calculated as a percentage of ME intake (P < 0.01). The difference in DIT calculated from the regression between HP and physical activity explained 75% of the difference in HP; the remaining 25% could be explained by activity-related HP. The results cannot be explained by differences in the plasma concentration of circulating thyroid hormones: plasma thyroxine concentration did not differ between genotypes, whereas plasma triiodothyronine concentration was lower in feed-deprived R+ than in R- birds and indistinguishable in fed birds of the two lines. Heat production, however, was higher in the R+ line. Propranolol decreased HP only in the R+ line, suggesting a beta-adrenergic control of DIT at least in cockerels of this line. Plasma triglyceride concentration was lower in the R+ than in the R- line in fed cockerels, and plasma nonesterified fatty acid concentration was higher in the R+ than in the R- line in feed-deprived cockerels. These results are consistent with the leanness of the R+ compared with the R- line. The R+ and R- lines constitute an original model of diet-induced thermogenesis (DIT), a process that is under genetic control of appetite and allows R+ birds to balance all of their excessive energy intake without any adipose tissue storage.

Animals↗
PubMedDate not supplied

[Production of potato cybrids without using genetic selection markers of the parental material: Solanum tuberosum L. plants (cv. Svitanok Kyivsky) with S. pinnatisectum Dun. plastids].

Interspecific cybrid plants of potato were created using the cell technology that does not require the presence of any genetic selectable markers in the parental material. Cybrid production was based on double inactivation of S. pinnatisectum Dun. protoplasts, served as the donors of organelles (by gamma-irradiation for the nuclei inactivation and by chemical mutagenesis for the efficient induction of chlDNA mutations), and the transfer of mutant plastids into S. tuberosum L. by protoplast fusion. Selection of cell colonies for streptomycin resistance was performed to identify the cybrid clones with mutant donor-type plastids. Restriction analysis of organelle DNA, chromosome counting, and isoenzyme analysis of the cybrids revealed the presence of nuclear material of S. tuberosum L. (cv. Svitanok kyivsky) and plastids from wild tuber-bearing S. pinnatisectum Dun. These plants enable the study of traits encoded by organelle DNA and to broaden the cytoplasmic diversity of cultivated potato.

Alkylating Agents↗
PubMed2005

Rapid detection of subtype-selective nuclear hormone receptor binding with bacterial genetic selection.

Subtype-selective nuclear hormone receptor modulators could potentially allow the development of valuable tissue-specific therapeutics. A simple biosensor that allows subtype-specific nuclear hormone receptor binding to be reflected by the growth phenotype of Escherichia coli cells has been constructed. This system will potentially enable the facile detection or evolution of subtype-selective hormone analogues.

Bacterial Adhesion↗
PubMed1997

A genetic selection for isolating cDNAs encoding secreted proteins.

We describe a simple, rapid technique for simultaneously isolating large numbers of cDNAs encoding secreted proteins. The technique makes use of a facile genetic selection performed in a strain of Saccharomyces cerevisiae deleted for its endogenous invertase gene. A cDNA cloning vector which carries a modified invertase gene lacking its leader sequence is used in conjunction with this strain. Heterologous secreted genes fused appropriately upstream of this defective invertase provide the necessary signals to restore secretion, allowing the yeast to grow on sugars such as sucrose or raffinose. This microbial growth selection facilitates scanning cDNA libraries containing millions of clones, enabling the wholesale identification of novel secreted proteins without the need for specific bioassays. The technique is similar to one previously described (Klein et al. (1996) Proc. Natl. Acad. Sci. USA 93, 7108-7113). We describe results using a cDNA library derived from activated human peripheral blood mononuclear cells (PBMC). Genes identified from this library encoded signal sequences of proteins of diverse structure, function, and cellular location such as cytokines, type 1 and type 2 transmembrane proteins, and proteins found in intracellular organelles. In addition, a number of novel secreted proteins were identified, including a chemokine and a novel G-protein-coupled receptor. Since signal sequences possess features conserved throughout evolution, the procedure can be used to isolate genes encoding secreted proteins from both eukaryotes and prokaryotes.

Amino Acid Sequence↗
PubMed1991

Differential expression of migration inhibitory and migration stimulatory factors in two lines of mice genetically selected for high or low responsiveness to phytohemagglutinin. 1. Migration stimulatory factor(s) from T and B cells of immune spleen.

Expression of the lymphokine migration inhibition factor in two lines of mice genetically selected for the high (Hi/PHA) or low (Lo/PHA) response of their lymph node cells to phytohemagglutinin was found to be modulated by concomitant expression of migration stimulation factor(s) [MStF(s)]. The expression of both lymphokines was dependent on genetic character and the immunizing dose of antigen. In mice immunized 5 days earlier with 50 micrograms ovalbumin in Freund's complete adjuvant (Ova in FCA immune), migration inhibition factor, assessed with a sensitive photoelectric method, was well expressed by male spleen or lymph node 24-hour culture supernatants of Lo/PHA but Hi/PHA, especially female, expressed marked MStF(s) instead. Immunization with 500 micrograms Ova in FCA markedly enhanced expression of MStF(s) in Lo/PHA but inhibited it in Hi/PHA. MStF(s) of Ova in FCA immune spleens of the two lines were found to derive from both T and B cells, B cell activity being greater. Lo/PHA were by far better expressors of both T- and B-cell-derived MStF(s) as compared to Hi/PHA (p less than 0.01). Spleen cells of mice immunized with FCA alone also expressed MStF(s) but to lesser extent than Ova in FCA immune spleens, expression by Lo/PHA B cells being significantly higher than in Hi/PHA (p less than 0.05). The MStF(s) of Ova in FCA immune spleens was found to be non-immunoglobulin in nature.

Animals↗
PubMed1985

Diurnal, premating, and postmating testosterone levels in roosters genetically selected for mating frequency.

Circulating testosterone (T) levels were measured in roosters from lines genetically selected for high (HM) and low (LM) mating frequency and the randombred control (AC) base population. Lights were provided from 0600 through 2000 hr. A dirunal profile was determined by sampling at 0600, 1200, 1800, and 2400 hr. Plasma T levels prior to mating, immediately after mating, and 1 and 3 hr after mating were measured. By 1800 hr, AC males had higher T levels than both HM and LM males, which had levels that were similar to each other. Testosterone was at a minimum at 0600 hr and peaked at 2400 hr. Lines differed significantly both before and after mating with AC males highest, HM males intermediate, and LM males lowest. Control males had higher than baseline values prior to and after mating. Testosterone levels of AC males were correlated with their mating frequencies.

Analysis of Variance↗
PubMed2005

Do mice genetically selected for resistance to oral tolerance provide selective advantage for Schistosoma mansoni infection?

A highly evolved relationship exists between the parasitic flatworm Schistosoma mansoni and its vertebrate hosts that include the use of host immune signals by parasites. The S. mansoni infection was studied in two strains of mice genetically selected, over 18 generations of assortative mating, for extreme phenotypes of susceptibility (TS) and resistance (TR) to immunological tolerance. The objective was to observe whether the different host genetic backgrounds affected the outcome of experimental schistosomiasis. Fecal egg excretion, tissue egg count, worm recovery, and adult worm morphology and morphometry were monitored throughout the period of infection. TR mice presented total fecal egg excretion and thickness of tegument in adult male worms significantly higher than TS mice. Therefore, the comparative analysis of mice with extreme phenotypes of immunological response turns out to be useful in host-parasite relationship studies. Our results suggest that the TR mouse immunological profile provides a more favorable environment for the development of S. mansoni.

Animals↗
PubMed1989

Genetic selection for novelty-induced rearing behavior in mice produces changes in hippocampal mossy fiber distributions.

Previous investigations in mice revealed the existence of a set of genes that influence variations in hippocampal anatomy as well as variations in behavioral responses to novelty. In particular, a positive genetic correlation was found between the size of the intra- and infrapyramidal mossy fiber (iip-MF) projection and rearing frequency in an open-field. On the basis of these findings, we hypothesized that genetic selection for rearing would entail correlated changes in hippocampal morphology. This was tested in the inbred selection lines SRH (selection for rearing: high) and SRL (selection for rearing: low). As expected, the SRH mice appeared to possess iip-MF terminal fields that were larger than those of the SRL mice. Because the behavioral difference between the two lines is most probably caused by a single genetic unit, these animals represent valuable material for molecular-genetic investigations into the mechanisms that control behavioral and neuroanatomical variation.

Animals↗
PubMed2001

Genetic selection of mice for high voluntary wheel running: effect on skeletal muscle glucose uptake.

Effects of genetic selection for high wheel-running activity (17th generation) and access to running wheels on skeletal muscle glucose uptake were studied in mice with the following treatments for 8 wk: 1) access to unlocked wheels; 2) same as 1, but wheels locked 48 h before glucose uptake measurement; or 3) wheels always locked. Selected mice ran more than random-bred (nonselected) mice (8-wk mean +/- SE = 8,243 +/- 711 vs. 3,719 +/- 233 revolutions/day). Body weight was 5-13% lower for selected vs. nonselected groups. Fat pad/body weight was ~40% lower for selected vs. nonselected and unlocked vs. locked groups. Insulin-stimulated glucose uptake and fat pad/body weight were inversely correlated for isolated soleus (r = -0.333; P < 0.005) but not extensor digitorum longus (EDL) or epitrochlearis muscles. Insulin-stimulated glucose uptake was higher in EDL (P < 0.02) for selected vs. nonselected mice. Glucose uptake did not differ by wheel group, and amount of running did not correlate with glucose uptake for any muscle. Wheel running by mice did not enhance subsequent glucose uptake by isolated muscles.

Animals↗
PubMed2003

Quantitative study of the response to genetic selection of the Wistar audiogenic rat strain (WAR).

To study the relationship between genetics and epilepsy, Wistar rats susceptible to audiogenic seizure were selected from the main breeding stock of the Ribeirão Preto School of Medicine at the University of São Paulo, Brazil, and inbred. The criteria for selection were the highest seizure severity index (SI) and shortest latencies of the first running fit (LI). Because behavioral response to sound stimulation (120 dB) at 70 to 78 days of age was very stable, SI and LI were evaluated within this age range. Analysis of 9450 observations in 1575 animals from the 3rd to 17th generations demonstrated significant effects of generation, parity, and litter on SI and of generation and litter on LI. The SI and LI averages were, respectively, 37.13 and 22.82 s in the 3rd generation and 83.06 and 7.84 in the 17th generation. Heritabilities of both characters were estimated, by maximum likelihood, as 0.37 +/- 0.066 and 0.44 +/- 0.059, respectively. Because a significant regression related individual breeding values for both SI and LI to generation number, we concluded that genetic selection has a positive impact on the traits analyzed. Therefore, the Wistar audiogenic rat (WAR) strain appears, as per the 17th to 20th generations of genetic selection to be an audiogenic rat strain suitable for epilepsy studies.

Acoustic Stimulation↗
PubMed2003

A genetic selection method for expression products that induce apoptosis in adherent mammalian cell lines.

The process of apoptosis is carefully controlled in cells, and different cell types display different sensitivities to pro-apoptotic stimuli. The prospect of exploiting such differences for treatment of diseases such as cancer, via novel therapeutic agents, is extremely attractive. Therefore, genetic selections for novel expression products that kill cells may have considerable value. However, such selections are difficult to devise and perform because the selected cells do not grow. We developed a selection scheme designed to enrich for genetic agents that kill cells. The selection is based on detachment of apoptotic cultured mammalian cells from adherent monolayers. We characterized the properties of these detached cells (floating cells), and various aspects of the selection process. This selection method is potentially applicable to many mammalian cell lines.

Animals↗
PubMed1987

Modification of muscle proteins IEF patterns in a fish Lebistes reticulatus, related to diet, immunization and genetic selection.

The effects of some environmental and genetic factors on the isoelectric focusing (IEF) patterns of muscle proteins from a fish Lebistes reticulatus (Cyprinodonts) were investigated. Of two diets applied the restricted one resulted in a loss of bands in the IEF patterns of the treated fishes muscle homogenate. Instead immunization gave an increase in both number and intensity of IEF bands in (4.0-6.0; 7.0-7.9) pH ranges which are typical of immunoglobulins. Lastly IEF patterns from random and assortative mating animals were compared, resulting in loss of IEF-bands in the inbred lines. Results showed that IEF of muscle proteins could be an excellent method to evaluate the response by fish phenotypes to environmental or genetic factors.

Animals↗