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PubMed1994

Genetic control of the antibody repertoire against excretory/secretory products and acetylcholinesterases of Dictyocaulus viviparus.

Outbred Dunkin-Hartley and inbred strain 2 and strain 13 guinea pigs were immunized with Dictyocaulus viviparus adult ES products prior to challenge with third stage larvae. Antibody responses of the three strains to adult ES products and the acetylcholinesterase (AChE) isoforms which they contain were examined. Using immunoprecipitation and ELISA, it was observed that responses in the three strains to adult ES products were distinct: considerable heterogeneity in the antibody repertoire was observed between outbred Dunkin-Hartley animals, with only slight variation occurring amongst the inbred individuals. Responses to the AChE isoforms were heterogeneous amongst individual outbred guinea pigs but were more consistent in inbred strain 2 and 13 animals in which strain-specific patterns of recognition were observed. Previous studies with nematode infections have indicated a role for the major histocompatibility complex in determining the nature and level of the immune response. As the inbred strains bear different alleles at the Class II region but are identical at the Class I region, the differences observed are likely to be due to genes mapping to the Class II locus. This is therefore the first report of genetic restriction of the antibody repertoire to secreted AChEs of a parasitic nematode.

Acetylcholinesterase↗

PubMed1999

Direct and correlated responses to selection for efficiency of lean gain in mice.

Improvement in feed efficiency when selection is based on gain:feed ratio has often been accompanied by a reduction in feed intake. The following four criteria were used in mass selection for improved lean gain efficiency in mice with an objective of evaluating changes in lean gain and intake: 1) gain deviation, animals selected had the greatest gain in fat-free mass (FFM) after adjustment to a constant intake; 2) intake deviation, mice selected had the least feed intake after adjustment to a constant gain in FFM; 3) intrinsic efficiency, similar to the second criterion except that adjustment was also made for average weight maintained during the period; and 4) a positive control that used the ratio of gain in FFM: feed intake as the selection criterion. A fifth line, in which a male and a female were selected at random from each litter, served as a negative control. Experimental animals were outbred mice of the CF1 strain. Two replicates of the five lines were included in the study. Twelve males and females were pair-mated within each line-replicate combination each generation. Feed disappearance was measured from 25 to 42 d. Mice were scanned to obtain an electrical conductivity measurement for prediction of FFM. After six generations of selection, realized heritabilities for gain:feed, gain deviation, intake deviation, and intrinsic efficiency were .00 +/- .04, .04 +/- .29, .35 +/- .08, and .28 +/- .06, respectively. There were no differences among lines for gain:feed ratio. The correlated response in feed intake reduction was significant in the intake deviation and intrinsic efficiency lines (-.17 +/- .05 and -.21 +/- .04 g x d(-1) x generation(-1), respectively). The realized genetic correlations between the ratio and gain deviation, intake deviation, and intrinsic efficiency were .83 +/- .15, .01 +/- .04, and .21 +/- .12, respectively. Litter size was depressed in all selected lines.

Animals↗

PubMed2005

Microsuction of blastocoelic fluid before vitrification increased survival and pregnancy of mouse expanded blastocysts, but pretreatment with the cytoskeletal stabilizer did not increase blastocyst survival.

OBJECTIVE: To ascertain effects of thermal change, size of blastocoele, artificial shrinkage, and cytoskeletal stabilizer on survival of blastocysts from vitrification. DESIGN: In vitro and in vivo study. SETTING: University infertility clinic and academic research laboratory. ANIMAL(S): Female mice of outbred ICR strain, aged 6 to 8 weeks. INTERVENTION(S): In experiment 1, various stages of mouse blastocysts were vitrified by using conventional straws or closed pulled straws (CPS). In experiment 2, microsuction was performed of blastocoelic fluid for blastocysts and expanded blastocysts before vitrification. In experiment 3, cytochalasin B (CCB) was used to treat embryos during vitrification. In experiment 4, vitrified expanded blastocysts were transferred to pseudopregnant mice. MAIN OUTCOME MEASURE(S): Survival and pregnancy. RESULT(S): The survival rates of early blastocysts were high and not different between the conventional straws and the CPS. The survival rates decreased for blastocysts and expanded blastocysts in both of the two methods. But the use of CPS achieved higher survival rates for blastocysts (83% vs. 70%) and expanded blastocysts (60% vs. 39%) than did the conventional straws. Microsuction before vitrification increased the survival rates for blastocysts (92% vs. 80%) and expanded blastocysts (89% vs. 59%). Survival of vitrified embryos was not distinct between CCB treatment and non-CCB treatment. The percentage of live young from vitrified expanded blastocysts using microsuction was greater than that from vitrified expanded blastocysts without microsuction (34% vs. 9%). CONCLUSION(S): The size of the blastocoeles influenced survival of blastocysts from vitrification. A rapid thermal change of CPS and effective reduction of blastocoelic fluid by microsuction may facilitate vitrification and reduce ice crystal damage for blastocysts and expanded blastocysts. The microfilament inhibitor of CCB treatment did not increase their survival rates.

Animals↗

PubMed1993

Variations in disease in inbred and outbred strains of rodents.

Risk assessment from toxicity testing is difficult because of interanimal variability and high levels of spontaneous pathology in rodents. Genetically defined animals might improve risk assessment by reducing the effects of these two factors. Data is presented on disease patterns in mice from an outbred colony of Swiss albino Alpk:APfCD-1 mice, and inbred mouse strain C57Bl/10JfCD-1/Alpk, and an outbred rat colony Alpk:APfSD (Wistar derived). All of the strains have been bred, maintained and used continuously, in one laboratory, for 20 to 30 years. The data is derived from control animals in toxicity tests completed between 1960 and 1990, chiefly from oncogenicity studies. The results indicate that the major diseases, particularly the most common tumours in each strain, liver tumours in the outbred mouse, lymphomas in the inbred strain, and pituitary adenomas in the outbred rat, increase in incidence with time. Renal disease also increases in the outbred rat. This increase has been shown to be due to overnutrition. The effects of overnutrition can override the genetic characteristics of strain, and risk assessment is, therefore, unlikely to be improved by the use of genetically defined animals alone. The environmental conditions of toxicity experiments must also be defined and controlled.

Animals↗

PubMed2004

Mouse model characterisation for anthrax vaccine development: comparison of one inbred and one outbred mouse strain.

In order to evaluate the immunogenicity and protective efficacy of anthrax vaccine candidates a suitable small animal model is required. The inbred A/J strain of mouse has been selected as a potential model, and its immune response to immunisation with recombinant protective antigen (rPA) vaccine characterised, by assessment of rPA specific antibody production, and protection against injected challenge, with the unencapsulated STI strain of Bacillus anthracis. Studies were conducted to determine the time required post immunisation to develop a protective immune response, to define the minimum protective dose of vaccine required and to assess the long-term immune response to immunisation. From the results of these studies it was possible to establish that the A/J mouse is a consistent and robust small animal model for rPA vaccine testing. A comparison of the immune response to rPA vaccine immunisation in the Turner Outbred (TO) mouse strain was also conducted. Both inbred and outbred mouse strains displayed a predominantly Th2 biased immune response and showed a comparable antibody response to rPA immunisation. An assessment of protection in the TO mouse against aerosol challenge with the fully virulent strain of B. anthracis, Ames, was also made.

Animals↗

PubMed1998

Strain differences in adrenal microsomal steroid metabolism in guinea pigs.

We recently reported that CYP2D16, a xenobiotic-metabolizing P450 isozyme, was expressed at higher levels in adrenal microsomes from inbred Strain 13 guinea pigs than in those from outbred English Short Hair (ESH) animals. Studies were done to determine if there also were strain differences in adrenal microsomal steroid metabolism. In both inner (zona reticularis) and outer (zona fasciculata plus zona glomerulosa) zone preparations of the adrenal cortex, 21-hydroxylase activities were greater in microsomes from ESH than from Strain 13 guinea pigs. By contrast, 17alpha-hydroxylase activities were similar in the two strains. In both strains, 21-hydroxylase activities were greater in inner than outer zone microsomes, but the opposite was found for 17alpha-hydroxylase activities (outer>inner). Northern and Western analyses revealed higher levels of CYP21 mRNA and protein in adrenals from ESH than Strain 13 guinea pigs, but there were no strain differences in CYP17 mRNA or protein concentrations. Despite the zonal differences in adrenal 17alpha-hydroxylase and 21-hydroxylase activities, CYP17 and CYP21 mRNA and protein levels were similar in the inner and outer zones within each strain of guinea pig. The results demonstrate strain differences in microsomal steroid metabolism that are explained by differences in CYP21 expression. By contrast, the zonal differences in steroid hydroxylase activities may be attributable to post-translational mechanisms.

Adrenal Glands↗

PubMed1983

In vivo assessment of 99mTc-labeled hematoporphyrin derivative in tumor-bearing animals.

The efficacy of 99mTc-labeled hematoporphyrin derivative in localizing neoplasms has been investigated with tumor-bearing animal models. Spontaneous mammary adenocarcinomas of outbred CFW strain Swiss-Webster white mice and chemical carcinogen induced breast tumors in female Sprague-Dawley white rats were clearly visualized in the scintigrams. Mice tissue distribution data demonstrate favourable tumor to organ ratios sufficiently high to permit tumor detection. [99mTc]HPD appears promising as a tumor imaging agent.

9,10-Dimethyl-1,2-benzanthracene↗

PubMed2004

Factors influencing therapeutic efficacy and the host immune response to helper-dependent adenoviral gene therapy in hemophilia A mice.

BACKGROUND: Adenoviral-based methods of gene therapy have been ineffective at providing sustained factor (F)VIII expression in outbred populations of large animal hemophilic models primarily due to the immunogenicity of these vectors. Improvements have been made in vector design leading to the development of the helper-dependent adenoviral (HD) system. Unfortunately, it remains unclear whether these modifications are sufficient to circumvent the induction of inhibitor formation associated with adenoviral gene transfer. OBJECTIVE: To develop an HD vector capable of mediating sustained FVIII expression and to determine the variables that influence inhibitor development. METHODS: HD vectors were constructed encoding the canine FVIII B-domain deleted transgene under the control of either the cytomegalovirus (CMV) promoter or a tissue-restricted hybrid element consisting of five HNF-1 binding sites, located upstream of the human FVIII proximal promoter. Inbred and outbred populations of hemophilic mice were treated, and monitored for vector-induced toxicity, therapeutic efficacy, and inhibitor formation. RESULTS: When HD vectors utilizing the CMV promoter were administered, all hemophilic mice developed high levels of FVIII inhibitors. In contrast, vectors under the control of the HNF/FVIII element were capable of achieving sustained elevations of FVIII for over 6 months. Strain-specific differences were also observed, with outbred animals showing a greater propensity towards inhibitor development in response to treatment. CONCLUSIONS: HD vectors can be used to provide long-term FVIII expression in hemophilic animals, but treatment outcome and the induction of inhibitors is dependent on a number of variables including the transgene promoter, the vector dose, and the genetic background of the host.

Adenoviridae↗

PubMed1990

Failure to induce resistance of Schistosoma japonicum to praziquantel.

In order to explore the possible occurrence of inducing resistance of Schistosoma japonicum to praziquantel (PZQ), a set of animal experiments were carried out. Outbred mice (NIH strain), Anhui isolates of S. japonicum and Oncomelania hupensis were used. In one protocol five weeks after being infected with 48-52 cercariae, mice were orally dosed with PZQ 300 mg/kg, and killed 82 days later to isolate eggs from the liver. Snails were exposed to miracidia released from egg-hatching. F1 progeny were thus obtained through cercarial inoculation. The same scheme was applied for the establishment of the F2 generation. In another protocol two weeks after infection, PZQ 50 mg/kg/day was given to mice for 5 days. Eggs were collected 26-27 days post treatment and the identical procedures were adopted for F1 and F2 generations successively. Analysis of total worm and female worm reduction rates indicated that there was no significant difference between the sensitivity to PZQ of F1 and F2 progenies of S. japonicum and the parent worms.

Animals↗

PubMed1982

Genetics of natural resistance to thermal injury.

The possible influence of genetic factors in conditioning the host's natural resistance to the lethal effects of severe thermal injury has been studied in 908 rats of comparable age and weight, originating from two outbred, eight inbred, and two congenic strains of animals of defined genetic background. Each animal was exposed to a standard, full-thickness, 40% body surface area skin burn by controlled contact with a heated metal plate. The 21-day postburn mortality was 100% in 217 Fisher (F-344) and 97 ACI male and female rats. The mortality was reduced to 49-63% in an intermediate group of 84 Lewis, 98 Wistar, 48 Sprague-Dawley, 96 Wistar-Furth (WF), and 48 Osborne-Mendel (OM) male rats; 48 female OM rats had a mortality of 86% at 21 days after injury. The same injury produced a mortality in 4% in 90 Buffalo (BUF) and 22% in 41 Brown-Norwegian (BN) males, while females of the same strains exhibited a 21-day mortality of 23% and 54%, respectively. Further studies of the effects of similar injury in two congenic strains of rats derived from some of the inbred lines of animals listed above yielded a 21-day mortality of 50% in 18 BN.1B(BUF) and 20% in 15 BN.1U(WF) male rats, and 65% and 36%, respectively, in females of the same lines. These data point to the importance of genetic factors as a key determinant of host resistance or susceptibility to the effects of severe thermal injury. The segregation of responses to thermal injury in inbred rats into susceptible, intermediate, and resistant groups on the basis of strain origin indicates that such natural resistance may be a quantitative trait. One of the genetic components affecting host resistance is sex-linked. The existence of genetically controlled variations in natural resistance to trauma may be an important determinant of survival and may be a source of guidelines for the triage and clinical care of injured patients. It may also be an important selective factor in evolution.

Animals↗

PubMed1983

Morphologic and phenotypic analysis of an outcross line of blotchy mouse.

Blotchy is an X-linked recessive mutation at the "Mottled" locus in the mouse. The affected blotchy male (Blo/Y) mouse from an inbred genetic background demonstrates morphologic and physiologic abnormalities consistent with emphysema in adult life. Breeding of Blo/Y mice has been difficult because the inbred Blo/Y males are sterile. We report the successful development of a line of outbred Blo/Y male and Blo/Blo female nice by the controlled outcross mating of the inbred heterozygous Blo/+ female with the Argonne hybrid B6CF1 male mouse. The subsequent outcross Blo/Y progeny breed vigorously with the outcrossed Blo/+ female. The lungs of the outbred Blo/Blo female and inbred Blo/Y male mice demonstrate mild to moderate panacinar emphysema with a significant decrease in internal surface area (p less than 0.005) and an increase in mean linear intercept (p less than 0.005). In contrast, the lungs of the outbred Blo/Y is structurally normal. Despite the absence of emphysema-like changes in the outbred Blo/Y males, there were phenotypic features that suggest inherited abnormalities in connective tissue proteins including 1) high incidence of aortitis leading to premature death from aneurysmal rupture, and 2) significant decrease in the morphometrically determined parenchymal elastic fiber length in the lung (p less than 0.01). The outbred blotchy strain may be a useful experimental animal model in determining the pathogenesis of emphysema.

Animals↗

PubMed1976

An IgG thymolytic autoantibody in rats which has specificity for a subpopulation of T cells.

A cytotoxic anti-thymocyte IgG auto-antibody is present in Lewis rats which, in the presence of autologous complement, destroys (in vitro) 12-28 per cent of isologous or autologous thymocytes, a smaller number of lymph node cells and splenocytes, but not bone marrow or circulating lymphocytes. The labile cells in the thymus represent a finite subpopulation which is autologous antithymocyte antibody (ATS) sensitive and steroid resistant. The presence of the autoantibody is randomly distributed in outbred animals whereas in inbred Lewis rats, a strain in which the induction of some autoimmune reactions is under genetic control, the antibody is always present. In this strain, the susceptible T cells and the quantity of circulating autoantibody is significantly depressed during the productive phase of a T-cell mediated disease (adjuvant polyarthritis) and returns to normal after the disease becomes stabilized. There is a direct relationship between the amount of susceptible cells in the thymus and the amount of antibody in circulation, suggesting that the antibody could serve as a marker for a specific subpopulation of thymocytes which may have a regulatory influence on T-cell reactivity.

Animals↗

PubMedDate not supplied

Increased mRNA for corticotrophin releasing hormone in the amygdala of fawn-hooded rats: a potential animal model of anxiety.

Compared to the outbred Wistar rat strain, the Fawn-hooded rat strain has several characteristics which suggest that the Fawn-hooded strain is hyperaroused. Fawn-hooded rats exhibit more freezing behavior in response to stress, have an increased preference for alcohol, develop adult onset hypertension, and have elevated urinary catecholamine levels. We used quantitative in situ hybridization to investigate central components of the corticotropin releasing hormone (CRH), arginine vasopressin (AVP) and noradrenergic stress response and arousal systems in these rats. We also measured basal corticosterone levels and adrenal weights to assess tonic hypothalamic-pituitary-adrenal axis activity. Compared to Wistar rats, Fawn-hooded rats had significantly increased CRH mRNA in the central nucleus of the amygdala and reduced CRH mRNA in the paraventricular nucleus of the hypothalamus. Fawn-hooded rats also bad reduced AVP mRNA expression in the parvocellular cells of the hypothalamic paraventricular nucleus. There were no differences between strains in glucocorticoid receptor mRNA in the hippocampus or the paraventricular nucleus or in mineralocorticoid receptor mRNA in the hippocampus. There was also no difference between strains in tyrosine hydroxylase mRNA in the locus ceruleus. Finally, adrenal weights were significantly reduced in the Fawn-hooded rats while basal corticosterone levels were similar in the two strains, which suggests central hypoactivity of the hypothalamic-pituitary-adrenal axis in Fawn-hooded rats compared to Wistar rats. Increased CRH mRNA in the central nucleus of the amygdala and reduced tonic hypothalamic-pituitary-adrenal axis activity may play a role in the unique behavioral and physiological characteristics of Fawn-hooded rats.

Amygdala↗

PubMed2002

The effects of chronic mild stress on male Sprague-Dawley and Long Evans rats: I. Biochemical and physiological analyses.

The chronic unpredictable mild stress (CMS) is a paradigm developed in animals to model the relatively minor and unanticipated irritants that lead to a state of anhedonia in some individuals. However, the effectiveness of CMS is sometimes difficult to establish, for which unique strain sensitivities has been attributed as one contributing factor. These considerations led us to design the present study, which was an investigation of the corticosterone response to CMS in two outbred rat strains--Sprague-Dawley and Long Evans. Animals were exposed to one of two conditions--control or CMS--for 3 weeks during which body weight and fecal count were regularly monitored. At the end of this period, blood was sampled at a variety of time intervals following induction of a brief restraint stressor. First, a significant effect of CMS on corticosterone levels was evident at time 0 (prior to the application of the acute restraint stressor) in both strains. Second, the typical quadratic pattern of stressor-elicited fluctuations in this measure was similar in both Sprague-Dawley and Long Evans rats, with consistently elevated levels for the first hour following exposure to the acute stressor; near baseline values were observed at 2 h. However, only in the Long Evans strain were CMS related values much less than that observed in the control group after restraint stress. Third, both strains showed a reduced weight gain in the CMS groups relative to control groups. Fourth, spleen and adrenal weights were similar across all groups. Fifth, fecal counts remained stable across weeks of treatment in all groups with the exception of the Long Evans rats exposed to CMS; in this group, average counts were systematically reduced over the treatment period. We conclude that a history of chronic stress significantly blunts corticosterone levels in Long Evans but not Sprague-Dawley rats following exposure to an acute stressor. Physiological indices however are less influenced by this experience, at least when the exposure is limited to 3 weeks.

Adrenal Glands↗

PubMed1991

Differences in the duration of sedative and anxiolytic effects of desmethyldiazepam in two outbred Wistar strains.

Different sensitivities to benzodiazepines have been described for various strains of both rats and mice suggesting that variations in biological features of the animals are responsible for these differences. Since all reports concern inbred strains, we studied two outbred Wistar strains which are used routinely in several research disciplines. The pharmacodynamics of desmethyldiazepam (DMD), the main active metabolite of diazepam in man, were compared for male rats of the Riv:TOX strain (from the National Institute of Public Health and Environmental Protection) and the Crl:(WI)BR strain. The duration of sedative action of DMD after oral administration, as derived from suppression of the nocturnal locomotor activity, was longer in the Riv:TOX strain than in the Crl:(WI)BR strain. Accordingly, suppression of novelty-induced corticosterone release as an index of anxiolytic action was observed 11 hours after DMD administration in Riv:TOX rats but not in Crl:(WI)BR rats. At that time, serum DMD concentration was shown to be higher in the Riv:TOX strain than in the Crl:(WI)BR strain. The data are discussed in relation with possible metabolic differences between the two strains.

Animals↗

PubMed1986

The case for isogenic strains in toxicological screening.

The fundamental principle of the controlled experiment is that treated and control groups should be identical, with minimal within-group variability. Toxicologists recognise this and control age, body weight, disease and the physical environment of the test animals. However most toxicological screening in done with genetically variable outbred stocks, even though isogenic inbred strains, F1 hybrids or identical siblings are usually available. The result is poor experiments with the inevitable genetic differences between groups resulting in increased false positive and negative results, and no indication that the response is under genetic control. It is also illogical to treat genetic variation differently from other types of variation. The argument that it is essential to use outbred animals to model outbred man is illogical. If bacteria can be used to model man (as in the Ames test), so can inbred animals. The uncontrolled variation present in an outbred stock can not be used efficiently to increase the range of phenotypes tested because it also introduces "noise" which obscures experimental effects. The use of two or more isogenic strains gives a much more efficient experimental design with low "noise" and an indication of whether the response is under genetic control. Inbred and F1 hybrid strains (but not identical siblings) have the added advantage of an immortal genotype which outlives any individual animal. Such immortal genotypes may be studied in detail to gather background information. Toxicologists should treat genetics like every other variable and control it, using several isogenic strains in cases where testing needs to be done on more than one genotype.

Animals↗

PubMed1991

Cytochrome P-450 isozyme pattern is related to individual susceptibility to diethylnitrosamine-induced liver cancer in rats.

Differences in susceptibility to chemical carcinogenesis between rodent strains and species have been linked to variations in genetically-determined mixed function oxidase activities. In order to verify whether such variations also determine the susceptibility of individual animals of the same strain to a chemical carcinogen, outbred male Wistar rats were administered diethylnitrosamine (DEN) (1, 2, or 3 mg/kg) five times a week for 20 weeks. The relationship was examined between the outcome (i.e., presence or absence of liver tumors, and latency period) and the hepatic activities of mixed function oxidases and conjugating enzymes, as well as of O6-methylguanine-DNA-methyltransferase, measured before the carcinogen treatment. In addition, the metabolic profiles of two model drugs, antipyrine and disopyramide, in the urine were analyzed and correlated with the carcinogen susceptibility. The length of the latency period of hepatocellular tumors in individual rats was negatively related to the activities of hepatic dimethylnitrosamine N-demethylase, aryl hydrocarbon hydroxylase and epoxide hydrolase and positively related to the amount of microsomal protein. Consistent relationships between the other 10 measured parameters and the susceptibility to DEN-induced carcinogenesis were not detected. Long-term treatment with DEN slightly decreased the proportion of metabolism of antipyrine into norantipyrine, and increased the share of 4-hydroxyantipyrine; a decrease in the metabolism of disopyramide to N-deisopropyldisopyramide was also detected. It is concluded that the pattern of cytochrome P-450 isoenzymes is related to differences in individual susceptibility to nitrosamine-induced carcinogenesis. The relationship was most marked at low dose levels, which are the levels at which nitrosamine exposures of humans are known to occur.

7-Alkoxycoumarin O-Dealkylase↗

PubMed1997

Differential lateral septal vasopressin in wild-type rats: correlation with aggression.

The vasopressin (VP)-containing projections from the cells of the bed nucleus of the stria terminalis to the lateral septum (LS) are sexually dimorphic and dependent on gonadal steroids. Recently, the difference in VP distribution found among both sexes was also demonstrated in male mice genetically selected for different levels of intermale aggression. In the present study we examined whether this differential VP distribution in males also exists in an outbred strain of wild-type rats. After the animals were tested for their level of aggression, the VP content and the fiber density of the LS were measured using radioimmunoassay and immunocytochemistry, respectively. In addition, basal levels of plasma testosterone (T) were measured. Both biochemical data and immunocytochemical data revealed a negative correlation between VP and intermale aggression. Aggressive rats exhibited low levels of VP whereas intermediate and nonaggressive animals showed higher levels. Differences in adult levels of T were not found. The results are in accordance with the observations previously found in male mice, reconfirming the correlation between lateral septal VP and aggression.

Aggression↗