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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↗

Interindividual variation in carcinogen metabolism, DNA damage and DNA repair.

A number of laboratory approaches are currently available to measure interindividual variation in carcinogen metabolism, amounts of carcinogen-DNA adducts and DNA repair capacity. Wide interindividual variations have been found among strains of experimental inbred animals and among individuals in outbred populations, including humans. The quantitative relationships, if any, between these parameters and the human cancer risk will be determined by biochemical and molecular epidemiological studies, of high and low cancer risk populations and of cancer-prone families.

Animals↗

Different degree of ileal colonization by segmented, filamentous bacteria in two strains of mice.

Segmented, filamentous bacteria (SFBs) are autochthonous, apathogenic inhabitants of the ileum of various animal species. Outbred Swiss (Cpb:SE) mice have significantly higher degrees of SFB colonization than do inbred BALB/c mice. The present studies were carried out to identify determinants of this strain difference. In a cross-fostering experiment it was shown that SFB colonization of the pups is determined by the strain of the pups themselves rather than by the strain of the nursing dam. Thus, maternal effects may not be involved in SFB colonization. In a cross-infecting experiment using germ-free and SFB-positive animals of the two mouse strains, it was found that ileal SFB colonization is determined by host characteristics rather than by origin of the SFBs. Thus, SFBs that are specific for a given mouse strain may not exist in the two strains of mice. It is concluded that the mouse strain difference in SFB colonization is determined by host characteristics, which probably have a genetic basis.

Animals↗

Rapid genetic analysis of Helicobacter pylori gastric mucosal colonization in suckling mice.

Previously described animal models for Helicobacter pylori infection have been limited by cumbersome host requirements (e.g., germ-free conditions or unusual species) or are applicable to only special subsets of H. pylori strains (e.g., fresh clinical isolates or animal-adapted derivatives). Here, we report that 5- to 6-day-old outbred CD-1 (ICR) suckling mice support 24-h colonization of all H. pylori strains tested (SS1, 26695 SmR-1, 43504 SmR-1, and G27 SmR-1), including lab-passaged strains that cannot be adapted for colonization of adult animals. Total colony-forming units (cfu) recovered from infection with lab-passaged strains did not differ from those with mouse-adapted SS1. We also tested this model's ability to detect colonization defects in strains carrying mutations in known virulence genes by coinfecting with wild-type H. pylori and measuring differential recovery. This competition assay identified colonization defects in several classes of known attenuated mutants, including those defective in acid resistance (ureA), metabolism (frdA), motility (motB), and chemotaxis (cheY). A mutant defective in copA (copper transporting P-type ATPase) is nonattenuated in adult and infant mice. Possibly because of the limited duration of infection, our model did not identify defects in vacuolating cytotoxin (vacA) or gamma-glutamyltranspeptidase (ggt) as attenuating, in contrast to results from other animal models. We also identified a new virulence gene (HP0507) encoding a conserved hypothetical protein, which is important for colonization in our model. The suckling mouse model offers a rapid method to identify colonization defects in any H. pylori strain and may have utility as a new tool for studying immunity to primary infection.

Aging↗

The choice of animal model and reduction.

Careful choice of the animal model is essential, if research is to be conducted efficiently, by using the minimum number of animals in order to provide the maximum amount of information. Inbred strains of rodents provide an excellent way of controlling and investigating genetic variation in characters of interest and in response to experimental treatments. Outbred stocks, in which genetic and non-genetic factors are inextricably mixed, are much less suitable, because random and uncontrolled genetic variation tends to obscure any treatment responses. In some cases, the use of inbred strains has led to major advances in scientific understanding. The specific example given here is in the understanding of host-parasite relationships but, more generally, inbred strains have been of critical importance in research which has resulted in the award of at least 17 Nobel prizes. And yet, despite the extensive literature on the properties and scientific value of inbred strains, many scientists continue to use outbred stocks in the mistaken belief that the use of such animals will, in some mysterious way, make their research more applicable to humans. There is really no evidence that this is so, and there is much evidence that the use of inbred strains has been highly successful in many disciplines.

Animal Use Alternatives↗

Chromosome mapping of Soa, a gene influencing gustatory sensitivity to sucrose octaacetate in mice.

Strain distribution patterns among recombinant inbred strains suggested that a locus influencing taste sensitivity to sucrose octaacetate was on chromosome 6. A location for Soa was established by linkage analysis of behavioral and electrophoretic data from outbred and congenic strains and from test-cross progeny. Haplotyping of 41 outbred CFW-Cr animals with a cDNA probe showed perfect cosegregation of Soa and Prp, a gene for salivary proline-rich proteins. Five of twelve B6.SW-Soaa strains were found to retain Ldr-1, lactate dehydrogenase regulator-1, on chromosome 6 as an allelic passenger from the SWR/J donor strain (source of the Soaa Taster allele). Centimorgan distance was estimated using the ABP/Le linkage-testing strain (non-Taster, Soab) and the SWR/J strain (Taster, Soaa) in a testcross breeding system. The data are consistent with a position for the Soa locus on mouse chromosome 6, 62 cM from the centromere.

Animals↗

Production of monoclonal antibodies defining guinea pig T-cell surface markers and a strain 13 Ia-like antigen: the value of immunohistological screening.

The production and characterization of eight monoclonal antibodies (MAbs) against surface markers of guinea pig T-cells is reported. MAbs CT5 and CT7 define putative pan-T-cell markers. CT5, however, also reacts with the B-cell leukemic line L2C. MAb CT6 is reactive with less than 30% of peripheral T-cells. MAbs CT1, 2, 3, 4, and 8 are reactive with lymphocytes, but not with germinal center B-cells. In addition to the CT's, a MAb (CI-13.1) has been prepared that reacts with an Ia-like antigen on cells of strain 13 and outbred guinea pigs, but not with cells of strain 2 animals. CI-13.1 cross-reacts with human tissue sections: About 30% of the OKIa-positive dendritic cells in the human dermis are recognized by CI-13.1. In the course of production and characterization, various binding assays and an immunohistological method were used for determining the antibody specificity. Immunohistological screening was found to be the most informative method.

Animals↗

Interaction between 6/94 virus, a parainfluenza type 1 strain, and mouse macrophages.

The 6/94 virus, a type 1 parainfluenza virus recovered from multiple sclerosis brain cells after lysolecithin-induced fusion of these cells with African green monkey kidney cells (CV-1), has been found to grow in splenic and peritoneal macrophages obtained from outbred and different strains of inbred mice. Macrophages from C57BL animals were least susceptible to infection, a resistance apparently partially age related. The virus also has been found to replicate in IC21 cells, a line of simian virus 40 virus-transformed mouse macrophages. Viral growth was detected by development of hemadsorption in infected cells, followed by the appearance of infectious virus. The growth of 6/94 virus had different kinetics in mouse macrophages, in the standard continuous cell lines L, 3T3, and CV-1, and in primary mouse kidney and mouse embryo cells. The virus produced in macrophages could be passed in series to other macrophage cultures. In addition, once infected, the cultures continued to produce virus, and permanently infected cell lines were thus obtained. Macrophages from immunized mice with high titers of humoral neutralizing antibodies were found variably able to support virus growth.

Animals↗

Resistance to myocardial ischemia in five rat strains: is there a genetic component of cardioprotection?

There is a need to develop new and more consistent animal models of cardioprotection. Traditionally, outbred dogs, rabbits, and rats have been studied. We determined resistance to ischemia in isolated hearts from inbred strains of rats. Hearts from inbred rats: SS/Mcw (Dahl S, Dahl salt-sensitive), DA/Hsd (Dark Agouti), LEW/Hsd (Lewis), and BN/SsN/Mcw (Brown Norway); and from an outbred rat: Hsd:WIST (Wistar) were subjected to 27 min of global, no-flow ischemia, followed by 3 h of reperfusion. Infarct size in the Brown Norway rat was 2.5 times less than that observed in the Dahl S rat, with the Dark Agouti, Lewis, and Wistar rats intermediate in response. Hearts from Brown Norway rats were also most resistant to ischemia in terms of postischemic enzyme leakage and contractile and vascular function compared with other strains. The average polymorphism rate between strains revealed that such strains were genetically diverse. This study demonstrates strain differences in resistance to myocardial ischemia, suggesting these rats could be used to study a genetic and/or environmental basis for these differences and to provide new animal models for the physiological study of cardioprotection.

Animals↗

Immune response gene control of determinant selection. I. Intramolecular mapping of the immunogenic sites on insulin recognized by guinea pig T and B cells.

T-cell DNA synthesis and T-helper cell function in response to isolated insulin chains and naturally occurring insulin variants was assessed in insulin immune guinea pigs. Two distinct antigenic determinants, recognized by T cells, were defined. One localized in the B chain and the other one constituted by amino acids A8, A9, and A10 of the insulin A-chain loop. Recognition of the B-chain determinant is under the control of Ir genes linked to the strain 13 major histocompatibility complex. This was shown by studying the response to isolated insulin B chain in F1(2 x 13) guinea pigs, as well as serologically defined backcrosses and outbred animals. Insulin recognition through the A-chain loop determinant is specific for strain 2 guinea pigs. These animals recognize this region of the molecule even when displaying different amino acid sequences. The strain differences observed in those antigenic sites eliciting T-cell recognition was not found at an antibody level. No differences could be detected in the ability of the different insulin variants to inhibit the binding of 125I-labeled pork insulin to strain 2 guinea pig anti-pork insulin or to strain 13 guinea pig anti-pork insulin.

Amino Acid Sequence↗

Attempts to establish experimental Cyclospora cayetanensis infection in laboratory animals.

Attempts were made to develop an animal model for Cyclospora cayetanensis to identify a practical laboratory host for studying human cyclosporiasis. Oocysts collected from stool of infected humans in the United States, Haiti, Guatemala, Peru, and Nepal were held in potassium dichromate solution to allow development of sporozoites. The following animal types were inoculated: 9 strains of mice, including adult and neonatal immunocompetent and immune-deficient inbred and outbred strains, rats, sandrats, chickens, ducks, rabbits, jirds, hamsters, ferrets, pigs, dogs, owl monkeys, rhesus monkeys, and cynomolgus monkeys. Most animals were inoculated by gavage, although some of the primates were fed oocysts on food items. The animals were examined for signs of infection, particularly diarrhea, and stool samples were examined for 4-6 wk after inoculation. None of the animals developed patent infections or signs of infection. We conclude that none of the animals tested is susceptible to infection with C. cayetanensis.

Animals↗

Immunogenicity testing of diphtheria and tetanus vaccines by using isogenic mice with possible implications for potency testing.

The antitoxin response of isogenic mice (inbred strains and F1 hybrids), immunized with diphtheria and tetanus reference vaccines, was compared with the response of animals from an NIH outbred stock. The variance in antitoxin response was smaller within the groups of isogenic mice than within the group of mice from the outbred strain. The case for estimating the potency of diphtheria and tetanus vaccines by using isogenic mice is discussed. It is concluded that the general introduction of one common inbred strain or F1 hybrid selected on the basis of sensitivity would increase the comparability of data between laboratories and would enable an estimated reduction in the use of animals of about 35% or an improvement in the precision of the estimate of potency.

Animals↗

Role of rat strain in the differential sensitivity to pharmaceutical agents and naturally occurring substances.

The development of drugs to combat diseases, chemicals to improve food production, or compounds to enhance the quality of life necessitates, by law, the use of laboratory animals to test their safety. In order to simulate the human condition it is necessary to choose a species in which pharmacokinetic and toxicokinetic mechanisms are established and resemble those of humans. The advantages of the use of the rat in drug and chemical toxicity testing include (a) metabolic pathway similarities to humans; (b) numerous similar anatomical and physiological characteristics; (c) a large database, which is extremely important for comparative purposes; and (d) the ease of breeding and maintenance of animals at relatively low cost. However, the choice of rat can be complicated, especially when over 200 different strains of rat are known to exist. The aim of this review is to summarize genetically determined differences in the responsiveness of rat strains to drugs and naturally occurring chemicals and to show that susceptibility is dependent on the target organ sensitivities, which may also be strain dependent. It is suggested that detailed studies of strain differences may help to clarify toxic mechanisms. Such studies are usually best conducted using inbred strains in which the genetic characteristics have been fixed, rather than in outbred stocks in which individual samples of animals may differ, the phenotype is variable, and the stocks are subject to substantial genetic drift. The fact that strains may differ also needs to be taken into account in assessing the potential hazard of the chemical, particularly when a study involves only a single strain and therefore provides no assessment of likely strain variation.

Animals↗

Strain differences in neuropathic hyperalgesia.

The purpose of this study was to investigate strain-related differences in the onset and maintenance of thermal hyperalgesia following the induction of peripheral nerve injury in two inbred strains of rats (Fischer 344 and Lewis) and two outbred strains of rats (Sprague-Dawley and Wistar). Neuropathic pain was induced via unilateral ligation of the left sciatic nerve with chromic gut sutures. A plantar analgesia meter was used to measure paw-withdrawal latency from the ligated vs. unligated hind paws of inbred vs. outbred strains of rats to investigate strain-related differences in nerve injury-induced thermal hyperalgesia. The results demonstrated no significant effects of animal strain on presurgical paw-withdrawal latency values. Following the sciatic nerve ligation (SNL) surgery, a significant hyperalgesic response was elicited from the Sprague-Dawley and Wistar rats (outbred strains) for at least 28 days. Conversely, data analyses from the inbred strains failed to demonstrate significant hyperalgesic responses to peripheral nerve injury, with the exception of postsurgical day 10. These data emphasize the importance of considering the strain of the rat being investigated before extrapolating the results from animals experiments to treatment strategies for humans with chronic neuropathic pain.

Animals↗

Worm burdens in outbred and inbred laboratory rats with morphometric data on Syphacia muris (Yamaguti, 1935) Yamaguti, 1941 (Nematoda, Oxyuroidea).

Syphacia muris worm burdens were evaluated in the rat Rattus norvegicus of the strains Wistar (outbred), Low/M and AM/2/Torr (inbred), maintained conventionally in institutional animal houses in Brazil. Morphometrics and illustration data for S. muris recovered from Brazilian laboratory rats are provided for the first time since its proposition in 1935.

Animals↗

Use of synthetic peptides corresponding to sequences of Mycobacterium leprae proteins to study delayed-type hypersensitivity response in sensitized guinea pigs.

In this work we report the synthesis of 10 peptides (P1-P10) corresponding to one or several segments of the amino acid sequence of proteins from Mycobacterium leprae: 65 kDa, 28 kDa, 18 kDa, and 28 kDa superoxide dismutase, recently renamed antigens 2L, 9L, 12L, and 14L, respectively. They were assayed in the guinea pig model for the induction of a delayed-type hypersensitivity response in M. leprae and BCG-sensitized animals. To sensitize the animals two schemes were used: either a single dose of 5 x 10(9) irradiated or autoclaved whole bacilli, or four weekly intramuscular injections each containing 500 micrograms of soluble extract of M. leprae (MLSE) in incomplete Freund's adjuvant. Because the second scheme used far too much antigen, we decided to use the first scheme for the experiments we report here. DTH reactions of sensitized animals were induced after 30 days with intradermal injections of 5 micrograms of MLSE and with each of the 10 peptides at three different concentrations: 250 micrograms, 100 micrograms, and 0.05 micrograms. All M. leprae-sensitized guinea pigs gave indurations of 10 mm or more with MLSE, which indicates that the animals were sensitized. None of them gave DTH indurations with 250 micrograms or 100 micrograms, but some of them had positive DTH reactions with the 0.05 micrograms doses of the synthetic peptides. This is most likely due to the fact that we have used an outbred strain of guinea pigs. The peptides were also tested at 0.05 micrograms in animals sensitized with BCG. P7 and P10 seem to be nonspecific peptides; the remaining peptides only induced DTH in the M. leprae-sensitized guinea pigs. P3 (segments 65-85 of the 65-kDa protein) induced a positive DTH in 58% of the tested animals. In other experiments, guinea pigs were sensitized with a single injection (500 micrograms) of each of the synthetic peptides. All animals, except those sensitized with P4 and P8, had positive DTH responses when the homologous peptide was used. Those sensitized with P2, P4, P5, P7, and P8 were able to produce indurations when MLSE was used for the induction of the DTH reaction.

Amino Acid Sequence↗