RESUMEN
The use of induced and spontaneous mutant mice and genetically engineered mice (and combinations thereof) to study cancers and other aging phenotypes to advance improved functional human life spans will involve studies of aging mice. Genetic background contributes to pathology phenotypes and to causes of death as well as to longevity. Increased recognition of expected phenotypes, experimental variables that influence phenotypes and research outcomes, and experimental design options and rationales can maximize the utility of genetically engineered mice (GEM) models to translational research on aging. This review aims to provide resources to enhance the design and practice of chronic and longevity studies involving GEM. C57BL6, 129, and FVB/N strains are emphasized because of their widespread use in the generation of knockout, transgenic, and conditional mutant GEM. Resources are included also for pathology of other inbred strain families, including A, AKR, BALB/c, C3H, C57L, C58, CBA, DBA, GR, NOD.scid, SAMP, and SJL/J, and non-inbred mice, including 4WC, AB6F1, Ames dwarf, B6, 129, B6C3F1, BALB/c,129, Het3, nude, SENCAR, and several Swiss stocks. Experimental strategies for long-term cross-sectional and longitudinal studies to assess causes of or contributors to death, disease burden, spectrum of pathology phenotypes, longevity, and functional healthy life spans (health spans) are compared and discussed.
Asunto(s)
Envejecimiento/patología , Longevidad/fisiología , Proyectos de Investigación , Animales , Ingeniería Genética , Humanos , Ratones , Ratones Endogámicos , Ratones Noqueados , Ratones Mutantes , Ratones Transgénicos , Fenotipo , Análisis de SupervivenciaRESUMEN
The phenotype of genetically engineered mice is a combination of both genetic and environmental factors that include the microflora of the mouse. The impact a particular microbe has on a mouse reflects the host-microbe interaction within the context of the mouse genotype and environment. Although often considered a confounding variable, many host-microbe interactions have resulted in the generation of novel model systems and characterization of new microbial agents. Microbes associated with overt disease in mice have been the historical focus of the laboratory animal medical and pathology community and literature. The advent of genetic engineering and the complex of mouse models have revealed previously unknown or disregarded agents that now oblige the attention of the biomedical research community. The purpose of this article is to describe and illustrate how phenotypes can be affected by microflora by focusing on the infectious diseases present in genetically engineered mouse (GEM) colonies of our collective institutions and by reviewing important agents that are rarely seen in most research facilities today. The goal is to introduce the concept of the role of microflora on phenotypes and in translational research using GEM models.
Asunto(s)
Enfermedades Transmisibles/veterinaria , Modelos Animales de Enfermedad , Ratones Transgénicos , Fenotipo , Enfermedades de los Roedores/microbiología , Enfermedades de los Roedores/parasitología , Animales , Enfermedades Transmisibles/microbiología , Enfermedades Transmisibles/parasitología , Ingeniería Genética , Humanos , Ratones , Enfermedades de los Roedores/genética , Organismos Libres de Patógenos EspecíficosRESUMEN
Spontaneous vestibular syndrome in mice, characterized clinically by head tilt, circling or rolling, can be caused by otitis media, arteritis or central nervous system lesions. Postmortem examination of eleven non-inbred Swiss mice submitted for necropsy due to acute onset of vestibular signs revealed lesions consistent with brainstem infarction. The lesions were characterized by unilateral, well-demarcated areas of necrosis, malacia, and gliosis, with variable amounts of hemorrhage, in the lateral aspect of the medulla and caudal pons. The affected area included the medial, lateral and superior vestibular nuclei, the facial nucleus and the spinal trigeminal nucleus. While vestibular disease secondary to otitis media, periarteritis, and central nervous system neoplasia has been reported in many mouse strains, these unilateral brainstem infarctions were only seen in Swiss mice. These lesions share features with Wallenberg's Lateral Medullary Syndrome, the most common type of brainstem infarct in humans.
Asunto(s)
Infartos del Tronco Encefálico/veterinaria , Enfermedades de los Roedores/patología , Animales , Infartos del Tronco Encefálico/patología , Femenino , Ciencia de los Animales de Laboratorio , RatonesRESUMEN
Mice deficient in CD18, which lack all four CD11 integrins, have leukocytosis and increased susceptibility to bacterial infection. To determine the effect of deficiencies in LFA-1 (CD11a/CD18) or Mac-1 (CD11b/CD18) on host defense against systemic bacterial infection, knockout mice were inoculated i.p. with Streptococcus pneumoniae. Increased mortality occurred in both LFA-1(-/-) (15 of 17 vs 13 of 35 in wild type (WT), p < 0.01) and Mac-1(-/-) (17 of 34 vs 6 of 25, p < 0.01) mice. All deaths in LFA-1(-/-) mice occurred after 72 h, whereas most deaths in Mac-1(-/-) mice occurred within 24-48 h. At 24 h, 21 of 27 Mac-1(-/-) mice were bacteremic, vs 15 of 25 WT (p = 0.05); no difference was observed between LFA-1(-/-) and WT. Increased bacteria were recovered from Mac-1(-/-) spleens at 2 h (p = 0.03) and 6 h (p = 0.002) and from livers (p = 0.001) by 6 h. No difference was observed at 2 h in LFA-1(-/-) mice, but by 6 h increased bacteria were recovered from spleens (p = 0.008) and livers (p = 0.04). Baseline and peak leukocyte counts were similar between Mac-1(-/-) and WT, but elevated in LFA-1(-/-). At 8 h, peritoneal neutrophils were increased in Mac-1(-/-), but not significantly different in LFA-1(-/-). Histopathologically, at 24 h Mac-1(-/-) animals had bacteremia and lymphoid depletion, consistent with sepsis. LFA-1(-/-) mice had increased incidence of otitis media and meningitis/encephalitis vs WT at 72 and 96 h. Both Mac-1 and LFA-1 play important but distinct roles in host defense to S. pneumoniae.
Asunto(s)
Antígeno-1 Asociado a Función de Linfocito/fisiología , Antígeno de Macrófago-1/fisiología , Infecciones Neumocócicas/inmunología , Animales , Líquido Ascítico/sangre , Bacteriemia/genética , Bacteriemia/inmunología , Bacteriemia/microbiología , Bacteriemia/mortalidad , Humanos , Recuento de Leucocitos , Antígeno-1 Asociado a Función de Linfocito/genética , Antígeno de Macrófago-1/genética , Meningitis Bacterianas/genética , Meningitis Bacterianas/inmunología , Meningitis Bacterianas/mortalidad , Meningitis Bacterianas/patología , Meningitis Neumocócica/genética , Meningitis Neumocócica/inmunología , Meningitis Neumocócica/mortalidad , Meningitis Neumocócica/patología , Meningoencefalitis/genética , Meningoencefalitis/inmunología , Meningoencefalitis/mortalidad , Meningoencefalitis/patología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Técnicas de Cultivo de Órganos , Otitis Media/genética , Otitis Media/inmunología , Otitis Media/mortalidad , Otitis Media/patología , Infecciones Neumocócicas/genética , Infecciones Neumocócicas/mortalidad , Infecciones Neumocócicas/patología , Streptococcus pneumoniae/inmunología , Streptococcus pneumoniae/aislamiento & purificación , Análisis de SupervivenciaRESUMEN
Osteogenesis imperfecta (OI), a heritable disease caused by molecular defects in type I collagen, is characterized by skeletal deformities and brittle bones. The heterozygous and homozygous oim mice (oim/+ and oim/oim) exhibit mild and severe OI phenotypes, respectively, serving as controlled animal models of this disease. In the current study, bone geometry, mechanics, and material properties of 1-year-old mice were evaluated to determine factors that influence the severity of phenotype in OI. The oim/oim mice exhibited significantly smaller body size, femur length, and moment of area compared with oim/+ and wild-type (+/+) controls. The oim/oim femur mechanical properties of failure torque and stiffness were 40% and 30%, respectively, of the +/+ values, and 53% and 36% of the oim/+ values. Collagen content was reduced by 20% in the oim/oim compared with +/+ bone and tended to be intermediate to these values for the oim/+. Mineral content was not significantly different between the oim/oim and +/+ bones. However, the oim/oim ash content was significantly reduced compared with that of the oim/+. Mineral carbonate content was reduced by 23% in the oim/oim bone compared with controls. Mineral crystallinity was reduced in the oim/oim and oim/+ bone compared with controls. Overall, for the majority of parameters examined (geometrical, mechanical, and material), the oim/+ values were intermediate to those of the oim/oim and +/+, a finding that parallels the phenotypes of the mice. This provides evidence that specific material properties, such as mineral crystallinity and collagen content, are indicative and possibly predictive of bone fragility in this mouse model, and by analogy in human OI.
Asunto(s)
Osteogénesis Imperfecta/patología , Osteogénesis Imperfecta/fisiopatología , Animales , Fenómenos Biomecánicos , Densidad Ósea/genética , Colágeno/genética , Colágeno/metabolismo , Modelos Animales de Enfermedad , Heterocigoto , Homocigoto , Humanos , Ratones , Ratones Mutantes , Osteogénesis Imperfecta/genética , FenotipoRESUMEN
The homozygous oim/oim mouse, a model of moderate-to-severe human osteogenesis imperfecta, contains a G-nucleotide deletion in the Cola-2 gene (the murine pro alpha(I) collagen gene) that results in accumulation of alpha1(I) homotrimer collagen. Although these mice have a distinctive phenotype that includes multiple fractures and deformities, genotyping is necessary to distinguish them from their wildtype (+/+) and heterozygote (oim/+) littermates. In this study, the dye primer and dye terminator chemistry methods, in combination with automated direct DNA sequencing, were compared for accuracy and ease in genotyping. A total of 82 mice from 14 litters were bred and genotyped; this resulted in 18 +/+, 35 oim/+, and 29 oim/oim mice. The dye primer and dye terminator chemistry methods worked equally well for identification of the deletion mutation and thus the genotype of all of the mice. However, the dye terminator method was found to be superior on the basis of the reduced amount of sample handling and reduced quantity of reagent required.
Asunto(s)
Colágeno/genética , Mutación , Osteogénesis Imperfecta/genética , Análisis de Secuencia de ADN , Animales , Colorantes , Genotipo , Ratones , Reacción en Cadena de la PolimerasaRESUMEN
We previously reported that heparin inhibits the proliferation of fibroblasts and vascular smooth muscle cells (SMC), in part, by binding to and increasing the antiproliferative activity of transforming growth factor-beta 1 (TGF-beta 1). We now report that certain other polyanions which are structurally distinct from heparin, such as fucoidan and polyinosinic acid, are more avid ligands for TGF-beta 1 and more potent antiproliferative agents than heparin. Fucoidan possessed more potent antiproliferative activity than heparin against rat and bovine aortic SMC in vitro, though possessing much lower anticoagulant activity than heparin. Furthermore, fucoidan suppressed in vivo intimal hyperplasia when continuously infused into rats subjected to balloon-catheter injury. Unlike heparin, which also suppressed intimal hyperplasia, fucoidan did not cause systemic anticoagulation. Thus, fucoidan may be useful as a non-anticoagulant inhibitor of post-angioplasty intimal hyperplasia.
Asunto(s)
Aorta/citología , Heparina/farmacología , Músculo Liso Vascular/citología , Polisacáridos/farmacología , Animales , Aniones , Aorta/efectos de los fármacos , Aorta/patología , Bovinos , División Celular/efectos de los fármacos , Células Cultivadas , Relación Dosis-Respuesta a Droga , Hiperplasia , Músculo Liso Vascular/efectos de los fármacos , Músculo Liso Vascular/patología , Ratas , Ratas Endogámicas F344 , Receptores de Superficie Celular/efectos de los fármacos , Receptores de Superficie Celular/metabolismo , Receptores de Factores de Crecimiento Transformadores beta , Factor de Crecimiento Transformador beta/metabolismoRESUMEN
The control of smooth muscle cell (SMC) proliferation is determined by the combined actions of mitogens, such as platelet-derived growth factor, and the opposing action of growth inhibitory agents, such as heparin and transforming growth factor-beta (TGF-beta). The present studies identify an interaction between heparin and TGF-beta in which heparin potentiates the biological action of TGF-beta. Using a neutralizing antibody to TGF-beta, we observed that the short term antiproliferative effect of heparin depended upon the presence of biologically active TGF-beta. This effect was observed in rat and bovine aortic SMC and in CCL64 cells, but not in human saphenous vein SMC. Binding studies demonstrated that the addition of heparin (100 micrograms/ml) to medium containing 10% plasma-derived serum resulted in a 45% increase in the specific binding of 125I-TGF-beta to cells. Likewise, heparin induced a twofold increase in the growth inhibitory action of TGF-beta at concentrations of TGF-beta near its apparent dissociation constant. Using 125I-labeled TGF-beta, we demonstrated that TGF-beta complexes with the plasma component alpha 2-macroglobulin, but not with fibronectin. Heparin increases the electrophoretic mobility of TGF-beta apparently by freeing TGF-beta from its complex with alpha 2-macroglobulin. Dextran sulfate, another highly charged antiproliferative molecule, but not chondroitin sulfate or dermatan sulfate, similarly modified TGF-beta's mobility. Relatively high, antiproliferative concentrations of heparin (1-100 micrograms/ml) were required to dissociate the TGF-beta/alpha 2-macroglobulin complex. Thus, it appears that the antiproliferative effect of heparin may be partially attributed to its ability to potentiate the biological activity of TGF-beta by dissociating it from alpha 2-macroglobulin, which normally renders it inactive. We suggest that heparin-like agents may be important regulators of TGF-beta's biological activity.
Asunto(s)
Heparina/farmacología , Factores de Crecimiento Transformadores/farmacología , alfa-Macroglobulinas/metabolismo , Animales , División Celular/efectos de los fármacos , Células Cultivadas , Sinergismo Farmacológico , Técnicas Inmunológicas , Técnicas In Vitro , Músculo Liso Vascular/citología , Unión Proteica/efectos de los fármacosRESUMEN
The goal of this study was to determine if experimental Mycoplasma pulmonis infection augmented splenic natural killer (NK) cell activity in mice. A 4 hour 51Cr-release in vitro assay using YAC-1 tumor target cells was employed to measure splenic NK cell activity in C57BL/6J mice infected intraperitoneally with M. pulmonis and in uninfected controls. Transient augmentation of the NK cells was observed, peaking at day 3 postinoculation (PI) and gradually returning to normal levels by day 10 PI. Selective depletion studies showed that the cells responsible for killing target cells were NK cells. They were nonadherent to nylon wool, not susceptible to Thy-1.2 antibody and susceptible to asialo GM1 ganglioside antibody. Inadvertent augmentation of the NK cell system due to M. pulmonis infection may complicate the interpretation of research data, especially in immunology and cancer studies.
Asunto(s)
Células Asesinas Naturales/fisiología , Infecciones por Mycoplasma/fisiopatología , Enfermedades Peritoneales/microbiología , Bazo/citología , Animales , Ratones , Ratones Endogámicos C57BL , Enfermedades Peritoneales/fisiopatologíaRESUMEN
Dimethyl sulfoxide (DMSO) is a very simple compound that has stimulated much controversy in the scientific and popular literature. Fig. 1 It is an aprotic solvent. Therapeutic and toxic agents that are not soluble in water are often soluble in DMSO. DMSO has a very strong affinity for water; on exposure to air, pure DMSO is rapidly diluted. DMSO's physiologic and pharmacologic properties and effects are incompletely understood. Properties that are considered to be particularly important to its therapeutic and toxic effects include: its own rapid penetration and enhanced penetration of other substances across biologic membranes; free radical scavenging; effects on coagulation; anticholinesterase activity; and DMSO-induced histamine release by mast cells. DMSO's systemic toxicity is considered to be low. Combinations of DMSO with other toxic agents probably constitute its greatest toxic potential. The scientific literature is reviewed with particular attention to mechanisms underlying DMSO's reported therapeutic and toxic effects. Currently approved, veterinary applications of DMSO are limited. DMSO's potential value in specific, approved and unapproved veterinary applications is discussed.