ABSTRACT
The "humanized mouse" is a mouse harboring functioning human tissues used as in vivo human models for both physiological and pathological conditions. The NOD/Shi-scid IL2rgamma(null) (NOG) mouse, an excellent immunodeficient mouse used as the basis for the humanized mouse, requires strict genetic and environmental control for production and use in experiments. Genetic control using marker-assisted selection is described. In addition, NOG mice are easily affected by microbiological and proximate environmental factors, which can cause severe damage to the mice in some cases. Therefore, rigorous microbiological and environmental controls are necessary to ensure reproducibility of experimental results. At the end of this chapter, future aspects of the application of "humanized mice" based on novel super-immunodeficient mice such as NOG mice and Rag2(null) IL2rgamma(null) mice in biomedical research and testing are briefly reviewed.
Subject(s)
Disease Models, Animal , Animals , DNA-Binding Proteins/deficiency , Humans , Interleukin Receptor Common gamma Subunit/deficiency , Mice , Mice, Inbred NOD , Mice, SCIDABSTRACT
Autosomal dominant polycystic kidney disease, a leading cause of end-stage renal disease in adults, is characterized by progressive focal cyst formation in the kidney. Embryonic lethality of Pkd1-targeted mice limits the use of these mice. Here we developed a floxed allele of Pkd1 exons 2-6. Global deletion mutants developed polyhydramnios, hydrops fetalis, polycystic kidney and pancreatic disease. Somatic Pkd1 inactivation in the kidney was achieved by crossing Pkd1(flox) mice with transgenic mice expressing Cre controlled by a gamma-glutamyltranspeptidase promoter. These mutants developed cysts in both proximal and distal nephron segments and survived for about 4 weeks. Somatic loss of heterozygosity was shown in a reporter mouse strain to cause cystogenesis. Some cysts in young mice are positive for multiple tubular markers and a mesenchymal marker, suggesting a delay in tubular epithelial differentiation. A higher cell proliferation rate was observed in distal nephron segments probably accounting for the faster growth rate of distal cysts. Although we observed an overall increase in apoptosis in cystic kidneys, there was no difference between proximal or distal nephron segments. We also found increased cyclic AMP, aquaporin 2 and vasopressin type 2 receptor mRNA levels, and apical membrane translocation of aquaporin 2 in cystic kidneys, all of which may contribute to the differential cyst growth rate observed. The accelerated polycystic kidney phenotype of these mice provides an excellent model for studying molecular pathways of cystogenesis and to test therapeutic strategies.
Subject(s)
Disease Models, Animal , Mice , Polycystic Kidney Diseases/genetics , Polycystic Kidney Diseases/pathology , Sequence Deletion , TRPP Cation Channels/metabolism , Alleles , Animals , Apoptosis , Base Sequence , Cell Proliferation , Cyclic AMP/metabolism , Disease Progression , Kidney Tubules, Distal/metabolism , Kidney Tubules, Distal/pathology , Mice, Knockout , Polycystic Kidney Diseases/metabolismABSTRACT
BACKGROUND: The risk of post-transfusion hepatitis B virus (HBV) infection has been reduced after the implementation of HBV nucleic acid amplification technology (NAT). However, the problem of HBV DNA-positive and HBV surface antigen (HBsAg)-negative occult HBV infections remains to be solved. This is in part due to the HBV DNA load being too low to detect these occult HBV infections using mini-pool NAT. In Japan, the assay for the antibody against the HBV core antigen (anti-HBc) has not completely excluded occult HBV infection. To solve this problem, we have developed a new method of concentrating HBV DNA and HBsAg simultaneously to increase the sensitivity of detection tests. METHODS: Virus concentration is achieved by the enhancement of the agglutination of viruses using poly-L-lysine in the presence of a bivalent metal. Poly-L-lysine-coated magnetic beads are used to shorten the time of each step of the concentration procedure. Seventy-seven anti-HBc-positive and HBsAg-negative donations were examined. HBsAg and anti-HBc were tested by enzyme immunoassay (EIA) (AxSYM; Abbott) and haemagglutination inhibition test (Japanese Red Cross), respectively. RESULTS: HBV surface antigen and HBV DNA levels were concentrated up to four- to sevenfold. Using this method, 35 of the 77 anti-HBc-positive and HBsAg-negative donors were HBV DNA-positive by individual NAT and a further five donors became HBV DNA-positive by HBV concentration. Twenty-seven of 40 occult HBV infections became HBsAg-positive by HBsAg concentration. CONCLUSION: Our new method of concentrating HBV and HBsAg increased the sensitivities of EIA and HBV NAT, and enabled us to detect 27 of 40 occult HBV infections by HBsAg EIA.
Subject(s)
Blood Transfusion , DNA, Viral/blood , Hepatitis B Surface Antigens/blood , Hepatitis B virus , Hepatitis B/blood , Hepatitis B/prevention & control , Nucleic Acid Amplification Techniques/methods , Blood Banking/methods , Female , Hepatitis B/transmission , Humans , MaleABSTRACT
Phox2b is required for development of the peripheral autonomic nervous system and a subset of cranial nerves and lower brainstem nuclei. Phox2b mutations in man cause diffuse autonomic dysfunction and deficits in the automatic control of breathing. Here we study the distribution of Phox2b in the adult rat hindbrain to determine whether this protein is selectively expressed by neurons involved in respiratory and autonomic control. In the medulla oblongata, Phox2b-immunoreactive nuclei were present in the dorsal vagal complex, intermediate reticular nucleus, dorsomedial spinal trigeminal nucleus, nucleus ambiguus, catecholaminergic neurons, and retrotrapezoid nucleus (RTN). Phox2b was expressed by both central excitatory relays of the sympathetic baroreflex (nucleus of the solitary tract and C1 neurons) but not by the inhibitory relay of this reflex. Phox2b was absent from the ventral respiratory column (VRC) caudal to RTN and rare within the parabrachial nuclei. In the pons, Phox2b was confined to cholinergic efferent neurons (salivary, vestibulocochlear) and noncholinergic peritrigeminal neurons. Rostral to the pons, Phox2b was detected only in the oculomotor complex. In adult rats, Phox2b is neither a comprehensive nor a selective marker of hindbrain autonomic pathways. This marker identifies a subset of hindbrain neurons that control orofacial movements (dorsomedial spinal trigeminal nucleus, pontine peritrigeminal neurons), balance and auditory function (vestibulocochlear efferents), the eyes, and both divisions of the autonomic efferent system. Phox2b is virtually absent from the respiratory rhythm and pattern generator (VRC and dorsolateral pons) but is highly expressed by neurons involved in the chemical drive and reflex regulation of this oscillator.
Subject(s)
Central Nervous System/metabolism , Homeodomain Proteins/metabolism , Transcription Factors/metabolism , Animals , Animals, Newborn , Central Nervous System/anatomy & histology , Choline O-Acetyltransferase/metabolism , Gene Expression Regulation, Developmental/physiology , Glutamate Decarboxylase/genetics , Glutamate Decarboxylase/metabolism , Isoenzymes/genetics , Isoenzymes/metabolism , Male , Motor Neurons/metabolism , Proto-Oncogene Proteins c-fos/metabolism , Rats , Rats, Sprague-Dawley , Stilbamidines/metabolism , Tyrosine 3-Monooxygenase/metabolism , Vesicular Glutamate Transport Protein 2/genetics , Vesicular Glutamate Transport Protein 2/metabolismABSTRACT
Small GTP-binding protein GDP dissociation stimulator (Smg GDS) regulates GDP/GTP exchange reaction of Ki-Ras and the Rho and Rap1 family members and inhibits their binding to membranes. In fibroblasts, Smg GDS shows mitogenic and transforming activities in cooperation with Ki-Ras. However, the physiological function of Smg GDS remains unknown. Here we show that mice lacking Smg GDS died of heart failure shortly after birth, not resulting from developmental heart defects but from enhanced apoptosis of cardiomyocytes triggered by cardiovascular overload. Furthermore, neonatal thymocytes and developing neuronal cells underwent apoptotic cell death. Smg GDS-/- thymocytes were susceptible to apoptotic inducers, such as etoposide and UV irradiation. Smg GDS-/- thymocytes were protected from etoposide-induced cell death by ex vivo transduction of the Smg GDS cDNA. These phenotypes partly coincide with those observed in Ki-Ras-deficient mice, suggesting that Smg GDS is involved in antiapoptotic cell survival signaling through Ki-Ras.
Subject(s)
Apoptosis/physiology , GTP-Binding Proteins/genetics , GTP-Binding Proteins/metabolism , Signal Transduction , Animals , Animals, Newborn , Apoptosis/drug effects , Apoptosis/radiation effects , Cell Survival/physiology , Cells, Cultured , Etoposide/pharmacology , Female , Genes, ras , Heart Defects, Congenital/pathology , Mice , Mice, Inbred C57BL , Mice, Transgenic , Myocardium/pathology , Neurons/pathology , Survival Rate , Thymus Gland/drug effects , Thymus Gland/growth & development , Thymus Gland/pathology , Thymus Gland/radiation effects , Ultraviolet RaysABSTRACT
Bisphosphonates (BPs) and teriparatide (TPTD) are both effective treatments for osteoporosis, but BP treatment prior to daily TPTD treatment has been shown to impair the effect of TPTD in some clinical studies. In contrast, the loss of bone mineral density (BMD) that occurs after withdrawal of TPTD can be prevented by BP treatment. Although various studies have investigated the combination and/or sequential use of BP and TPTD, there have been no clinical studies investigating sequential treatment with zoledronic acid (ZOL) and TPTD (or vice versa). In this study, we evaluated the effects of sequential treatment with TPTD followed by ZOL, and ZOL followed by TPTD, using ovariectomized (OVX) rats. Two months after OVX, osteopenic rats were treated with ZOL, TPTD, or vehicle for a period of 4 months (first treatment period), and then the treatments were switched and administered for another 4 months (second treatment period). The group treated with ZOL followed by TPTD showed an immediate increase in BMD of the proximal tibia and greater BMD and bone strength of the lumbar vertebral body, femoral diaphysis, and proximal femur than the group treated with ZOL followed by vehicle. Serum osteocalcin, a marker of bone formation, increased rapidly after switching to TPTD from ZOL. The group treated with TPTD followed by ZOL did not lose BMD in the proximal tibia after TPTD was stopped, while the group treated with TPTD followed by vehicle did lose BMD. The BMD and bone strength of the lumbar vertebral body, femoral diaphysis, and proximal femur were greater in the group treated with TPTD followed by ZOL than in the group treated with TPTD followed by vehicle. The increase in serum osteocalcin and urinary CTX after withdrawal of TPTD was prevented by the switch from TPTD to ZOL. In conclusion, our results demonstrate that switching from ZOL to TPTD resulted in a non-attenuated anabolic response in the lumbar spine and femur of OVX rats. In addition, switching from TPTD to ZOL caused BMD to be maintained or further increased. If these results can be reproduced in a clinical setting, the sequential use of ZOL followed by TPTD or vice versa in the treatment of osteoporosis patients would contribute to increases in BMD that, hopefully, would translate into a corresponding decrease in the incidence of vertebral and non-vertebral fractures.
ABSTRACT
AIM: Recent evidence suggests that adenosine triphosfate (ATP)-mediated purinergic signalling at the level of the rostral ventrolateral medulla contributes to both central and peripheral chemoreceptor control of breathing and blood pressure: neurones in the retrotrapezoid nucleus (RTN) function as central chemoreceptors in part by responding to CO2 -evoked ATP release by activation of yet unknown P2 receptors, and nearby catecholaminergic C1 neurones regulate blood pressure responses to peripheral chemoreceptor activation by a P2Y1 receptor-dependent mechanism. However, potential contributions of purinergic signalling in the RTN to cardiorespiratory function in conscious animals have not been tested. METHODS: Cardiorespiratory activity of unrestrained awake rats was measured in response to RTN injections of ATP, and during exposure to hypercapnia (7% CO2 ) or hypoxia (8% O2 ) under control conditions and after bilateral RTN injections of P2 receptor blockers (PPADS or MRS2179). RESULTS: Unilateral injection of ATP into the RTN increased cardiorespiratory output by a P2-receptor-dependent mechanism. We also show that bilateral RTN injections of a non-specific P2 receptor blocker (pyridoxal-phosphate-6-azophenyl-2',4'-disulfonate (PPADS) reduced the ventilatory response to hypercapnia (7% CO2 ) and hypoxia (8% O2 ) in unanesthetized rats. Conversely, bilateral injections of a specific P2Y1 receptor blocker (MRS2179) into the RTN had no measurable effect on ventilatory responses elicited by hypercapnia or hypoxia. CONCLUSION: These data exclude P2Y1 receptor involvement in the chemosensory control of breathing at the level of the RTN and show that ATP-mediated purinergic signalling contributes to central and peripheral chemoreflex control of breathing and blood pressure in awake rats.
Subject(s)
Chemoreceptor Cells/metabolism , Medulla Oblongata/metabolism , Receptors, Purinergic P2Y1/metabolism , Respiratory Physiological Phenomena , Adenosine Triphosphate/pharmacology , Animals , Male , Medulla Oblongata/drug effects , Purinergic P2Y Receptor Antagonists/pharmacology , Rats , Rats, Wistar , WakefulnessABSTRACT
The Rho small G protein family members regulate various actin cytoskeleton-dependent cell functions. The Rho GDI (GDP dissociation inhibitor) family, consisting of Rho GDIalpha, -beta, and -gamma, is a regulator that keeps the Rho family members in the cytosol as the GDP-bound inactive form and translocates the GDP-bound form from the membranes to the cytosol after the GTP-bound form accomplishes their functions. Rho GDIalpha is ubiquitously expressed in mouse tissues and shows GDI activity on all the Rho family members in vitro. We have generated mice lacking Rho GDIalpha by homologous recombination to clarify its in vivo function. Rho GDIalpha -/- mice showed several abnormal phenotypes. Firstly, Rho GDIalpha -/- mice were initially viable but developed massive proteinuria mimicking nephrotic syndrome, leading to death due to renal failure within a year. Histologically, degeneration of tubular epithelial cells and dilatation of distal and collecting tubules were readily detected in the kidneys. Secondly, Rho GDIalpha -/- male mice were infertile and showed impaired spermatogenesis with vacuolar degeneration of seminiferous tubules in their testes. Thirdly, Rho GDIalpha -/- embryos derived from Rho GDIalpha -/- female mice were defective in the postimplantation development. In addition, these morphological and functional abnormalities showed age-dependent progression. These results suggest that the signaling pathways of the Rho family members regulated by Rho GDIalpha play important roles in maintaining the structure and physiological function of at least kidneys and reproductive systems in adult mice.
Subject(s)
Guanine Nucleotide Dissociation Inhibitors/physiology , Renal Insufficiency/etiology , Age Factors , Animals , Epithelial Cells/pathology , Female , Guanine Nucleotide Dissociation Inhibitors/deficiency , Guanine Nucleotide Dissociation Inhibitors/genetics , Infertility, Male/etiology , Infertility, Male/genetics , Infertility, Male/pathology , Kidney Tubules/pathology , Male , Mice , Mice, Inbred C57BL , Mice, Transgenic , Nephrotic Syndrome/etiology , Nephrotic Syndrome/genetics , Renal Insufficiency/genetics , Renal Insufficiency/metabolism , Testis/pathology , rho Guanine Nucleotide Dissociation Inhibitor alpha , rho-Specific Guanine Nucleotide Dissociation InhibitorsABSTRACT
Parkinson's disease (PD) is a neurodegenerative disease characterized by loss of the dopaminergic nigrostriatal pathway. In addition to deficits in voluntary movement, PD involves a disturbance of breathing regulation. However, the cause and nature of this disturbance are not well understood. Here, we investigated breathing at rest and in response to hypercapnia (7% CO2) or hypoxia (8% O2), as well as neuroanatomical changes in brainstem regions essential for breathing, in a 6-hydroxydopamine (6-OHDA) rat model of PD. Bilateral injections of 6-OHDA (24µg/µl) into the striatum decreased tyrosine hydroxylase (TH(+))-neurons in the substantia nigra pars compacta (SNpc), transcription factor phox2b-expressing neurons in the retrotrapezoid nucleus and neurokinin-1 receptors in the ventral respiratory column. In 6-OHDA-lesioned rats, respiratory rate was reduced at rest, leading to a reduction in minute ventilation. These animals also showed a reduction in the tachypneic response to hypercapnia, but not to hypoxia challenge. These results suggest that the degeneration of TH(+) neurons in the SNpc leads to impairment of breathing at rest and in hypercapnic conditions. Our data indicate that respiratory deficits in a 6-OHDA rat model of PD are related to downregulation of neural systems involved in respiratory rhythm generation. The present study suggests a new avenue to better understand the respiratory deficits observed in chronic stages of PD.
Subject(s)
Corpus Striatum/drug effects , Disease Models, Animal , Parkinson Disease/complications , Respiration Disorders/etiology , Adrenergic Agents/toxicity , Animals , Cell Count , Hydrogen-Ion Concentration/drug effects , Lactic Acid/blood , Locomotion/drug effects , Male , Neurons/drug effects , Neurons/metabolism , Oxidopamine/toxicity , Parkinson Disease/etiology , Psychomotor Performance , Pulmonary Ventilation/drug effects , Rats , Rats, Wistar , Receptors, Neurokinin-1/metabolism , Respiratory Center/drug effects , Respiratory Center/metabolism , Respiratory Center/pathology , Substantia Nigra/drug effects , Substantia Nigra/metabolism , Time FactorsABSTRACT
We investigated the role of the autonomic nervous system to cardiovascular responses to obstructive apnea in awake, unrestrained rats, and measured expression of Fos induced by apnea in the brainstem. We implanted a tracheal balloon contained in a rigid tube to allow the induction of apnea without inducing pain in the trachea. During bouts of 15s of apnea, heart rate fell from 371±8 to 161±11bpm (mean±SEM, n=15, p<0.01) and arterial pressure increased from 115±2 to 131±4mmHg (p<0.01). Bradycardia was due to parasympathetic activity because it was blocked by the muscarinic antagonist, methylatropine. The pressor response was due to vasoconstriction caused by sympathetic activation because it was blocked by the α1 antagonist, prazosin. Apnea induced Fos expression in several brainstem areas involved in cardiorespiratory control such as the nucleus of the solitary tract (NTS), ventrolateral medulla (VLM), and pons. Ligation of the carotid body artery reduced apnea-induced bradycardia, blocked heart rate responses to i.v. injection of cyanide, reduced Fos expression in the caudal NTS, and increased Fos expression in the rostral VLM. In conclusion, apnea activates neurons in regions that process signals from baroreceptors, chemoreceptors, pulmonary receptors, and regions responsible for autonomic and respiratory activity both in the presence and absence of carotid chemoreceptors.
Subject(s)
Apnea/pathology , Apnea/physiopathology , Brain Stem/physiopathology , Wakefulness , Analysis of Variance , Animals , Atropine Derivatives/pharmacology , Blood Pressure/drug effects , Brain Stem/drug effects , Carotid Body/cytology , Chemoreceptor Cells/drug effects , Heart Rate/drug effects , Male , Oncogene Proteins v-fos/metabolism , Parasympatholytics/pharmacology , Prazosin/pharmacology , Rats , Rats, Wistar , Tyrosine 3-Monooxygenase/metabolismABSTRACT
Although cholinergic agonists such as pilocarpine injected peripherally can act directly on salivary glands to induce salivation, it is possible that their action in the brain may contribute to salivation. To investigate if the action in the brain is important to salivation, we injected pilocarpine intraperitoneally after blockade of central cholinergic receptors with atropine methyl bromide (atropine-mb). In male Holtzman rats with stainless steel cannulas implanted into the lateral ventricle and anesthetized with ketamine, atropine-mb (8 and 16 nmol) intracerebroventricularly reduced the salivation induced by pilocarpine (4 micro mol/kg) intraperitoneally (133 + 42 and 108 + 22 mg/7 min, respectively, vs. saline, 463 + 26 mg/7 min), but did not modify peripheral cardiovascular responses to intravenous acetylcholine. Similar doses of atropine-mb intraperitoneally also reduced pilocarpine-induced salivation. Therefore, systemically injected pilocarpine also enters the brain and acts on central muscarinic receptors, activating autonomic efferent fibers to induce salivation.
Subject(s)
Pilocarpine/pharmacology , Receptors, Muscarinic/physiology , Salivation/drug effects , Acetylcholine/administration & dosage , Acetylcholine/pharmacology , Animals , Atropine Derivatives/pharmacology , Autonomic Nervous System/drug effects , Blood Pressure/drug effects , Brain/drug effects , Heart Rate/drug effects , Injections, Intraperitoneal , Injections, Intraventricular , Male , Muscarinic Antagonists/pharmacology , Nerve Fibers/drug effects , Neurons, Efferent/drug effects , Parasympatholytics/pharmacology , Pilocarpine/administration & dosage , Rats , Rats, Sprague-Dawley , Receptors, Cholinergic/drug effects , Receptors, Muscarinic/drug effectsABSTRACT
To examine the susceptibility of the epithelial cell line to rat coronavirus (RCV), we inoculated sialodacryoadenitis virus and Parker's RCV into five cell lines; JTC-19, rat L2, LLC, RCN-9 and LBC cells originating in the lungs, intestines and mammary tumors of rodents. Both RCVs were replicated in LBC and RCN-9 cells, but not in the others. The infectivity titers of both RCVs grown in RCN-9 cells were significantly higher than those in LBC cells in every passage (2.5-3.9 log rate). Both RCVs replicated in LBC cells showed higher tropism to RCN-9 cells than to LBC cells, suggesting that RCN-9 cells are more suitable for the replication of RCVs than LBC cells. The RCN-9 cell line would be useful for the investigation of RCV infection in rodents.
Subject(s)
Coronavirus, Rat/growth & development , Virus Replication/physiology , Animals , Cell Line , Coronavirus, Rat/isolation & purification , Coronavirus, Rat/pathogenicity , Epithelial Cells , Giant Cells , Intestines/cytology , Mice , Rats , Rats, Inbred F344 , Rodent Diseases/virology , Tumor Cells, CulturedABSTRACT
Cilia-associated respiratory (CAR) bacillus was detected by means of the reverse transcription (RT)-polymerase chain reaction (PCR), and the results were compared with those of indirect immunofluorescence test (IFAT) for the detection of the organism. In the experimental infections, 15 mice were in contact with mice previously inoculated with CAR bacillus. Three mice each were tested at days 3, 5, 7, 12 and 20 postexposure. On day 3 postexposure, CAR bacillus was detected in oral swab samples from all 3 mice by RT-PCR, but was not detected in any sampling sites from the mice by IFAT. Total numbers of positive samples from nasal, oral and tracheal swabs obtained through the test were 6/15, 14/15 and 8/15, respectively, by RT-PCR, and 2/15, 6/15 and 3/15, respectively by IFAT. For the detection of CAR bacillus in samples from 52 rats, 34 serum antibody negative rats by enzyme-linked immunosorbent assay were also negative by RT-PCR and IFAT except for one sample from the oral cavity, and all serum antibody positive rats were positive for the organism by RT-PCR but it could not be detected in five of them by IFAT. By means of RT-PCR, no differences in the positive rates depending on sampling sites were observed except in one rat. The RT-PCR was found to be a specific, highly sensitive and reliable procedure for detecting CAR bacillus in mice and rats. The oral cavity was the most suitable site for the diagnosis of the early stage of this infection by RT-PCR.
Subject(s)
Cilia/microbiology , Gram-Negative Bacteria/isolation & purification , Mice/microbiology , Rats/microbiology , Respiratory Tract Diseases/veterinary , Rodent Diseases/microbiology , Animals , Base Sequence , Fluorescent Antibody Technique, Indirect , Gram-Negative Bacteria/genetics , Molecular Sequence Data , Polymerase Chain Reaction , RNA, Bacterial/analysis , RNA-Directed DNA Polymerase , Respiratory Tract Diseases/microbiologyABSTRACT
A polymerase chain reaction with new primers (new PCR) designed from Pasteurella pneumotropica 16S rDNA as an identification system for this organism was compared with the PCR reported by Wang et al. (Wang's PCR) by using 15 bacterial reference species and 70 clinical isolates with the conventional identification system. For the 15 reference strains, both PCRs were identical. For the 70 clinical isolates, the new PCR and Wang's PCR showed consistency with the conventional system in 62.9% (44/70) and 51.4% (36/70), respectively. Twenty-six isolates were inconsistent with the conventional system and the new PCR with respect to morphology and serology. These findings suggested that the new PCR was more sensitive than Wang's PCR, and the new PCR in combination with morphology and serology is useful for P. pneumotropica identification.
Subject(s)
Pasteurella Infections/diagnosis , Pasteurella/isolation & purification , Polymerase Chain Reaction/methods , Animals , Animals, Laboratory , Base Sequence , DNA, Bacterial/analysis , DNA, Bacterial/genetics , DNA, Ribosomal/analysis , DNA, Ribosomal/genetics , Molecular Sequence Data , Pasteurella/classification , Pasteurella/genetics , Pasteurella Infections/microbiology , RNA, Ribosomal, 16S/genetics , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
To quarantine human tumor samples for transplantation into immune deficient mice or tumor xenograft lines established and introduced from other institutions, we performed isolated implantation and passaging of tumors in a vinyl isolator, and microbiological examinations of sentinel mice kept together with tumor bearing mice. We examined 105 pairs of sentinel mice used to quarantine 907 tumors, and found six cases of contamination or infection with Staphylococcus aureus, 20 cases with Pseudomonas aeruginosa and one case with mouse hepatitis virus (MHV). It was, however, possible that Mycoplasma pulmonis contamination was overlooked because the microbe had been isolated from tumors passaged after quarantine, even though the results of the quarantine of these tumors showed no sign of pathogens. Direct culture of tumors for the microbe was recommended to improve the quarantine system.
Subject(s)
Neoplasms, Experimental/microbiology , Neoplasms, Experimental/virology , Animals , Female , Humans , Male , Mice , Mice, Inbred BALB C , Mice, Nude , Mice, SCID , Microbiological Techniques , Murine hepatitis virus/isolation & purification , Mycoplasma/isolation & purification , Neoplasm Transplantation , Pseudomonas aeruginosa/isolation & purification , Quarantine , Staphylococcus aureus/isolation & purification , Transplantation, Heterologous , Tumor Cells, CulturedABSTRACT
A human tumor xenograft contaminated with mouse hepatitis virus (MHV) was implanted in a nude rat in order to decontaminate the tumor line. The decontamination failed in the first trial, but succeeded in the second trial. The difference between the two trials was the duration of implantation of the tumor in the nude rat, i.e., 12 days in the first and 24 days in the second trial. Duration of implantation might be a factor in the decontamination of transplantable tumors infected with MHV by passaging in the nude rat.
Subject(s)
Coronavirus Infections/prevention & control , Murine hepatitis virus , Neoplasm Transplantation , Neoplasms, Experimental/virology , Transplantation, Heterologous , Animals , Coronavirus Infections/transmission , Decontamination , Female , Humans , Mice , Mice, Nude , Rats , Rats, Inbred F344 , Rats, Nude , Specific Pathogen-Free Organisms , Tumor Cells, CulturedABSTRACT
"Orphan" parvovirus (OPV) infection in laboratory mice and rats was serologically surveyed for 465 mouse sera and 271 rat sera collected from 1986 to 1987 and from 1993 to 1996 in Japan. The results suggest that parvovirus infection is rare in mice but common in rats (positive rate: 13-22%) and that most putative viruses were OPVs. OPV is therefore considered to already have been harbored for at least ten years in Japan.
Subject(s)
Animals, Laboratory , Mice , Parvoviridae Infections/veterinary , Rats , Rodent Diseases/epidemiology , Animals , Japan/epidemiology , Parvoviridae Infections/epidemiology , PrevalenceABSTRACT
Murine pathogenic Escherichia coli O115a,c:K(B) (MPEC) is the causative agent of mouse megaenteron, the pathology of which resembles that of transmissible murine colonic hyperplasia caused by Citrobacter rodentium. We compared their genetic and pathological features to reveal the relationship between these two bacteria. To evaluate the genetic distances, 16S rDNA genes were sequenced and biochemical reactions were tested. Mouse strain susceptibility tests, using CF1 MPEC-susceptible germfree mice and BALB/cA(Jic) resistant mice were performed. MPEC strains and C. rodentium showed more than 99.6% identity by comparison of 16S rDNA gene sequences. All results from biochemical reactions and the mouse strain susceptibility tests were identical. It is proposed that MPEC should be reclassified as C. rodentium.
Subject(s)
Citrobacter freundii/genetics , Escherichia coli/genetics , Rodent Diseases/microbiology , Animals , Citrobacter freundii/classification , Citrobacter freundii/pathogenicity , Colon/pathology , Colonic Diseases/microbiology , DNA, Bacterial/chemistry , Disease Susceptibility , Escherichia coli/classification , Escherichia coli/pathogenicity , Female , Germ-Free Life , Hyperplasia/microbiology , Male , Mice , Mice, Inbred BALB C , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Sequence HomologyABSTRACT
Protein A was applied as a reagent for the secondary reaction in ELISA (protein A-ELISA). Mouse hepatitis virus antibody in 6 prevalent mouse strains or stocks reared in a MHV-contaminated room was effectively detected by protein A-ELISA, whereas significant strain differences in the antibody detection rate were demonstrated using the complement fixation test. C57BL/6 mice were particularly reactive in the protein A-ELISA test.
Subject(s)
Antibodies, Viral/analysis , Hepatitis, Viral, Animal/diagnosis , Mice, Inbred Strains/microbiology , Mice/microbiology , Murine hepatitis virus/immunology , Animal Husbandry , Animals , Animals, Laboratory , Enzyme-Linked Immunosorbent Assay , Hepatitis, Viral, Animal/immunology , Murine hepatitis virus/isolation & purification , Species Specificity , Staphylococcal Protein AABSTRACT
The rat retrotrapezoid nucleus (RTN) contains neurons that have a well-defined phenotype characterized by the presence of vesicular glutamate transporter 2 (VGLUT2) mRNA and a paired-like homeobox 2b (Phox2b)-immunoreactive (ir) nucleus and the absence of tyrosine hydroxylase (TH). These neurons are important to chemoreception. In the present study, we tested the hypothesis that the chemically-coded RTN neurons (ccRTN) (Phox2b(+)/TH(-)) are activated during an acute episode of running exercise. Since most RTN neurons are excited by the activation of perifornical and lateral hypothalamus (PeF/LH), a region that regulates breathing during exercise, we also tested the hypothesis that PeF/LH projections to RTN neurons contribute to their activation during acute exercise. In adult male Wistar rats that underwent an acute episode of treadmill exercise, there was a significant increase in c-Fos immunoreactive (c-Fos-ir) in PeF/LH neurons and RTN neurons that were Phox2b(+)TH(-) (p<0.05) compared to rats that did not exercise. Also the retrograde tracer Fluoro-Gold that was injected into RTN was detected in c-Fos-ir PeF/LH (p<0.05). In summary, the ccRTN neurons (Phox2b(+)TH(-)) are excited by running exercise. Thus, ccRTN neurons may contribute to both the chemical drive to breath and the feed-forward control of breathing associated with exercise.