Status Of Mice Research Articles

Gene expression profiling using a mouse full-length 20 K cDNA microarray

Most public-founded large-scale sequencing projects that use cDNAs are primarily using cDNA libraries which are not enriched for full-length cDNAs. Consequently, only a fraction of the resulting ESTs matches the 5' end of the original transcript. The target of the Genome Science Laboratory of RIKEN is to clone and sequence the largest number possible of full-length mouse cDNAs in two phases. The first phase is to classify the cDNAs and the second is to complete full-length sequencing and functional annotations. We have developed two original methods to construct full-length cDNAs efficiently: 'cap-trapper', which preferentially recognizes the Cap site of mRNA; and the 'trehalose-thermoactivated reverse transcriptase', which allows the reverse transcriptase reaction at higher (60°C) temperatures. We have constructed over 80 libraries from embryonic tissues of different developmental stages and adult tissues to ensure the greatest possible coverage of the expressed mRNA. More than 200,000 successful sequencing passes have been performed with the use of two tools developed in-house: a high-throughput plasmid preparation system and the RISA 384 capillary sequencer. Most of the sequences were performed from the 3' end to select individual cDNAs. We have selected more than 30,000 different cDNAs. Using these sets of RIKEN full-length cDNA, we have established gene expression microarrays containing a 20 K set of RIKEN full-length cDNA unique mouse genes (http://genome.rtc.riken.go.jp). This set has been used to profile expression patterns of various adult and embryonic tissues. Target DNAs were PCR amplified and printed on Poly-L-lysine coated glass slides. Target DNAs were blocked by excess amounts of Cot1DNA. Probes were labelled by two-colour fluorescent dye using random primer and reverse transcriptase. Normalization has been achieved using a global normalization method. We have also developed a program to filter the noise. The experiment was done twice and reproducible results were extracted and clustered. We will present a large set of data that show the spatial and temporal expression patterns of mice. These mouse full-length 20 K cDNA microarrays are widely applicable to analyse the global expression profiling of normal and diseased status of mice.

Effects of Lactic Acid Bacteria Ingestion on Basal Cytokine mRNA and Immunoglobulin Levels in the Mouse

An increasing number of functional foods and pharmaceutical preparations containing lactic acid bacteria are being promoted with health claims based on the potential probiotic characteristics and on their capacity for stimulating the host immune system. However, the specific immune effects of oral administration of these microbes remain undefined. In this study, we tested the hypothesis that basal gastrointestinal immune status in mice is affected by orally administered lactic acid bacteria. The specific objective of this research was to evaluate the effects of repeated oral exposure to viable and nonviable lactic acid bacteria (Lactobacillus acidophilus, L. bulgaricus, L. casei, and Streptococcus thermophilus) in mice on basal cytokine mRNA expression in mucosal (Peyer's patches), systemic (spleen), and lymphoid tissue and on immunoglobulin levels. The results indicated that oral exposure to 109 CFU/day for up to 14 days did not significantly affect basal interferon-γ, tumor necrosis factor-α, or interleukin-6 mRNA expression or total serum and intestinal immunoglobulins.

Pharmacological Studies of Mentha longifolia Phenolic Extracts. II. Hepatoprotective Activity

The effects of Mentha longifolia crude ethanolic extract (F 1), rich in luteolin glycosides (F 2), apigenin glycosides (F 3) and phenolic acids (F 4), were studied on CCl 4 -induced liver injury in mice. Administration of CCl 4 -induced significant impairment in hepatic antioxidant status, decreasing the glutathione content and superoxide dismutase activity, and stimulating lipid peroxidation. Pretreatment with Mentha longifolia extracts significantly improved hepatic antioxidant status in mice, which was most pronounced in the reduction of CCl 4 -mediated lipid peroxidation. In addition, in pretreated animals, an increase in hepatic glutathione and superoxide dismutase activity, as well as a decrease of cytochrome P450, was found.

Copper metabolism in metallothionein-null mice fed a high-zinc diet

Humans and animals develop low copper (Cu) status when fed diets containing large amounts of zinc (Zn) for an extended period. Current theory states that Zn-induced metallothionein (MT) in the intestinal mucosa binds Cu and prevents its absorption. We tested this theory by using a mouse model with a disruption in the MT gene that renders it incapable of producing functional MT-I and MT-II (MT-null). If the theory is true, then the MT-null mouse should not develop low Cu status when fed a high Zn diet. For 1 week, groups of 4-week-old MT-null and control mice were fed a diet that contained 35 mg Zn and < 1 mg Cu/kg. Each genotype was then divided into two groups each. One group was fed a diet containing 35 mg Zn and 1.5 mg Cu/kg and the other was fed a diet containing 400 mg Zn and 1.5 mg Cu/kg. After 14 days, plasma was harvested and plasma ceruloplasmin amine oxidase (CPAO) activity, a good indicator of Cu status, was determined. The plasma CPAO activity of control mice fed 400 mg Zn/kg diet was 50% of that in similar mice fed 35 mg Zn/kg. Likewise, plasma CPAO activity in MT-null mice fed 400 mg Zn/kg diet was 40% of that in MT-null mice fed 35 mg Zn/kg. These data suggest that MT induction is not required for the development of low Cu status in mice fed a high Zn diet and that the actual mechanism may involve the modulation or inhibition of a Cu transporter protein by Zn.

Evaluation of coagulation tests in mouse plasma

The coagulation variables thrombin time (TT), activated partial thrombin time (APTT) and prothrombin time (PT), were investigated in mouse plasma. TT and APTT clotting times were determined using a KC10 coagulation analyser and test kits Dia Thrombin (bovine thrombin diluted to a concentration of 2.5 U/l) and Dia Celin-L (rabbit brain cephaloplastin, activated with complexed Kaolin), respectively. PT was determined with IL Test™ Hepatocomplex (rabbit brain thromboplastin and calcium ions) using an ACL 200. Furthermore, the test procedures were also used to assess the anticoagulant status of mice treated orally with Melagatran, a thrombin inhibitor. The results showed that the kits could be used successfully on mouse plasma to measure the effect of a thrombin inhibitor on haemostasis.

Methylenedioxy group as determinant of schisandrin in enhancing hepatic mitochondrial glutathione in carbon tetrachloride-intoxicated mice

As a preliminary approach to exploring whether the methylenedioxy group of the dibenzocy-clooctadiene skeleton of schisandrins plays an important role in hepatic mitochondrial-reduced glutathione (GSH) stimulatory activity, we examined the effects of three schisandrins, namely schisandrin A (Sch A), schisandrin B (Sch B), and schisandrin C (Sch C), on carbon tetrachloride (CCl 4) hepatotoxicity and hepatic mitochondrial GSH status in mice. Pretreating mice with Sch A at a daily oral dose of 1 mmol/kg for 3 days did not protect against CCl 4 hepatotoxicity, whereas pretreatment with Sch B or Sch C at the same dosage regimen produced almost complete protection. The hepatoprotection afforded by Sch B or Sch C pretreatment was associated with significant increases in the hepatic mitochondrial GSH level and glutathione reductase (EC 1.6.4.2) activity. Our results indicate that the methylenedioxy group of the dibenzocyclooctadiene skeleton of schisandrin is an important structural determinant in the stimulation of hepatic mitochondrial GSH, particularly under conditions of CCl 4 intoxication.

Enhanced depletion of glutathione and increased liver oxidative damage in aflatoxin-fed mice infected with Plasmodium berghei

The effect of dietary aflatoxins b 1 and g 1 and Plasmodium berghei infection on glutathione (GSH) levels and liver status in mice was investigated. Three days after intraperitoneal injection of 0.1 × 10 6 parasitized red blood cells into the mice, there was a significant fall in blood glutathione levels accompanied by a significant increase in serum cholinesterase and liver malonic dialdehyde levels in the mice fed aflatoxin compared with those in the control group. The results suggested that malaria parasites can enhance depletion of host glutathione and oxidative damage of the liver in mice fed low levels of aflatoxins.

Contrasting Effects of a Dietary Copper Deficiency in Male and Female Mice

Female rats are protected from the lethal effects of a dietary copper (Cu) deficiency, but female mice fed a Cu-deficient diet develop atrial thromboses and die. To further investigate the effect of sex on Cu status in mice (n = 16), male and female adult Swiss-Webster mice were fed Cu-supplemented (8.4 mg Cu/kg) or Cu-deficient (0.3 mg Cu/kg) diets with deionized water for 43-49 days. Six female mice, but only one male mouse, fed the Cu-deficient diet died during the experiment. Both male and female mice fed the Cu-deficient diet exhibited typical features of deficiency. The severity of anemia and the values observed for several indicators of Cu status (plasma ceruloplasmin [EC 1.16.3.1.] and erythrocyte copper-zinc superoxide dismutase [EC 1.15.1.1.] activities, cardiac Cu) were similar in both male and female Cu-deficient mice. However, cardiac enlargement (0.97 vs 0.73 g/100 g body wt, P < 0.05), cardiac edema (79.9% vs 78.2% cardiac water, P < 0.05) and depletion of renal Cu (10.4 vs 12.5 micrograms/g dry weight, P < 0.05) were more severe in female compared with male, Cu-deficient mice. Furthermore, although hepatic Cu was significantly (P < 0.05) lower in female Cu-deficient compared with Cu-supplemented mice, it was not significantly decreased by deficiency in male mice. These data indicate that the female mice experienced a more extreme form of Cu deficiency than the males.

Male-male interference in a reproductive context: effect of social status in mice

Direct competition between males in a sexual context was tested in stable male pairs. Albino male mice were exposed to their male antagonist interacting with a female on the other side of a transparent plexiglas partition provided with small holes. The behavioural response of males varied according to their social status, dominant males being more activated by the experimental procedure than subordinate males. In addition, the stimuli that elicited the dominant male's attention seemed unrelated to the female, but to the simultaneous presence of the female and the male antagonist. In a second experiment, the behavioural performance of males of different rank was evaluated when only one female was introduced into their home-cage, or when an additional female was introduced into the home-cage of the male antagonists, with the males separated by a partition. This experiment indicated that the dominant male's interest in the female was markedly reduced and exploration of the partition was enhanced when two fem...

Courtship ultrasonic vocalizations and social status in mice

A series of experiments was conducted to investigate whether the poorer sexual performance of subordinate than dominant male mice, Mus domesticus, was linked to lower sexual motivation. Ultrasonic calls uttered by a male in the presence of a female were used as an index of sexual interest. Males were housed in pairs for 5 days and dominant/subordinate roles were assigned. Subordinates, when tested in their home cage immediately after the removal of the dominant male, uttered more ultrasounds than the latter. When the dominant male was tested before the subordinate, there was no difference in the number of ultrasounds uttered and the subordinates' performance was consistently poorer. The fewer calls recorded when subordinate males were tested after the dominant partner was not associated with less defence/escape behaviour, nor could it be explained as habituation to female odour, as a consequence of being tested after the dominant partner. Within sexually experienced pairs, the urine of dominant males interacting with a female for 3 min reduced the number of ultrasounds uttered by the subordinate in the presence of a female. It is suggested that an inhibitory factor in the dominant male's urine functions as an indirect competitive mechanism when direct competition is prevented by removing the dominant subject.

Depression of Phagocytic Activity of The Immune System Following Traditional Cautery in Experimental Animals

Traditional cautery, which is practiced widely in Saudi Arabia and some other countries, has not been exposed to detailed scientific investigation. In an attempt to elucidate some of its physiological aspects, we assessed the effect of cautery on a normal animal's nonspecific immune system. Male Wistar rats and guinea pigs were cauterized to simulate traditional cautery in size and percentage of cauterized area. Using radioactive sulfur colloid uptake and clearance, the effect of cautery on the mononuclear phagocyte system was evaluated in rats. The respiratory burst in peripheral polymorphonuclear leukocytes (PMNs) after cautery was measured in the guinea pigs using the chemiluminescence technique. The results showed marked reduction in the intravascular clearance of the colloid with prolonged clearance time following cautery. In addition, the liver uptake of the colloid was reduced in cauterized animals compared with control animals and the white cell count was also significantly reduced. The study showed marked inhibition of phagocytic function in guinea pigs in both whole blood and isolated PMNs. These results indicate that traditional cautery reduces the physiological function of the phagocyctic system in normal experimental animals. The influence of traditional cautery on infected animals deserved further investigation.

Photocarcinogenicity of psoralens used in PUVA treatment: Present status in mouse and man

There is good evidence that 8-methoxypsoralen is photocarcinogenic in psoriatic patients undergoing long-term photochemotherapy (PUVA) in the U.S.A. However, this conclusion has not been supported by two major European studies which have indicated that PUVA is a tumour promoter of damage initiated by other agents. Variation in PUVA treatment protocols in the U.S.A. and Europe may partly account for the different conclusions. There is much interest in the therapeutic potential of monofunctional psoralens. It is hoped that these may reduce long-term risk. Monofunctional and cross-linking psoralens have been shown to be photocarcinogenic in mouse skin. The relative risk of different compounds may be assessed in the mouse, but it is important to base comparisons on dose protocols that have been shown to be therapeutically effective.

Copper status and adriamycin treatment: effects on antioxidant status in mice

The biochemical response of Cu-sufficient and Cu-deficient mice to adriamycin (ADR) treatment was evaluated. Mice were injected intraperitoneally with ADR (17 mg/kg body wt.) or saline (0.9% w/v) and killed 4 d after injection. There was no effect of ADR on cardiac Superoxide dismutase (SOD) activity in Cu-sufficient of Cu-deficient mice. ADR injection resulted in higher cardiac glutathione (GSH) concentrations in Cu-sufficient mice while it resulted in lower GSH concentrations in Cu-deficient mice relative to their saline-injected controls. The effects of ADR in Cu-deficient mice were tissue-specific as its administration resulted in lower hepatic SOD activity and higher hepatic GSH concentrations relative to saline-injected controls.

The moral status of mice and humans.

The moral status of mice and humans.

Changes in cellular immunity and nutritional status in mice after thermal injury

A 13 per cent body surface area (BSA), full skin thickness burn was inflicted on LACA male mice and the changes in cellular immunity and nutritional status were observed. The results showed that thymus, spleen and circulating lymphocytes were significantly involved. A diminished mitogen responsiveness of spleen cells and altered peritoneal macrophage function were confirmed. Ear swelling tests indicated that the cellular immunity of burned mice was most severely depressed in week 2 postburn. The present study also showed that the dramatic change in nutritional status occurred earlier than that in cellular immunity and suggested the importance of early nutritional support after thermal injury.

Severe or Marginal Copper Deficiency Results in a Graded Reduction in Immune Status in Mice

From birth mice received diets containing copper at 0.5, 1, 2 or 6 mg/kg diet. At 8 wk of age they were killed and copper status and immune responsiveness were determined. Only the groups that received copper at 0.5 or 1 mg/kg showed signs of copper deficiency, such as reduced serum ceruloplasmin, hemoglobin, hematocrit and red blood cell counts and characteristic changes in organ pathology. Body and lymphoid organ weights were altered in the groups that received copper at 0.5 or 1 mg/kg. Males were more severely affected than females. A dose-related reduction in splenic T-cell subpopulations was noted in the 0.5 and 1 mg/kg groups. Responses to lipopolysaccharide challenge were reduced, and an increase in spontaneous cycling cells was noted in the groups receiving copper at 0.5 or 1 mg/kg. Only the group receiving copper at 0.5 mg/kg had increased stem cell activity; this increase was probably due to increased erythropoiesis to meet increased demands for red blood cells in this group. These data indicate that only groups receiving copper at 0.5 or 1 mg/kg in the diet were depleted and marginally depleted in copper, respectively, and that immune hyporesponsiveness differs between the depleted and marginally depleted groups.

The moral status of mice.

The moral status of mice.

Humoral and cell-mediated immune status of mice exposed to 1,1,2-trichloroethane.

The purpose of this study was to assess the immunological effects of 1,1,2-trichloroethane (TCE) on random-bred CD-1 mice following 14 and 90 days of oral exposure. A toxicological evaluation conducted at the same time revealed the target organs to be the liver of both sexes and the erythroid elements of the females. The 14-day immunological range-finding study in males exposed to doses 1/10 and 1/100 the LD50 (38 and 3.8 mg/kg) revealed no alterations in either humoral or cell-mediated immune status. Following 90 days of exposure in the drinking water (4.4., 46, and 305 mg/kg for males and 3.9, 44, and 384 mg/kg for females), a more detailed series of immunological parameters was assessed. Cell-mediated immunity was unaltered in both sexes, while humoral immune status was depressed in both sexes, particularly when determined by hemagglutination titers. Macrophage function was depressed only in the males as indicated by the ability of thioglycolate-recruited peritoneal exudate cells (PEC) to phagocytize sheep erythrocytes (sRBC).

Humoral and cell-mediated immune status of mice exposed to trans-1,2-dichloroethylene.

This study assessed possible adverse immunological effects of trans-1,2-dichloroethylene (DCE) on random-bred CD-1 mice following 14 and 90 days of exposure. A 14-day range-finding study was performed on male mice by gavage at doses 1/10 and 1/100 the LD50 (210 and 21 mg/kg). No alterations in either humoral or cell-mediated immunity were observed following this exposure. A 90-day study was conducted in which DCE was administered in the drinking water of male and female mice. The levels of DCE in the drinking water were calculated to deliver levels equivalent to, and higher than, those delivered for 14 days (17, 175, and 387 mg/kg for males and 23, 224, and 452 mg/kg for females). No changes were observed in the cell-mediated immune status of either sex or in the humoral immune status of females. However, a marked suppression in humoral immune status was observed in male mice exposed to all three levels of DCE, as indicated by a decreased ability of spleen cells to produce antibody against sheep erythrocytes (sRBC). Macrophage function was depressed only in females, as indicated by the decreased ability of thioglycollate-recruited peritoneal exudate cells (PEC) to phagocytize sRBC.

Humoral and cell-mediated immune status in mice exposed to chloral hydrate.

Chloral hydrate has been found in our drinking water supplies at levels up to 5 micrograms/1. The purpose of this study was to evaluate the functional status of the immune system in random-bred CD-1 mice exposed to chloral hydrate for 14 and 90 days. Male mice, following 14 or 90 days of exposure to 1/10 and 1/100 the actual oral LD50, exhibited no alterations in either humoral or cell-mediated immunity. However, female mice exposed for 90 days to chloral hydrate in the drinking water demonstrated a significant depression in humoral immune function. This depression was observed when spleen cells from exposed mice were evaluated for their ability to produce antibody against sheep erythrocytes. These females did not demonstrate any changes in cell-mediated immune status.