Stereotypies in Laboratory Mice — Quantitative and Qualitative Description of the Ontogeny of ‘Wire-gnawing’ and ‘Jumping’ in Zur:ICR and Zur:ICR nu

The ontogeny of two stereotypic patterns, wire-gnawing and jumping, was studied in 24 laboratory mice: six males and six females each of two closely related outbred strains, kept under standard housing conditions, a conventional albino strain (ICR) and a nude, athymic mutant (ICR nu; hereafter: NU). All 24 individuals developed wire-gnawing after weaning at 20 d of age. In ICR one female and in NU five males and three females additionally developed jumping. ICR developed wire-gnawing between the age of 20 and 30 d, in NU jumping started at the age of 20 d, but intense jumping and wire-gnawing comparable to that of ICR did not develop in NU before the age of 40–50 d. Within each strain there was no significant difference between males and females with respect to the development of stereotypic behaviour. By contrast, ICR showed significantly more wire-gnawing but less jumping than NU. Stereotypy level increased with age up to a mean of 10.7 % of total activity in ICR and up to 7.4 % in NU at 100 d of age. However, there was huge inter- and intra-individual variability with respect to all parameters assessed in this study, i.e. total duration, number of bouts and bout length of the two stereotyped patterns. Wire-gnawing developed from outside-directed explorative climbing at the cage lid, whereas the source behaviour pattern (Mason 1991 a, Anim. Behav. 41, 1015–1037) of jumping was outside-directed explorative rearing at the cage wall. At 20 d of age, before the onset of stereotypy development, ICR showed significantly more climbing but less rearing than NU. Physical retardation of NU at weaning may account for decreased climbing ability during early ontogeny, and hence for the retarded development of wire-gnawing. The difference in early experience with either of the two patterns rather than genetic effects may be responsible for the qualitative difference between the strains with respect to the form of later stereotypy.     

Age and strain dependence of O6-methylguanine DNA methyltransferase activity in mice

The activity of the DNA repair protein O6-methylguanine DNA methyltransferase (MT) was compared in liver extracts from female ICR and male C57BL/6 mice at various ages (3-130 weeks old). Similar patterns of overall enzyme activity were observed in both strains with O6-MT activity being relatively low in young mice (3 or 8 weeks old). However, the activity significantly increased after adolescence (middle age), thereafter decreasing with old age (over 100 weeks old) to a level equivalent to that found in young mice. In an additional strain difference study, O6-MT activities in liver extracts from 4 strains of mice were compared at 5 and 30 weeks of age. Although a similar age-associated increase of enzyme activity in adolescence was confirmed in all 4 strains investigated, the closed-colony ICR mice differed from the inbred strains in demonstrating significantly higher levels of O6-MT activity in females than in males. However, the same tendency was also observed in a comparison of the sexes in 30-week-old C3H/HeN, C57BL/6 and BALB/c mice.     

Development of nephrotic ICGN mice--the origin, reproductive ability, and incidence of glomerulonephritis

ICGN is a strain of spontaneous nephrotic mice with nonproliferative glomerular lesions. It was derived from an outbred Yok: ICR colony in our laboratory. The renal disease constantly occurred in animals of the first to the tenth generations (greater than 13.0%; 70 days of age). When affected males were mated with unaffected females, the incidence of the disease in their offspring was 38.8% (n = 49) at 70 days after birth. When both parents were affected, their offspring were all affected (n = 12). The disease evenly progressed in both sexes. It usually began 40 to 150 days after birth and death occurred within two months after onset. The animals usually showed sufficient reproductive ability as long as unaffected females were used for mating.     

Estimation of structural changes in the cataractous rat lens using Raman spectroscopy.

For quantitative evaluation of cataract-related changes in lens proteins and lens water, the relative contents of water and SH residues and changes in the microenvironments of aromatic amino acid residues were quantitatively examined in cataract of the rat lens which had been induced by sodium selenite. Using Raman spectroscopy, results were compared with those of age-matched control lenses. The relative contents of water and SH residues decreased with increasing age in normal lenses from 3 to 8 weeks of age. In the cataractous lens, the relative water content increased constantly as compared with that of age-matched controls. The relative SH residue content continued to decline in the cataractous lenses of animals at every age. The microenvironments of tyrosine residues in cataractous lenses also changed progressively.     

The M16 Mouse: An Outbred Animal Model of Early Onset Polygenic Obesity and Diabesity

Objective: To characterize the phenotypic consequences of long‐term selective breeding for rapid weight gain, with an emphasis on obesity and obesity‐induced diabetes (diabesity). Research Methods and Procedures: M16 is the result of long‐term selection for 3‐ to 6‐week weight gain from an ICR base population. Experiment 1 characterized males from both lines for body weights (3, 6, and 8 weeks), feed (4 to 8 weeks) and H₂O (6 to 8 weeks) consumption, and heat loss, body composition, and levels of several plasma proteins at 8 weeks of age. Experiment 2 characterized differences between lines for both sexes at three ages (6, 8, and 16 weeks) and fed two diets (high and normal fat). Body weight, composition, blood glucose, and plasma insulin and leptin levels were evaluated after an 8‐hour fast. Results: At all ages measured, M16 mice were heavier, fatter, hyperphagic, hyperinsulinemic, and hyperleptinemic relative to ICR. M16 males and females were hyperglycemic relative to ICR, with 56% and 22% higher fasted blood glucose levels at 8 weeks of age. Discussion: M16 mice represent an outbred animal model to facilitate gene discovery and pathway regulation controlling early onset polygenic obesity and type 2 diabetic phenotypes. Phenotypes prevalent in the M16 model, with obesity and diabesity exhibited at a young age, closely mirror current trends in human populations.     

Characterization of dominant hereditary cataract in DDD/1 mice

We found a female cataractous DDD/1-nu/+ mouse and established a hairy mutant strain (DDD/1-Cti/Cti) with 100% incidence of cataract from it by repeating sibmating. Genetic studies demonstrated that a single autosomal semidominant gene controls cataractogenesis. This gene was named Cti. In homozygotes, DDD/1-Cti/Cti, the lenses began to opacify at 14 days of fetal life and were recognized clinically as cataract at 13-14 days of age when the eyes first open. The opacification became more and more intense with age and looked like mature cataract at 28-42 days of age. However, clarification of the opacified lenses commenced at the periphery after 56 days of age and expanded to the inside with time, and only an opaque spot was left at the center at 140 days of age. In heterozygotes, DDD/1-Cti/+, the lenses were recognizable as cataract after 28 days and became like mature cataract around 35 days of age. The opacity began to be lightened at 42 days and the lenses appeared normal at 56 days of age. Both lenses and eyeballs developed in similar courses in DDD/1(-)+/+, -Cti/+ and -Cti/Cti, although slightly retarded in the last. Microphthalmia was not accompanied even in DDD/1-Cti/Cti. The lens water content remained higher during the time when intense lens opacity continued in DDD/1-Cti/Cti and -Cti/+. Background genes appeared to affect the expression of Cti. DDD/1-Cti(-)+ mice may provide a model for researches into clarification of opaque lenses. A discussion concerning the possible allelism of Cti and Cts with Lop was made based on their phenotypic characteristics.     

Effect of the energy density of non-purified diets on reproduction, obesity, alopecia and aging in mice.

Four diets with graded levels of energy at 24% crude protein were fed to C57BL/6J mice for approximately 3 years to develop pelleted non-purified diets. The nitrogen-corrected metabolizable energy (MEn) of the diets ranged from 2.86 to 3.73 kcal per g of dry matter (DM). Fifteen males and 30 females were assigned to each diet. The mice were housed together for 1 week at 7 week intervals, experiencing 5 matings. After the reproduction stage, they were allowed to complete their life span. Moribund mice after 60 weeks of age were subjected to histopathological examination. The highest energy diet showed the following results in comparison with the lowest energy diet: (1) weaning weight of pups increased by 31.6%; (2) males showed slight obesity even during the reproduction stage, but females did not; (3) both sexes developed remarkable obesity after 50 weeks of age with 41.2% (males) and 49.6% (females) increasing in maximum body weight; (4) although daily feed intake decreased by approximately 18%, the MEn was slightly over consumed; (5) females showed higher incidence of alopecia with age; (6) the survival rate after 50 weeks of age decreased earlier and life span was shortened; (7) histopathological changes associated with aging developed earlier. On the other hand, the early death rate of dams at parturition increased with a decrease in dietary energy. It was concluded that at least 2 types of diets are needed throughout the life span of C57BL/6J mice; a high energy diet with an MEn value of 3.73 kcal/g DM for maximum reproduction and a low energy diet with an MEn value of 2.86 kcal/g DM for the long term stage after reproduction to retard obesity and aging most effectively.     

Breeding and growth of the inbred strain of seizure-prone Mongolian gerbils (Meriones unguiculatus)

Recently a Mongolian gerbil has become a useful laboratory animal, while little of the basic data are known about the breeding and growth of this laboratory rodent. In our animal center, an inbred strain of the seizure prone gerbil has been produced and kept these ten years. The data on the breeding and growth of the animal were obtained and described as following in the present paper. The mean life span was 26 +/- 14 month old for males and 27 +/- 13 month old for females. The mean body weight of the Mongolian gerbils at 16 weeks of age was 72 +/- 5.6 g for males and 62 +/- 7.3 g for females. The mean litter size at birth was 4.8 +/- 1.7 head. (range; 1-9) More newborn pups were born in the warm seasons (March-October) than the cold seasons (November-February). The mean rate of weaning was 80.2%. The man litter size was relatively constant in each generation.     

Life span and spontaneous tumors incidence of the Wistar Mishima (WM/MsNrs) rat.

The life spans and spontaneous tumors in a total of 1960 Wistar Mishima (WM/MsNrs) rats, inbred strain, from the 80-130th generations were examined. The average life span (mean +/- SD) was 731 +/- 173 days (n = 1053) in the males and 813 +/- 214 days (n = 907) in the females (p < 0.0001). The average life span of tumor-afflicted females was significantly longer than that of the non-tumor group (p < 0.0001), while no such difference was observed in males. Tumors were observed in 33 males (3.1%) and 246 females (27.1%). In the males, tumors were often observed under the skin (2.2%). Frequencies of tumors in lung and liver, bones and intestine were less than 0.5%. In the females, incidence of mammary tumor was 20.1%, and various organs such as ovaries, uterus, bones, lung, and liver had tumor incidence frequencies of less than 3.5%. It was concluded that WM/MsNrs rats might be suitable for life span and age-related studies because of their characteristics of length of longevity and the low incidence of spontaneous tumors in both sexes.     

Comparison of post-translational modifications of alpha A-crystallin from normal and hereditary cataract rats

Summary. In order to investigate the relationship between lens opacities and the various modifications of lens proteins, we analyzed and compared the properties of lens proteins of 85-day old normal Wistar rats and the hereditary cataract model, ICR/f rats. The present study identified many differences between normal and mutant lens proteins. In the ICR/f mutant rats, the relative amounts of gamma-crystallin decreased and high molecular weight (HMW) protein increased. Racemization and isomerization of Asp-151 of alpha A-crystallin was observed in the mutant ICR/f rats, and Met-1 of alpha A-crystallin was oxidized to methionine sulfoxide. These modifications were not found in the age-matched normal rats. These tendencies are consistent with aged and cataractous human lenses.     

先天性与后天性大鼠白内障晶体中游离氨基酸含量变化的研究

用药物诱发大鼠先天性白内障动物模型,检测了晶体中１８种游离氨基酸（ＦＡＡ）含量,并与大鼠硒性和半乳糖性白内障晶体中ＦＦＡ含量进行了比较,发现：仔一代和仔二代先天性白内障晶体中各ＦＡＡ含量接近;先天性白内障晶体内,酪氨酸和羟脯氨酸含量明显高于正常对照组（Ｐ＜０．０５）;谷氨酸,甘氨酸等９种ＦＡＡ明显低于硒性白内障（Ｐ＜０．０５）;而酪氨酸高于半乳糖性白内障组,半胱氨酸等５种ＦＡＡ含量均低于半乳糖性白内障组。同时还发现：硒性和半乳糖性白内障晶体内一些ＦＡＡ变化与以往报道不同,这些现象提示先天性与后天性白内障病变过程可能不同。     

Background pathology in BDF1 mice allowed to live out their life-span

Fifty male and 50 female BDF1 mice were observed allowing them to live out their life-span. Mortality up to 104 weeks of age was higher in males (42%) than in females (34%), and the 50% survival age was 112 weeks for males and 118 weeks for females. Body weight reached the peak at 82 weeks of age in males and 92 weeks of age in females, showing the mean body weight of 54.3 g for males and 48.0 g for females. The incidence of calculi and proteinaceous casts in the kidneys, that were not associated with exposure to chloroform, cell-alteration in the adrenal cortex, and islet cell hyperplasia in the pancreas was significantly higher in males than in females. On the other hand, hyaline droplet degeneration of the renal tubular epithelium, spindle cell proliferation in the adrenal cortex and milk-retention in the mammary glands occurred at a significantly higher incidence in females than in males. Cerebral mineralization in both sexes, atrophy and calcification of the testes and enlargement of the seminal vesicles of males, as well as cyst-formations in the ovary and endometrium of females developed at a very high incidence. Frequent neoplasms in males were hepatocellular adenomas and carcinomas, blood vessel tumours, pulmonary adenomas and carcinomas, and malignant lymphomas. In females, malignant lymphomas were the most common neoplasm, followed by blood vessel tumours, chromophobe pituitary adenomas and hepatocellular adenomas. Hepatocellular carcinomas developed only in males, whereas the histiocytic and lymphocytic types of malignant lymphomas and chromophobe cell adenomas arose solely or at a significantly higher incidence in females than in males.     

Characteristics of spontaneous erythema appeared in hairless rats.

The hairless rat (WBN/Kob-Ht), a dominant mutant rat derived from the Wistar strain, rarely develops spontaneous erythema of a progressive nature on its skin. Erythema was first observed at 8 weeks of age and the incidence at 20 weeks of age was about 4% in both males and females. Histopathologically, erythema was characterised by dermatitis induced by an immunological reaction. Areas of erythema in the skin were decreased by treatment with dexamethasone (1 mg/kg) or ciclosporin (25 or 50 mg/kg). These results suggested that erythema on the hairless rat could be used as an animal model of spontaneous dermatitis.     

Analysis of the chromosome complement in outbred mouse sperm fertilizing in vitro

The chromosome complements in a population of mouse sperm from random-bred ICR donors were analyzed at first-cleavage metaphase after in vitro fertilization (IVF) of oocytes from females of the same strain. The sperm were aged as donations occurred within an average of 31 days, either since last mating or at arrival at the animal facility in the case of virgin males. Of a total of 598 sperm complements studied from 22 sexually mature males aged 10–26 weeks old, there was one diploid complement (0.17%). The frequencies of hyperhaploidy and structural aberrations that were studied in 338 complements were 4.4% and 3.6%, respectively, giving an overall frequency of 8.0%. The hyperhaploid complements consisted of n + 1, n + 2, n + 3, and n + 7 counts, while the structural abnormalities were of the chromosome type and included large and small fragments and a possible translocation. This is the highest frequency of sperm chromosome abnormalities reported for mouse sperm obtained from males under physiological conditions and fertilized in vitro or in vivo. Sperm aging, strain, and/or technique differences are among the factors that may be responsible for this high frequency. Since the 8.0% frequency of hyperhaploidy and structural abnormalities is similar to the frequency reported for human sperm after IVF, the outbred murine in vitro fertilization system may be a useful model to study the origin of human sperm chromosome abnormalities.     

Mortality, hemodynamics, and aortic properties among male and female turkeys fed beta-aminopropionitrile

Broad-Breasted White turkeys were randomized into four treatment groups at 4 weeks of age in each of three trials. Group 1, males, and Group 2, females, were fed an unsupplemented (control) diet from 4 to 10 weeks of age. Group 3, males, and Group 4, females, were fed the control diet supplemented with 0.07% beta-aminopropionitrile (BAPN). The experiments were terminated at 10 weeks of age. There was no mortality among male and female turkeys fed the control diet or significant differences in blood pressure, heart rate, aortic tensile strength, or aortic hydroxyproline between these males and females. Sixty-five percent of the males and 21% of the females fed BAPN died of dissecting aneurysms; blood pressure, heart rate, and dp/dt max were not altered as a result of feeding BAPN. Aortic tensile strength was higher in control turkeys than those fed BAPN, but males fed BAPN had the lowest value. Males fed BAPN also had the lowest aortic hydroxyproline content. Ultrastructural alterations of aortic elastic and collagen fibers were more severe in males than females fed BAPN. Aortic changes did not occur in control turkeys.     

Inherited hydrocephalus in Csk: Wistar-Imamichi rats; Hyd strain: a new disease model for hydrocephalus

Hydrocephalic rats were found in a breeding colony of Csk: Wistar-Imamichi strain rats. In males, the hydrocephalus were serious and could be detected from 7 days after birth. Survival was 3-4 weeks. In females, the hydrocephalus was moderate, there was no abnormal external appearance, and the rats were able to mature. Ventricular dilatation was excessive in males but moderate in females. The total frequency of hydrocephalus was 34.3% in both males and females. Breeding data indicated that this disease is heritable and is single dominant and X-linked (symbol, Hyd). The female moderate hydrocephalics could be detected by progeny tests without examining brain sections. No evidence of developmental anomaly was observed in the ventricles. This hydrocephalus was classified as being of the communicating type, and this strain was named the Hyd strain as an animal model for human hydrocephalus.     

Accumulation of the hydroxyl free radical markers meta-, ortho-tyrosine and DOPA in cataractous lenses is accompanied by a lower protein and phenylalanine content of the water-soluble phase

Post-translational modifications of lens proteins play a crucial role in the formation of cataract during ageing. The aim of our study was to analyze protein composition of the cataractous lenses by electrophoretic and high-performance liquid chromatographic (HPLC) methods. Samples were obtained after extracapsular cataract surgery performed by phacoemulsification technique from cataract patients with type 2 diabetes mellitus (DM CAT, n = 22) and cataract patients without diabetes (non-DM CAT, n = 20), while non-diabetic non-cataractous lenses obtained from cadaver eyes served as controls (CONTR, n = 17). Lens fragments were derived from the surgical medium by centrifugation. Samples were homogenized in a buffered medium containing protease inhibitor. Soluble and insoluble protein fractions were separated by centrifugation. The electrophoretic studies were performed according to Laemmli on equal amounts of proteins and were followed by silver intensification. Oxidized amino acid and Phe content of the samples were also analyzed by HPLC following acid hydrolysis of proteins. Our results showed that soluble proteins represented a significantly lower portion of the total protein content in cataractous lenses in comparison with the control group (CONTR, 71.25%; non-DM CAT, 32.00%; DM CAT, 33.15%; p < 0.05 vs CONTR for both). Among the proteins, the crystallin-like proteins with low-molecular weight can be found both in the soluble and insoluble fractions, and high-molecular weight aggregates were found mainly in the total homogenates. In our HPLC analysis, oxidatively modified derivatives of phenylalanine were detected in cataractous samples. We found higher levels of m-Tyr, o-Tyr and DOPA in the total homogenates of cataractous samples compared to the supernatants. In all three groups, the median Phe/protein ratio of the total homogenates was also higher than that of the supernatants (total homogenates vs supernatants, in the CONTR group 1102 vs 633 micromol/g, in the DM CAT group 1187 vs 382 micromol/g and in the non-DM CAT group 967 vs 252 micromol/g; p < 0.05 for all). In our study we found that oxidized amino acids accumulate in cataractous lenses, regardless of the origin of the cataract. The accumulation of the oxidized amino acids probably results from oxidation of Phe residues of the non-water soluble lens proteins. We found the presence of high-molecular weight protein aggregates in cataractous total homogenates, and a decrease of protein concentration in the water-soluble phase of cataractous lenses. The oxidation of lens proteins and the oxidative modification of Phe residues in key positions may lead to an altered interaction between protein and water molecules and thus contribute to lens opacification.     

Dynamics of energy utilization in male and female turkeys during growth

Determining energy utilization in growing animals enables to adjust the nutritional constraints to nutrient requirements while maximizing the ratio between lean retention and fat retention to improve feed efficiency. In turkey production, the important sexual dimorphism and differences between strains may contribute to differences in basal energy metabolism and the partitioning of energy retention between protein and lipid. The objective of this study was to determine the dynamics of energy utilization in males and females of a heavy strain of turkeys fed ad libitum from 1 to 23 weeks of age. Heat production (HP) was determined by indirect calorimetry and retained energy (RE) was calculated as the difference between metabolizable energy (ME) intake and HP. The RE as protein was determined by a nitrogen balance, while the remaining RE was assumed to be lipid. A modeling procedure allowed partitioning HP between fasting HP (FHP), activity-related HP and thermic effect of feeding. A multiple regression analysis was used to estimate the maintenance energy expenditure (ME(m)) and the energy efficiencies of protein and lipid retention (k(p) and k(f), respectively). Results were expressed either per day or per kg BW(0.75) per day. In comparison with females, males consumed more feed (440 v. 368 g/day), grew faster (163 v. 147 g/day) and retained more protein (38 v. 28 g/day) during the experimental period. Expressed per kg BW(0.75) per day, ME intake decreased linearly with increasing age and was not affected by gender. Similarly, RE as protein decreased with increasing age and tended to be greater in males than in females, whereas RE as lipid increased with increasing age and was lower in males than in females. In addition, HP decreased with increasing age and was greater in males than in females, because of greater activity-related HP and FHP (47% and 9% greater in males compared with females). The FHP averaged 417 kJ/(kg BW)(0.75) per day during the first 3 weeks of age and decreased to 317 and 277 kJ/(kg BW)(0.75) per day in males and females, respectively, from 20 weeks of age onwards. Similar to FHP, ME(m) was lower in females than in males ((586 to 12 × BW) and (586 to 5 × BW) kJ/(kg BW)(0.75) per day, respectively) and the k(p) and k(f) were estimated at 0.63 and 0.87, respectively. This study shows that the partitioning of RE and HP differs between genders in growing turkeys, which likely results in differences in nutrient requirements.     

Study on the ophthalmic diseases in ICR mice and BALB/c mice.

In pharmaceutical companies and research institutes, many toxicity tests are performed with laboratory animals. This study was performed to produce reference data for eye toxicity tests and to investigate the ophthalmic diseases of 408 ICR mice and 119 BALB/c mice, which are commonly used as subjects in toxicity tests. The experimental animals without clinical disorders were selected regardless of sex. The ophthalmic diseases were examined by using special ophthalmic instruments: direct ophthalmoscope, indirect ophthalmoscope, slit-lamp biomicroscope and focal illuminator. The most prevalent ocular variation within normal limits was hyaloid vessel remnant (ICR mice, 28.2%; BALB/c mice, 31.9%) and the incidence gradually decreased with age. The ocular diseases found in ICR mice were retinal degeneration (9.8%), corneal scar (4.2%), focal cataract (2.2%), anisocoria (1.2%), corneal ulcer (0.2%) and uveitis (0.2%). In BALB/c mice, corneal scar (9.2%), focal cataract (1.7%) and corneal ulcer (0.8%) were the ocular diseases found.     

Identification of compatibility between ooplasmic factor and sperm gene in the intersubspecific crosses involving DDK and PWK mice strains

The DDK strain (Mus musculus domesticus) of inbred mouse has a unique peculiarity known as DDK syndrome.The DDK females are mostly infertile when crossed with males of other inbred strains,while DDK males exhibit normal fertility in the reciprocal crosses,as intrastrain matings.This DDK syndrome has been demonstrated to be caused by an incompatibility system between DDK ooplasmic factor and the sperm gene of other strains owing to the ovum mutant (Om) locus on mouse Chromosome 11.Recently,it was reported that DDK females are fully fertile when crossed to males of MOM (M.m.molossinus) and CASP (M.m.castaneus) strains,indicating that no incompatibilities exist between DDK ooplasmic factor and sperm gene of MOM or CASP males.In the present study,DDK females were found to be also fully fertile when crossed to the males of PWK wild-derived inbred strain (originated from Czech Republic wild mice,M.m.musculus).The crosses of DDK females × F1 (DDK♀ × PWK♂) males also resulted in normal fertility.Furthermore,the transmission ratios of Om alleles from these F1 males to their backcross N2 offspring are 50％∶50％ as genotyped by microsatellite markers closely linked to Om locus.Moreover,it was demonstrated that PWK females are also fully fertile when crossed to DDK males.All above results indicated that no incompatibility exists between ooplasmic factor and sperm gene in the intersubspecific crosses with DDK and PWK strains.PWK strain would also be useful for further investigations on the DDK syndrome,and DDK strain can be used more widely for various studies in the mouse.