Circadian wheel-running activity rhythms are unaltered by acute cage enrichment with nests and shelters

C57BL/6J inbred male mice were assessed for the effects of cotton nesting material on circadian running-wheel assays at 60, 157 and 222 days of age. The oldest mice were also tested for effects of access to plastic igloo shelters and the combination of cotton plus igloos. All mice were assayed for mean activity level, acrophase and entrained circadian period in 12:12 LD, and mean activity level and free-running period in DD for circadian wheel-running activity. There were no significant effects, at any age, of either nesting material or shelters, or the combination of both, on any measures of circadian running-wheel activity. We conclude that circadian rhythm assessment is unlikely to be significantly altered by the inclusion of enrichment materials in running wheel cages.     

一种新型叶螨饲养器的设计与制作

分析了现有的叶螨个体饲养或定量饲养的方法或器具的缺陷。根据酢浆草如叶螨Tetranychina harti(Ewing)及其寄主植物红花酢浆草(Oxalis corymbosa DC)的特点,设计、制作了一种新型饲养器,解决了常用的离体湿基叶片法中叶片表面温湿度与设定的环境条件不一致、叶螨生活状态不自然和活体叶片法中镜检观察不方便的问题,取得了较好的效果。本设计在其他叶螨或微小昆虫的饲养中也可以作为借鉴。     

A model for studies of the deformable rib cage.

An earlier model for the study of rib cage mechanics was modified so that rib deformity in scoliosis could be better represented. The rigid ribs of that model were replaced by five-segment deformable ribs. Literature data on cadaver rib mechanical behavior were used to assign stiffnesses to the new individual model ribs so that experimental and model rib deflections agreed. Shear and tension/compression stiffnesses had little effect on individual rib deformation, but bending stiffnesses had a major effect. Level-to-level differences in mechanical behavior could be explained almost exclusively by level to level differences in the rib shape. The model ribs were then assembled into a whole rib cage. Computer simulations of whole rib cage behaviors, both in vivo and in vitro, showed a reasonable agreement with the measured behaviors. The model was used to study rib cage mechanics in two scolioses, one with a 43 degrees and the other with a 70 degrees Cobb angle. Scoliotic rib cage deformities were quantified by parameters measuring the rib cage lateral offset, rib cage axial rotation, rib cage volume and rib distortion. Rib distortion was quantified both in best-fit and simulated computer tomography (CT) scan planes. Model rib distortion was much smaller in best-fit planes than in CT planes. The total rib cage volume changed little in the presence of the scolioses, but it became asymmetrically distributed.     

Intracerebroventricular neuropeptide Y suppresses open field and home cage activity in the rat

Effects of intracerebroventricular administration of neuropeptide Y on open field behaviour, behavioural habituation and corticosterone response to open field testing, and on home cage activity have been investigated in the rat. In the open field, NPY reduced activity in a dose-dependent manner. Behavioural habituation was not influenced. After 5 days of recovery, NPY-treated animals did not differ from non-treated in any of the measured parameters. Peripheral corticosterone levels were not significantly affected, although there was a strong tendency towards an increase. Injection of 2 nmol NPY did not produce any gross neurological deficits. At this dose, NPY greatly suppressed home cage activity. The effect lasted throughout the recording period of 22 h, abolishing the normal circadian variation in activity. After 5 days of recovery, the effect was no longer present. Our interpretation of these findings is, that NPY is a highly potent endogenous agent capable of producing certain important aspects of behavioural sedation in a reversible manner. Since NPY did not decrease the corticosterone response to a novel stimulus, its pattern of actions seems to differ from synthetic sedative drugs.     

A thermoelectrically regulated multipurpose cuvette for simultaneous time-dependent measurements

An improved design of a thermostatically controlled reaction cuvette for time-dependent biochemical measurements is described. The design is such that a multiple choice of single and simultaneous spectroscopic and electrode analyses can be performed in a sample of about 1.8 ml. This choice is quite flexible due to the use of exchangeable tapered plugs suited with either optical quartz-rod windows for absorption or fluorescence measurements or selective electrodes for changes of O₂, H₂, H⁺, etc. Temperature is accurately controlled by a thermoelectric (Peltier) module. An overhead constant-stirring device includes solute addition and gasflow ports. A bottom window allows actinic illumination for photobiological and photochemical experiments. Some examples of application in combination with commercial or laboratorymade instruments are presented.     

A new technique for simultaneous and in situ measurements of Ca2+ signals in arteriolar smooth muscle and endothelial cells

We report here the first local and global Ca(2+) measurements made from in situ terminal arterioles. The advantages of the method are that there is minimal disturbance to the vessels, which retain their relationship to the tissue they are supplying (rat ureter) and the small size of vessel that can be studied. Good loading with the Ca(2+) indicator, Fluo-4 was obtained, and confocal sectioning through the tissue enabled vascular smooth muscle and endothelial cells to be clearly seen, along with red blood cells, nerve endings and the ureteric smooth muscle cells. We find the terminal arterioles to be extremely active, both spontaneously and in response to nor-adrenaline stimulation, with Ca(2+) sparks occurring in the vascular myocytes and Ca(2+) puffs in the endothelial cells. Even under resting conditions, endothelial cells produced oscillations and waves, which could pass from cell to cell, whereas the vascular myocytes only produced waves in response to agonist stimulation, and with no increase in the frequency of Ca(2+) sparks, and no spread from cell to cell. We compare our data to those obtained in dissected intact vessels and single cells. We conclude that this approach is a convenient and useful method for studying inter- and intracellular Ca(2+) signalling events and communication between cell types, particularly in very small vessels.     

Apparatus for simultaneous spectrophotometric and polarographic measurements

We have recently constructed a 10-mm, light path quartz cuvet which will accept a Clark oxygen electrode; it is temperature controlled and is suitable for use in a Unicam (Cambridge, England) SP 800 recording spectrophotometer. Several enquiries have prompted this publication, although such an apparatus was mentioned much earlier by Chance and Williams,¹ and has been used extensively. Figure 1a, b, c, and d and their legends provide sufficient detail for the construction of the cuvet and provision of the commercially available electrode, quartz faces, stirring motor and disk magnet. Circuit diagrams for temperature control (range 22–38°C., ± 0.2°C.) and stirrer speed control are shown in Figure 2a and b. The cuvet is shown situated in the spectrophotometer cell housing in Figure 3, and the cuvet with its associated equipment is shown in Figure 4.     

Wheel-running activity and energy metabolism in relation to ambient temperature in mice selected for high wheel-running activity

Interrelationships between ambient temperature, activity, and energy metabolism were explored in mice that had been selectively bred for high spontaneous wheel-running activity and their random-bred controls. Animals were exposed to three different ambient temperatures (10, 20 and 30°C) and wheel-running activity and metabolic rate were measured simultaneously. Wheel-running activity was decreased at low ambient temperatures in all animals and was increased in selected animals compared to controls at 20 and 30°C. Resting metabolic rate (RMR) and daily energy expenditure (DEE) decreased with increasing ambient temperature. RMR did not differ between control and selected mice, but mass-specific DEE was increased in selected mice. The cost of activity (measured as the slope of the relationship between metabolic rate and running speed) was similar at all ambient temperatures and in control and selected mice. Heat generated by running apparently did not substitute for heat necessary for thermoregulation. The overall estimate of running costs was 1.2 kJ/km for control mice and selected mice.     

Lessons from the Eker rat model: from cage to bedside

Rodent models of human diseases serve a vital role in translating bench observations to bedside therapies. In vivo manipulation of these animals allows us to explore the biologic significance of the underlying molecular and biochemical pathways. The study of human cancers has been highly enriched by the observations made from numerous transgenic mouse models. Long before the techniques of genetic engineering were discovered, Dr. Reidar Eker described one of the earliest examples of an autosomal dominant model of renal tumors in a unique strain of rats. They were used in the 1980's by Alfred Knudson to validate the "two-hit" hypothesis and to study the multi-step process of carcinogenesis. Following the identification of the Tsc2 germline mutation in the Eker rat, it became the first rodent model of tuberous sclerosis and has since been exploited in many areas of tumor biology as illustrated in the content of this issue. The focus of our review is to highlight the contribution of the Eker rat towards understanding the Tsc2 signaling pathways in tumorigenesis and evaluating potential therapeutics in the pre-clinical setting.     

A rat restraining cage for IV infusion in epigastric flap studies

A rat restraining cage is described for use in epigastric flap studies requiring IV infusions of 24 hours or more. The device is simple, effective, and reliable; it appears to be less stressful for the rat than other methods of restraint.     

A model for studies of mechanical interactions between the human spine and rib cage

A three-dimensional mathematical model useful for studies of the mechanics of the human skeletal thorax is described. To construct this model, rib cage elements are incorporated into a previously reported model of the thoracolumbar spine. The vertebrae and bony portions of the ribs and sternum are idealized as rigid bodies. The behavior of the discs, ligaments and costal cartilages are modelled by deformable elements. Appropriate geometric and stiffness property data are assigned to the elements of the model. In constructing the model, it was found that the mechanical response of the costo-vertebral joint is strongly influenced by articulation geometry. Although rigid bodies were used to model calcified portions of the ribs, the model predicted rib cage deformations in close agreement with those measured experimentally. These studies indicate that the rigid body motion of calcified portions of the rib makes a major contribution to the deformation of the rib cage in response to certain types of loadings. Quantitative results are also reported on the roles the rib cage plays in bending responses of the spine, the lateral stability of the spine, and the production and correction of several scoliotic deformities.     

Effects of repeated blood samplings on locomotor activity, evasion and wheel-running activity in mice

The effects of serial blood sampling on nocturnal locomotor activity, evasion, wheel-running activity and body mass were studied in male NMRI mice aged 7-8 weeks. The erythrocyte count, haematocrit and haemoglobin concentration at the beginning and end of the study showed no difference in group 1 (two samples per week, 0.08 ml each) while there was a significant decrease in the group 2 values (three samples per week, 0.08 ml each). The total amount of nocturnal locomotor activity decreased in the animals bled repeatedly while the periods with locomotor activity increased. These alterations appeared particularly after bleeding. In the test-group animals evasion showed a decrease compared with the untreated control animals, but there was no evidence of a relation to the timing of the bleedings.     

A fully automated and computerized system for simultaneous measurements of motility and phototaxis in Chlamydomonas

A fully automated and computerized method for simultaneous measurements of motility and phototaxis of unicellular flagellates is described. Both systems are directly coupled with a homocontinuous culture. The motility measuring apparatus is equipped with a video camera and recorder for simultaneous single cell behaviour studies. First results of studies on the effects of the phototaxis inhibitor sodium azide and the Ca²⁺ conducting ionophore A23 187 on motility and phototaxis of Chlamydomonas are reported and correlated with video observations. These results demonstrate that the described systems give informations of whether phototaxis or motility or both are inhibited by chemicals.     

Increased wheel-running activity in the genetically skeletal muscle fast-twitch fiber-dominant rats.

The purpose of the present study was to investigate whether genetic differences in muscle histochemical characteristics were related to the voluntary wheel-running activity level by using genetically fast-twitch fiber-dominant rats (FFDR) and control rats (CR). The rats were divided into four groups; sedentary CR (Sed-CR), wheel-running CR (WR-CR), sedentary FFDR (Sed-FFDR), and wheel-running FFDR (WR-FFDR). Wheel access was started at age 9 wk and lasted for 7 days. The FFDR showed a lower percentage of type I fibers of the deep portion of gastrocnemius and soleus muscles and a higher percentage of both type IIX fibers of the gastrocnemius muscle and type IIA fibers of the soleus muscle compared with CR. A higher capillary density and smaller fiber cross-sectional area were also observed in FFDR. The daily running distance in WR-FFDR was higher than in WR-CR for each 7 days. The total running distance for 7 days in WR-FFDR was 3.2-fold higher than in WR-CR. On day 7 of the 7-day test, the total number of active 1-min intervals for 24 h, the average rpm when they were active, and the maximum rpm for any single 1-min period in the WR-FFDR were significantly higher than in the WR-CR (1.5-, 2.9-, and 2.0-fold, respectively). These results suggest that mechanical or physiological muscle characteristics may thus affect the wheel-running activity level.     

A simplified microwave-based motion detector for home cage activity monitoring in mice

Background

Locomotor activity of rodents is an important readout to assess well-being and physical health, and is pivotal for behavioral phenotyping. Measuring homecage-activity with standard and cost-effective optical methods in mice has become difficult, as modern housing conditions (e.g. individually ventilated cages, cage enrichment) do not allow constant, unobstructed, visual access. Resolving this issue either makes greater investments necessary, especially if several experiments will be run in parallel, or is at the animals’ expense. The purpose of this study is to provide an easy, yet satisfying solution for the behavioral biologist at novice makers level.

Results

We show the design, construction and validation of a simplified, low-cost, radar-based motion detector for home cage activity monitoring in mice. In addition we demonstrate that mice which have been selectively bred for low levels of anxiety-related behavior (LAB) have deficits in circadian photoentrainment compared to CD1 control animals.

Conclusion

In this study we have demonstrated that our proposed low-cost microwave-based motion detector is well-suited for the study of circadian rhythms in mice.

     

Pharmacological manipulation of the dopaminergic system affects wheel-running activity in differentially active mice

The genetic factors involved in the regulation of physical activity are not well understood. The dopamine system has been implicated in the control of voluntary locomotion and wheel running (WR) in mice and is thus a likely candidate as a genetic/biological system important to the regulation of physical activity. This study evaluated the effects of four different dopaminergic acting drugs on WR in differentially active inbred strains of mice. High active C57L/J (n=7, 3 controls, 4 experimental) and low active C3H/HeJ (n=8, 3 controls, 5 experimental) were analyzed for baseline wheel-running indices of distance (km/day), duration (mins/day), and speed (m/min) for 21 days. Experimental mice received increasing doses over four days of each of the following drugs: SKF 81297 (D1 agonist), SCH 23390 (D1 antagonist), GBR 12783 (DAT inhibitor), and AMPT (tyrosine hydroxylase inhibitor). Each drug dose response treatment was separated by three days of recovery (no drug injections). WR indices were monitored during drug treatments and during drug wash-out phases. SKF 81297 significantly reduced (p=0.0004) WR in the C57L/J mice, but did not affect WR in the C3H/HeJ mice. GBR 12783 significantly increased (p=0.0005) WR in C3H/HeJ mice, but did not affect WR in C57L/J mice. Only duration (not overall WR) was significantly reduced in C57L/J mice in response to SCH 23390 (p=0.003) and AMPT (p=0.043). SCH 23390 (p=0.44) and AMPT (p=0.98) did not significantly affect WR in C3H/HeJ mice. These results suggest that genetic differences in dopamine signaling may play a role in the WR response to dopaminergic-acting drugs in inbred strains of mice. The high activity in the C57L/J strain appears most responsive to D1-like receptor acting drugs, while in the C3H/HeJ strain, dopamine re-uptake appears to have an influence on activity level.