Flow cytometric analysis of thymocyte subpopulations in mice after whole-body X-irradiation.

To determined the cellular kinetics of thymocyte subpopulations in DBA1 mice after whole-body 6.8 Gy X-irradiation, they were analyzed for the expression of several cell surface antigens using flow cytometry. The results show that i) The majority of thymocytes rapidly depleted by irradiation was CD4+8+ cells. ii) radioresistant CD4+8- and CD4-8+ survived 18-48 hr after X-irradiation were considered to be relatively mature type, since they expressed high levels of CD3 and LECAM-1. iii) CD3-positive cells were detected in CD4-8- cells at 72 hr after irradiation.     

Usefulness of infrared thermometry in determining body temperature in mice

The rectal temperature obtained using a standard electronic thermometer was compared with ear, back skin, tail skin, and sole skin temperatures obtained using an infrared thermometer in B6C3F1 mice. Using both methods, we investigated baseline temperatures, diurnal and 2-week variations in temperatures, and ethanol-induced hypothermia in these body locations. Ear and back temperatures were shown to be close to and consistent with rectal temperatures in various situations, and measured temperatures at these sites were almost constant, with very similar diurnal variation. Conversely, tail and sole temperatures were lower and much more variable. These results indicate that ear and back skin temperatures obtained using a convenient and non-invasive infrared thermometer are as reliable, and should be safer and less stressful to animal subjects, compared to standard rectal temperature measurements.     

B220 is expressed on apoptotic thymocytes induced by X-irradiation.

CD45 is cell surface glycoprotein and expressed on all haematopoietic cells except mature erythrocytes and platelets. Eight isoforms of CD45 are generated by alternative splicing of exons 4-6. B220 including all three exons is expressed specifically on pan-B cell lineage. Recently, it was reported that B220 was expressed on apoptotic T cells induced by staphylococcal enterotoxin B (SEB). In the present study, we investigated the expression of B220 on murine thymocytes after whole-body X-irradiation. We used the forward light scattering of flow cytometry as a parameter of cell size, and defined two populations; FSChigh (normal cell size) and FSClow (correspond to apoptotic cell in size) fraction. B220+ cells in FSChigh fraction reached a maximum value (35%) at 18 hr after irradiation. In FSClow fraction, 40-60% cells were positive for B220 at any time points. These results suggest that B220 is expressed on thymocytes in the pre-apoptotic stage, because B220 was expressed on not only FSClow cells but also FSChigh cells.     

Estimation of the proportion of body fat in mice from the proportion of body water

We compared the proportion of body fat in mice as measured by chemical analysis with that estimated from the proportion of body water. First, we measured the proportion of fat by chemical analysis in 78 mice that had a proportion of body fat in the range from approximately 5 to 20%. Then, we constructed a regression line that described the relationship between the proportion of body water and the proportion of body fat by using data from several other published studies in mice (% body fat = -1.20 x % body water + 88.07, r = 0.9597, sy.x = 2.75, p < 0.001). With this regression line, we estimated the proportion of body fat from the proportion of body water that was measured by drying the carcasses at 60 degrees C for 3 days. Body fat data obtained from this regression line were similar to those obtained by chemical analysis. Thus, these results suggest that reliable values for the proportion of body fat can be derived from the proportion of body water and this method provides a tool to rapidly measure the proportion of body fat in mice.     

Genetic variation in liver mass, body mass, and liver:body mass in mice.

Genetic variation for liver mass (LM), body mass (BM), and liver:body mass (LM/BM) was examined for outbred populations of laboratory mice. Liver mass and body mass data were collected on 170 pureline sires at 12 wk of age, representing four outbred stocks of laboratory mice; 523 of their male and female two-way-cross progeny at 9 or 12 wk; and 214 four-way-cross offspring at 12, 14, or 16 wk. Genetic differences for LM, BM, and LM/BM were found among the base sire lines and between two-way crosses. Heritabilities and genetic correlations for LM, BM, LM/BM, and LM/MBM (MBM = BM.75) were estimated using offspring-sire regression within and across characteristics. Estimates of heritabilities and genetic correlations were also derived from full-sib covariances in the two-way-cross generation. Heritability estimates pooled over all analyses were .53, .54, .36, and .40 for LM, BM, LM/BM, and LM/MBM, respectively. Body mass was highly genetically correlated (.87) with LM and lowly correlated with LM/BM. Previous research has indicated possible positive relationships between LM/BM and maintenance energy requirements in mature, nonlactating, nonpregnant animals. A selection index was developed for increasing BM but restricting genetic change in LM to zero. Selection using this index would be 40% as efficient in increasing BM as selection on BM alone but may hold maintenance energy requirements at a stable level.     

Reproductive performance in mice selected for divergence in body fat content

SUMMARY: Correlated responses in female reproductive performance were studied in lines of mice selected for high (HF) and low (LF) fat content relative to a randomly selected control line (RC). Selection was based on an indicator trait of body fat percentage in males (epididymal fat pad weight as a percentage of body weight at 12 weeks of age). LF mice showed a positive correlated response (P < 0.05) in number of pups born (NB), whereas HF showed no significant change. The increase in NB could be explained partially by an increase in ovulation rate (adjusted for body weight at mating; P < 0.01), measured as number of corpora lutea (CL) 16 days postcoitus (P < 0.05) or number of ova (OR) shed 2 days postcoitus (P < 0.01). Asymmetry was observed for CL (P < 0.05) and OR (P < 0.01); the major correlated response was observed in the LF line. Preimplantation and total embryo survival were higher (P < 0.01) in LF than in HF mice, whereas no significant divergence was found for post-implantation embryo survival. Reciprocal crosses of HF and LF lines allowed partitioning of divergence in OR into maternal (P < 0.10) and individual (P < 0.10) effects with LF being larger than HF. No significant heterosis was found for OR or mating weight. Body weight at mating was larger in HF females than in RC controls, while LF females were larger than controls in one experiment and not different from controls in a second experiment. Therefore, body weight at mating did not explain the positive correlated response in ovulation rate observed in the LF line. ZUSAMMENFASSUNG: Reproduktion in gegens?tzlich auf K?rperfett selektierten M?usen Korrelierte Selektionswirkungen auf weibliche Reproduktion wurden in Linien untersucht, die auf hohen (HF) oder niedrigen (LF) Fettgehalt selektiert worden waren, relativ zu einer zufallsgepaarten Kontrolle. Selektion beruhte auf Nebenhodenfett, korrigiert auf 12 Wochengewicht, als Indikator-merkmal. LF M?use hatten signifikant h?here Zahl geborener Junge (NB), HF zeigten keine signifikanten Wirkungen. Die Steigerung von NB kann teilweise durch hochsignifikante Steigerung der gewichtsadjustierten Ovulationsrate, gemessen als Zahl von Gelbk?rper (CL) 16 Tage postcoitus oder Zahl Eier (OR) 2 Tage pc, erkl?rt werden. Asymmetrie zeigte sich für CL (p < .01) und OR (p < .1), aber keine signifikanten Unterschiede zeigten sich für Post-Implantations überleben. Reziproke Kreuzungen von HF und LF erlaubten Aufteilung der OR Unterschiede in maternal (p < .1) una individuell verursachte (p < .1) Wirkungen, wobei LF gr??er als HF waren, aber sich keine Heterosis für OR oder Gewicht bei der Paarung ergab. Dieses war in HF Tieren h?her als in RC Kontrollen, was bei LF Tieren in einem, aber nicht im zweiten Versuch zutraf, so da? K?rpergewicht nicht die korrelierte Wirkung auf OR erkl?ren kann.     

High sensitivity of thymocytes of LEC strain rats to induction of apoptosis by X-irradiation

It is known that physical disruption of cell contacts induces apoptosis of thymocytes. When thymocytes from LEC and WKAH rats were incubated in vitro at 37 degrees C for 0-6 hr and then the proportion of apoptotic cells was determined using a flow cytometer, it was found that the percentages of apoptotic thymocytes from both LEC and WKAH rats increased with incubation time and that the proportion of apoptotic cells from LEC rats was significantly higher than that from WKAH rats at each incubation time. The fact that cycloheximide, an inhibitor of protein synthesis, did not show significant inhibitory effects on induction of apoptosis of thymocytes indicates that induction of apoptosis during in vitro cultivation did not require de novo protein synthesis. When thymocytes from LEC and WKAH rats were X-irradiated in vitro at 4 and 8 Gy, the percentages of radiation-induced apoptotic cells increased with post-incubation time after X-irradiation in both LEC and WKAH rat thymocytes and the proportions of apoptotic cells from LEC rats were significantly higher than those from WKAH rat cells at 2 and 4 hr post-incubation after X-irradiation. When thymocytes from LEC and WKAH rats were X-irradiated in the presence of cycloheximide, the induction of apoptosis was substantially inhibited, indicating that radiation-induced apoptosis of thymocytes from LEC and WKAH rats required de novo protein synthesis. The present results showed high sensitivities of thymocytes of LEC rats to induction of apoptosis during in vitro cultivation and by X-irradiation.     

Reallocation of body resources in lactating mice highly selected for litter size

The present study investigated differences in the allocation patterns of body stores in lactating female mice from a line selected for high litter size at birth (S-line, average litter size of 20) and dams from a nonselected control line (C-line, average litter size of 10). Body weight, litter size, litter weight, and absolute and relative lipid and protein mass were measured at peak lactation (2 wk in lactation) and at weaning (3 wk in lactation). Body size in S-line females has been increased as a correlated effect of selection for high litter size at birth, allowing for larger litters and higher absolute milk production. However, these dams produce larger litters relative to their own body weight. At peak lactation, lipid and protein percentage did not differ between lines. At weaning, S-line females had a higher protein percentage (P < 0.001) and lower lipid percentage (P < 0.05) than C-line females. Apparently, S-line females produce more offspring but at a greater cost to their own metabolism. This process was insufficient to supply the offspring with adequate resources, resulting in reduced (P < 0.0001) pup development and increased (P < 0.0001) preweaning mortality rates.     

Correlated responses in body composition based on selection for different indicator traits in mice

Correlated responses in whole-body composition were determined in 12-wk-old male mice from replicate lines selected for 12 generations for high (HF) or low (LF) epididymal fat pad weight as a percentage of body weight (EPID) and high (HL) or low (LL) hind carcass weight as a percentage of body weight. The HF and LF lines diverged (P less than .01) in body fat percentage (FAT) and subcutaneous depot fat by 93 and 71%, respectively, of the control line (RC) mean. EPID increased (P less than .01) proportionately more than FAT in the HF line; EPID decreased (P less than .01) proportionately less than FAT in LF. Protein, fat and water as a percentage of empty body weight showed negative correlated responses (P less than .01) due to selection for EPID, but lean body mass, body weight and body length had positive correlated responses (P less than .01). Correlated responses of fat-free protein and ash percentage were minor. Correlated responses in HL and LL were the mirror images of those in HF and LF, but they generally were of smaller magnitude. The results indicate that, although there are high positive genetic correlations between fat depots in mice, local control of lipogenesis and(or) lipolysis exists at different sites of fat deposition. Further, the lack of correlated responses in fat-free percentage of protein (and percentage of ash) suggests that additive genetic variances are low for these traits and(or) the genetic correlations of these traits with the selection criteria are low.     

SPF级近交系BALB/c小鼠的体重、体长、尾长和脏器系数的测定

本中心于2002年从国家啮齿类种子中心上海分中心引进5公5母近交系BALB/c小鼠进行饲养繁殖,目前本中心已经建立起核心群、血缘扩大群和生产群,至今已达到F5代,已经取得广东省实验动物微生物监测所颁发的SPF级微生物检测合格证(合格证号2005A 013)。为了了解近交系BALB/c小鼠幼龄期3周龄、性成熟期6周龄、体成熟期9周龄和老年期72周龄等不同阶段雌雄小鼠的体重、身长、尾长、脏器重量和脏器系数的情况。我们分别对生产群小鼠不同阶段雌雄小鼠的体重、身长、尾长、脏器重量进行测量,为科研工作者提供基础数据。1材料与方法1.1动物核心群…     

Correlated responses to selection for large body size in oMt1a-oGH mice: growth, feed efficiency and body composition

Correlated responses were determined for growth, feed consumption, feed efficiency and body composition following short‐term selection for large 8‐week body weight in transgenic and nontransgenic mice. Replicate lines which either carried or did not carry the sheep metallothionein 1a‐sheep growth hormone transgene (oMt1a‐oGH) were established. The lines carrying the transgene at an initial frequency of 0.5 came from a high‐growth (TM) and a randomly selected (TC) background. The respective nontransgenic lines were identified as NM and NC. Control replicates (CC) came from the randomly selected background. During the selection process the transgene was activated by adding 25 mm ZnSO₄ to the drinking water of all mice. Correlated responses were measured with (Z) and without (C) the addition of zinc. After seven and eight generations of selection, the frequency of transgenic mice in line TM had fallen sharply, whereas transgene frequency had risen moderately in TC. The reduced frequency of oMt1a‐oGH in the high‐growth genetic background may have been caused by a lower additive effect compared with the randomly selected background combined with a fitness disadvantage of the transgene. Therefore, the utility of a transgene in improving a quantitative trait may depend in part on genetic background. Correlated responses for most traits in NC were similar for Z and C. In contrast, correlated responses in TC showed marked differences in C compared with Z. For example, daily gain and feed efficiency showed no significantly correlated responses under C and positive responses (p < 0.001) under Z, and the reverse was found for indicators of body fat percentage. These line by environment (Z versus C) interactions may represent a genetic correlation of less than one between a trait expressed in two distinct environments. Thus, in developing lines with a transgene that can be regulated, a critical question is whether selection for quantitative trait(s) should be conducted when the transgene is activated or not activated.