To drive or be driven: the path of a mouse model of recurrent pregnancy loss.

This review is an example of the use of an animal model to try to understand the immune biology of pregnancy. A well-known model of recurrent spontaneous pregnancy loss is put in clinical, historical, and theoretical context, with emphasis on T cell biology.

Serial depletion and regeneration of the murine hematopoietic system. Implications for hematopoietic organization and the study of cellular aging.

The mouse hematopoietic system was subjected to repeated depletion and regeneration either by serial transfer of bone marrow cells through lethally irradiated recipients or by repeated treatment with the cycle-active drug hydroxyurea (HU). The capacity of surviving stem cells to proliferate and self-renew was assayed at intervals by two methods: (a) the spleen colony method; and (b) competitive repopulation of irradiated recipients using chromosome markers, with normal bone marrow cells as an internal control. The progressive decline in stem cell function that occurred during serial transfer of bone marrow and that had already begun after a single transfer was not seen during HU treatment; up to 25 pairs of HU injections given over more than 1 yr had no discernible effect on the number of stem cells present 3 wk after the final injection or on their capacity to self-renew. Within 2 d after exposure to HU, the average self-renewal capacity of surviving stem cells was enhanced. This implies that the drug selectively eliminates poorly self-renewing stem cells and hence that these enter cycle more readily than stem cells with a high self-replicative potential. However, the fact of being in cycle at the time of injection did not of itself affect self-renewal. The results show that serial transfer of bone marrow is not a valid method for studying clonal aging phenomena because it does not fulfill the assumptions on which such studies are based. No evidence was obtained for any intrinsic limitation in the capacity of bone marrow populations for repeated regeneration after HU-induced depletion. However, this does not necessarily imply that individual hematopoietic clones are capable of indefinite expansion because hematopoiesis may (as suggested by the relative resistance of highly self-replicative stem cells to mitogenic signals) proceed on the basis of clonal succession.

Circadian variation of mouse aqueous humor protein.

PURPOSE: The first aim was to document the 24 h pattern of total protein concentration in aqueous humor of the C57BL/6J mouse strain under various environmental light-dark conditions. The second aim was to determine the diurnal/nocturnal change of aqueous humor protein concentration in the CBA/CaJ mouse strain reported to show a unique absence of diurnal/nocturnal elevation of intraocular pressure (IOP). METHODS: Mice of both strains were entrained to a daily 12 h diurnal/light (6 AM to 6 PM) and 12 h nocturnal/dark cycle. Total protein concentrations of aqueous humor were determined using specimens collected from C57BL/6J mice every 4 h under standard light-dark, acute constant dark, and acute constant light conditions. Aqueous humor protein concentrations in CBA/CaJ mice were determined at three diurnal times and three nocturnal times. RESULTS: The C57BL/6J mouse strain showed a higher aqueous humor protein concentration during the diurnal period than during the nocturnal period under standard light-dark conditions. This pattern persisted under acute constant dark and was modulated under acute constant light. The CBA/CaJ mouse strain showed a similar diurnal/nocturnal decrease of total aqueous humor protein concentration. CONCLUSIONS: Endogenous circadian variation of aqueous humor protein concentration occurs in the C57BL/6J mouse strain. The nocturnal concentration is significantly lower than the diurnal concentration. A similar diurnal/nocturnal decrease of aqueous humor protein concentration occurs in the CBA/CaJ mouse strain, indicating that this change of aqueous humor protein is unrelated to the diurnal/nocturnal pattern of IOP.

Susceptibility of various strains of mice to urethan-induced lung tumors and depressed natural killer cell activity.

Inbred A/J, CBA/J, and C57BL/6 mice at age 7-8 weeks were inoculated with 1 mg urethan/g body weight. Three months later, 13.2, 1.8, and 0.1 tumors per lung were found in these mice, respectively. Cytotoxicity by spleen cells of normal and urethan-treated mice was analyzed in a 4-hour 51Cr release assay against YAC-1 cells. During the first 2 weeks after treatment, urethan strongly suppressed the cytotoxicity by spleen cells of A/J mice but had relatively little effect on the reactivity of spleen cells of CBA/J mice. Natural killer (NK) activity was not suppressed in urethan-treated C57BL/6 mice. The effect of urethan on natural in vivo antitumor resistance in the lungs was studied by measurement of elimination from the lungs of [125I]-deoxyuridine-labeled YAC-1 cells. Urethan caused profound and sustained suppression of in vivo NK reactivity in the lungs of A/J mice, had only a transient effect on CBA/J mice, and had no detectable effect on C57BL/6 mice. The differences in the effects of urethan on the natural antitumor resistance in the three strains of mice could not be attributed to a generally greater susceptibility of A/J mice to inhibition of NK activity, since cyclophosphamide (0.15 mg/g) equally suppressed the clearance from the lungs of the radiolabeled tumor cells in A/J, CBA/J, and C57BL/6 mice. These results indicate a positive correlation between susceptibility of A/J, CBA/J, and C57BL/6 mice to urethan-induced lung carcinogenesis and inhibition of in vitro and in vivo natural cell-mediated cytotoxicities and support the hypothesis that NK cells play a role in resistance to urethan-induced lung carcinogenesis.

Resistance to erythroleukemia in immunodeficient xid- CBA/N mice with susceptibility after immune stimulation.

CBA/N (xid-) mice failed to develop erythroleukemia when inoculated with an NB-tropic, anemia-causing Friend virus stock (FVA), while Fv-2ss mouse strains succumbed rapidly to the characteristic Friend disease, even after a virus dose 30-fold lower than that given to CBA/N mice. Immunization with bacterial antigens or with spleen cell allografts prior to FVA inoculation rendered CBA/N mice highly susceptible to FVA. Transplantation studies confirmed that non-immunized CBA/N mice were able to both support erythroleukemic cells and permit erythroleukemic transformation, thus arguing against host defense mechanisms as a cause of resistance. On the basis of early erythroid progenitor cell sensitivity to hydroxyurea in vivo, the CBA/N strain appeared to have the FVA sensitive genotype (Fv-2ss). These results imply that CBA/N mice are not intrinsically resistant to FVA and that an as yet unknown type of immunological activity, evoked both by various immunizations and allogeneic transplantation, is required for Friend leukemogenesis in this immunodeficient inbred strain. These findings further suggest that the erythroid target cells transformed by Friend viruses are influenced by immunological activity.

[Induction of meiotic disturbances in spermatocytes I by pheromones as an inhibiting mechanism of male reproductive function in house mice].

The influence of pheromons on reproduction and other important physiological characteristics has been reported for many mammalian species. However, mechanisms of this action at the level of target cells still remain unclear. A study was made of the influence of non-identified pheromones from adult males and a female pheromone 2,5-dimethylpyrazine on germ cells of CBA inbred strain mice. Cytogenetic analysis shows a significant increase in such meiotic disturbances as multivalent associations and autsomal univalents 24 h after exposure to pheromonal cues. Results of in situ hybridization show that the level of c-fos and c-jun expression is significantly higher 3.5 h after exposure to pheromones of adult males. It is likely that destabilization of chromosomal apparatus in dividing meiotic cells forms the basis of some reproductive effects of murine pheromones. Possible mechanisms of pheromone influence on reproduction are discussed.

Hematopoietic stem cell proliferative behavior as revealed by bromodeoxyuridine labeling.

Hematopoietic stem cells (CFUS) are thought to represent a heterogeneous population with different probabilities of self-renewal and different rates of proliferation. As an approach to further characterization of this population, we determined the disappearance of bromodeoxyuridine (BrdUrd)-induced CFUS sensitization to ultraviolet light in normal mice infused with BrdUrd for three weeks and in hydroxyurea-pretreated mice regenerating their CFUS pool during a one-week BrdUrd infusion. the same exponential disappearance with a T1/2 of six days was found in each case. From this and from the apparent absence of a secondary slope on the sensitization decay curve for the recovered hydroxyurea-treated bone marrow, we conclude that fewer than 10% of CFUS may be mitotically quiescent for prolonged periods and that the age structure of the CFUS population reflects a relatively short proliferative history.

Measurement of 3-methoxytyramine by in vivo voltammetry: evidence for differences in central dopamine function in BALB/c and CBA mice.

Differential pulse voltammetry (DPV) combined with carbon fibre electrodes allows selective detection of electroactive dopamine and serotonin metabolites in vivo. While usually employed in rats, we have now applied this in vivo technique in two inbred strains of mice: BALB/c and CBA. Three distinct oxidation peaks were recorded in vivo in the striatum of either BALB/c or CBA mice with a small shoulder occurring after the third peak at approximately +400 mV. Pargyline (150 mg/kg i.p.) potentiated this voltammetric shoulder into an easily measurable peak (Peak 4). In addition, Peak 4 was 2-3 times larger in BALB/c than in CBA mice. Homovanillic acid (HVA) and 3-methoxytyramine (3-MT), both catabolites of dopamine, oxidised at approximately +400 mV in vitro. Brain tissue levels of HVA and 3-MT, measured by high-performance liquid chromatography (HPLC) with electrochemical detection, demonstrated that pargyline treatment reduced striatal HVA, but increased 3-MT. These results support the view that Peak 4 recorded in the striatum of pargyline-treated mice in vivo is due to the oxidation of extracellular 3-MT. Thus, Peak 4 may be a useful index of dopamine release in situations where dopamine itself cannot be detected. Local infusion of KCl (2 microliters, 0.1 M) further increased the size of Peak 4 in the striatum of both BALB/c and CBA mice. However, the increase was approx. 3 times greater in BALB/c mice, supporting previous evidence of greater dopaminergic function of BALB/c compared with CBA mice. In addition these two inbred strains of mice provide model systems for investigating the comparative functional roles of nigrostriatal pathways.

The influence of different light intensities on pineal melatonin content in the retinal degenerate C3H mouse and the normal CBA mouse.

The sensitivity of light-induced suppression of pineal melatonin content was compared between C3H mice with hereditary retinal degeneration and CBA mice with normal retinas. At 2 h before lights on of light-dark (LD) cycles, when pineal melatonin content is the highest in both strains, groups of mice were exposed to different intensities of white fluorescent light (100, 0.14, 0.017 lux in both strains and 0.0021 and 0.00026 lux in CBA mice). For each intensity, pineals were collected just before and 5, 15 and 30 min after exposure to light. In C3H mice, the threshold of light intensity to suppress pineal melatonin content was between 0.14 and 0.017 lux, whereas that in CBA mice was between 0.0021 and 0.00026 lux. These results suggest that both rods and cones mediate photic information to the pineal gland in mice.

Interstrain variation in murine aerobic capacity.

PURPOSE: The contribution of genetic factors to aerobic capacity is unknown. The purpose of this study was to measure maximal aerobic performance among inbred strains of mice to provide basic heritability estimates. METHODS: Eight female mice, 8 to 10 wk old, in 10 inbred strains (A/J, AKR/J, Balb/cJ, C(3)H/HeJ, C57Bl/6J, C57L/J, C(3)Heb/FeJ, CBA/J, DBA/2J, and SWR/J) were run on a treadmill until exhaustion. The protocol started at 22 m.min(-1) and increased in speed approximately 6 m.min(-1) every 4 min. After 4 min at 42.4 m.min(-1), the grade was increased 2% every 4 min thereafter until the mouse could not run off of the shock grid (150 V; 1.5 mA). RESULTS: There were significant differences between inbred strains in maximal duration of exercise accomplished (P < 0.0001). The order of strain-specific exercise duration was Balb/cJ > SWR/J > CBA/J > C57L/J > C3H/HeJ > C3Heb/FeJ > C57Bl/6J > AKR/J > DBA/2J > A/J. Two measures of heritability in the broad sense, intraclass correlation (0.73), and the coefficient of genetic determination (0.58) were both significant. CONCLUSION: These data indicate that there is a strong genetic contribution to aerobic capacity in mice.

Vulnerability to noise-induced hearing loss in 'middle-aged' and young adult mice: a dose-response approach in CBA, C57BL, and BALB inbred strains.

Vulnerability of the cochlea to noise-induced permanent threshold shifts (NIPTS) was examined in young adult (1-2 months) and 'middle-aged' (5-7 months) CBA/CaJ, C57BL/6J, and BALB/cJ inbred mice. For each age and strain, a dose-response paradigm was applied, whereby groups of up to 12 animals were exposed to intense broadband noise (110 dB SPL) for varying durations. Exposure durations reliably associated with <10% and >90% probability of a criterion amount of NIPTS (determined 2 weeks post-exposure) were identified, and the minimum NIPTS exposure and the slope of the dose-response relation were then derived by numerical modeling. For all three strains, young adult mice were more susceptible to NIPTS than older adults; That is, a shorter exposure was able to cause NIPTS in the younger mice. Strain comparisons revealed that C57 mice were more susceptible than CBAs in the older age group only. At both ages examined, however, BALB mice were most susceptible to NIPTS. When animals with a similar amount of NIPTS were compared, outer hair cell loss in the cochlear base was more widespread in the younger animals. BALB mice appear particularly susceptible to noise-induced outer hair cell loss throughout life. Our data suggest that the mechanism or site of noise injury differs between young adults and older adults, and may depend on genetic background. The finding that both BALB and C57 mice, which show pronounced age-related hearing loss, are also especially vulnerable to noise supports the notion that genes associated with age-related hearing loss often act by rendering the cochlea susceptible to insults.

Effects of exposure to an augmented acoustic environment on auditory function in mice: roles of hearing loss and age during treatment.

The effects of exposure to an augmented acoustic environment (AAE) on auditory function were evaluated in mouse strains that exhibit various degrees and time courses of progressive hearing loss (BXD-22, BXD-12, BXD-16, BXD-14, BALB/cJ), and in normal-hearing CBA/CaJ mice. Beginning at age 25 days, mice were exposed 12 h every night to a 70 dB SPL broadband noise AAE. The AAE was maintained for at least 30 days in each strain. Same-strain control mice were age-matched and maintained under normal vivarium acoustic conditions. The auditory brainstem response (ABR), acoustic startle response amplitude, and prepulse inhibition (PPI) were used to assess the auditory system. Exposure to the AAE resulted in improved auditory performance (better PPI, lower ABR thresholds) when hearing impairment was present, but not when hearing was normal. The ameliorative effects occurred irrespective of a mouse's age at the onset of hearing loss, as long as initiation of AAE treatment preceded the occurrence of severe hearing loss. If AAE treatment was delayed beyond such a point, loss of threshold sensitivity progressed as usual, although PPI could still benefit. Finally, AAE treatment can slow, but not prevent, the occurrence of severe genetically determined hearing loss.

Sensorineural hearing loss alters recovery from short-term adaptation in the C57BL/6 mouse.

Several strains of laboratory mouse (Mus musculus) have a pattern of hearing loss which resembles that found in humans. The C57BL/6 strain of mouse has a genetic defect that results in degeneration of the organ of Corti, originating in the basal, high-frequency region and then proceeding apically over time. The end result is a severe-to-profound sensorineural hearing loss (SNHL) by 14 months of age. In contrast, auditory function of the CBA strain remains normal through its early life span then slowly declines later in life, much like that typified by human presbycusis. The purpose of the present study was to compare ABR (peak 5) forward masking recovery functions in young, normal-hearing CBA and C57BL/6 mice to hearing-impaired C57BL/6 mice. ABR audiograms were obtained prior to collecting the tone-on-tone forward masking data. Masking was defined as a 50% reduction in the P5 component of the ABR, elicited and masked by 12 kHz tone bursts, using masker/probe time delays from 0 to 100 ms. Time constants were computed from an exponential model fit to the recovery functions (masker level vs. time delay). In hearing-impaired animals there was a significant increase in recovery from short-term adaptation as measured by the time constants, as well as a significant latency shift in the P5 component. The effects of SNHL on the recovery of the P5 component from short-term adaptation was comparable to that reported behaviorally for human hearing-impaired listeners and physiologically from the inferior colliculus (IC) of chinchillas suffering permanent threshold shifts.

Carcinogenic effects of cytostatic protocols in CBA/Ca mice.

An in vivo mouse model was developed in order to study the characteristics of secondary tumor induction by cytostatic drug combinations used in human anticancer treatment. In this model we have proved the carcino-leukemogenic effects of widely used chemotherapeutical drug combinations (CHOP, COPP, COPBLAM, VAM). The carcinogenic hazards of cyclophosphamide and other alkylating drugs could also be demonstrated in our model.

Variation among inbred and linecross mice in response to fescue toxicosis.

Variation in response to fescue toxicosis was examined in inbred and linecross mice. In Exp. 1, exposure to a 50% endophyte-infected tall fescue diet (E+) reduced ADG of males from six inbred lines, but ADG of males from one line was modestly higher on E+. Lines differed (P < .01) for reproductive organ weight, but the diet x line interaction was not significant. In Exp. 2, an apparently susceptible (C57) and an apparently resistant line (FVB) were mated to produce inbred and linecross offspring. The reduction in weight gain caused by the E+ diet did not differ significantly among the genetic groups. In Exp. 3, C57 and C57 backcrosses had smaller reductions in ADG during E+ vs control feeding periods than FVB and FVB backcrosses (P < .10). In Exp. 4, the E+ diet reduced litter size of mates of C57 males by one pup, whereas litter size of mates of FVB males was four pups larger (interaction P = .07). Neither diet, line, nor their interaction affected male reproductive organ weights or tissue proportions in testis cross-sections. In Exp. 5, the E+ diet did not affect weight gain of C57 or FVB males, but effects of the E+ diet on litter size of mates were similar to those in Exp. 4. Percentage of abnormal sperm was increased in C57 males on the E+ diet but decreased in FVB males (Exp. 5). Differences among inbred lines in susceptibility to fescue toxicosis may depend on severity of the challenge and life cycle stage when the challenge is imposed.

The response of transgenic mice to beta-adrenergic agonist administration is different from that of normal mice.

Eighteen transgenic mice carrying an ovine metallothionein la-ovine growth hormone (oMTla-oGH) transgene and 18 littermate normal mice were used to investigate the effects of transgene expression and clenbuterol administration on growth performance and skeletal muscle characteristics. The oGH transgene was activated from 21 d of age, and half of the mice were fed 15 ppm clenbuterol from 42 to 70 d of age. All mice were killed at 70 d of age after 4 wk of treatment, and organs and muscles were dissected, weighted, and analyzed. Transgenic mice (TM) gained 2.6 times more than normal mice (NM). However, TM had a significantly lower (-20%, P < .01) proportion of muscle, expressed as percentages of body weights, and a higher percentage of heart (+10%), liver (+26%, P < .01) and spleen (+64%, P < .01) than NM. Clenbuterol improved the weight gain of TM by 20%, compared with 10% for NM. The growth-promoting effect of clenbuterol was almost exclusively confined to skeletal muscle (24% increase) in NM, in contrast to a more generalized growth increase in all tissues including skeletal muscle (11% increase) in TM. The skeletal muscles of TM were longer but smaller in diameter due to 30% smaller muscle fiber cross-sectional area. Clenbuterol increased the muscle fiber size of all fiber types by 60% in NM, compared to 30% in TM. Muscle DNA concentrations and content were higher (P < .05) in TM than in NM, and clenbuterol administration decreased DNA concentrations but not total DNA content for both genotypes. Cathepsin B, C, and H activities were higher (P < .01) in TM muscle, but the significance is not clear at the present time, although it points to a potential for greater protein degradation and(or) turnover rates as suggested by smaller muscle weights.

Different response of maturing oocytes from two inbred strains of mice to sperm penetration.

Oocytes from two inbred strains of mice, CBA and KE, were inseminated at prometaphase of the first meiotic division. Pronuclei were formed in 21-36% of inseminated oocytes from the CBA strain. In the KE strain the formation of pronuclei occurred in 2-5% of oocytes and was at the same level as in the non-inseminated control. These results show that in oocytes of the CBA strain maturity of the cytoplasm is acquired earlier than in those of the KE or other so far studied strains of mice. This fact is discussed in the context of different maturation rates of oocytes from different strains. In oocytes which remained non-activated after penetration, transformation of sperm heads into separate chromosomes was observed. An interstrain difference in efficiency of this process was also found.

[The role of the sex hormones in regulating the spermatogenesis process in different strains of mice irradiated at low doses of gamma quanta]. / Rol' polovykh gormonov v reguliatsii protsessa spermatogeneza u myshei raznykh linii, obluchennykh v malykh dozakh gamma-kvantami.

Intact BALB/c and CBA mice distinct in the total number of germ cells and testosterone level in blood plasma were exposed to doses of 0.1 and 0.25 Gy. Being tested 4-8 days after irradiation the BALB/c mice display compensatory-protective reaction which promote the maintenance of the germ cell number by active division of all spermatogonia types including the reserve ones. The CBA mice use the reserve later and only when the cells have reduced their proliferation activity or died. Testosterone plays a significant role in the process as the increase in its concentration stimulates proliferation activity and promotes mitosis block.

[The role of testosterone in regulating the aggressive behavior of male laboratory mice of different strains]. / Rol' testosterona v reguliatsii agressivnogo povedeniia samtsov laboratornykh myshei razlichnykh linii.

The influence of testosterone on aggressive behaviour, on urinal pheromonal activity, on weight of the preputial gland, seminal vesicles and coagulating glands of male mouse CBA, C57BL/6 and hybrids CBAB6F1 was studied. The interstrain differences in reactivity to testosterone in aggressive behaviour tests and in seminal vesicles and coagulating gland weight were found. The urinal pheromonal characteristics were not changed by testosterone influence. No significant correlations between the aggressive behaviour parameters, urinal pheromonal activity, testosterone plasma levels and reactivity to its action were discovered.

[The characteristics of the manifestation of hereditarily induced anxiety in male C57Bl/6J and CBA/Lac mice]. / Osobennosti proiavleniia nasledstvenno-obuslovlennoi trevozhnosti u samtsov myshei linii C57Bl/6J i CBA/Lac.

Behavior of male mice of C57Bl/6J and CBA/Lac strains was tested in the elevated plus-maze and open field in order to estimate state anxiety in novel conditions. The cube and partition tests were used to reveal trait anxiety in the familiar conditions of the home cage. It is concluded that genetically defined state anxiety is more pronounced in CBA/Lac mice and trait anxiety in C57Bl/6J strain.