Heritable translocation test and dominant-lethal assay in mice with methyl methanesulfonate.

A dominant-lethal test and a heritable translocation test were performed with methyl methanesulphonate (MMS) at 40 mg/kg by treating the sensitive periods of post-meiotic spermatogenesis i.e. spermatozoa and spermatids. In the dominant-lethal test 25 to 60% dominant-lethal mutations were obtained depending on the mating intervals. In the heritable translocation test 11% sterile and partially sterile F1 males were observed in 250 offspring of the MMS group. All of the 14 partially sterile and 6 of the 14 sterile F1 males were demonstrated to be translocation carriers. Fertility of the partial steriles was about 40% of normal fertility. The translocation frequencies in the primary spermatocytes of the partially sterile F1 males varied between 2 and 99%. Transmission of partial sterility and translocations was confirmed in the F2 generation. There were no partially sterile or sterile males among the 245 controls.     

An evaluation of the micronuclei test using triethylenemelamine,trimethylphosphate, hycanthone and niridazole

To determine the feasibility of the micronuclie procedure for cytogenetic studies, a comparatively weak chromosome breaking agent, trimethylphosphate (TMP) and the potent alkylating agent, triethylenemelamine (TEM) were evaluated. The procedure followed was that of Matter and Schmid with the following modifications: (a) direct flushing of bone marrow with 0.2 ml calf fetal serum, (b) air drying slides for a period of only 1 h, and (c) the use of pH 6.0 phosphate buffer to dilute both the Wright and Giemsa stains.With this technique a dose-response curve was generated for both TMP and TEM, using mice as the experimental animal. With TMP, a doubling over background was found when a concentration of 0.5 g/kg per day for five days was administered. To establish a statistically significant doubling dose over the control, a minimum of five animals must be used woth 2000 polychromatic cells being analyzed per animal.Of the two antischistosomal agents tested, hycanthone yielded an increase of 20-fold in the number of mircronuclei over control at 40 mg/kg administered i.p. for five days, while with niridazole no increase in micronuclei at several concentrations tested both by single and multiple injection was found.The results obtained with these compounds compare favorably woth what has been reported for the standard in vivo metaphase analysis.     

Heritable translocation study in male mice with trimethyl phosphate

Dominant lethal and heritable translocation studies were performed in male mice receiving a single intraperitoneal injection of trimethyl phosphate (TMP). The germ cell stage investigated was the spermatid. Methyl methanesulfonate (MMS) was used as a positive control in the latter study. A dominant lethal assay gave marked dose-dependent increases in early fetal deaths. Heritable translocations were detected at 1000 or 1500 mg of TMP/kg in F1 male progeny when screening for semi-sterility and cytogenetically analyzing the meiotic or mitotic chromosomes. Translocation induction was higher at the higher TMP dose (14.3%) than at the lower dose (5.3%) and the yield from the higher dose was similar to that induced by 50 mg of MMS/kg (11.0%). Most of the translocation carriers were semi-sterile or sterile. The data confirm conclusions from other dominant lethal studies showing TMP to be capable of causing chromosomal damage in mouse spermatids and show that certain types of damage result in heritable translocations.     

H-gene (histocompatibility) mutations induced by triethylenemelamine in the mouse

Based on a simple dermal-graft procedure, the H-test for histo-compatibility mutations screens the mouse “H-system” of not less than 29 and perhaps considerably more than 100 loci that are scattered throughout the genome. Graft-tests with normally heterozygous or hemizygous loci permit scoring mutations as gains, losses, and gains+losses. Tests with normally homozygous loci screen only for gains, but further analysis can detect accompanying losses. Assayed by the H-test, triethylenemelamine (TEM) increased the spontaneous mutation rate per generation by approximately four- to five-fold in the case of BALB/c spermatogonia and F₁ hybrid oocytes (BALB/c females × C57BL/6 males), but had a much smaller effect, if any, on the rate of C57BL/6 spermatogonia. Single intraperitoneal doses of 2.4–4.0 mg/kg were given.     

Characterization of small lymphocytes in the bone marrow of mice after prolonged treatment with anti-IgM antibodies.

In mice treated with anti-IgM antibodies from birth, small lymphocytes in the bone marrow and spleen have been characterized by their incidence, surface markers, and turnover. Essentially complete elimination of IgM-bearing small lymphocytes followed injections of anti-IgM but this treatment did not deplete the IgM-negative small lymphocytes in the marrow. These IgM-negative cells also lacked other markers of B lymphocytes, such as receptors for Fc and complement; they failed to bind anti-mouse T-lymphocyte serum and they formed a rapidly renewing population within the marrow. This population may represent cells whose normal differentiation has been aborted by the anti-IgM antibodies or, alternatively, they may be “null” cells, distinct from B lymphocytes.     

Heritability of directional and fluctuating asymmetry for mandibular characters in random-bred mice

In bilateral characters, two kinds of asymmetries are common: fluctuating asymmetry (FA), or nondirectional variation between left and right sides, and directional asymmetry (DA), in which one side is consistently larger than the other. FA has been extensively used as a measure of developmental stability because of its presumed environmental basis whereas DA has not typically been recommended because it has been presumed to have at least some genetic basis. To test these two hypotheses, heritabilities were calculated via parent–offspring regression for both DA and FA in 10 triply measured mandible characters in random-bred mice. Midparent estimates of heritabilities of DA in the 10 characters were quite low (mean = 0.06), but significant for one character as well as the sum of the DA values over all characters (0.21). Midparent estimates of heritability of FA in the 10 characters also were low (mean = 0.03), but not significant for any individual character or the sum of the FA values over all characters. Heritabilities of developmental stability calculated from heritabilities and repeatabilities of FA in the mandible characters were higher in magnitude (mean of midparent estimates = 0.45), but all still were not statistically significant. It was concluded that both hypotheses were supported, but that genetic variation in DA was so small that the potential for DA as an indicator of developmental stability should be explored.     

Ganglioside content and composition in rat cerebellum after prolonged diazepam treatment.

The main purpose of this study was to determine the content and composition of cerebellar gangliosides after prolonged diazepam treatment and their possible recovery after diazepam withdrawal. Male Wistar rats were administered diazepam in a dose of 10 mg/kg/day in drinking water for 3, 5 or 6 months. A additional group of rats had a one-month recovery period after five months of diazepam treatment. Control animals were age-matched and pair-fed. At the end of the experiment, the animals were sacrificed and the total cerebellar contents of ganglioside-NeuAc as well as its content in particular ganglioside fractions were estimated. After three months of diazepam consumption, no changes of ganglioside-NeuAc in investigated fractions (G(Q1b), G(T1b), G(D1b), G(D1a), G(M1), G(M2), and G(M3)) were observed. Five months of diazepam treatment caused a significant decrease in the total amount of gangliosides, which was evident in most of the investigated fractions, with the exception of the monosialoganglioside G(M2). Six months of treatment induced a generalized decrease in all the investigated ganglioside fractions. The diazepam-induced ganglioside reduction found after five months of treatment was also present after a one-month recovery period. The only fraction, which recovered and reached its control value, was monosialoganglioside G(M3).     

Beta-adrenoceptor activity change after prolonged treatment with growth hormone and somatostatin.

1. The effect of 10 days treatment with growth hormone (GH) (1 mg/kg body wt/day) and somatostatin (SRIF) (0.25 mg/kg body wt/day) subcutaneously on the activity of beta-adrenoceptors in rat hypothalamic, pituitary and cerebral cortical membrane fractions was studied using [3H]dihydroalprenolol ([3H]DHA) as radioligand. 2. The administration of GH significantly increased the beta-adrenoceptor binding affinity and the administration of SRIF decreased the beta-adrenoceptor binding capacity in the hypothalamus. 3. In the pituitary the beta-adrenoceptor binding affinity was significantly decreased after both hormonal applications. 4. In the cerebral cortex the beta-adrenoceptor binding affinity was significantly decreased after the GH treatment and increased after the SRIF treatment. 5. The present study provides direct evidence for GH and SRIF effects on the activity of rat beta-adrenoceptors and supports the view about the involvement of beta-adrenergic mechanisms in the neurotransmitter regulation of GH secretion in the rat.     

Differences in sperm function in vitro but not in vivo between inbred and random-bred mice

Genetic variation in spermatozoa was used to examine mechanisms important for fertilization in the mouse. A significantly greater proportion of cauda epididymal sperm from C57BL/6 (inbred) males were motile than from random-bred (CFW) males. Random-bred sperm, however, were able to fertilize a significantly greater percentage of eggs in vitro than were inbred sperm. When sperm of these two genotypes were used for insemination in vivo, and the penetrated eggs cultured through the first cleavage, the levels of cleavage were similar, suggesting that neither levels of sperm motility nor sperm penetration in vitro accurately reflect the ability of the same sperm populations to penetrate eggs in vivo.     

Genetic control of immune responses to parasites: selection for responsiveness and non-responsiveness to Trichuris muris in random-bred mice.

Populations of Schofield strain, random-bred mice were shown to have a bimodal variation in ability to bring about immune expulsion of the nematode Trichuris muris. This variation was genetically determined and independent of the size of infection experienced. The proportion of mice unable to achieve worm expulsion (non-responders) was relatively constant in various populations of the strain but was increased by selective breeding from mice of known status. Crosses made between non-responder and responder mice produced progeny that were almost all (92%) of responder phenotype, showing that the ability to achieve worm expulsion was inherited as a dominant characteristic. It is suggested that the genetic control involves a small number of genes; the possible immunological mechanisms by which control is mediated are briefly discussed.     

Pathogenesis of wasting disease after cyclophosphamide treatment of neonatal mice.

Cyclophosphamide is a potent immunosuppressive agent and is being widely used in organ transplantation. The effects of this anti-rejection drug on lymphoid organs are poorly understood. Newborn Swiss mice injected with various doses of cyclophosphamide suffered from wasting disease at 4 weeks post treatment. The incidence of wasting disease was dose dependent. Haematological picture of the wasting animals revealed leukocytosis of variable degree. Lymphocyte/granulocyte ratio was not inhibited. The cyclophosphamide treatment caused shrinkage of lymphoid organs. Bone marrow showed degeneration of haematopoietic cells. The failure to sustain lymphopoiesis by the potential lymphoid sites following cyclophosphamide treatment and the associated immunological insufficiency resulted in the fatal wasting disease.