Effect of the estrus cycle stage on sensitivity to pheromone 2,5-dimethylpyrazine in the house mouse <Emphasis Type="Italic">Mus musculus</Emphasis>

Frequency of cytogenetic disturbances was estimated in mitotically dividing bone marrow cells of CBA strain female mice after 24-h long action of pheromone 2,5-dimethylpyrazine (2,5-DMP). The stage of estrus cycle of each animal was taken into acount at the moment of the end of the pheromone action. The analysis was performed using the anatelophase method that allows evaluating frequencies of various types of disturbances—bridges, fragments, delayed chromosomes. The spontaneous level of the mitotic disturbances revealed by the anatelophase method in animals of the control group amounts to 5.4%. Action of pheromone 2,5-dimethylpyrazine induced the mitosis disturbances detected in the dividing bone marrow cells at the anaphase-telophase stage in the females at the di-+ postestrus stage. The corresponding frequency of disturbances after the pheromone action was equal to 9.2%. In the female in estrus, the mitotic disturbance level amounted 6.7%, which not differed statistically significantly from control. It is suggested that differences in female mouse hormonal state at different estrus cycle stages affect sensitivity to olfactory signals. Mechanisms of the revealed effect and significance of the differences in sensitivity to pheromone for reproductive processes are discussed.     

The role of metabolic activation of promutagens in the genome destabilization under pheromonal stress in the house mouse (<Emphasis Type="Italic">Mus musculus</Emphasis>)

The hypothesis on a relationship between the high frequency of mitotic disturbances in bone marrow cells and the change in the activity of the S9 liver fraction containing promutagen-activating enzymes under olfactory stress in the house mouse Mus musculus has been tested. For this purpose, the effect of the pheromone 2,5-dimethylpyrazine on the frequency of mitotic disturbances in mouse bone marrow cells has been measured by the anaphase-telophase assay. In paralled, we compared the capacities of the S9 liver fractions from stressed and intact mice for activating the promutagen 2-aminofluorene in the Ames test utilizing Salmonella typhimurium. It has been demonstrated that the increased frequency of mitotic disturbances in bone marrow cells induced by the pheromonal stressor in male house mice is accompanied by an increased ability of the S9 liver fraction to activate the promutagen. The model system used in the study allowed the genetic consequences of the exposure to the olfactory stressor to be estimated and the possible mechanisms of genome destabilization to be assumed.     

Stress,chemocommunication, and the physiological hypothesis of mutation

The review considers stress as a physiological state of the organism, affecting the cellular, genomic, and population levels. Literature data and cytogenetic studies by the author support basic statements of the physiological hypothesis of mutation, which was advanced as early as in the 1940s. Studies of pheromonal effects in germline and somatic cells of the house mouse demonstrated the role of olfactory stressors in generating genetic variation in microevolutionary changes.     

DNA damage in bone marrow cells of mouse males in vivo after exposure to the pheromone: Comet assay

It is shown by single-cell gel electrophoresis that the exposure of CBA mouse males with pheromone 2,5-dimethylpyrazine for 4 or 24 h increases DNA damage level in interphase nuclei of bone marrow cells. The results may reflect the work of a mechanism of formation of chromosomal aberrations and other mitotic disturbances, detected at the stage of ana-telophase, the frequency of which in dividing bone marrow cells increased after similar pheromonal exposure. The comparison of the damaged cell distribution types is proposed as an approach to analysis of comet assay data. The significance of the revealed effects on the immune system in the recipient organism is discussed.     

Induction of dominant lethals in progeny of male CBA mice after pheromonal action

The inhibiting effect of pheromone 2,5-dimethylpyrazine of house mouse females on the reproductive function of the CBA male mice was studied. The mutagenic effect of six-day pheromonal effect was assessed by dominant lethal test. Analysis for the frequency of dominant lethals showed that the pheromonal effect results in an increased death rate of the progeny of the treated males. This is probably explained by implantation failure and is expressed in a reduced average number of the implantation sites and low live embryos per female. The proportion of females with live embryos decreased significantly. The implication of the effect of female mouse pheromone 2,5-dimethylpyrazine on the genetic processes in germ cells of male mice is discussed.     

The female pheromone 2,5-dimethylpyrazine induces sperm head abnormalities in male CBA mice

The effect of the house mouse female pheromone 2,5-dimethylpyrazine (2,5-DMP) on sperm differentiation in male CBA mice has been studied. For this purpose, mature males were treated with a 0.01% aqueous solution of the pheromone for six days. Control mice were similarly treated with physiologic saline. The mice were sacrificed 23 days after the treatment, and material for the analysis of sperm-head abnormalities was sampled from the caudal portion of the epididymis. Analysis of the frequency of abnormal sperms has demonstrated that the pheromonal treatment significantly increases the frequencies of various sperm-head abnormalities. Apparently, this results from disturbances in sex-cell differentiation germline cells caused by the induction of genetic damage at stages immediately preceding meiosis, as well as during the first and second meiotic divisions. The relationship between the effect of 2,5-DMP and the decrease in the fertility of male CBA mice that was earlier observed after a similar treatment is discussed.     

The role of metabolic activation of promutagens in the genome destabilization under pheromonal stress in the house mouse (Mus musculus)

The hypothesis on a relationship between the high frequency of mitotic disturbances in bone marrow cells and the change in the activity of the S9 liver fraction containing promutagen-activating enzymes under olfactory stress in the house mouse Mus musculus has been tested. For this purpose, the effect of the pheromone 2,5-dimethylpyrazine on the frequency of mitotic disturbances in mouse bone marrow cells has been measured by the anaphase-telophase assay. The Ames test using Salmonella typhimurium has been employed to compare the capacities of the S9 liver fractions from stressed and intact mice for activating the promutagen 2-aminofluorene. It has been demonstrated that the increased frequency of mitotic disturbances in bone marrow cells induced by the pheromonal stressor in male house mice is accompanied by an increased promutagen-activating capacity of the S9 liver fraction. The model system used in the study allowed the genetic consequences of the exposure to the olfactory stressor to be estimated and the possible mechanisms of genome destabilization to be assumed.     

Action of two pyrazine-containing chemosignals on cells of bone marrow and testes in male house mouse Mus musculus L

Evolutionary conservative chemosignal 2,5-dimethylpyrazin that is pheromone in female mice has been shown to increase frequency of mitotic aberrations analyzed with aid of metaphasic and ana-telophasic analysis in bone marrow cells. Replacement of one of methyl radicals in the pheromone molecule by the carboxyl radical reveals specificity of action of the used derivative: the frequency of disturbances revealed only by the ana-telophasic analysis increases, whereas by the metaphasic analysis, no induction of disturbance is detected. In the sperm head abnormality test there is shown a rise of the anomalies by both compounds. Possible mechanisms of specific action of the tested substances on stability of genetic apparatus of the bone marrow dividing cells in the house mouse are discussed.     

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.     

Induction of Dominant Lethals in Progeny of CBA Male Mice after Pheromonal Action

The inhibiting effect of pheromone 2,5-dimethylpyrazine of house mouse females on the reproductive function of the CBA male mice was studied. The mutagenic effect of six-day pheromonal treatment was assessed by dominant lethal test. Analysis for the frequency of dominant lethals showed that the pheromonal effect results in an increased death rate of the progeny of the treated males. This is probably explained by implantation failure and is expressed in a reduced average number of the implants and low live embryos per female. The proportion of females with live embryos decreased significantly. The implication of the effect of female mouse pheromone 2,5- dimethylpyrazine on the genetic processes in germ cells of male mice is discussed.