Spermatogenesis parameters and testosterone production at puberty as predictors of testicular functional activity in mice (<Emphasis Type="Italic">Mus musculus</Emphasis>)

The aim of this study was to investigate fertility-associated parameters of spermatogenesis and androgenic status in male laboratory mice at puberty and to assess their prognostic significance in the realization of the definitive testicular function. In three inbred murine strains, BALB/cLac, CBA/Lac and PT, the serum testosterone level, its testicular concentration, epididymal sperm count (sperm reserve) and portion of sperm with abnormal head morphology were evaluated on days 45 (puberty) and 90 (adulthood) of postnatal development. CBA/Lac males were characterized by a lower epididymal sperm count vs. other strains at both ages indicative of poorer spermatogenesis. At the same time, CBA/Lac males had a lower portion of sperm with abnormal head morphology, and this could be considered as a compensatory reaction aimed at improving sperm fertility. Distinct inter-strain differences in the portion of sperm with morphologically abnormal heads were established at both ages, while the inter-strain ratio remained invariable (BALB/cLac > PT > CBA/Lac). Thus, the level of abnormal spermatogenesis in the pubertal period may have a predictive significance for the definitive testicular activity in adult mice. No inter-strain and age-dependent changes were found in serum and testicular testosterone levels except for the PT strain, in which both testosterone levels rose from puberty to adulthood, suggesting a shift of the pubertal testosterone peak towards later times. Our data show that in male laboratory mice the genetic peculiarities of the testicular function manifest themselves during puberty and persist until adulthood.     

Interstrain differences in testicular responses to hCG: effects of a female exposition

It is known that among many animal species, including laboratory mice, a short-time exposition to a female causes activation of the pituitary-testicular axis in males, which then rapidly decreases. Effects of a prolonged female exposition on the testicular testosterone output in response to hCG injections were investigated in adult males of two inbred mice strains CBA/Lac and PT. The males of both genotypes kept with females for 5 days were injected subcutaneously with 10 IU of hCG 120 minutes before decapitation. Males of the same genotype and similar age keeping alone were served as control. The serum testosterone concentration and its testicular content were measured with immune-enzyme assay. It has been shown that the hCG increased the testosterone concentration and its testicular content in control males of both strains, but the testosterone output was expressed more significantly in PT males in comparison with CBA/Lac. The prolonged exposition of a female itself did not influence the testosterone output in males of both strains. However, it increased significantly the testicular response to hCG in PT strain. The data obtained suggest that a long-term exposition of a female could reinforce the testicular reactivity to hCG in genotype-dependent manner.     

Interlinear differences in generative function formation in pubescence of male mice

The formation of generative function in pubescence is studied in males of three inbred mice lines BALB/cLac, CBA/Lac, and PT. From days 35 through 60 of life every 5 days the amount of sperm cells and the quantity of abnormal heads of spermatozoa in males are calculated in both epididymises and the morphometry of the testicles, epididymises, and seminal vesicles is carried out. Interlinear deviations in the pubertal dynamics of the parameters of spermatogenesis and the morphometric indexes are determined, indicating a weak spermatogenesis process in males of CBA/Lac in comparison with males from the other lines. Males from the line CBA/Lac are characterized by a low amount of epididymal spermatozoa combined with low frequency of abnormal spermatozoon heads; these traits can be considered as a compensatory process that increases fertility. By the end of the period, i.e., on days 55–60 of life, the males of all three inbred mice lines have not reached the definitive level in the number of epididymal spermatozoa; the weight of the testicles, epididymise, and seminal vesicles; and body weight. Thus, in laboratory mice, the beginning of reproductive activity is not connected with these reproductive indexes reaching the definitive level. The results of the study show that in adult mature males of laboratory mice the interlinear deviations in generative function emerge in the pubertal period and persist thereafter.     

Features of adrenal gland and gonadal reactions in mice with a genetic predisposition to dominance under zoo-social stress]

Intensity and duration of endocrine responses of adrenals and gonads in QT, CBA/Lac male mice and heir first generation reciprocal hybrids (F1) mice, social stress were studied. PT males had higher level of corticosterone in 1 h after stress, the adrenals and gonads reactions being some hrs shorter, as compared with CBA/Lac mice. No differences were observed in dynamics among reciprocal hybrids. They inherited not only high capacity to dominance in micropopulations but also reactions of adrenal activation and gonadal inhibition. Similar results were obtained, when these features were studied in dominant and subordinate males of PT strain mice.     

Role of Genotype, Social Stress, and Season in the Regulation of Hormonal Function of Testes Incubated in Vitro in Mice

Micropopulations consisting of six male mice of different genotypes were studied (each of lines A/He, CBA/Lac, C57BL/6J, DD, YT, and PT was represented by one male). Interlinear differences in the level of social dominance and the effects of genotype, social hierarchy, and season on in vitro testosterone production by testes were examined under different incubation conditions. The testosterone production was estimated under control conditions and under stimulation with human chorionic gonadotropin (CG). Significant genetic differences in the initial and CG-stimulated testosterone production by testes incubated in vitro were found. By the control production, the genotypes fell into two groups: lines C57BL/6J, A/He, and CBA/Lac had low production of the hormone; lines YT, PT, and DD, high production. By responsiveness of gonads to CG, the genotypes fell into three groups: line CBA/Lac had low testosterone production by testes; lines C57BL/6J, A/He, YT, and DD, line PT, intermediate production; and line PT, high production. The obtained data indicate stability of genetic polymorphism for the responsiveness of testes to gonadotropins, because neither season nor the formation of social hierarchy could significantly change the interlinear differences. In line PT characterized by high hormonal activity of gonads in the control and under stimulation with gonadotropins, males became dominant in a significantly greater number of cases studied during the formation of hierarchy in micropopulations. The dynamics of both control production of a male sex hormone and responsiveness of testes to CG was established in vitro during the formation of social hierarchy; the effects of season on this dynamics were revealed. Specific characteristics of secretory activity of testes were detected in the control and under stimulation with gonadotropins, depending on incubation conditions. Seasonal and genotypic characteristics of the responsiveness of testes to CG were revealed under different incubation conditions. Genotypic characteristics indicate interlinear differences in the degree of inertia of testosterone biosynthesis on exposure to gonadotropins.     

The role of genotype, social stress, and season of the year in regulation of hormonal function of murine testis in vitro

Micropopulations consisting of six male mice of different genotypes were studied (each of lines A/He, CBA/Lac, C57BL/6J, DD, YT, and PT was represented by one male). Interlinear differences in the level of social dominance and the effects of genotype, social hierarchy, and season on in vitro testosterone production by testes were examined under different incubation conditions. The testosterone production was estimated under control conditions and under stimulation with human chorionic gonadotropin (CG). Significant genetic differences in the initial and CG-stimulated testosterone production by testes incubated in vitro were found. By the control production, the genotypes fell into two groups: lines C57BL/6J, A/He, and CBA/Lac had low production of the hormone; lines YT, PT, and DD, high production. By responsiveness of gonads to CG, the genotypes fell into three groups: line CBA/Lac had low testosterone production by testes; lines C57BL/6J, A/He, YT, and DD, line PT, intermediate production; and line PT, high production. The obtained data indicate stability of genetic polymorphism for the responsiveness of testes to gonadotropins, because neither season nor the formation of social hierarchy could significantly change the interlinear differences. In line PT characterized by high hormonal activity of gonads in the control and under stimulation with gonadotropins, males became dominant in a significantly greater number of cases studied during the formation of hierarchy in micropopulations. The dynamics of both control production of a male sex hormone and responsiveness of testes to CG was established in vitro during the formation of social hierarchy; the effects of season on this dynamics were revealed. Specific characteristics of secretory activity of testes were detected in the control and under stimulation with gonadotropins, depending on incubation conditions. Seasonal and genotypic characteristics of the responsiveness of testes to CG were revealed under different incubation conditions. Genotypic characteristics indicate interlinear differences in the degree of inertia of testosterone biosynthesis on exposure to gonadotropins.     

Phenogenetic analysis of testicular responsiveness to chorionic gonadotropin in inbred mouse strains

Genetic differences in the testicular hormonal responsiveness to in vivo administration of chorionic gonadotropin (CG) between adult male mice of eight inbred strains (A/Sn, CBA/Lac, CC57Br, C57Bl/6J, DBA/2J, GR, PT, and YT) were determined. In addition, the genetic variation of the body and testis weights was estimated as related to the responsiveness to stimulation of steroidogenesis with CG. Adult males were subcutaneously injected with 10 IU of CG or physiological saline 120 min before decapitation. It was found that the baseline testosterone level in the blood serum and its content in the testes only slightly varied in males of the strains studied. Administration of CG increased these parameters by a factor of 3–45, depending on the strain. The results of the study indicate genetic differences in the testicular reactivity to CG. In addition, it has been found that the response to administration of CG, as compared to the baseline levels, provides the most reliable information on the genetic characteristics of the hormonal potential of the testes. The given set of inbred mouse strains may be a promising genetic model for studying the physiological and hereditary variations of testicular steroidogenesis.     

Maturation of sexual behavior in laboratory male mice: a role of genotype

Sexual behaviour and testosterone output in response to a receptive female were investigated in male mice of three inbred strains BALB/cLac, CBA/Lac and PT at puberty (45 days of age) and in adulthood (90 days of age). The animals were exposed for 10 min to a receptive female separated by a plastic grill, which would not allow contact between male and female. Male and female behaviour was recorded by measuring the time the male or female spent at the grill and the number of approaches to it (sexual motivation). The grill was then removed and the number of mounts and chemoinvestigatory behavior towards a female (nasal and anogenital sniffing) was recorded for each male. An increase in serum concentration and testicular content of testosterone was used as an endocrine index of the sensitivity to female pheromones. It has been shown the significant genotype and developmental effects on sexual behaviour and the hormonal response to sexual stimuli. The pubertal BALB/cLac males were characterised by the adult pattern of sexual motivation, chemoinvestigatory behaviour and the evident testosterone respond to a female. Males of the strain PT showed the lowest sexual motivation, chemoinvestigatory behavior towards a receptive female and no testosterone responses at both ages. This is a very different situation with the CBA/Lac's who showed the developmental increase in the sexual motivation, sniffing behaviour and the endocrine reflex, and the highest level of sexual behaviour but the moderate testosterone respond to a female at adulthood. The data obtained suggest genotype related asynchrony in maturation of the olfactory system, pituitary-gonadal axis and neural circuits of sexual behavior, and their independent genetic control. So, the set of mice strains investigated represents a useful tool for genetic and endocrine study of sexual behavior and the chemosensory control of testicular steroidogenesis.     

Tyrosine hydroxylase activity and expression of its gene in mice with contrasting capacity to dominate in social stress]

Changes of tyrosine hydroxylase (TH) activity and level of mRNA of TH gene in PT and CBA/Lac mouse strains, which are contrast by ability to dominate in heterogenous populations, were investigated. It was established, that the activity of TH both in dominate PT and subordinate CBA/Lac mice in hypothalamus, hippocampus and brain stem elevated in one hour after forming of micropopulations. But the appearance of this increase was different: activation of TH in hypothalamus and brain stem of PT mice was stronger then one in CBA/Lac mice. Moreover, the beginning of the reaction in brain stem of PT mice was earlier then that of CBA/Lac mice. MRNA level of TH gene in hypothalamus and brain stem in one hour was elevated only in PT mice for 50% and 200%, respectively. No changing in expression TH gene was found in hippocampus. In conclusion, it was suggested that the activation of catecholamine biosynthesis under social stress in hypothalamus and brain stem of male mice was due to the TH activation and increase of its gene expression.     

Effect of genotype on hormonal function formation of Leydig cells during mouse postnatal development

Changes in in vitro testosterone production by Leydig cells induced by chorionic gonadotropin, dibutyryl-cAMP, and pregnenolone have been studied during postnatal development of four inbred mouse strains BALB/c, PT, CBA/Lac, and A/He, with contrast hormonal activity of testes in sexually mature males. The interlinear differences significantly change with age of the males by all studied indices indicating genotype-dependent formation of hormonal activity of Leydig cells during postnatal development. Coordinated interlinear variability between all indices of Leydig cells reactivity has been established for each studied period of postnatal development. Hence, we have established coordinated interlinear genetic variability of hormonal function of Leydig cells, which was confirmed by considerable changes in it during postnatal development at puberty. Definitive genotypic differences in hormonal activity of Leydig cells appeared by late pubertal and early postpubertal development (day 60) and coincided with termination of morphological differentiation of Leydig cells and appearance of the differentiated cell population.     

Effect of Genotype on Formation of Hormonal Function of Leydig Cells during Mouse Postnatal Development

Changes in in vitro testosterone production by Leydig cells induced by chorionic gonadotropin, dibutyryl-cAMP, and pregnenolone have been studied during postnatal development of four inbred mouse strains BALB/c, PT, CBA/Lac, and A/He, with contrast hormonal activity of testes in sexually mature males. The interlinear differences significantly change with age of the males by all studied indices indicating genotype-dependent formation of hormonal activity of Leydig cells during postnatal development. Coordinated interlinear variability between all indices of Leydig cells reactivity has been established for each studied period of postnatal development. Hence, we have established coordinated interlinear genetic variability of hormonal function of Leydig cells, which was confirmed by considerable changes in it during postnatal development at puberty. Definitive genotypic differences in hormonal activity of Leydig cells appeared by late pubertal and early postpubertal development (day 60) and coincided with termination of morphological differentiation of Leydig cells and appearance of the differentiated cell population.     

Genetic-neurochemical analysis of the role of brain catecholamines in the manifestation of dominance in male mice in micropopulations

The ability of male mice to dominate in micropopulations, in connection with brain catecholamines was studied. It was shown that the F1 obtained from breeding PT and CBA/Lac which have respectively large and small numbers of high rank males were of high level of dominance in micropopulations. Moreover, they had high levels of noradrenaline and dopamine in hippocampus, striatum and brain stem, and low--in hypothalamus.     

Social domination and reproductive success in male laboratory mice (Mus musculus)

The task of the present study was to evaluate effect of social status and of genotype on success of reproduction of male laboratory mice with use of ethological model of social hierarchy of “minimal socium”. The model represents the paired maintenance of two male mice of different genotypes (PT and CBA/Lac). The mice resided for 30 days in one experimental cage and established between them the dominant-subordinant relations. After this, each pair of males was supplemented by two DD/He females. Regardless of their genotype, the dominants became fathers more often than the subordinants and left more offspring. It has been established that the complete suppression of fertility did not occur. It is suggested that this allows them to make genetic contribution to the next generation and to remain in the genetic pool of population.     

Interstrain differences in the maturation of congenital reflexes in 101/HY and CBA mice in early postnatal ontogeny]

We have identified interstrain differences in the rate of formation of certain reflexes and parameters of physical development in CBA/LacSto and 101/HY mice. We have shown that in young 101/HY mice, the maturation of reflexes reflecting the development of the vestibular, neurosensory, and neuromuscular systems, such as surface righting, bar holding, negative geotaxis, and auditory startle response, takes place later during postnatal life. Opening of eyes and auditory canals in mice of all studied groups takes place at the same time. In CBA females, maturation of cliff avoidance reflex occurs later than in 101/HY females; hair also appeared later. Body weight of young 101/HY mice of both sexes is significantly higher during the period of postnatal development from day 2 to day 20 than in the CBA strain. However, the relative brain weight was lower in the 101/HY strain. CBA males had a higher brain weight and also showed a faster rate of formation of inborn reflexes. We discuss possible factors underlying the observed difficulties in the rate of formation of reflexes in 101/HY and CBA young mice in relation to general background information about their genetic and neurobehavioral features. The results provide evidence that these differences are genetically determined; the rate of reflex formation does not depend on the overall physical development of mice and is rather due to a delay and/or abnormalities in nervous system development.     

Experience of defeat increases the susceptibility to catatonic - like state in mice

The ability to develop the catatonic-like state (reflex immobility reaction - RIR) due to stimulation of the skin at the scruff was investigated in mice of two strains — C57BL/6J and CBA/Lac. The total time of immobility and number of paroxysms during test were measured. It has been shown that the number of paroxysms was significantly fewer and the total time of immobility was significantly longer in CBA/Lac strain than in C57BL/6J. In each strain group housed animals as well as submissive ones with successive experience of defeats demonstrated a more expressed immobility than individually housed or aggressive males with successive experience of victories, respectively. Changing the social status in aggressive animals as a consequence of confrontation with aggressive males resulted in the increased immobility in CBA/Lac but not in C57BL/6J mice. The results suggest that the experience of defeat in submissive males is connected with increased ability to develop RIR.     

Comparative characteristics of the parameters of an aggressive reaction in 2 mouse genotypes

The parameters have been studied of the aggressive reaction of male mice of CBA/Lac and C57BL/6J lines differing by olfactory sensitivity to zoosocial pheromone stimuli. It has been shown that CBA males, characterized by a high olfactory sensitivity, have lower latency of the first attack than C57BL males with low olfactory sensitivity. A prolonged distant exposition to an unknown litter and male appearance lowers the latency of the first attack in mice of the studied lines proportionally to their meanings demonstrated after short time exposition. The number of attacks and total time of attacking is considerably higher in C57BL mice during the whole test period (15 min) than in CBA mice in which aggressivity is already sharply lowered after 5 min of agonistic interactions. The factors are discussed, influencing the parameters of mice aggressive reaction.     

Effect of intranasal administration of dopamine on anxiety and locomotor activity of two lines of mice

Intranasal administration of dopamine (0.3; 3 and 30 microg/kg) on anxious behaviour of mice was studied using elevated plus-maze and open fields tests and the pinch-induced catalepsy on parallel bars test. Dopamine was introduced as nose drops or inhalation of nanoparticles of the compound solution in C57B1/6J and CBA/Lac mice with differences of dopaminergic function features. In our experiment, dopamine had anxiolytic and elevated motor activity effects in C57B1/6J, but not in the CBA/Lac mice. Nose drops were more effective than inhalation; perhaps, it was a more stressful manipulation. Apparently dopamine increased the number of CBA/Lac mice who demonstrated catalepsy and the reflex duration. Indeed, the neurotransmitter is active in different psycho-emotional phenomena.     

Immunological,Cytogenetical, and Behavioral Changes in CBA and C57BL/6 Male Mice after Pheromonal Action

Immunological and cytogenetical changes were studied in mice-recipients of CBA and C57BL/6 lines as well as modifications of their behavior appearing as a result of action of various stressors. Post-stress volatile excretions of CBA males cause led to a pronounced immunosuppression in males-recipients of both lines. This resulted in a decrease of capability for immune response of the stressed mice to sheep erythrocytes. Besides, in young males of the CBA line, frequency of chromosome aberrations in bone marrow cells increased statistically significantly after stressing by syngenec volatile excretions of mature animals. Attractive properties of volatile excretions of the CBA and C57BL/6 line males were studied. In norm, animals of both lines prefer syngenic odors of intact males donors. However, for the CBA line males, post-stress excretions from males-donors of both lines became more attractive than excretions from intact animals. Also revealed were interline differences in incidence of choice of syngenic and allogenic post-stress excretions. This can be explained both by different stress-reactivity of animals-donors and differences of recipients of the studied genotypes in selectivity to olfactory stimuli. An importance of olfactory stresses and of the revealed interline differences for explanation of some micro-and macroevolutionary synecological processes in the domestic mouse is discussed.     

Effects of repeated experience of aggression on aggressive motivation and development of anxiety in male mice

The sensory contact technique allows an aggressive type of behavior to be formed as a result of repeated experience of social victories in daily agonistic confrontations. In mice of the low-aggressive and high-emotional CBA/Lac strain repeated positive fighting experience increases plus-maze anxiety. Behavioral reactivity of males to other conspecifics in the partition test (which measures aggressive motivation) significantly rises. It is concluded that repeated experience of aggression provokes the development of anxiety in male mice. The results suggest that level of anxiety and its behavioral realization depend on duration of aggressive experience and genetic strain.     

Effect of a single severe stress on behaviour of males and females in the mouse inbred strains CBA/Lac and C57BL/6J

The effect of single severe stress in the form of forced swimming on the behavior of males and females in the mouse inbred strains CBA/Lac and C57BL/6J were examined in the open field test. Measurements were carried out within two hours after the stress exposure (Trial 1) and repeated 2 hours thereafter (Trial 2). Intact males and females of the both mouse strains which tested in the open field twice too were used as control. An increased latency was found until first escape from the center both in males and females of the CBA/Lac strain within two hours after the end of forced swimming. This parameter was still high in females in the Trial2. Four out of seven behavior parameters were changed in females of the C57BL/6J strain two hours after the stress exposure, but their behavior was similar to control in the Trial 2. The males of the C57BL/6J strain demonstrated the least changed behavior in the open field test after the stress exposure with the exception of increased number of grooming in the Trial 1. Further on, a detailed analysis of repeated testing in the open field within intact and stressed mice of both strains was performed. This comparison allowed revealing hereditary and gender peculiarities in the mouse behavior after single severe stress exposure. The results are discussed in respect to the possible genetically inherent increased traitanxiety in females of C57BL/6J strain and the state of anxiety in females of CBA/Lac strain.