Doklady Biological Sciences, Vol. 387, 2002, pp. 508–509. Translated from Doklady Akademii Nauk, Vol. 387, No.

5, 2002, pp. 697–698. Original Russian Text Copyright © 2002 by Novikov, Tsapygina, Babalyan.

PHYSIOLOGY

The Period of the Sensitivity to the Pheromone Inhibiting Spermatogenesis in Laboratory CBAB6F1 Mice
S. N. Novikov, R. I. Tsapygina†, and V. V. Babalyan
Presented by Academician A.D. Nozdrachev July 2, 2001 Received August 12, 2002

In 1979, Russian researchers were the first to publish experimental data on a pheromone that inhibits spermatogenesis in mice [1]. This pheromone is currently known to be a complex mixture of strongly and weakly polar volatile compounds [2] contained in urine of sexually mature males [3]. In 30-day-old animals, this pheromone causes various meiotic abnormalities at the stage of diakinesis–metaphase [4] to enhance the frequency of abnormal sperm cells in the caudal part of the epididymis [5]. Animals of different genotypes differ in the sensitivity to this pheromone [6], which causes a dramatically increased frequency of dominant lethals in the progeny of young male CBAB6F1 mice [7]. Here, we report a cytogenetic analysis of meiotic abnormalities observed during the pre- and postpubertal periods in male laboratory CBAB6F1 mice aged 30, 42, and 56 days. The experimental groups consisting of four to six animals of the corresponding age were exposed to volatile components of urine in an olfactometer with the supply of the odorant in an air flow [2, 3, 5]. The time of exposure was 2 h; Vb = 4 l/min; Vo = 1 l/min, where Vb and Vo are the rates of the carrying and odorant flows, respectively. The evaporation surface in the cuvette containing urine was 5 cm2; t = 20°C. The urine was freshly collected from five- to eight-month-old CBA/LacSto males that were kept singly for two weeks before the experiment. In control groups, the animals were exposed to water in an olfactometer. Eight hours from the start of exposure, the animals were decapitated, their gonads were fixed, and the squash preparations of the seminiferous tubules were examined using a modified Evens’ technique [4]. The cells with meiotic abnormalities such as multivalent association (MA) and univalent autosome (UA) were examined at the stage of

diakinesis–metaphase I. The experiments were conducted in autumn. The figure shows that the frequency of the pheromone-induced meiotic abnormalities increased only in 30-day-old males. The control and experimental variants did not differ significantly in sexually mature animals, i.e., in six- and eight-week-old males. This testifies to a relative insensitivity of the mature recipient reproductive system to the pheromone that causes various abnormalities of meiosis and spermatogenesis [4, 5] in the generative cells of sexually immature animals. Thus, in the laboratory CBAB6F1 mice, we have found a differential spermatocyte I ontogenetic sensitivity to the genotoxic effect of the pheromone contained in the urine of sexually mature male CBA/LacSto mice.
Chromosome abnormalities, % 12 * 1 10 8 6 4 2 0 30 42 56 Age of CBAB6F1 males, days *

2

3

4

†Deceased.

Pavlov Institute of Physiology, Russian Academy of Sciences, nab. Makarova 6, St. Petersburg, 199034 Russia St. Petersburg State University, Universitetskaya nab. 7/9, St. Petersburg, 199164 Russia

Meiotic abnormalities, including autosomal univalents (AUs) and multivalent associations (MAs) in the generative cells of CBAB6F1 males of different ages exposed to volatile components of the urine of sexually mature CBA males: 1, AU (experiment); 2, AU (control); 3, MA (experiment); 4, MA (control). * Significant difference from the control (p < 0.05; the χ2 test).

0012-4966/02/1112-0508$27.00 © 2002 MAIK “Nauka /Interperiodica”

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In general, the pheromone-induced neurophysiological mechanisms responsible for the inhibition of spermatogenesis [5] and animal reproduction [7] have not been studied sufficiently. According to the assumption made in [8], the pheromone-caused inhibition of spermatogenesis in mice is directly related to neither adrenocortical activation nor a pronounced stress response of the recipient animals [9]. Although prolactin may also be involved in the regulation of this effect, we assume that the pheromone effect on 30-day-old animals leads to a selective inhibition of the gonadotropin secretion (luteinizing and follicle-stimulating hormones) [10], which results in a reduced testicular 5α-reductase activity [11], which is the highest at this ontogenetic stage [12, 13]. The imbalance between testosterone, its 5α-dihydroderivatives, and gonadotropins is presumed to result in abnormal chromosome conjugation during meiosis and inhibition of spermatogenesis in general at this extremely sensitive stage of ontogeny [14]. Based on our results and their interpretation, the phenomenon of the pheromone-induced inhibition of spermatogenesis may be assigned to the ontogenetic facultative primer effects, with the prepubertal period of the life of laboratory mice being “critical.” Hence, the concept of the pheromone primer effect may be significantly extended based on the above considerations. It is still difficult to determine whether the pheromone-induced inhibition of spermatogenesis [1–5] and the testicular [15] and gonadotropic [10] functions in the house mouse is important for the biology of the Mus musculus L., because the phenomenon has been discovered under laboratory conditions. As shown previously, the pheromone effect leads to a long-term decrease in stress resistance [4], fertility [7], and behavioral changes [8] in the recipient animals. Taking into account the important role of hierarchy in the polygamic system of mating, which is observed by the fifth to sixth week of the ontogeny of the synanthropic house mouse, as well as the genotype-dependent differential sensitivity of a recipient to the pheromone effect [6, 8], the ontogenetic facultative pheromone-induced inhibition of spermatogenesis shown in this study seems to be an important factor of the formation of the hierarchic relationships in a deme structure of either linear or partial type. Thus, the phenomenon described may determine the selective value of the dominant individual and,

hence, the efficient population size (NÂ) and serve as a vector of natural selection. ACKNOWLEDGMENTS This work was supported in part by the State Program “Frontiers in Genetics” (project no. 2.152) and by the FDS Charitable Trust (United Kingdom). REFERENCES
1. Tsapygina, R.I., Daev, E.V., and Novikov, S.N., in Issledovanie biologicheskogo deistviya antropogennykh faktorov, zagryaznyayushchikh vodoemy (Study of the Biological Effects of Anthropogenic Factors Contaminating Water Bodies), Irkutsk: Irk. Gos. Univ., 1979, pp. 157−162. 2. Novikov, S.N. and Babalyan, V.V., Dokl. Akad. Nauk SSSR, 1988, vol. 302, no. 2, pp. 470–472. 3. Babalyan, V.V. and Novikov, S.N., Dokl. Akad. Nauk, 2001, vol. 378, no. 3, pp. 407–409. 4. Novikov, S.N., Tsapygina, R.I., Daev, E.V., and Togo, E.F., Dokl. Akad. Nauk SSSR, 1982, vol. 262, no. 3, pp. 746–748. 5. Aref’ev, A.A., Daev, E.V., Kaidanov, L.Z., et al., Dokl. Akad. Nauk SSSR, 1986, vol. 291, no. 5, pp. 1257–1259. 6. Daev, E.V., Genetika (Moscow), 1994, vol. 30, pp. 1105–1112. 7. Daev, E.V., Tsapygina, R.I., and Lopatina, N.G., Genetika (Moscow), 1988, vol. 24, pp. 2015–2021. 8. Novikov, S.N., Feromony i razmnozhenie mlekopitayushchikh (Pheromones and Reproduction in Mammals), Leningrad: Nauka, 1988. 9. Pavlova, M.B., Garina, I.A., Dyuzhikova, N.A., et al., Fiziol. Zh. im. I.M. Sechenova, 1989, vol. 75, pp. 138−142. 10. Clancy, A.N., Singer, A.G., Macrides, F., et al., Biol. Reprod., 1988, vol. 38, pp. 183–191. 11. Murono, E.P. and Payne, A.H., Biol. Reprod., 1979, vol. 20, pp. 911–917. 12. Murono, E.P., Acta Endocrinol., 1989, vol. 121, pp. 477−483. 13. Weis, D.B., Rodriques, L.J., Smith, K.D., and Steinberger, A., Gen. Comp. Endocrinol., 1982, vol. 46, p. 399. 14. McLachlan, R.I., Wreford, N.G., O’Donnell, L., et al., J. Endocrinol., 1996, vol. 148, pp. 1–9. 15. Pandey, S.C. and Pandey, S.D., Arch. Biol., 1986, vol. 97, pp. 129–138.

DOKLADY BIOLOGICAL SCIENCES

Vol. 387

2002