A significant aspermatogenic activity, ascertained by microscopic studies of seminiferous tubules and interstitial tissue, and by the observation of the entrance of immunity and fibrocytic cells in mice injected with polyspermine (PS) or polyspermine conjugated to monomeric or dimeric RNase A (PS-RNase A or PS-dimeric RNase A, respectively), was found either in mice injected or in non-injected testes. Polyspermine and its complexes with RNase A destroyed all spermatogenic and intertestitial tissue, including Leydic cells, as well as their ability to secrete testosterone. The total loss of spermatogenic activity in injected testes is irreversible because spermatogonia cells also were destroyed. The injection of PS into both mice testes determined the total degeneration of testicle tissue in 50% of injected testes. The second half of testes was also partly degenerated, and if they were re-injected, almost all testes were fully destroyed. PS-dimeric RNase A injected once into both testicles produced a stronger degeneration and also the interruption of testosterone secretion in comparison with the effects due to injection of mice with PS or PS-RNase A. In all mice treated with these substances, as well as in rats in which PS was injected twice into their testes, we detected polymorfonucleates, monocytes, plasma cells, lymphocytes and fibrocytic cells. Antibodies against PS, PS-RNase A or PS-dimeric RNase A did not influence the aspermatogenic activity. Animals in which a repeated intra-peritoneal injection was carried out did not lose body mass and remained in good condition, with the exception of mice injected with spermine.
aspermatogenesis; cell degeneration; testes; spermine; polyspermine; polyspermine-RNase A
Agostinelli E, Arancia G, Vedova LD, Belli F, Marra M, Salvi M, Toninello A: The biological functions of polyamine oxidation products by amine oxidases: Perspectives of clinical applications. Amino Acids 27:347-358, 2004.
Dudley HW, Rosenheim O, Starling WW: The constitution and synthesis of spermidine, a newly discovered base isolated from animal tissues. Biochem J 21:97-101, 1927.
Gotte G, Testolin L, Costanzo C, Sorrentino S, Armato U, Libonati M: Cross-linked trimers of bovine ribonuclease A: Activity on doublestranded RNA and antitumour action. FEBS Lett 415:308-312, 1997.
Jensen DE, von Hippel PH: DNA “melting“ proteins. I. Effects of bovine pancreatic ribonuclease binding on the conformation and stability of DNA. J Biol Chem 251:7198-7214, 1976.
Libonati M, Beintema JJ: Basic charges on mammalian ribonuclease molecules and activity towards double-stranded polyribonucleotids. Biochem Soc Trans 5:470-474, 1977.
Libonati M, Gotte G: Oligomerization of bovine ribonuclease A: Structural and functional features of its multimers. Biochem J 380:311-327, 2004.
Libonati M, Sorrentino S: Degradation of doublestranded RNA by mammalian pancreatic-type ribonucleases. Methods Enzymol 341:234-248, 2001.
Libonati M, Malorni MC, Parente A, D´Alessio G: Degradation of double-stranded RNA by a monomeric derivative of ribonuclease BS-1. Biochim Biophys Acta 402:83-87. 1975a.
Libonati M, Sorrentino S, Galli R, La Montagna R, Di Donato A: Degradation of DNA-RNA hybrids by aggregates of pancreatic ribonuclease. Biochim Biophys Acta 407:292-298, 1975b.
Libonati M, Furia A, Beintema JJ: Basic charges on mammalian ribonuclease molecules and the ability to attack double-stranded RNA. Eur J Biochem 69:445-451, 1976.
Lindsay GS, Wallace HM: Changes in polyamine catabolism in HL-60 human promyelogenous leukaemic cells in response to etoposideinduced apoptosis. Biochem J 337:83-87, 1999.
Mann T, Lutwak-Mann C: Male Reproductive Function and Semen. Springer-Verlag, Berlin, Heidelberg, New York 1981, pp. 291-295.
Matousek J: Aspermatogenic effect of the bull seminal ribonuclease (BS-RNase) in the presence of anti-BS-RNase antibodies in mice. Anim Genet 25(Suppl. 1):45-50, 1994.
Matousek J, Gotte G, Pouckova P, Soucek J, Slavik T, Vottariello F, Libonati M: Antitumour activity and other biological actions of oligomers of ribonuclease A. J Biol Chem 278:23817-23822, 2003.
Pegg AF: Recent advances in the biochemistry of polyamines in eukaryotes. Biochem J 234:249-262, 1986.
Pouckova P, Morbio M, Vottariello F, Laurents DV, Matousek J, Soucek J, Gotte G, Donadelli M, Costanzo C, Libonati M: Cytotoxicity of Polyspermine-Ribonuclease A and Polyspermine-Dimeric Ribonuclease A. Bioconjug Chem - in press.
Schreiner P: Neber eine organische Basis in thierischen Organismem. Liebigs Annalen, 94:68-72, 1878.
Seiler N, Delcros JG, Moulinoux JP: Polyamine transport in mammalian cells. An update. Int J Biochem Cell Biol 28:843-861, 1996.
Sorrentino S, Libonati M: Human pancreatic-type and nonpancreatic-type ribonucleases: a direct side-by-side comparison of their catalytic properties. Arch Biochem Biophys 312:340-348, 1994.
Soucek J, Marinov J, Benes J, Hilgert J, Matousek J, Raines RT: Immunosuppressive activity of bovine seminal ribonuclease and its mode of action. Immunobiology 195:271-285, 1996.
Wang D, Wilson G, Moore S: Preparation of crosslinked dimers of pancreatic ribonuclease. Biochemistry 15:660-665, 1976.
Wang D, Moore S: Polyspermine-ribonuclease prepared by cross-linkage with dimethyl suberimidate. Biochemistry 16:2937-2941, 1977.
Yatin M, Venkataraman GM, Marcinek R, Ain KB: Polyamine synthesis and transport inhibition in a human anaplastic thyroid carcinoma cell line in vitro and as xenograft tumours. Thyroid 9:805-814, 1999.