El proceso de congelación-descongelación altera genéticamente el esperma del jabalí

Title: Freezing-thawing induces alterations in histone H1-DNA binding and the breaking of protein-DNA disulfide bonds in boar sperm

Abstract: The main aim of this work is to gain insight into the mechanisms by which freezing-thawing alters the nucleoprotein structure of boar sperm. For this purpose, the freezing-thawing-related changes of structure and location of histones-DNA domains in the boar sperm head were analyzed through Western blot and immunocytochemistry. Afterwards, it was analyzed whether freezing-thawing induced changes in tyrosine phosphorylation levels of both protamine 1 and histone HI, through Western blot analyses in samples previously subjected to immunoprecipitation. This analysis was completed with the determination of the changes induced by freezing-thawing on the overall levels of sperm-head disulfide bonds through analysis of free-cysteine radicals levels. Freezing-thawing induced significant changes in the histones-DNA structures, which were manifested in the appearance of a freezing-thawing-linked histone HI-DNA aggregate of about a 35-kDa band and in the spreading of histone Hi-positive markings from the caudal area of the sperm head to more cranial zones. Freezing-thawing did not have any significant effect on the tyrosine phosphorylation levels of either protamine I or histone HI. However, thawed samples showed a significant (P < 0.05) increase in the free cysteine radical content (from 3.1 +/- 0.5nmol/mu g protein in fresh samples to 6.7 +/- 0.8 nmol/mu g protein). In summary, our results suggest that freezing-thawing causes significant alterations in the nucleoprotein structure of boar sperm head by mechanism/s linked with the rupture of disulfide bonds among the DNA. These mechanisms seem to be unspecific, affecting both the protamines-DNA unions and the histones-DNA bonds in a similar way. Furthermore, results suggest that the boar-sperm nuclear structure is heterogeneous suggesting the existence of a zonated pattern, differing in their total DNA density and the compactness of the precise nucleoprotein structures present in each zone.

Authors: Flores, E (Flores, E.)1; Ramio-Lluch, L (Ramio-Lluch, L.)1; Bucci, D (Bucci, D.)2; Fernandez-Novell, JM (Fernandez-Novell, J. M.)3,4; Pena, A (Pena, A.)1; Rodriguez-Gil, JE (Rodriguez-Gil, J. E.)1

Addresses:
1. Autonomous Univ Barcelona, Dept Anim Med & Surg, Sch Vet Med, E-08193 Barcelona, Spain
2. Univ Bologna, DIMORFIPA, I-40064 Bologna, Italy
3. Univ Barcelona, Dept Biochem & Mol Biol, E-08028 Barcelona, Spain
4. Univ Barcelona, IRRB, E-08028 Barcelona, Spain