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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Detection of protamine 2 in bovine spermatozoa and testicles

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Author(s):
Hamilton, T. R. S. [1] ; Simoes, R. [2] ; Mendes, C. M. [1] ; Goissis, M. D. [1] ; Nakajima, E. [3] ; Martins, E. A. L. [3] ; Visintin, J. A. [1] ; Assumpcao, M. E. O. A. [1]
Total Authors: 8
Affiliation:
[1] Univ Sao Paulo, Dept Anim Reprod, Sch Vet Med, Ave Prof Doutor Orlando Marques de Paiva, Sao Paulo - Brazil
[2] Fed Univ ABC, Ctr Nat & Human Sci, Santo Andre - Brazil
[3] Butantan Inst, Proc Dev Lab, Inovat Div, Sao Paulo - Brazil
Total Affiliations: 3
Document type: Journal article
Source: ANDROLOGY; v. 7, n. 3, p. 373-381, MAY 2019.
Web of Science Citations: 0
Abstract

Background Sperm DNA integrity is crucial for transmission of genetic information to future generations and DNA damage can occur during chromatin packaging. Chromatin packaging involves the replacement of somatic nucleosomal histones by nuclear proteins called protamines. Protamine 1 (PRM1) is transcribed and translated in spermatids of all mammals; however, protamine 2 (PRM2) is transcribed in low levels in spermatids and it is not yet described in bull mature spermatozoa. Objectives The aim of this study was to assess gene and protein expression of PRM2 and corroborate gene and protein expression of PRM1 in bull spermatozoa and testis. Materials and methods For this purpose, absolute q-RT-PCR was performed to calculate the number of copies of PRM1 and PRM2 mRNAs in bovine epididymal spermatozoa and testicular tissue. Western blot and mass spectrometry were performed to identify PRM1 and PRM2 in samples of bovine epididymal spermatozoa. Samples of bovine testicular tissue were collected to identify PRM1 and PRM2 by immunohistochemistry. Results We evaluated that the number of PRM1 mRNA copies was about hundred times higher than PRM2 mRNA copies in sperm and testicular samples (p < 0.0001). In addition, we estimated the PRM1: PRM2 ratio based on mRNA number of copies. In spermatozoa, the ratio was 1: 0.014, and in testicle, the ratio was 1: 0.009. We also evaluated the immunolocalization for PRM1 and PRM2 in bovine testis, and both proteins were detected in spermatids. Western blot and mass spectrometry in bovine epididymal spermatozoa confirmed these results. Conclusion Our work identifies, for the first time, PRM2 in bovine epididymal spermatozoa and in testis. Further studies are still needed to understand the role of PRM2 on the chromatin of the spermatozoa and to verify how possible changes in PRM2 levels may influence the bull fertility. (AU)

FAPESP's process: 11/19551-1 - Relationship between protamine type and endonuclease activity in the protection of the bovine sperm DNA
Grantee:Mayra Elena Ortiz D' Avila Assumpcao
Support Opportunities: Regular Research Grants