An improved acetic acid-urea polyacrylamide electrophoresis method to evaluate bovine sperm protamines

Autor(es)

dos Santos Hamilton, Thais Rose ^[1] ; Simoes, Renata ^[2] ; Ortiz D'Avila Assumpcao, Mayra Elena ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Sch Vet Med & Anim Sci, Dept Anim Reprod, Av Prof Orlando Marques de Paiva 87, Cidade Univ, Sao Paulo - Brazil

^[2] Fed Univ ABC, Ctr Nat & Human Sci, Santo Andre, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: REPRODUCTION IN DOMESTIC ANIMALS; v. 56, n. 7, p. 1050-1056, JUL 2021.
• Citações Web of Science: 0

Resumo

The acetic acid-urea polyacrylamide gel electrophoresis system could separate very similar basic proteins on differences in size and effective charge. This system has been used for many years to analyse histones and their post-translational modifications and widely used in the study of mammal protamines. Two types of protamine have been described, the protamine 1 (P1) and the protamine 2 (P2) family members, which are synthetized by PRM1 and PRM2 genes. The ratio of P1 and P2 is important for predicting fertility in humans and mice. Therefore, the quantification of protamines is a fundamental step in order to establish the ratio between P1 and P2 in these species. In other mammals, studies linking sperm protamination and the protamine ratio with fertility are increasing. So, the use of an effective technique to separate and quantify protamines is important to study sperm P1/P2 ratio. Therefore, this article describes in detail a feasible and useful procedure to isolate bovine sperm protamines, to perform pre-electrophoresis with PEG solution and finally to carry out acid-urea polyacrylamide gel electrophoresis in reverse polarity. This technique allows a clear separation and efficient detection of bovine sperm protamines. (AU)

• Processo FAPESP: 11/11231-8 - Estudo dos fatores envolvidos na fragmentação de DNA dos espermatozóides em ovinos.
• Beneficiário: Thais Rose dos Santos Hamilton
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 10/09145-3 - Expressão e localização das protaminas e das proteínas de transição em espermatozóides de bovinos
• Beneficiário: Renata Simões
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado