Cholesterol-loaded cyclodextrin is efficient in preserving sperm quality of cryopreserved ram semen with low freezability

Author(s)

Batissaco, Leonardo ^[1] ; de Arruda, Rubens Paes ^[2] ; Rodrigues Alves, Maira Bianchi ^[1] ; Torres, Mariana Andrade ^[3] ; Lemes, Kleber Menegon ^[2] ; Prado-Filho, Roberto Romano ^[1] ; de Almeida, Tamie Guibu ^[1] ; Cesar de Andrade, Andre Furugen ^[3] ; Carvalho Celeghini, Eneiva Carla ^[1]

Affiliation

^[1] Univ Sao Paulo, Ctr Biotechnol Anim Reprod, Lab Teaching & Res Pathol Reprod, Dept Anim Reprod, Sch Vet Med & Anim Sci, Pirassununga, SP - Brazil

^[2] Univ Sao Paulo, Sch Vet Med & Anim Sci, Ctr Biotechnol Anim Reprod, Lab Semen Biotechnol & Androl, Dept Anim Reprod, Pirassununga, SP - Brazil

^[3] Univ Sao Paulo, Sch Vet Med & Anim Sci, Dept Anim Reprod, Lab Androl & Embryo Transfer Swine, Pirassununga, SP - Brazil

• Document type: Journal article
• Source: REPRODUCTIVE BIOLOGY; v. 20, n. 1, p. 14-24, MAR 2020.
• Web of Science Citations: 1

Abstract

Semen freezability is positive correlated with the cholesterol content in the sperm cell. Freeze-thawing mainly cause temperature chock and change on media osmolarity, which can modify plasma membrane lipids content and sperm conformation, resulting in decreased fertility. Therefore, the aim of this study is to investigate the effect of adding cholesterol-loaded cyclodextrin (CLC) to the cryopreservation process of ram semen with low freezability. For that, two experiments were performed using 5 ejaculates of 6 rams, totalizing 30 samples. For experiment 1 the following treatments were tested: in natura (IN), Tris solution (CON), CLC + Tris solution (CLC), and pure methyl-beta-cyclodextrin + Tris solution (MCD). For experiment 2 treatments CON and CLC were tested in samples subdivided into three freezability classes: high (n = 10), intermediate (n = 10) and low (n = 10). Freezability classes were based on the variation of sperm motility between IN and CON groups from the first experiment. Sample analyzes included sperm motility, sperm morphology, plasma and acrosome membrane integrity, mitochondrial membrane potential, reactive oxygen species content, lipid peroxidation, and fluidity of plasma membrane. Results showed that CLC treatment was more efficient in maintaining sperm motility, integrity of plasma membrane, integrity of acrosome, and mitochondria membrane potential. In addition, CLC treatment in the groups with low and intermediate freezability showed improvement on progressive motility and percentage of rapid cells. In contrast, no difference was noted between CLC and CON treatments in the high freezability group. Therefore, the addition of CLC to semen extender improved sperm cryopreservation, especially in rams with low freezability. (AU)

• FAPESP's process: 12/00040-0 - Testicular degeneration treatment in ram with vitamin and mineral supplementation or low intensity laserterapy
• Grantee: Eneiva Carla Carvalho Celeghini
• Support Opportunities: Regular Research Grants
• FAPESP's process: 12/09927-7 - Cryopreservation of ram semen with incorporation of cholesterol through cyclodextrin
• Grantee: Leonardo Batissaco
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 13/13438-4 - Semen freezeability of ram supplemented with vitamin A or submitted to low intensity laser terapy
• Grantee: Tamie Guibu de Almeida
• Support Opportunities: Scholarships in Brazil - Scientific Initiation