Production of F₁ offspring with vitrified sperm from a live-bearing fish, the green swordtail Xiphophorus hellerii.

Abstract:

:This study reports the first production of offspring with vitrified sperm from a live-bearing fish Xiphophorus hellerii. The overall goal of this study was to develop streamlined protocols for integration into a standardized approach for vitrification of aquatic species germplasm. The objectives were to (1) estimate acute toxicity of cryoprotectants, (2) evaluate vitrification solutions, (3) compare different thawing methods, (4) evaluate membrane integrity of post-thaw sperm vitrified in different cryoprotectants, and (5) evaluate the fertility of vitrified sperm. Nine cryoprotectants and two commercial vitrification additives were tested for acute toxicity and glass forming ability, alone and in combination. Two vitrification solutions, 40% glycerol (Gly) and 20% Gly+20% ethylene glycol (EG) in 500 mOsmol/kg Hanks' balanced salt solution (HBSS), were selected for vitrification of 10 μL sperm samples using inoculating loops plunged into liquid nitrogen. Samples were thawed at 24°C (one loop in 5 μL of HBSS or three loops in 500 μL of HBSS). Samples thawed in 500 μL were concentrated by centrifugation (1000 g for 5 min at 4°C) into 5 μL for artificial insemination. Offspring were produced from virgin females inseminated with sperm vitrified with 20% Gly+20% EG and concentrated by centrifugation.

journal_name

Zebrafish

journal_title

Zebrafish

authors

Cuevas-Uribe R,Yang H,Daly J,Savage MG,Walter RB,Tiersch TR

doi

10.1089/zeb.2011.0704

subject

Has Abstract

pub_date

2011-12-01 00:00:00

pages

167-79

issue

4

eissn

1545-8547

issn

1557-8542

journal_volume

8

pub_type

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