Establishment of a Practical Sperm Cryopreservation Pathway for the Axolotl (<i>Ambystoma mexicanum</i>): A Community-Level Approach to Germplasm Repository Development
The axolotl (<i>Ambystoma mexicanum</i>) draws great attention around the world for its importance as a biomedical research model, but housing and maintaining live animals is increasingly expensive and risky as new transgenic lines are developed. The goal of this work was to develop an i...
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MDPI AG
2024-01-01
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| Online Access: | https://www.mdpi.com/2076-2615/14/2/206 |
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| author | Nicholas Coxe Yue Liu Lucía Arregui Rose Upton Sarah Bodenstein Steven Randal Voss Maria T. Gutierrez-Wing Terrence R. Tiersch |
| author_facet | Nicholas Coxe Yue Liu Lucía Arregui Rose Upton Sarah Bodenstein Steven Randal Voss Maria T. Gutierrez-Wing Terrence R. Tiersch |
| author_sort | Nicholas Coxe |
| collection | DOAJ |
| description | The axolotl (<i>Ambystoma mexicanum</i>) draws great attention around the world for its importance as a biomedical research model, but housing and maintaining live animals is increasingly expensive and risky as new transgenic lines are developed. The goal of this work was to develop an initial practical pathway for sperm cryopreservation to support germplasm repository development. The present study assembled a pathway through the investigation of axolotl sperm collection by stripping, refrigerated storage in various osmotic pressures, cryopreservation in various cryoprotectants, and in vitro fertilization using thawed sperm. By the stripping of males, 25–800 µL of sperm fluid was collected at concentrations of 1.6 × 10<sup>6</sup> to 8.9 × 10<sup>7</sup> sperm/mL. Sperm remained motile for 5 d in Hanks’ Balanced Salt Solution (HBSS) at osmolalities of 100–600 mOsm/kg. Sperm cryopreserved in 0.25 mL French straws at 20 °C/min in a final concentration of 5% DMFA plus 200 mM trehalose and thawed at 25 °C for 15 s resulted in 52 ± 12% total post-thaw motility. In six in vitro fertilization trials, 20% of eggs tested with thawed sperm continued to develop to stage 7–8 after 24 h, and a third of those embryos (58) hatched. This work is the first report of successful production of axolotl offspring with cryopreserved sperm, providing a general framework for pathway development to establish <i>Ambystoma</i> germplasm repositories for future research and applications. |
| first_indexed | 2024-03-08T11:08:41Z |
| format | Article |
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| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-03-08T11:08:41Z |
| publishDate | 2024-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Animals |
| spelling | doaj.art-a862d83300114ce8b51fd14afadaf13f2024-01-26T14:30:48ZengMDPI AGAnimals2076-26152024-01-0114220610.3390/ani14020206Establishment of a Practical Sperm Cryopreservation Pathway for the Axolotl (<i>Ambystoma mexicanum</i>): A Community-Level Approach to Germplasm Repository DevelopmentNicholas Coxe0Yue Liu1Lucía Arregui2Rose Upton3Sarah Bodenstein4Steven Randal Voss5Maria T. Gutierrez-Wing6Terrence R. Tiersch7Aquatic Germplasm and Genetic Resources Center, School of Renewable Natural Resources, Louisiana State University Agricultural Center, Baton Rouge, LA 70820, USAAquatic Germplasm and Genetic Resources Center, School of Renewable Natural Resources, Louisiana State University Agricultural Center, Baton Rouge, LA 70820, USAAquatic Germplasm and Genetic Resources Center, School of Renewable Natural Resources, Louisiana State University Agricultural Center, Baton Rouge, LA 70820, USAAquatic Germplasm and Genetic Resources Center, School of Renewable Natural Resources, Louisiana State University Agricultural Center, Baton Rouge, LA 70820, USAAquatic Germplasm and Genetic Resources Center, School of Renewable Natural Resources, Louisiana State University Agricultural Center, Baton Rouge, LA 70820, USADepartment of Neuroscience, Ambystoma Genetic Stock Center and Spinal Cord and Brain Injury Research Center, University of Kentucky, Lexington, KY 40536, USAAquatic Germplasm and Genetic Resources Center, School of Renewable Natural Resources, Louisiana State University Agricultural Center, Baton Rouge, LA 70820, USAAquatic Germplasm and Genetic Resources Center, School of Renewable Natural Resources, Louisiana State University Agricultural Center, Baton Rouge, LA 70820, USAThe axolotl (<i>Ambystoma mexicanum</i>) draws great attention around the world for its importance as a biomedical research model, but housing and maintaining live animals is increasingly expensive and risky as new transgenic lines are developed. The goal of this work was to develop an initial practical pathway for sperm cryopreservation to support germplasm repository development. The present study assembled a pathway through the investigation of axolotl sperm collection by stripping, refrigerated storage in various osmotic pressures, cryopreservation in various cryoprotectants, and in vitro fertilization using thawed sperm. By the stripping of males, 25–800 µL of sperm fluid was collected at concentrations of 1.6 × 10<sup>6</sup> to 8.9 × 10<sup>7</sup> sperm/mL. Sperm remained motile for 5 d in Hanks’ Balanced Salt Solution (HBSS) at osmolalities of 100–600 mOsm/kg. Sperm cryopreserved in 0.25 mL French straws at 20 °C/min in a final concentration of 5% DMFA plus 200 mM trehalose and thawed at 25 °C for 15 s resulted in 52 ± 12% total post-thaw motility. In six in vitro fertilization trials, 20% of eggs tested with thawed sperm continued to develop to stage 7–8 after 24 h, and a third of those embryos (58) hatched. This work is the first report of successful production of axolotl offspring with cryopreserved sperm, providing a general framework for pathway development to establish <i>Ambystoma</i> germplasm repositories for future research and applications.https://www.mdpi.com/2076-2615/14/2/206axolotl<i>Ambystoma</i>spermcryopreservationpathwayprocess map |
| spellingShingle | Nicholas Coxe Yue Liu Lucía Arregui Rose Upton Sarah Bodenstein Steven Randal Voss Maria T. Gutierrez-Wing Terrence R. Tiersch Establishment of a Practical Sperm Cryopreservation Pathway for the Axolotl (<i>Ambystoma mexicanum</i>): A Community-Level Approach to Germplasm Repository Development Animals axolotl <i>Ambystoma</i> sperm cryopreservation pathway process map |
| title | Establishment of a Practical Sperm Cryopreservation Pathway for the Axolotl (<i>Ambystoma mexicanum</i>): A Community-Level Approach to Germplasm Repository Development |
| title_full | Establishment of a Practical Sperm Cryopreservation Pathway for the Axolotl (<i>Ambystoma mexicanum</i>): A Community-Level Approach to Germplasm Repository Development |
| title_fullStr | Establishment of a Practical Sperm Cryopreservation Pathway for the Axolotl (<i>Ambystoma mexicanum</i>): A Community-Level Approach to Germplasm Repository Development |
| title_full_unstemmed | Establishment of a Practical Sperm Cryopreservation Pathway for the Axolotl (<i>Ambystoma mexicanum</i>): A Community-Level Approach to Germplasm Repository Development |
| title_short | Establishment of a Practical Sperm Cryopreservation Pathway for the Axolotl (<i>Ambystoma mexicanum</i>): A Community-Level Approach to Germplasm Repository Development |
| title_sort | establishment of a practical sperm cryopreservation pathway for the axolotl i ambystoma mexicanum i a community level approach to germplasm repository development |
| topic | axolotl <i>Ambystoma</i> sperm cryopreservation pathway process map |
| url | https://www.mdpi.com/2076-2615/14/2/206 |
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