Evaluation of Female Recipient Infertility and Donor Spermatogonial Purification for Germ Cell Transplantation in <i>Paralichthys olivaceus</i>

Since the advent of germ cell transplantation (GCT), it has been widely used in shortening the fish breeding cycle, sex-controlled breeding and the protection of rare and endangered fish. In this study, the effectiveness of female sterile recipient preparation and donor stem cell isolation and purif...

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Main Authors: Yuqin Ren, Yuehong Tao, Zhaohui Sun, Yufen Wang, Weidong Li, Zhongwei He, Guixing Wang, Yucong Yang, Jilun Hou
Format: Article
Language:English
Published: MDPI AG 2024-10-01
Series:Animals
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Online Access:https://www.mdpi.com/2076-2615/14/19/2887
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author Yuqin Ren
Yuehong Tao
Zhaohui Sun
Yufen Wang
Weidong Li
Zhongwei He
Guixing Wang
Yucong Yang
Jilun Hou
author_facet Yuqin Ren
Yuehong Tao
Zhaohui Sun
Yufen Wang
Weidong Li
Zhongwei He
Guixing Wang
Yucong Yang
Jilun Hou
author_sort Yuqin Ren
collection DOAJ
description Since the advent of germ cell transplantation (GCT), it has been widely used in shortening the fish breeding cycle, sex-controlled breeding and the protection of rare and endangered fish. In this study, the effectiveness of female sterile recipient preparation and donor stem cell isolation and purification were comprehensively evaluated for spermatogonial stem cell transplantation (SSCT) in <i>Paralichthys olivaceus</i>. The best way to prepare sterile recipients was found to be giving three-year-old fish four intraovarian injections of busulfan (20 mg/kg body weight) combined with exposure to a high temperature (28 °C) after the spawning season compared with the two other ways, which induced apoptosis of most of the endogenous germ cells, resulting in shrinkage of the spawning plate and enlargement of the ovarian lumen. Further analysis showed that both the gonadosomatic index and germ-cell-specific <i>vasa</i> expression were significantly lower than those of the natural-temperature group before treatment (<i>p</i> < 0.05). A high percentage (>60.00%) of spermatogonial stem cells (SSCs) were obtained after isolation and purification and were transplanted into the prepared recipients. After three weeks of SSCT, the numbers of PKH26-labeled SSCs were increased in the ovaries of the recipients. These findings provide a basis for the establishment of an ideal SSCT technique using <i>P. olivaceus</i> females as the recipients, ultimately contributing to the efficient conservation of male germplasm resources and effective breeding.
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spelling doaj-art-37372f575fac4fad82e76bcbb78e96f52025-08-20T01:47:44ZengMDPI AGAnimals2076-26152024-10-011419288710.3390/ani14192887Evaluation of Female Recipient Infertility and Donor Spermatogonial Purification for Germ Cell Transplantation in <i>Paralichthys olivaceus</i>Yuqin Ren0Yuehong Tao1Zhaohui Sun2Yufen Wang3Weidong Li4Zhongwei He5Guixing Wang6Yucong Yang7Jilun Hou8Hebei Key Laboratory of the Bohai Sea Fish Germplasm Resources Conservation and Utilization, Beidaihe Central Experiment Station, Chinese Academy of Fishery Sciences, Qinhuangdao 066100, ChinaHebei Key Laboratory of the Bohai Sea Fish Germplasm Resources Conservation and Utilization, Beidaihe Central Experiment Station, Chinese Academy of Fishery Sciences, Qinhuangdao 066100, ChinaHebei Key Laboratory of the Bohai Sea Fish Germplasm Resources Conservation and Utilization, Beidaihe Central Experiment Station, Chinese Academy of Fishery Sciences, Qinhuangdao 066100, ChinaHebei Key Laboratory of the Bohai Sea Fish Germplasm Resources Conservation and Utilization, Beidaihe Central Experiment Station, Chinese Academy of Fishery Sciences, Qinhuangdao 066100, ChinaTangshan Haidu Aquatic Food Co., Ltd., Tangshan 063000, ChinaHebei Key Laboratory of the Bohai Sea Fish Germplasm Resources Conservation and Utilization, Beidaihe Central Experiment Station, Chinese Academy of Fishery Sciences, Qinhuangdao 066100, ChinaHebei Key Laboratory of the Bohai Sea Fish Germplasm Resources Conservation and Utilization, Beidaihe Central Experiment Station, Chinese Academy of Fishery Sciences, Qinhuangdao 066100, ChinaHebei Key Laboratory of the Bohai Sea Fish Germplasm Resources Conservation and Utilization, Beidaihe Central Experiment Station, Chinese Academy of Fishery Sciences, Qinhuangdao 066100, ChinaHebei Key Laboratory of the Bohai Sea Fish Germplasm Resources Conservation and Utilization, Beidaihe Central Experiment Station, Chinese Academy of Fishery Sciences, Qinhuangdao 066100, ChinaSince the advent of germ cell transplantation (GCT), it has been widely used in shortening the fish breeding cycle, sex-controlled breeding and the protection of rare and endangered fish. In this study, the effectiveness of female sterile recipient preparation and donor stem cell isolation and purification were comprehensively evaluated for spermatogonial stem cell transplantation (SSCT) in <i>Paralichthys olivaceus</i>. The best way to prepare sterile recipients was found to be giving three-year-old fish four intraovarian injections of busulfan (20 mg/kg body weight) combined with exposure to a high temperature (28 °C) after the spawning season compared with the two other ways, which induced apoptosis of most of the endogenous germ cells, resulting in shrinkage of the spawning plate and enlargement of the ovarian lumen. Further analysis showed that both the gonadosomatic index and germ-cell-specific <i>vasa</i> expression were significantly lower than those of the natural-temperature group before treatment (<i>p</i> < 0.05). A high percentage (>60.00%) of spermatogonial stem cells (SSCs) were obtained after isolation and purification and were transplanted into the prepared recipients. After three weeks of SSCT, the numbers of PKH26-labeled SSCs were increased in the ovaries of the recipients. These findings provide a basis for the establishment of an ideal SSCT technique using <i>P. olivaceus</i> females as the recipients, ultimately contributing to the efficient conservation of male germplasm resources and effective breeding.https://www.mdpi.com/2076-2615/14/19/2887ovarian depletionspermatogonia stem celltransplantation<i>Paralichthys olivaceus</i>busulfan
spellingShingle Yuqin Ren
Yuehong Tao
Zhaohui Sun
Yufen Wang
Weidong Li
Zhongwei He
Guixing Wang
Yucong Yang
Jilun Hou
Evaluation of Female Recipient Infertility and Donor Spermatogonial Purification for Germ Cell Transplantation in <i>Paralichthys olivaceus</i>
Animals
ovarian depletion
spermatogonia stem cell
transplantation
<i>Paralichthys olivaceus</i>
busulfan
title Evaluation of Female Recipient Infertility and Donor Spermatogonial Purification for Germ Cell Transplantation in <i>Paralichthys olivaceus</i>
title_full Evaluation of Female Recipient Infertility and Donor Spermatogonial Purification for Germ Cell Transplantation in <i>Paralichthys olivaceus</i>
title_fullStr Evaluation of Female Recipient Infertility and Donor Spermatogonial Purification for Germ Cell Transplantation in <i>Paralichthys olivaceus</i>
title_full_unstemmed Evaluation of Female Recipient Infertility and Donor Spermatogonial Purification for Germ Cell Transplantation in <i>Paralichthys olivaceus</i>
title_short Evaluation of Female Recipient Infertility and Donor Spermatogonial Purification for Germ Cell Transplantation in <i>Paralichthys olivaceus</i>
title_sort evaluation of female recipient infertility and donor spermatogonial purification for germ cell transplantation in i paralichthys olivaceus i
topic ovarian depletion
spermatogonia stem cell
transplantation
<i>Paralichthys olivaceus</i>
busulfan
url https://www.mdpi.com/2076-2615/14/19/2887
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