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13. Addition of Cryoprotectant DMSO Reduces Damage to Spermatozoa of Yellow Catfish (Pelteobagrus fulvidraco) during Cryopreservation: Ultrastructural Damage, Oxidative Damage and DNA Damage.

Author

Zhang, Yuxin, Liu, Dongqing, Wang, Qinghua, Ruan, Qingxin, Hua, Sijie, Zhang, Weiwei, Yang, Sen, and Meng, Zining

Subjects
FLATHEAD catfish, GERMPLASM conservation, SPERM motility, GERMPLASM, ELECTRON microscopes, FROZEN semen, CRYOPROTECTIVE agents
Abstract

Simple Summary: Spermatozoa cryopreservation technology has been widely used to preserve high-quality male spermatozoa and has to some extent solved the problem of spermatozoa supply in yellow catfish (Pelteobagrus fulvidraco). However, cryopreservation can still cause cellular damage and affect spermatozoa viability and fertility. Therefore, it is necessary to study cryo-injury, which is beneficial for further optimizing the spermatozoa cryopreservation protocol. In this experiment, we investigated the effects of adding or not adding 10% DMSO on spermatozoa motility, ultrastructural damage, oxidative damage, and DNA damage of thawed yellow catfish. Using 10% DMSO as a cryoprotectant can improve spermatozoa motility (75.64 ± 1.12), significantly alleviate injuries to organelles and membrane structures, improve SOD (7.99 × 104 ± 5.24 nmol/mgprot) and T-AOC (3.7 × 105 ± 0.06 nmol/mgprot) enzyme activity, reduce MDA (1.09 ± 0.45 nmol/mgprot) enzyme activity, and mitigate the degree of oxidative damage. It also greatly reduces the DNA fragmentation rate (59 ± 1.56%). The research results provide data support for the optimization of the cryopreservation protocol for the spermatozoa of yellow catfish, which is beneficial for the preservation of the germplasm resources of yellow catfish and the effective utilization of resources during large-scale production. Spermatozoa cryopreservation protocols have been established for yellow catfish (Pelteobagrus fulvidraco), but cryopreservation can still cause cellular damage and affect spermatozoa viability and fertility. Therefore, the aim of this paper was to evaluate the effects of adding or not adding cryoprotectants during low-temperature storage on the ultrastructural damage, oxidative damage, and DNA damage of thawed yellow catfish spermatozoa. The mixed semen of three male yellow catfish was divided into a fresh spermatozoa group, a frozen spermatozoa group (DMSO+) with a cryoprotectant (10% DMSO), and a frozen spermatozoa group without a cryoprotectant (DMSO−). Ultrastructural of the spermatozoa after thawing were observed under an electron microscope and the spermatozoa were assayed for SOD, MDA, and T-AOC enzyme activities, as well as for DNA integrity. In terms of movement parameters, compared with DMSO−, the addition of DMSO has significantly improved sperm motility, curve line velocity (VCL), and straight line velocity (VSL). The ultrastructural results showed that although thawed spermatozoa exhibited increased damage than fresh spermatozoa, 10% DMSO effectively reduced the damage to the plasma membrane, mitochondria, and flagellum of spermatozoa by cryopreservation, and most of the spermatozoa were preserved with intact structure. The results of oxidative damage showed that compared with frozen spermatozoa, 10% DMSO significantly increased the activities of SOD and T-AOC enzymes and clearly reduced the activity of the MDA enzyme. The antioxidant capacity of spermatozoa was improved, lipid peroxidation was reduced, and the oxidative damage caused by cryopreservation was mitigated. The DNA integrity of spermatozoa showed that 10% DMSO clearly reduced the DNA fragmentation rate. In conclusion, 10% DMSO can effectively reduce the ultrastructural damage, oxidative damage, and DNA damage of yellow catfish spermatozoa during cryopreservation; it can also further optimize the cryopreservation protocol for yellow catfish spermatozoa. Meanwhile, it also provides a theoretical basis for the future optimization of the cryopreservation protocols. [ABSTRACT FROM AUTHOR]

Published
2024
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25. Microsatellite records for volume 8, issue 1

Author

Arias, Maria Cristina, Aulagnier, Stéphane, Baerwald, Erin F., Barclay, Robert M. R., Batista, Jacqueline Silva, Beasley, Rochelle R., Bezerra, Rodrigo A., Blanc, Frédéric, Bridge, Eli S., Cabria, Maria Teresa, Caputo, Lívia Izabela, Chen, Jie, Chen, Jian-Hua, Chen, Zhi, Chen, Ching-Hung, Chu, Pengfei, Contina, Andrea, da Silva Carmo, Fausto Moreira, de Carvalho, Leiliane Campos, de Freitas Viana, Priscila Maria, de Macêdo, Jeferson Luis Vasconcelos, de Menezes Yazbeck, Gabriel, de Oliveira Teixeira, Zélia, de Souza Araujo, Natália, Dequigiovanni, Gabriel, Ding, Shaoxiong, Dos Santos, Priscila Karla Ferreira, Ernest, Holly B., Feng, Yan-Wei, Formiga, Kyara Martins, Fournier-Chambrillon, Christine, Françoso, Elaine, Fu, Yaru, Gagne, Roderick B., Gaiotto, Fernanda A., Gao, Tian-xiang, Ge, Hongzi, Ge, Taiming, Gillet, François, Gong, Yun, Gong, Hong-mei, Guo, Qiqiang, Guo, Liang, Guo, Liang, Harrigan, Ryan J., He, Libo, House, Carrie J., Der Hsiao, Chung-, Huang, Rong, Iyengar, Arati, Jiang, Xiayun, Jiang, Hai-Lin, Johnson, Tania A., Jones, Kenneth L., Kageyama, Paulo Yoshio, Kelly, Jeffrey F., Lam, Lydia, Lance, Stacey L., Li, Hong, Li, Cheng, Li, Fugui, Li, Huie, Li, Shaoke, Li, Yongming, Li, Mingming, Li, Mingming, Li, Fuhua, Li, Wei-guo, Li, Jitao, Li, Jian, Li, Xinru, Liang, Kai, Liang, Yaosi, Liao, Fuchu, Liao, Lanjie, Lin, Haoran, Lin, Haoran, Lin, Haoran, Lin, Qiang, Liu, Hong, Liu, Honggao, Liu, Lu, Liu, Xiang-Quan, Liu, Ping, Liu, Chen, Liu, Xin, Lopes, Maria Teresa Gomes, Lu, Danqi, Luan, Ming-Bao, Luo, Wei, Meng, Zining, Meng, Zining, Michaux, Johan R., Némoz, Mélanie, Pollinger, John P., Qiao, Ye, Qu, Hongyue, Ramos, Santiago Linorio Ferreyra, Le Roux, Stephné, Santiago, Leandro Rodrigues, Shao, Xiaoyan, Shen, Kang-Ning, Shen, Heding, Shen, Hao, Smith, Thomas B., Song, Dang-yu, Sourp, Eric, Sun, Yiwen, Tang, Huiyuan, Tang, Qing, Tong, Yiqin, van den Einden, Marion, van Hoppe, Moniek J. C., Veasey, Elizabeth Ann, Vial-Novella, Corinne, Wan, Li, Wang, Xiaojun, Wang, Yiming, Wang, Yu, Wang, Yaping, Wang, Ruoran, Wang, Xin, Wang, Rifang, Wang, Quanchao, Wang, Guo-dong, Wang, Jiajia, Wang, Dongfeng, Wang, Peipei, Wei, Xiu-Mei, Wen, Yanfei, Wen, Haishen, Wu, Xue-Ping, Wu, Xin, Xiang, Jianhai, Xie, Zhenzhen, Xu, Zhanning, Xu, Pao, Yang, Zhi, Yang, Sen, Yang, Sen, Yin, Cece, Yin, Shaowu, Yin, Shaowu, Yu, Yang, Yuan, Xiping, Yue, Gen Hua, Zang, Gong-Gu, Zardoya, Rafael, Zhang, Li-hui, Zhang, Xiaojun, Zhang, Yong, Zhang, Guosong, di Zhang, Yan-, Zhang, Hongye, Zhang, Guosong, Zhao, Jie, Zhao, Cheng, Zhu, Fei, Zhu, Hongchai, Zhu, Shilong, and Zou, Shuming

Published
2016
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31. Transposon-induced epigenetic silencing in the X chromosome as a novel form of dmrt1 expression regulation during sex determination in the fighting fish

Author

Temasek Life Sciences Laboratory, Agencia Estatal de Investigación (España), Wang, Le, Su, Fei, Wan, Zi Yi, Yang, Zituo, Tay, Yi Xuan, Lee, May Shu Qing, Ye, Baoqing, Wen, Yanfei, Meng, Zining, Fan, Bin, Alfiko, Yuzer, Shen, Yubang, Piferrer, Francesc, Meyer, Axel, Schartl, Manfred, Yue, Gen Hua, Temasek Life Sciences Laboratory, Agencia Estatal de Investigación (España), Wang, Le, Su, Fei, Wan, Zi Yi, Yang, Zituo, Tay, Yi Xuan, Lee, May Shu Qing, Ye, Baoqing, Wen, Yanfei, Meng, Zining, Fan, Bin, Alfiko, Yuzer, Shen, Yubang, Piferrer, Francesc, Meyer, Axel, Schartl, Manfred, and Yue, Gen Hua

Abstract

Background. Fishes are the one of the most diverse groups of animals with respect to their modes of sex determination, providing unique models for uncovering the evolutionary and molecular mechanisms underlying sex determination and reversal. Here, we have investigated how sex is determined in a species of both commercial and ecological importance, the Siamese fighting fish Betta splendens. Results. We conducted association mapping on four commercial and two wild populations of B. splendens. In three of the four commercial populations, the master sex determining (MSD) locus was found to be located in a region of ~ 80 kb on LG2 which harbours five protein coding genes, including dmrt1, a gene involved in male sex determination in different animal taxa. In these fish, dmrt1 shows a male-biased gonadal expression from undifferentiated stages to adult organs and the knockout of this gene resulted in ovarian development in XY genotypes. Genome sequencing of XX and YY genotypes identified a transposon, drbx1, inserted into the fourth intron of the X-linked dmrt1 allele. Methylation assays revealed that epigenetic changes induced by drbx1 spread out to the promoter region of dmrt1. In addition, drbx1 being inserted between two closely linked cis-regulatory elements reduced their enhancer activities. Thus, epigenetic changes, induced by drbx1, contribute to the reduced expression of the X-linked dmrt1 allele, leading to female development. This represents a previously undescribed solution in animals relying on dmrt1 function for sex determination. Differentiation between the X and Y chromosomes is limited to a small region of ~ 200 kb surrounding the MSD gene. Recombination suppression spread slightly out of the SD locus. However, this mechanism was not found in the fourth commercial stock we studied, or in the two wild populations analysed, suggesting that it originated recently during domestication. Conclusions. Taken together, our data provide novel insights into the rol

Published
2022

34. Additional file 1 of Transposon-induced epigenetic silencing in the X chromosome as a novel form of dmrt1 expression regulation during sex determination in the fighting fish

Author

Wang, Le, Sun, Fei, Wan, Zi Yi, Yang, Zituo, Tay, Yi Xuan, Lee, May, Ye, Baoqing, Wen, Yanfei, Meng, Zining, Fan, Bin, Alfiko, Yuzer, Shen, Yubang, Piferrer, Francesc, Meyer, Axel, Schartl, Manfred, and Yue, Gen Hua

Abstract

Additional file 1: Figures S1-S11 and Tables S1 & S2. Figure S1. Mapping of the sex determining locus in family P_xx��xy of fighting fish. Figure S2. Mapping of the sex determining locus in F2 populations including BM1 and RM2 families. Figure S3. Mapping of sex determining locus in a family, P_xx��yy, generated by crossing a putative XX female and a YY male. Figure S4. Male-biased expression of dmrt1 in fighting fish. Figure S5. Knockout of dmrt1 using the CRISPR/Cas9 system in fighting fish. Figure S6. Sequence coverage for X- and Y-specific reads on either the X or Y putative dmrt1 containing the SD locus. Figure S7. Predicted conserved noncoding elements flanking transposon drbx1 enhance reporter GFP expression. Figure S8. Difference in methylation profile in the genomic regions flanking the transposon drbx1 insertion site between X and Y alleles. Figure S9. Restricted differentiation between X and Y chromosomes in fighting fish. Figure S10. Transposon drbx1 is not associated with phenotypic sex in wild species of B. splendens complex. Figure S11. Original uncropped gels for figures and supplementary figures used in this study, where how the gels were cropped is indicated with red box. Table S1. Samples including commercial stocks and wild Betta fish used in this study. Table S2. Sequences of primers used in analyzing sex determination in the fighting fish.

Published
2022
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39. A single intronic single nucleotide polymorphism in splicing site of steroidogenic enzyme hsd17b1 is associated with phenotypic sex in oyster pompano, Trachinotus anak

Author

Fan, Bin, primary, Xie, Dizhi, additional, Li, Yanwei, additional, Wang, Xulei, additional, Qi, Xin, additional, Li, Shuisheng, additional, Meng, Zining, additional, Chen, Xinghan, additional, Peng, Junyao, additional, Yang, Yongjian, additional, Li, Yuanyou, additional, and Wang, Le, additional

Published
2021
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46. Supplementary information for the manuscript entitled ���A single intronic SNP in splicing site of steroidogenic enzyme hsd17b1 is associated with phenotypic sex in oyster pompano, Trachinotus anak��� from A single intronic SNP in splicing site of steroidogenic enzyme hsd17b1 is associated with phenotypic sex in oyster pompano, Trachinotus anak

Author

Fan, Bin, Xie, Dizhi, Li, Yanwei, Wang, Xulei, Qi, Xin, Li, Shuisheng, Meng, Zining, Chen, Xinghan, Peng, Junyao, Yang, Yongjian, Li, Yuanyou, and Wang, Le

Abstract

Teleosts show varied master sex determining (MSD) genes and sex determination (SD) mechanisms, with frequent turnovers of sex chromosomes. Tracing the origins of MSD genes and turnovers of sex chromosomes in a taxonomic group is of particular interest in evolutionary biology. Oyster pompano (Trachinotus anak), a marine fish, belongs to the family Carangidae, in which 17b-hydroxysteroid dehydrogenase 1 (hsd17b1) has repeatedly evolved to an MSD gene. Whole-genome resequencing identified a single SNP at Chromosome 24 to be strictly associated with phenotypic sex, with females being the heterozygous sex. This SNP is located in a splicing site at the first exon/intron boundary of hsd17b1. The Z-linked SNP results in malfunction of all spliced isoforms, whereas the W-linked isoforms were predicted to have open reading frames (ORF) that are conserved among vertebrates, suggesting that hsd17b1 is a female-determining gene. The differential alternative splicing patterns of ZZ and ZW genotypes were consistently observed both in undifferentiated stages and differentiated gonads. We observed elevated recombination around the SD locus and no differentiation between Z and W chromosomes. The extreme diversity of mutational mechanisms that hsd17b1 evolves to an MSD gene highlights frequent in situ turnovers between sex chromosomes in the Carangidae.

Published
2021
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47. Genomic Basis of Striking Fin Shapes and Colors in the Fighting Fish

Author

Temasek Life Sciences Laboratory, Agencia Estatal de Investigación (España), Wang, Le, Su, Fei, Wan, Zi Yi, Ye, Baoqing, Wen, Yanfei, Liu, Huiming, Yang, Zituo, Pang, Hongyan, Meng, Zining, Fan, Bin, Alfiko, Yuzer, Shen, Yubang, Bai, Bin, Lee, May Shu Qing, Piferrer, Francesc, Schartl, Manfred, Meyer, Axel, Yue, Gen Hua, Temasek Life Sciences Laboratory, Agencia Estatal de Investigación (España), Wang, Le, Su, Fei, Wan, Zi Yi, Ye, Baoqing, Wen, Yanfei, Liu, Huiming, Yang, Zituo, Pang, Hongyan, Meng, Zining, Fan, Bin, Alfiko, Yuzer, Shen, Yubang, Bai, Bin, Lee, May Shu Qing, Piferrer, Francesc, Schartl, Manfred, Meyer, Axel, and Yue, Gen Hua

Abstract

Resolving the genomic basis underlying phenotypic variations is a question of great importance in evolutionary biology. However, understanding how genotypes determine the phenotypes is still challenging. Centuries of artificial selective breeding for beauty and aggression resulted in a plethora of colors, long-fin varieties, and hyper-aggressive behavior in the air-breathing Siamese fighting fish (Betta splendens), supplying an excellent system for studying the genomic basis of phenotypic variations. Combining whole-genome sequencing, quantitative trait loci mapping, genome-wide association studies, and genome editing, we investigated the genomic basis of huge morphological variation in fins and striking differences in coloration in the fighting fish. Results revealed that the double tail, elephant ear, albino, and fin spot mutants each were determined by single major-effect loci. The elephant ear phenotype was likely related to differential expression of a potassium ion channel gene, kcnh8. The albinotic phenotype was likely linked to a cis-regulatory element acting on the mitfa gene and the double-tail mutant was suggested to be caused by a deletion in a zic1/zic4 coenhancer. Our data highlight that major loci and cis-regulatory elements play important roles in bringing about phenotypic innovations and establish Bettas as new powerful model to study the genomic basis of evolved changes

Published
2021

49. Genomic Basis of Striking Fin Shapes and Colors in the Fighting Fish

Author

Wang, Le, primary, Sun, Fei, additional, Wan, Zi Yi, additional, Ye, Baoqing, additional, Wen, Yanfei, additional, Liu, Huiming, additional, Yang, Zituo, additional, Pang, Hongyan, additional, Meng, Zining, additional, Fan, Bin, additional, Alfiko, Yuzer, additional, Shen, Yubang, additional, Bai, Bin, additional, Lee, May Shu Qing, additional, Piferrer, Francesc, additional, Schartl, Manfred, additional, Meyer, Axel, additional, and Yue, Gen Hua, additional

Published
2021
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