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宿主、细胞密度及注射时间对鲶鱼科鲶鱼外源基因生产效率的影响

Impacts of Host, Cell Density, and Timing of Injection on Efficiency for Xenogen Production in Ictalurid Catfish.

作者信息

Pottle Kate B, Hettiarachchi Darshika U, Shang Mei, Su Baofeng, Al-Armanazi Jacob, Wang Jinhai, Soman Misha, Dilawar Hamza, Butts Ian A E, Dunham Rex A

机构信息

School of Fisheries, Aquaculture and Aquatic Sciences, Auburn University, Auburn, Alabama, 36849, USA.

出版信息

Mar Biotechnol (NY). 2025 May 27;27(3):90. doi: 10.1007/s10126-025-10466-5.

Abstract

Optimization of xenogenesis for hybrid catfish (♀ channel catfish, Ictalurus punctatus × ♂ blue catfish, I. furcatus) embryo production was the goal. The effect of density of unsorted gonadal cells (80,000, 100,000, or 120,000 cells/fry) from blue catfish (BGCs) injected into triploid channel catfish surrogates, and BGCs or channel catfish (CGCs) into triploid white catfish (Ameiurus catus) surrogates on proliferation and colonization rates in surrogates injected at 4-, 5-, or 6-days post-hatch (DPH) was evaluated. At 45 and 90 DPH, survival and size of surrogates, and colonization/proliferation of donor cells (cell area < 150 μm and cluster area > 150 μm) were evaluated. Survival and size of all surrogate species were not impacted by cell density or donor. All surrogate species injected with 100,000 cells/fry had larger cluster cell areas than those injected with 80,000 cells/fry. White catfish surrogates with BGCs and CGCs had larger cell areas when injected with 100,000 cells/fry than those injected with 80,000 cells/fry. Both cell and cluster area increased by 90 DPH for all surrogates. PCR and PKH26 red fluorescence analysis confirmed that > 89% and > 86% of surrogates were positive xenogens at 45 and 90 DPH, respectively. No surrogate type or donor was superior to the others regarding colonization and proliferation, survival or growth, thus, channel catfish or white catfish were equally effective surrogates. Potential advantages of white catfish are small size, early sexual maturity, and spawning early in the season. These findings enhance the efficiency of germ cell transplantation for commercial hybrid catfish production.

摘要

目标是优化杂交鲶鱼(♀斑点叉尾鮰,Ictalurus punctatus × ♂蓝鲶,I. furcatus)胚胎生产的异种生殖。评估了将未分选的蓝鲶性腺细胞(80,000、100,000或120,000个细胞/鱼苗)注射到三倍体斑点叉尾鮰代孕鱼体内,以及将蓝鲶性腺细胞或斑点叉尾鮰细胞(CGCs)注射到三倍体白鲶(Ameiurus catus)代孕鱼体内,对孵化后4、5或6天(DPH)注射的代孕鱼体内增殖和定殖率的影响。在45和90 DPH时,评估代孕鱼的存活和大小,以及供体细胞的定殖/增殖情况(细胞面积<150μm,细胞簇面积>150μm)。所有代孕鱼的存活和大小不受细胞密度或供体的影响。所有注射100,000个细胞/鱼苗的代孕鱼的细胞簇面积均大于注射80,000个细胞/鱼苗的代孕鱼。注射100,000个细胞/鱼苗的含有蓝鲶性腺细胞和斑点叉尾鮰细胞的白鲶代孕鱼的细胞面积大于注射80,000个细胞/鱼苗的代孕鱼。所有代孕鱼的细胞和细胞簇面积在90 DPH时均增加。PCR和PKH26红色荧光分析证实,在45和90 DPH时,分别有>89%和>86%的代孕鱼为阳性异种。在定殖和增殖、存活或生长方面,没有哪种代孕类型或供体优于其他类型,因此,斑点叉尾鮰或白鲶作为代孕鱼同样有效。白鲶的潜在优势在于体型小、性成熟早以及在季节早期产卵。这些发现提高了商业杂交鲶鱼生产中生殖细胞移植的效率。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d150/12117003/cf09f0a6b461/10126_2025_10466_Fig1_HTML.jpg

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