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热带大西洋肺鱼成鱼和幼鱼中胰蛋白酶基因的表达。

Trypsin gene expression in adults and larvae of tropical gar Atractosteus tropicus.

机构信息

Laboratorio de Acuicultura Tropical, DACBiol-UJAT, Carretera Villahermosa-Cárdenas Km 0.5, C.P. 86139, Villahermosa, Tabasco, Mexico.

Laboratorio de Biología Molecular y Biotecnología, DACS-UJAT, Avenida Gregorio Méndez 2838-A Col. Tamulté, C.P. 86100, Villahermosa, Tabasco, Mexico.

出版信息

Fish Physiol Biochem. 2020 Feb;46(1):145-155. doi: 10.1007/s10695-019-00704-8. Epub 2019 Nov 9.

Abstract

Trypsin gene (try) expression levels were quantified in different organs of wild and captive tropical gar (Atractosteus tropicus) adults, and changes in expression during initial ontogeny of the species were determined. RNA was extracted from the pancreas, and cDNA was synthesized and later amplified by endpoint PCR using oligonucleotides designed from different try sequences of fish registered in GenBank. Subsequently, specific oligonucleotides were designed from the partial sequences. Gene expression was measured after RNA extraction and synthesis of the cDNA of 11 organs (liver, pancreas, stomach, esophagus, intestine, pyloric caeca, brain, muscle, gills, gonad, and kidney) of captive and wild adults. Likewise, samples of A. tropicus larvae were taken on days 0 (embryo), 5, 10, 15, 20, 25, and 30 days after hatching (DAH), the RNA was extracted, and the synthesis of cDNA was carried out to measure real-time gene expression (qPCR). The results showed that the highest relative try expression occurred mainly in the esophagus, liver, stomach, and pancreas of both wild and captive adult fish; however, captive organisms had a higher try expression level than wild fish. Although try expression during initial ontogeny was high in embryos (0 DAH), it did not reach the maximum value until 15 DAH. It was concluded that try expression levels in captive adults are due to the high protein content in the balanced feed (trout diet). The highest try expression level during larviculture was detected at 15 DAH, which indicates that A. tropicus larvae have a mature digestive system and can efficiently hydrolyze proteins from feed at this developmental stage.

摘要

从野生和圈养的热带大西洋肺鱼(Atractosteus tropicus)成鱼的不同器官中提取 RNA,并使用从 GenBank 中注册的鱼类不同胰蛋白酶基因(try)序列设计的寡核苷酸通过终点 PCR 合成和扩增 cDNA,以定量测定其基因表达水平,并确定该物种在初始发育过程中的表达变化。随后,从部分序列中设计出特定的寡核苷酸。从圈养和野生成鱼的 11 个器官(肝、胰脏、胃、食道、肠、幽门盲囊、脑、肌肉、鳃、性腺和肾)中提取 RNA 并合成 cDNA 后,测量基因表达。同样,在孵化后第 0(胚胎)、5、10、15、20、25 和 30 天(DAH)时采集 A. tropicus 幼虫样本,提取 RNA 并合成 cDNA,以测量实时基因表达(qPCR)。结果表明,野生和圈养成鱼的食道、肝、胃和胰脏中相对 try 表达水平最高;然而,圈养生物的 try 表达水平高于野生鱼类。尽管胚胎期(0 DAH)初始发育过程中的 try 表达水平较高,但直到 15 DAH 才达到最大值。研究结论认为,圈养成鱼的 try 表达水平是由于均衡饲料(鳟鱼饲料)中的高蛋白含量所致。在幼鱼期,try 表达水平最高出现在 15 DAH,这表明 A. tropicus 幼虫具有成熟的消化系统,能够在该发育阶段有效地从饲料中水解蛋白质。

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