Roles of <italic style="font-style: italic">try</italic> and <italic style="font-style: italic">amy</italic> in feeding, digestion, growth, and development of the Japanese medaka (<italic style="font-style: italic">Oryzias latipes</italic>): insight from a comparative gene knockout study

Xiaodan JIA; Shulin TANG; Hexiong FENG; Dimei XU; Chenyuan ZHU; Ke LU; Xufang LIANG

doi:10.1631/jzus.B2400246

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Roles of try and amy in feeding, digestion, growth, and development of the Japanese medaka (Oryzias latipes): insight from a comparative gene knockout study

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Research Article | Updated：2026-03-18

Roles of try and amy in feeding, digestion, growth, and development of the Japanese medaka (Oryzias latipes): insight from a comparative gene knockout study
Roles of try and amy in feeding, digestion, growth, and development of the Japanese medaka (Oryzias latipes): insight from a comparative gene knockout study
try和amy在日本青鳉（Oryzias latipes）摄食、消化、生长和发育中的作用：基因敲除比较研究的见解
Journal of Zhejiang University-SCIENCE B Vol. 27, Issue 3, Pages: 280-294(2026)
Affiliations：

1.College of Fisheries, Chinese Perch Research Center, Huazhong Agricultural University, Wuhan 430070, China
2.Engineering Research Center of Green Development for Conventional Aquatic Biological Industry in the Yangtze River Economic Belt, Ministry of Education, Wuhan 430070, China
Funds：
DOI：10.1631/jzus.B2400246
CLC：
CSTR：
Received：16 May 2024，

Revised：2024-12-02，

Online First：22 December 2025，

Published：18 March 2026
Accepted：

贾晓丹,汤树林,冯贺雄等.try和amy在日本青鳉（Oryzias latipes）摄食、消化、生长和发育中的作用：基因敲除比较研究的见解[J].浙江大学学报（英文版）（B辑：生物医学和生物技术）,2026,27(03):280-294.

Xiaodan JIA, Shulin TANG, Hexiong FENG, et al. Roles of try and amy in feeding, digestion, growth, and development of the Japanese medaka (Oryzias latipes): insight from a comparative gene knockout study[J]. Journal of Zhejiang University-SCIENCE B, 2026, 27(3): 280-294.
贾晓丹,汤树林,冯贺雄等.try和amy在日本青鳉（Oryzias latipes）摄食、消化、生长和发育中的作用：基因敲除比较研究的见解[J].浙江大学学报（英文版）（B辑：生物医学和生物技术）,2026,27(03):280-294. DOI： 10.1631/jzus.B2400246.

Xiaodan JIA, Shulin TANG, Hexiong FENG, et al. Roles of try and amy in feeding, digestion, growth, and development of the Japanese medaka (Oryzias latipes): insight from a comparative gene knockout study[J]. Journal of Zhejiang University-SCIENCE B, 2026, 27(3): 280-294. DOI： 10.1631/jzus.B2400246.

摘要

消化酶在鱼类仔鱼阶段对食物消化至关重要，然而其具体的功能和必要性目前仍缺乏有力证据。为探索此问题，本研究首次利用CRISPR/Cas9技术，在日本青鳉（

Oryzias latipes

）中构建了胰蛋白酶基因（

try

）和淀粉酶基因（

amy

）的敲除模型。研究结果显示，

try

基因缺失显著降低了

try

-/-

青鳉仔鱼中该基因的表达水平及其消化能力；投喂卤虫8 h后，在孵化后4天(4 dph)和15 dph的

try

-/-

青鳉仔鱼的消化道中观察到未被完全消化的卤虫。此外，15 dph的

try

-/-

青鳉仔鱼的肠绒毛高度和体长均显著降低。与之相反，

amy

基因的缺失所产生的影响较为有限。在4 dph时，

amy

仔鱼的消化过程未受明显影响；在15 dph时，其消化道内只观察到少量不完全消化的卤虫。通过进一步分析

try

-/-

成鱼青鳉的生长、氮代谢和肠道微生物，发现成鱼青鳉体长和体重显著下降，而血液中天冬氨酸转氨酶（AST）和丙氨酸转氨酶（ALT）活性显著升高。肝肠组织病理学观察发现，

try

敲除导致肝细胞出现空泡变性、肠壁变薄、绒毛排列稀疏以及高度降低。16S rRNA高通量测序结果显示，

try

敲除降低了肠道微生物的多样性。综上所述，

try

对青鳉仔鱼不可或缺，且对其生长、氮代谢以及肠道发育具有持续性影响。

Abstract

Digestive enzymes of fish are critical to food digestion at the larval stage

but convincing evidence proving t

he function and necessity of the associated digestive enzymes remains lacking. In this study

we generated the trypsin (

try

) gene and amylase (

amy

) gene in the Japanese medaka (

Oryzias latipes

) using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) for the first time.

try

deletion significantly decreased the expression of

try

and

digestive capacity in

try

-/-

medaka larvae; after 8.5 h of digestion

incompletely digested brine shrimp was observed in the digestive tract at 4 and 15 d post-hatching (dph) of

try

-/-

medaka larvae. Furthermore

the height of intestinal villi and total body length decreased significantly within 15-dph

try

-/-

medaka larvae. However

amy

deletion did not influence the digestion of medaka larvae at 4 dph. Only a small amount of incompletely digested brine shrimp was observed in 15-dph

amy

-/-

medaka larvae. Further analysis of the growth

nitrogen metabolism

and intestinal microbes of

try

-/-

adult medaka showed that the body length and weight of adult medaka decreased significantly

while the activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the blood significantly increased. Pathological observation of the liver and intestinal tissues showed that

try

knockout resulted in vacuolar degeneration of liver cells

thinning of the intestinal wall

sparse arrangement of villi

and lower villi height. High-throughput 16S ribosomal RNA (rRNA) sequencing revealed that

try

knockout reduced the diversity of intestinal microbes. These findings demonstrated that

try

was indispensable for medaka larvae because it continuously affects their growth

nitrogen metabolism

and intestinal development.

关键词

Keywords

references

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Related Author

Xufang LIANG

Ke LU

Xiaodan JIA

Szymon CHOWAŃSKI

Paweł MARCINIAK

Karolina WALKOWIAK-NOWICKA

Arkadiusz URBAŃSKI

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College of Fisheries, Chinese Perch Research Center, Huazhong Agricultural University

Department of Animal Physiology and Developmental Biology, Adam Mickiewicz University, Poznań

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Roles of try and amy in feeding, digestion, growth, and development of the Japanese medaka (Oryzias latipes): insight from a comparative gene knockout study

Roles of try and amy in feeding, digestion, growth, and development of the Japanese medaka (Oryzias latipes): insight from a comparative gene knockout study

DOI：10.1631/jzus.B2400246

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