基础医学与临床2026，Vol.46Issue(4)：478-483,6.DOI:10.16352/j.issn.1001-6325.2026.04.0478
条件性敲除小鼠十二指肠黏膜上皮多巴胺D5受体的类器官培养和鉴定
Establishment and characterization of intestinal organoids from dopamine D5 receptor conditional knockout mice in duodenal mucosal epithelium
摘要
Abstract
Objective To establish a mouse duodenal mucosal epithelial organoid model with conditional knockout of dopamine D5 receptor(D5R)in intestinal mucosal epithelium.Methods Villin-CreERT2+/+D5R-flox+/+C57BL/6J mice were utilized to isolate duodenal crypts for organoid culture.D5R conditional knockout(CKO)was induced via the Cre-loxP system by 4-hydroxytamoxifen administration.The structural characteristics of duodenal mucosal epithelial organoids and the knockout efficiency of D5R were evaluated through morphological observation,immuno-fluorescence staining,quantitative PCR(qPCR),and Western blot assays.Results A duodenal mucosal epithelial organoid model with conditional D5R knockout was successfully established.Morphological examination revealed that the organoids exhibited structural features similar to the in vivo duodenal mucosal epithelium.Immunofluorescence staining confirmed a significant reduction in D5 R expression following 4-hydroxytamoxifen induction,with fluores-cence intensity decreasing from 0.904±0.061 to 0.385±0.020(n=6,P<0.001).qPCR analysis demonstrated a marked decrease in D5 R mRNA levels from 2.483±0.204 to 0.241±0.048,and Western blot showed a significant reduction in protein expression from 0.923±0.075 to 0.569±0.032(n=6,P<0.001).Conclusions A conditional D5R knockout organoid model derived from duodenal mucosal epithelium is successfully developed,which faithfully recapitulates the physiological characteristics of duodenal mucosa.This model provides a reliable in vitro platform for elucidating the regulatory mechanisms of D5R in duodenal epithelial functions.关键词
类器官/十二指肠/肠黏膜上皮/多巴胺D5受体Key words
organoid/duodenum/intestinal mucosal epithelium/dopamine D5 receptor分类
医药卫生引用本文复制引用
朱愔喆,马雪睿,汪国庆,王益嘉,冯小燕..条件性敲除小鼠十二指肠黏膜上皮多巴胺D5受体的类器官培养和鉴定[J].基础医学与临床,2026,46(4):478-483,6.基金项目
国家自然科学基金(32071126) （32071126）
北京市自然科学基金(7222010) （7222010）
