条件性子宫细胞剔除的研究

doi:10.16431/j.cnki.1671-7236.2020.04.019

Abstract

Abstract: In order to study the function of cells and the interaction between various cells in vivo,a new method of conditional cell depletion which could specifically eliminate specific cells was studied.Uterine-specific diphtheria toxin receptor transgenic mice were obtained by mating PR^Cre/+ tool mice with pCAG-loxp-STOP-loxp-DTR-2A-EGFP transgenic mice (DTR mice).The genes were identified by PCR and agarose gel electrophoresis and PR^Cre/+/pCAG-loxp-STOP-loxp-DTR-2A-EGFP female mice (PR-DTR mice)were screened out.The DTR mice and wild type mice (WT mice)were used as experimental control groups.Immunohistochemistry and Western blotting were used to detect the expression of GFP protein in the uterus.1 ng/d diphtheria toxin was injected intraperitoneally to observe the differences in external morphology of the uterus of PR-DTR, DTR and wild type mice.The internal morphology of the uterus was observed by HE staining.Immunohistochemistry was used to detect the expression of FOXA2 protein in PR-DTR, DTR and WT female mice after injecting diphtheria toxin.It was found that epithelium,stromal and muscle cells of mouse expressed GFP protein.The uterus of PR-DTR mice was redness and swelling after injection of diphtheria toxin,and there were no significant changes in the uterus of DTR mice and WT mice.Abnormal cavities in the uterus of PR-DTR mice were observed by HE staining.There were no significant differences in the uterus of DTR and WT mice,the detection of FOXA2 expression in the uterus of PR-DTR mice injected with diphtheria toxin showed that these abnormal cavities were not glands,indicating that diphtheria toxin caused cell death.It was confirmed that cells expressing progesterone receptors in mouse uterus expressed DTR.Experiments showed that diphtheria toxin played a role in the uterus of PR-DTR mice,and could conditionally eliminate cells expressing progesterone receptors in the uterus.It provided a new method for studying the function of uterine cells and the interaction between uterine cells.

Key words: cell depletion; diphtheria toxin; diphtheria toxin receptor; PR^Cre/+ mice; pCAG-loxp-STOP-loxp-DTR-2A-EGFP mice

CLC Number:

Q95-33

WU Linlin, ZHANG Heng, LI Shijie, MA Xinghong. Preliminary Study of Conditional Cell Depletion in Uterus[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(4): 1140-1147.

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Preliminary Study of Conditional Cell Depletion in Uterus

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