用于研究输卵管分泌上皮细胞的输卵管糖蛋白 1 Cre 小鼠模型的产生

IF 3.8 3区 医学 Q2 ENDOCRINOLOGY & METABOLISM Endocrinology Pub Date : 2024-05-27 DOI:10.1210/endocr/bqae070
Emily A McGlade, Jiude Mao, Kalli K Stephens, Andrew M Kelleher, Lisette A Maddison, Miranda L Bernhardt, Francesco J DeMayo, John P Lydon, Wipawee Winuthayanon
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引用次数: 0

摘要

输卵管上皮细胞层在卵母细胞拾取、精子迁移、着床前胚胎发育和胚胎运输中发挥着重要作用。输卵管上皮细胞层由纤毛和非纤毛分泌细胞组成。纤毛细胞的功能已被证明可支持配子和胚胎在输卵管中的移动,但分泌细胞的功能尚未得到很好的描述。因此,我们的目标是建立一个分泌细胞特异性 Cre 重组酶小鼠模型,以研究输卵管分泌细胞的作用。通过从内源性Ovgp1(输卵管糖蛋白1)基因座表达Cre,并以增强型绿色荧光蛋白(eGFP)作为报告基因,我们建立了一个基因敲入小鼠模型--Ovgp1Cre:eGFP。成年 Ovgp1Cre:eGFP 小鼠发情时,在输卵管的分泌上皮细胞中能强烈检测到 EGFP 信号。在卵巢基质、子宫基质、阴道上皮细胞、附睾上皮细胞和伸长的精子中也检测到了信号。为了验证重组酶的活性,使用 Ovgp1Cre:eGFP; Pgrf/f 小鼠模型消减了孕酮受体(PGR)的表达。令人惊讶的是,这种缺失仅限于 Ovgp1Cre:eGFP; Pgrf/f 输卵管的子宫管交界(UTJ)区域的上皮细胞。在UTJ区域的上皮细胞中删除Pgr对雌性繁殖力没有影响。总之,我们发现 eGFP 信号可能是输卵管所有区域分泌上皮细胞的特异性信号。然而,由于Cre具有潜在的靶标特异性活性,因此在使用Ovgp1Cre:eGFP品系产生条件性基因敲除小鼠时,绝对需要验证适当的重组和相关基因的表达,以确认有效的基因缺失。
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Generation of Oviductal Glycoprotein 1 Cre Mouse Model for the Study of Secretory Epithelial Cells of the Oviduct.

The epithelial cell lining of the oviduct plays an important role in oocyte pickup, sperm migration, preimplantation embryo development, and embryo transport. The oviduct epithelial cell layer comprises ciliated and nonciliated secretory cells. The ciliary function has been shown to support gamete and embryo movement in the oviduct, yet secretory cell function has not been well characterized. Therefore, our goal was to generate a secretory cell-specific Cre recombinase mouse model to study the role of the oviductal secretory cells. A knock-in mouse model, Ovgp1Cre:eGFP, was created by expressing Cre from the endogenous Ovgp1 (oviductal glycoprotein 1) locus, with enhanced green fluorescent protein (eGFP) as a reporter. EGFP signals were strongly detected in the secretory epithelial cells of the oviducts at estrus in adult Ovgp1Cre:eGFP mice. Signals were also detected in the ovarian stroma, uterine stroma, vaginal epithelial cells, epididymal epithelial cells, and elongated spermatids. To validate recombinase activity, progesterone receptor (PGR) expression was ablated using the Ovgp1Cre:eGFP; Pgrf/f mouse model. Surprisingly, the deletion was restricted to the epithelial cells of the uterotubal junction (UTJ) region of Ovgp1Cre:eGFP; Pgrf/f oviducts. Deletion of Pgr in the epithelial cells of the UTJ region had no effect on female fecundity. In summary, we found that eGFP signals were likely specific to secretory epithelial cells in all regions of the oviduct. However, due to a potential target-specific Cre activity, validation of appropriate recombination and expression of the gene(s) of interest is absolutely required to confirm efficient deletion when generating conditional knockout mice using the Ovgp1Cre:eGFP line.

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来源期刊
Endocrinology
Endocrinology 医学-内分泌学与代谢
CiteScore
8.10
自引率
4.20%
发文量
195
审稿时长
2-3 weeks
期刊介绍: The mission of Endocrinology is to be the authoritative source of emerging hormone science and to disseminate that new knowledge to scientists, clinicians, and the public in a way that will enable "hormone science to health." Endocrinology welcomes the submission of original research investigating endocrine systems and diseases at all levels of biological organization, incorporating molecular mechanistic studies, such as hormone-receptor interactions, in all areas of endocrinology, as well as cross-disciplinary and integrative studies. The editors of Endocrinology encourage the submission of research in emerging areas not traditionally recognized as endocrinology or metabolism in addition to the following traditionally recognized fields: Adrenal; Bone Health and Osteoporosis; Cardiovascular Endocrinology; Diabetes; Endocrine-Disrupting Chemicals; Endocrine Neoplasia and Cancer; Growth; Neuroendocrinology; Nuclear Receptors and Their Ligands; Obesity; Reproductive Endocrinology; Signaling Pathways; and Thyroid.
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