A multiplex PCR method to determine the sex of fetal rat tissues.

IF 2.6 4区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Molecular Biology Reports Pub Date : 2025-03-13 DOI:10.1007/s11033-025-10406-5

Cristine Camp, Paige Drotos, Adrian Courville, Miranda Reed, Rachel West

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引用次数: 0

Abstract

Background: Fetal and placental sex influence a variety of developmental processes during prenatal life; including metabolism, growth, and the response to in utero insults. Additionally, the National Institute of Health's requirement that sex as a biological variable be included into proposal design necessitates the development of tools to investigate sex during embryonic and fetal life. Rodent models are insightful models in the study of sexual dimorphism due to large litter sizes, short gestation period, and frequency of use as an animal model. In this methods paper, we demonstrate a multiplex PCR method to determine sex in fetal rat tail snips and placentas.

Methods and results: We designed primers for X-chromosome and Y-chromosome homologs, DDX3X and DDX3Y, and developed a single-step PCR protocol that can determine the presence of both genes in one reaction. We performed PCR on fetal tail snips and placentas to amplify DDX3X only in females or DDX3X and DDX3Y in males. The multiplex PCR and subsequent gel electrophoresis revealed that the presence of only DDX3X or both DDX3X and DDX3Y could be detected in fetal tissues. We used adult male rat testis as a positive control and confirmed that both DDX3X and DDX3Y could be detected in adult male tissues as well.

Conclusion: This protocol provides an important method to determine genetic sex in tissues before the ability to visually determine sex, allowing for sex to be used as a biological variable in prenatal research using the rat model.

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背景：胎儿和胎盘的性别会影响产前的各种发育过程，包括新陈代谢、生长和对子宫内损伤的反应。此外，美国国家卫生研究院要求将性别作为生物变量纳入方案设计中，这就需要开发工具来研究胚胎和胎儿期的性别。由于啮齿类动物的产仔数多、妊娠期短，而且经常被用作动物模型，因此是研究性双态性的有价值的模型。在这篇方法论文中，我们展示了一种多重 PCR 方法来确定胎鼠尾剪和胎盘中的性别：我们设计了 X 染色体和 Y 染色体同源物 DDX3X 和 DDX3Y 的引物，并开发了一种单步 PCR 方案，可在一个反应中确定这两个基因的存在。我们对胎儿尾剪和胎盘进行了 PCR 扩增，只扩增雌性的 DDX3X 或扩增雄性的 DDX3X 和 DDX3Y。多重 PCR 和随后的凝胶电泳显示，在胎儿组织中可以检测到 DDX3X 或 DDX3X 和 DDX3Y 的存在。我们用成年雄性大鼠睾丸作为阳性对照，证实在成年雄性大鼠组织中也能检测到 DDX3X 和 DDX3Y：结论：该方案提供了一种重要的方法，可在目测性别之前确定组织中的遗传性别，从而使性别成为大鼠模型产前研究中的一个生物变量。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Molecular Biology Reports 生物-生化与分子生物学

CiteScore

5.00

自引率

0.00%

发文量

1048

审稿时长

5.6 months

期刊介绍： Molecular Biology Reports publishes original research papers and review articles that demonstrate novel molecular and cellular findings in both eukaryotes (animals, plants, algae, funghi) and prokaryotes (bacteria and archaea).The journal publishes results of both fundamental and translational research as well as new techniques that advance experimental progress in the field and presents original research papers, short communications and (mini-) reviews.