Department of Molecular Biosciences, Northwestern University, Evanston, IL, 60208, USA.
The Chemistry of Life Processes Institute, Northwestern University, Evanston, IL, 60208, USA.
J Assist Reprod Genet. 2022 Jun;39(6):1261-1276. doi: 10.1007/s10815-022-02495-z. Epub 2022 May 1.
The requirement of zinc for the development and maturation of germ lines and reproductive systems is deeply conserved across evolution. The nematode Caenorhabditis elegans offers a tractable platform to study the complex system of distributing zinc to the germ line. We investigated several zinc importers to investigate how zinc transporters play a role in the reproductive system in nematodes, as well as establish a platform to study zinc transporter biology in germline and reproductive development.
Previous high throughput transcriptional datasets as well as phylogenetic analysis identified several putative zinc transporters that have a function in reproduction in worms. Phenotypic analysis of CRISPR-generated knockouts and tags included characterization of offspring output, gonad development, and protein localization. Light and immunofluorescence microscopy allowed for visualization of physiological and molecular effects of zinc transporter mutations.
Disruption of two zinc transporters, ZIPT-2.4 and ZIPT-15, was shown to lead to defects in reproductive output. A mutation in zipt-2.4 has subtle effects on reproduction, while a mutation in zipt-15 has a clear impact on gonad and germline development that translates into a more pronounced defect in fecundity. Both transporters have germline expression, as well as additional expression in other cell types.
Two ZIP-family zinc transporter orthologs of human ZIP6/10 and ZIP1/2/3 proteins are important for full reproductive fecundity and participate in development of the gonad. Notably, these zinc transporters are present in gut and reproductive tissues in addition to the germ line, consistent with a complex zinc trafficking network important for reproductive success.
锌在生殖细胞系和生殖系统的发育和成熟中的需求在进化过程中是高度保守的。秀丽隐杆线虫（Caenorhabditis elegans）为研究将锌分配给生殖细胞系的复杂系统提供了一个易于处理的平台。我们研究了几种锌转运体，以研究锌转运体在线虫生殖系统中的作用，并建立一个平台来研究生殖细胞系和生殖发育中的锌转运体生物学。
之前的高通量转录数据集和系统发育分析确定了几种在蠕虫生殖中具有功能的锌转运体。CRISPR 生成的敲除和标记的表型分析包括后代产量、性腺发育和蛋白质定位的特征描述。荧光和免疫荧光显微镜允许可视化锌转运体突变的生理和分子效应。
两种锌转运体（ZIPT-2.4 和 ZIPT-15）的破坏被证明导致生殖输出缺陷。zipt-2.4 的突变对生殖有轻微影响，而 zipt-15 的突变对性腺和生殖细胞系发育有明显影响，从而导致繁殖力更明显的缺陷。这两种转运体都具有生殖细胞系表达，以及其他细胞类型的额外表达。
两种人类 ZIP6/10 和 ZIP1/2/3 蛋白的 ZIP 家族锌转运体同源物对于完整的生殖生育力很重要，并参与了性腺的发育。值得注意的是，这些锌转运体除了生殖细胞系外，还存在于肠道和生殖组织中，这与一个复杂的锌运输网络对于生殖成功很重要相一致。
