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The impact of sperm-expressed transcription factors on fate-mapping models

机译：精子表达的转录因子对命运映射模型的影响

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Genetic lineage tracing has been used extensively in developmental biology. Many transcription factors expressed in sperm may induce Cre-mediated loxP recombination during early zygote development. In this study, we investigated the effect of sperm-expressed Cre on cell type-specific Cre-mediated loxP recombination in fate-mapping models of Tbx18+ progenitor cells. We found the recombination frequency in a reverse mating (RM) lineage was inconsistent with a normal Mendelian distribution. However, the recombination frequency in a positive mating (PM) lineage agreed with a Mendelian distribution. In the PM lineage, LacZ and EYFP were expressed in specific locations, such as the limb buds, heart, and hair follicles. Therefore, the reporter genes accurately and reliably traced cell differentiation in the PM lineage. In contrast, EYFP and LacZ were expressed throughout the embryo in the RM lineage. Thus, the reporter genes did not trace cell differentiation specifically in the RM lineage. Furthermore, Tbx18 mRNA and protein were expressed in the testicles of male mice, but almost no Tbx18 expression was detected in the ovaries of female mice. Similarly, reporter genes and Tbx18 were coexpressed in the seminiferous tubules and sperm cells of testicles. These results revealed that Cre-loxP-mediated pre-recombination in zygotes is due to Tbx18 expressed in testicle sperm cells when Cre is transmitted paternally. Our results indicate that Cre-mediated specific recombination in fate-mapping models of sperm-expressed genes may be influenced by the paternal origin of Cre. Therefore, a careful experimental design is critical when using the Cre-loxP system to trace spatial, temporal or tissue-specific fates.
机译：遗传谱系追踪已广泛用于发育生物学中。精子中表达的许多转录因子可能在合子早期发育期间诱导Cre介导的loxP重组。在这项研究中，我们研究了在Tbx18 +祖细胞的命运映射模型中，精子表达的Cre对细胞类型特异性Cre介导的loxP重组的影响。我们发现反向交配（RM）谱系中的重组频率与正态孟德尔分布不一致。但是，阳性交配（PM）谱系中的重组频率与孟德尔分布一致。在PM谱系中，LacZ和EYFP在特定位置表达，例如肢芽，心脏和毛囊。因此，报告基因准确，可靠地跟踪PM谱系中的细胞分化。相反，EYFP和LacZ在RM谱系的整个胚胎中表达。因此，报道基因没有在RM谱系中明确追踪细胞分化。此外，Tbx18 mRNA和蛋白在雄性小鼠的睾丸中表达，但在雌性小鼠的卵巢中几乎未检测到Tbx18表达。同样，报告基因和Tbx18在睾丸的生精小管和精子细胞中共表达。这些结果表明，由Cre介导的Cre-loxP介导的预重组是由于父系传播Cre时，睾丸精子细胞中表达的Tbx18所致。我们的结果表明，Cre介导的在精子表达基因的命运映射模型中的特异性重组可能受Cre父系的影响。因此，使用Cre-loxP系统追踪空间，时间或组织特定命运时，仔细的实验​​设计至关重要。

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