Department of Biochemistry, Cellular and Molecular Biology, University of Tennessee, Knoxville, Tennessee, United States of America.
Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, Minnesota, United States of America.
PLoS One. 2024 Jun 24;19(6):e0305696. doi: 10.1371/journal.pone.0305696. eCollection 2024.
In Drosophila coordinated proliferation of two neural stem cells, neuroblasts (NB) and neuroepithelial (NE) cells, is pivotal for proper larval brain growth that ultimately determines the final size and performance of an adult brain. The larval brain growth displays two phases based on behaviors of NB and NEs: the first one in early larval stages, influenced by nutritional status and the second one in the last larval stage, promoted by ecdysone signaling after critical weight checkpoint. Mutations of the baboon (babo) gene that produces three isoforms (BaboA-C), all acting as type-I receptors of Activin-type transforming growth factor β (TGF-β) signaling, cause a small brain phenotype due to severely reduced proliferation of the neural stem cells. In this study we show that loss of babo function severely affects proliferation of NBs and NEs as well as conversion of NEs from both phases. By analyzing babo-null and newly generated isoform-specific mutants by CRISPR mutagenesis as well as isoform-specific RNAi knockdowns in a cell- and stage-specific manner, our data support differential contributions of the isoforms for these cellular events with BaboA playing the major role. Stage-specific expression of EcR-B1 in the brain is also regulated primarily by BaboA along with function of the other isoforms. Blocking EcR function in both neural stem cells results in a small brain phenotype that is more severe than baboA-knockdown alone. In summary, our study proposes that the Babo-mediated signaling promotes proper behaviors of the neural stem cells in both phases and achieves this by acting upstream of EcR-B1 expression in the second phase.
在果蝇中,两个神经干细胞,神经母细胞(NB)和神经上皮细胞(NE)的协调增殖对于适当的幼虫大脑生长至关重要,这最终决定了成年大脑的最终大小和性能。幼虫大脑生长根据 NB 和 NEs 的行为表现出两个阶段:第一个阶段在早期幼虫阶段,受营养状况的影响,第二个阶段在最后幼虫阶段,在关键体重检查点后由蜕皮激素信号促进。产生三种同工型(BaboA-C)的狒狒(babo)基因的突变,所有同工型都作为激活素型转化生长因子 β(TGF-β)信号的 I 型受体,导致由于神经干细胞增殖严重减少而出现小脑表型。在这项研究中,我们表明 babo 功能的丧失严重影响了 NB 和 NE 的增殖,以及 NE 从两个阶段的转化。通过以细胞和阶段特异性的方式分析 CRISPR 诱变产生的 babo 缺失和新生成的同工型特异性突变体以及同工型特异性 RNAi 敲低,我们的数据支持同工型对这些细胞事件的不同贡献,其中 BaboA 起主要作用。大脑中 EcR-B1 的阶段特异性表达也主要受 BaboA 以及其他同工型的调节。在神经干细胞中阻断 EcR 功能会导致小脑表型,其严重程度比单独敲低 baboA 更严重。总之,我们的研究表明,Babo 介导的信号转导促进了两个阶段神经干细胞的适当行为,并通过在第二阶段之前作用于 EcR-B1 的表达来实现这一目标。