Using Bacterial Extract along with Differential Gene Expression in Acropora millepora Larvae to Decouple the Processes of Attachment and Metamorphosis
Using Bacterial Extract along with Differential Gene Expression in Acropora millepora Larvae to Decouple the Processes of Attachment and Metamorphosis
复制标题
使用细菌提取物和鹿角珊瑚幼虫的差异基因表达来解耦附着和变态过程
作者：
N. Siboni;D. Abrego;F. Seneca;C. Motti;N. Andreakis;Jan Tebben;L. Blackall;T. Harder
假交替单胞菌细菌的生物膜诱导顶孔珊瑚幼虫的变态。从假交替单胞菌提取物中分离出细菌代谢物四溴吡咯 (TBP)。与甲壳珊瑚藻 (CCA) Neogoniolithon fosliei 相关，诱导珊瑚幼虫变态 (100%)，几乎没有或没有附着 (0–2%)。为了更好地了解支持千叶鹿角珊瑚幼虫变态诱导的分子事件和机制，包括细胞增殖、凋亡、分化、迁移、粘附和生物矿化，进行了两种新型珊瑚基因表达测定。这些涉及使用逆转录酶定量 PCR (RT-qPCR) 并采用了 47 个感兴趣的基因 (GOI)，这些基因是根据在定居和变态过程中的假定作用进行选择的。将千花幼虫与阈值浓度和浓度提高 10 倍的假交替单胞菌提取物中的 TBP 一起孵育后，检测到 GOI 转录组反应存在显着差异。变态过程中幼虫身体结构的显着且相对突然的变化在分子水平上与暴露后 12 小时 24 个 GOI 的基因表达谱的显着差异（p<0.05）相关。其中 14 个 GOI 在暴露于阈值浓度的含 TBP 的细菌提取物后也表现出表达差异 (p<0.05)。使用稳健、低成本、准确、生态相关且高度可重复的 RT-qPCR 检测，含 TBP 的细菌提取物在不附着的情况下诱导千花幼虫变态阶段的特异性，使得附着和变态的转录组过程部分解耦。含有 TBP 的细菌提取物提供了一个独特的机会来监测专门参与变态过程的基因的调节，这与之前利用线索的基因表达研究形成鲜明对比，例如硬壳珊瑚藻、生物膜或刺激幼虫变态和附着整个开始的 GLW-酰胺神经肽。
Biofilms of the bacterium Pseudoalteromonas induce metamorphosis of acroporid coral larvae. The bacterial metabolite tetrabromopyrrole (TBP), isolated from an extract of Pseudoalteromonas sp. associated with the crustose coralline alga (CCA) Neogoniolithon fosliei, induced coral larval metamorphosis (100%) with little or no attachment (0–2%). To better understand the molecular events and mechanisms underpinning the induction of Acropora millepora larval metamorphosis, including cell proliferation, apoptosis, differentiation, migration, adhesion and biomineralisation, two novel coral gene expression assays were implemented. These involved the use of reverse-transcriptase quantitative PCR (RT-qPCR) and employed 47 genes of interest (GOI), selected based on putative roles in the processes of settlement and metamorphosis. Substantial differences in transcriptomic responses of GOI were detected following incubation of A. millepora larvae with a threshold concentration and 10-fold elevated concentration of TBP-containing extracts of Pseudoalteromonas sp. The notable and relatively abrupt changes of the larval body structure during metamorphosis correlated, at the molecular level, with significant differences (p<0.05) in gene expression profiles of 24 GOI, 12 hours post exposure. Fourteen of those GOI also presented differences in expression (p<0.05) following exposure to the threshold concentration of bacterial TBP-containing extract. The specificity of the bacterial TBP-containing extract to induce the metamorphic stage in A. millepora larvae without attachment, using a robust, low cost, accurate, ecologically relevant and highly reproducible RT-qPCR assay, allowed partially decoupling of the transcriptomic processes of attachment and metamorphosis. The bacterial TBP-containing extract provided a unique opportunity to monitor the regulation of genes exclusively involved in the process of metamorphosis, contrasting previous gene expression studies that utilized cues, such as crustose coralline algae, biofilms or with GLW-amide neuropeptides that stimulate the entire onset of larval metamorphosis and attachment.
影响因子：
4.6
作者：
B. Davidson;B. Swalla
通讯作者：
B. Davidson;B. Swalla
影响因子：
2.7
作者：
Haag, ES;Sly, BJ;Raff, RA
通讯作者：
Raff, RA
隐头基因 (ATF4) 编码多种控制果蝇蜕皮和变态的碱性亮氨酸拉链蛋白。
影响因子：
3.3
作者：
Hewes,RS;Schaefer,AM;Taghert,PH
通讯作者：
Taghert,PH