Kawamura Kaz, Sekida Satoko, Nishitsuji Koki, Satoh Noriyuki
Department of Applied Science, Kochi University, Kochi, Japan.
Kuroshio Science Unit, Multidisciplinary Science Cluster, Kochi University, Kochi, Japan.
Dev Growth Differ. 2025 Apr;67(3):119-135. doi: 10.1111/dgd.70000. Epub 2025 Feb 21.
In previous studies, we have established approximately 15 cultured cell-lines derived from planula larvae of Acropora tenuis. Based on their morphology and behavior, these cells were classified into three types, flattened amorphous cells (FAmCs), vacuolated adherent cells (VAdCs), and small smooth cells (SSmCs). FAmCs include fibroblast-like cells and spherical, brilliant brown cells (BBrCs), which are transformable to each other. To examine the larval origin of the three cell types, we raised antibodies: anti-AtMLRP2 that appears to recognize FAmC, anti-AtAHNAK for BBrC, anti-AtSOMP5 and anti-AtEndoG for SSmC, and anti-AtGal and anti-AtFat4 for VAdC, respectively. Anti-AtMLRP2 antibody stained in vivo stomodeum and neuroblast-like cells embedded in larval ectoderm around the aboral pole. Anti-AtAHNAK antibody stained neuron-like and neuroblast-like cells, both of which were also stained with neuron-specific tubulin β-3 antibody. These results suggest that in vitro BBrCs and in vivo neuroblast-like cells share neuronal properties in common. Two antibodies for SSmCs, anti-AtSOMP5 and anti-AtEndoG, stained larval ectoderm cells, suggesting that SSmCs have larval ectoderm properties. Two antibodies for VAdCs, anti-AtGal and anti-AtFat4, stained larval endoderm cells, suggesting that VAdCs have larval endoderm properties. Therefore, the in vitro cell lines appear to retain properties of the stomodeum, neuroblast, ectoderm, or endoderm. Each of them may be used in future investigations to reveal cellular and molecular properties of cell types of coral larvae, such as the potential for symbiosis.
在之前的研究中，我们已经建立了约15种源自细枝鹿角珊瑚浮浪幼虫的培养细胞系。根据它们的形态和行为，这些细胞被分为三种类型，扁平无定形细胞（FAmCs）、空泡化贴壁细胞（VAdCs）和小光滑细胞（SSmCs）。FAmCs包括成纤维细胞样细胞和球形、明亮的棕色细胞（BBrCs），它们可相互转化。为了研究这三种细胞类型的幼虫起源，我们制备了抗体：分别用于识别FAmC的抗AtMLRP2、用于BBrC的抗AtAHNAK、用于SSmC的抗AtSOMP5和抗AtEndoG，以及用于VAdC的抗AtGal和抗AtFat4。抗AtMLRP2抗体在体内对口道以及位于反口极周围幼虫外胚层中的神经母细胞样细胞进行了染色。抗AtAHNAK抗体对神经元样和神经母细胞样细胞进行了染色，这两种细胞也都被神经元特异性微管蛋白β-3抗体染色。这些结果表明，体外的BBrCs和体内的神经母细胞样细胞具有共同的神经元特性。两种针对SSmCs的抗体，抗AtSOMP5和抗AtEndoG，对幼虫外胚层细胞进行了染色，表明SSmCs具有幼虫外胚层特性。两种针对VAdCs的抗体，抗AtGal和抗AtFat4，对幼虫内胚层细胞进行了染色，表明VAdCs具有幼虫内胚层特性。因此，体外细胞系似乎保留了口道、神经母细胞、外胚层或内胚层的特性。它们中的每一种都可用于未来的研究，以揭示珊瑚幼虫细胞类型的细胞和分子特性，例如共生潜力。
