Dynamic Regulation of Intracellular Labile Cu(I)/Cu(II) Cycle in Microalgae Chlamydomonas reinhardtii: Disrupting the Balance by Cu Stress.

The labile metal pool involved in intracellular trafficking and homeostasis is the portion susceptible to environmental stress. Herein, we visualized the different intracellular distributions of labile Cu(I) and Cu(II) pools in the alga Chlamydomonas reinhardtii. We first demonstrated that labile Cu(I) predominantly accumulated in the granules within the cytoplasmic matrix, whereas the labile Cu(II) pool primarily localized in the pyrenoid and chloroplast. The cell cycle played an integral role in balancing the labile Cu(I)/Cu(II) pools. Specifically, the labile Cu(II) pool primarily accumulated during the SM phase following cell division, while the labile Cu(I) pool dynamically changed during the G phase as cell size increased. Notably, the labile Cu(II) pool in algae at the SM stage exhibited heightened sensitivity to environmental Cu stress. Exogenous Cu stress disrupted the intracellular labile Cu(I)/Cu(II) cycle and balance, causing a shift toward the labile Cu(II) pool. Our proteomic analysis further identified a putative cupric reductase, potentially capable of reducing Cu(II) to Cu(I), and four putative multicopper oxidases, potentially capable of oxidizing Cu(I) to Cu(II), which may be involved in the conversion between the labile Cu(I) pool and labile Cu(II) pool. Our study elucidated a dynamic cycle of the intracellular labile Cu(I)/Cu(II) pools, which were accessible and responsive to environmental changes.

Transcriptional regulation analysis reveals the complexity of metamorphosis in the Pacific oyster (Crassostrea gigas).

Many marine invertebrate phyla are characterized by indirect development. These animals transit from planktonic larvae to benthic spats via settlement and metamorphosis, which contributes to their adaption to the marine environment. Studying the biological process of metamorphosis is, thus, key to understanding the origin and evolution of indirect development. Although numerous studies have been conducted on the relationship between metamorphosis and the marine environment, microorganisms, and neurohormones, little is known about gene regulation network (GRN) dynamics during metamorphosis. Metamorphosis-competent pediveligers of the Pacific oyster Crassostrea gigas were assayed in this study. By assaying gene expression patterns and open chromatin region changes of different samples of larvae and spats, the dynamics of molecular regulation during metamorphosis were examined. The results indicated significantly different gene regulation networks before, during and post-metamorphosis. Genes encoding membrane-integrated receptors and those related to the remodeling of the nervous system were upregulated before the initiation of metamorphosis. Massive biogenesis, e.g., of various enzymes and structural proteins, occurred during metamorphosis as inferred from the comprehensive upregulation of the protein synthesis system post epinephrine stimulation. Hierarchical downstream gene networks were then stimulated. Some transcription factors, including homeobox, basic helix-loop-helix and nuclear receptors, showed different temporal response patterns, suggesting a complex GRN during the transition stage. Nuclear receptors, as well as their retinoid X receptor partner, may participate in the GRN controlling oyster metamorphosis, indicating an ancient role of the nuclear receptor regulation system in animal metamorphosis. Supplementary Information: The online version contains supplementary material available at 10.1007/s42995-023-00204-y.

A surge of copper accumulation in cell division revealed its cyclical kinetics in synchronized green alga Chlamydomonas reinhardtii.

Trace transition metal uptake is tightly associated with cellular biological processes. Herein, we demonstrated that copper (Cu) bioaccumulation and uptake were controlled by the cell cycle. A cyclical kinetics of Cu bioaccumulation and surge in S/M phase were observed in the synchronized green algae Chlamydomonas reinhardtii. The labile Cu(I) content also increased in the S/M phase, although the increase was moderate. Based on the comparative analysis of bioaccumulation and transcriptome data, we found the CRR1-mediated Cu uptake pathway, and CTR1 and CTR2 transporters were regulated by the intracellular Cu quota and suppressed during cell division with high Cu content. In contrast, we hypothesized a novel intracellular Cu-quota-independent Cu(I) uptake pathway in which the transporter COPT1 might be responsible for the Cu influx during cell division. Besides, a plunge of ATX1 expression level was also observed during cell division, which indicated an inhibition of the secretory pathway of Cu with the participation of ATX1 in terms of transcriptome level, probably resulting in reduced Cu efflux. Additionally, both fluorometric probe staining and transcriptomic data demonstrated that mitochondria were the dominant destination for the extra Cu content in S/M phase. Finally, some cytotoxic responses were also observed in S/M phase. Pathways related to reactive oxygen species and glutamine metabolic process were enriched in GO term and KEGG enrichment analysis, and glutathione content and cell membrane permeability determined by fluorometric probes also increased during cell division. This study showed a sharp increase of Cu uptake in cell division and revealed the genetic regulation mechanisms for the cell cycle control of Cu uptake.