- Proteome of larval metamorphosis induced by epinephrine in the Fujian oyster. - Calmodulin and MAPK pathways were involved in the regulation of settlement of the oyster. - Expression levels of proteins that were enriched in the epinephrine signaling pathway may reflect the developmental stage of the larvae, that may reflect whether or not larvae were directly involved in metamorphosis when the larvae were treated with epinephrine.. - Transcriptional regulation is involved in the process of settlement and metamorphosis.. - In the Fujian oyster, Cacaspase-2, Cacaspase-3 mRNA, and settlement and metamorphosis-related protein (SMRP1) are highly expressed in larvae during settlement and metamorphosis [12, 13].. - proposed that adrenergic re- ceptors are involved in the regulation of settlement and metamorphosis in oysters [20, 32]. - The relative molecular weights of the identified proteins were mainly distributed in the range of 10–60 kDa, of which 20–30 kDa was the largest group (Supplementary Fig. - There were 117 co-expressed proteins in the PL-PA and PA-MET groups (Fig. - The results of a cluster analysis showed that expression levels of most proteins had a consistent trend in the PL-PA-MET developmental process (Fig. - There were 246, 341, and 230 DEPs in the PL-eSEN group, the PL-einSEN group, and the eSEN-einSEN groups, respectively (Fig. - There were 35 DEPs present in all 3 groups, of which 22 were down-regulated in the PL-eSEN and PL-einSEN groups, and 3 proteins were down-regulated in the eSEN group (Fig. - 2a) and up- regulated in the einSEN group (Cluster 1 of Fig. - A total of 101 proteins were differentially expressed only in the PL-einSEN group and eSEN-einSEN group (Fig. - there were 26 proteins that were down-regulated in the eSEN group but up-regulated in the einSEN group. - There were 560, 539, and 443 DEPs in the PL-MET, PL-eMET and MET-eMET groups, respectively, of which 286, 235, and 194 proteins were up-regulated and. - There were 132 DEPs observed only in the PL-MET and MET-eMET groups, and 116 DEPs found only in the PL-eMET and MET-eMET group (Fig. - In addition, 197 proteins were differentially expressed in the PL- MET and PL-eMET groups but not in the MET-eMET group. - 2 Summary analysis of differentially expressed proteins in the PL-eSEN-einSEN groups. - 3 The summary analysis of differentially expressed proteins in the PL-MET-eMET groups. - Supplementary Table 7 shows the ex- tremely significant GO enrichment in the PL-PA and PA-MET groups. - The statistical analysis of GO enrichment in the PL- eSEN, PL-einSEN, and eSEN-einSEN groups is showed in Table 2. - Supplementary Table 8 shows that seven ex- tremely significant GO terms were only expressed in the PL-eSEN and eSEN-einSEN groups. - GO entries that were extremely expressed only in the PL-einSEN and eSEN-einSEN groups are shown in Supplementary Table 8.. - The significantly enriched and extremely significantly enriched GO terms were involved in the molecular process among three groups. - the most extremely signifi- cant GO terms were in the PL-MET group, which had 37 terms. - 0.05 was used as a determiner of significant enrich- ment in the KEGG pathway analysis. - Pathways were classified in the PL-PA group, the PA-MET group, the PL-eSEN group, the PL-einSEN group, the eSEN-eMET Table 2 Summary of GO enrichment of DEPs. - During the process of oyster pediveliger settlement and metamorphosis, 22 DEPs were significantly enriched in 32 pathways in the PL-PA group, and these were mainly involved in organismal systems pathways (Fig. - 23 DEPs were significantly enriched in 38 Pathways in the PL-MET group, 29 of which were related to metab- olism (Fig. - From the pediveliger of the oyster to post-epinephrine- induced metamorphosis, in the PL-eSEN group, 14 DEPs were enriched in 17 pathways, of which 7 were related to organismal systems, and 4 were related to metabolism (Fig. - Compared with PL-eSEN group, 15 of 32 DEPs in the eSEN-eMET group were related to organismal sys- tems, and 12 were related to metabolism. - Twenty-three DEPs were significantly enriched in 50 pathways in the PL-einSEN group. - In the MET-eMET group, 26 DEPs were enriched in 38 pathways, 7 of which were re- lated to organismal systems pathways. - three genes (PSMB, HYOU, and GST) were differentially expressed at the transcriptional level but showed no sig- nificant changes in the protein levels.. - Some larval tissues degenerate during settlement and metamorphosis, e.g., the anterior adductor, the cili- ated velum, and the ventral retractors [53]. - There were 246 DEPs in the PL-eSEN group. - the num- ber of DEPs in the PL-einSEN group was nearly one- third more, at about 341 proteins. - In addition, down- regulated proteins accounted for 67.1 and 66.6% of the differentially expressed proteins in the PL-eSEN group and PL-einSEN group, respectively. - The number of significantly enriched KEGG pathways in the PL-einSEN group was three times that in the PL- eSEN group. - Metabolism mediated by cytochrome P450 enzymes can decompose lipophilic toxic substances into aqueous solution [56], and this was up-regulated in eSEN and down-regulated in the einSEN group.. - In this study, compared with the PL group, the expression levels of GSTs were low in the einSEN and eSEN groups (Supplementary Table 6). - The unique signaling pathways in the PL-einSEN group included adrenergic signaling in cardiomyocytes, dopaminergic synapse, insulin signaling pathway, and. - Although the expression of these proteins was down-regulated in the eSEN group, only three proteins (calmodulin, ryanodine receptor, calcium/calmodulin-dependent protein kinase). - We speculated that the expression levels of proteins related to the epinephrine pathway in the larvae of the eSEN group were higher than in the einSEN group before epinephrine treatment. - There were 29 KEGG enrichment results related to metabolic pathways in the PL-MET group, including carbohydrate and lipid metabolism. - Glycolysis-related proteins play a key role in the embryonic development of zebra- fish and ascidians [59]. - Most proteins involved in carbo- hydrate metabolic processes were up-regulated in the PL group and down-regulated in the MET group (Supple- mentary Table 6). - [63] found that cal- modulin (CaM) and calmodulin-determined myosin light chain kinase (MLCK) played important roles in the settlement process of barnacles. - In this study, in comparing the PL and PA groups, calcium/calmodulin-dependent protein kin- ase type II delta chain (GI and calmodulin (GI20137620, GI762161385) were more highly expressed in the PL group. - This result indicated that calmodulin was involved in the regulation of settlement of oysters.. - In comparisons of the PL and PA groups, mitogen-activated protein kinase 2-like was expressed at a higher level in the PL group. - The NOTCH signaling pathway was involved in the regulation of somatic division of Capitella sp. - In this study, the expression levels of neurogenic locus Notch protein in the PL, MET and eMET groups were and 13.89, respectively. - The protein was up-regulated in the MET and eMET groups (Supple- mentary Table 6). - This result indicated that MAPK and NOTCH signaling pathways were also involved in the regulation of settlement and metamorphosis of oysters.. - Those pathways are involved in the regulation of settlement and metamorphosis of marine invertebrates . - Pre- vious study reported up-regulation of innate immune- related genes during metamorphosis in the ascidian Bol- tenia villosa [69].. - [77] dis- cussed how lectin-like factors were involved in larval settlement and metamorphosis in the abalone H. - These results suggested that hepatic lectin-like proteins are involved in the metamorphosis of C. - In this study, cadherins, calmodulin and calreticulin were up-regulated in the MET and eMET groups (Supplementary Table 6). - on the expression of calmodulin in the processes of settlement and metamorphosis of the polychaete Hydro- ides elegans [80]. - Comparing the PA and MET groups, apoptosis- inducing factor 3-like isoform X1 was expressed at a lower level in the MET group. - Changes in shell formation are important in the develop- ment of mollusks. - In this study, cal- modulin and calreticulin were up-regulated in the MET and eMET groups (Supplementary Table 6). - Calmodulin and calreticulin were also identified in the larvae of the snail P. - canaliculata during shell development [43] and in the Pacific oyster C. - The up- regulation of both proteins suggests that they may be in- volved in the calcification of larval shells. - In this study, comparison of PA and MET group, colla- gen alpha (GI was up-regulated in the MET group. - In shell matrix, collagen-like VWA do- main containing proteins, fibronectin-like proteins and laminins were involved in the regulation of shell formation [85, 86]. - In this study, comparison of MET and eMET group, chitinase-3-like protein 1 isoform X3 was up-regulated in the eMET group.. - In the present study, 14–3-3 and SCO-spondin were identified. - Both proteins were signifi- cantly up-regulated in the eMET group (Supplementary Table 6). - In this study, there were no significant dif- ferences among PL, PA, and MET groups, but the 14–3- 3 protein was up-regulated in the eMET group (Supple- mentary Table 6). - The overexpression of 14–3-3 in the body plays an important role in nerve cell differentiation [92]. - The 14–3-3 proteins also play important roles in left–right patterning in the axial dir- ection of the embryo during amphibian embryogenesis [94]. - In the present study, SCO-spondin was expressed at a high level in the PA group and was up-regulated in the eMET group. - The up-regulation of 14–3-3 and calmodulin in the MET and eMET groups may indicate their roles in neuronal function and may indicate the en- hanced development of the nervous system in C. - In this study, neuronal acetylcho- line receptor subunit alpha-10 (c88012_g1) and ryano- dine receptor 44F (c96209_g1) were upregulated post epinephrine stimulation in the eSEN group. - Up-regulation of skeleton proteins is often found in the study of settlement and metamorphosis of marine invertebrates [40, 41]. - Studies have shown that skeleton proteins play an important role in the metamorphic development of amphibians [101] and insects [102]. - In this study, the proteome of larval settlement and meta- morphosis induced by epinephrine in the Fujian oyster C.. - The results identified the DEPs involved in the epinephrine signaling pathway, and the levels of these proteins may reflect the developmental stage of oyster larvae. - After filtration, more than 95% of the larvae could be identified under a microscope as being in the pediveliger stage. - For each developmental stage, almost all of the collected individuals were in the same period as confirmed by microscopic examination.. - The supernatant was filtered with a 0.22-μm microporous membrane, and the polypeptide solution was mixed in the sample bottle using an oscillator. - Identification of differentially expressed proteins involved in the early larval development of the Pacific oyster Crassostrea gigas. - Proteomic analysis of larvae during development, attachment, and metamorphosis in the fouling barnacle, Balanus amphitrite.. - Phosphoproteome analysis during larvae development and metamorphosis in the spionid polychaete Pseudopolydora vexillosa. - Quantitative proteomics study of larvae settlement in the barnacle Balanus amphitrite. - Identification of thyroid hormones and functional characterization of thyroid hormone receptor in the Pacific oyster, Crassostrea gigas, provide insight into evolution of the thyroid hormone system. - Proteomic analysis of trochophore and veliger larvae development in the small abalone Haliotis diversicolor. - Transcriptome and quantitative proteome analysis reveals molecular processes associated with larvae metamorphosis in the polychaete Pseudopolydora vexillosa. - Proteomic profiles of embryonic development in the ascidian Ciona intestinalis. - Notch signaling during larvae and juvenile development in the polychaete annelid Capitella sp I. - Developmental expression of Hsp90, Hsp70 and HSF during morphogenesis in the vetigastropod Haliotis asinina. - A complement response may activate metamorphosis in the ascidian Boltenia villosa. - Involvement of lectins in the settlement and. - Involvement of lectin-like factors in larval settlement and metamorphosis in the abalone, Haliotis discus hannai.. - Structure of the nacreous organic matrix of a bivalve mollusk shell examined in the hydrated state using Cryo-TEM. - 14-3-3 ε and ζ regulate neurogenesis and differentiation of neuronal progenitor cells in the developing brain
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