Click
here to close Hello! We notice that
you are using Internet Explorer, which is not supported by Echinobase
and may cause the site to display incorrectly. We suggest using a
current version of Chrome,
FireFox,
or Safari.
Mar Drugs
2022 Sep 06;209:. doi: 10.3390/md20090568.
Show Gene links
Show Anatomy links
Characterization and Expression Analysis of Regeneration-Associated Protein (Aj-Orpin) during Intestinal Regeneration in the Sea Cucumber Apostichopus japonicus.
???displayArticle.abstract???
Apostichopus japonicus achieves intestinal regeneration in a short period after evisceration, and multiple genes are involved in this process. The transcriptome of A. japonicus was screened for regeneration-associated protein (Aj-Orpin), a gene that is specifically upregulated during intestinal regeneration. The expression and function of Aj-Orpin were identified and investigated in this study. The 5' and 3' RACE polymerase chain reaction (PCR) was used to clone the full-length cDNA of Aj-Orpin. The open reading frame codes for a 164 amino-acid protein with an EF-hand_7 domain and overlapping signal peptides and transmembrane regions. Moreover, Aj-Orpin mRNA and protein expression during intestinal regeneration was investigated using real-time quantitative PCR and Western blot. The expression pattern of Aj-Orpin in the regenerating intestine was investigated using immunohistochemistry. The results showed that Aj-Orpin is an exocrine protein with two EF-hand-like calcium-binding domains. Expression levels were higher in the regenerating intestine than in the normal intestine, but protein expression changes lagged behind mRNA expression changes. Aj-Orpin was found to play a role in the formation of blastema and lumen. It was primarily expressed in the serosal layer and submucosa, suggesting that it might be involved in proliferation. These observations lay the foundation for understanding the role of Orpin-like in echinoderm intestinal regeneration.
Figure 1. Sequence analysis of Aj-Orpin in A. japonicus. (a) Nucleotide sequence (1136 bp) of the Aj-Orpin gene (accession No. ARI48335.1) aligned with the predicted amino acid (AA) sequence (capital single-letter code). The full-length nucleotide sequence includes 22 bp 5â²-untranslated region (UTR), a 619 bp 3â²-UTR, and a 495 bp open-reading frame (164 AA). The start and stop codons are included in the black box. The stop codon is indicated by an asterisk. The signal peptide and the transmembrane region are denoted by the red box and the red underline respectively, and the cleavage site is shown with the red arrow. (b) Multiple alignment of the deduced amino acid sequences of Aj-Orpin. The EF-hand domain pair is denoted by the red box. (c) Phylogenetic trees based on Aj-Orpin amino acid sequences from A. japonicus and other species. The tree was constructed based on the multiple sequences generated by MUSCLE algorithm and aligned using the Maximum Likelihood method with 100 bootstraps.
Figure 2. Relative expression levels of Aj-Orpin mRNA and protein during intestinal regeneration. (a) Real-time quantitative PCR analysis of Aj-Orpin expression at different intestinal regeneration time points: control, 0, 30 min, 1, 2, 6, 12 h and 1, 3, 7, and 14 days post-evisceration. Normal intestine was treated as the control group. (b) Western blot analysis showed Aj-Orpin protein expression at different time points during intestinal regeneration. β-actin served as the reference protein. (c) The protein expression level was evaluated by calculating the gray value of the bands. Data are shown as the mean ± SE. The lowercase letters indicate statistically significant differences (p < 0.05).
Figure 3. (a) Diagram to show the location of the slices. (b) Immunohistochemical staining for Aj-Orpin at different regeneration time points. (A1âF1) showed the pattern of Aj-Orpin protein expression from time 0 to day 7; (A2âF2) are the 4-fold magnification images of (A1âF1). ser: serosal layer, mus: muscle layer, sub: submucosal layer, muc: mucosal layer, lum: lumen. The reddish-brown particles pointed to by the arrows indicate positive Aj-Orpin expression. Aj-Orpin was mainly expressed in the serosal layer and submucosal layer and weakly in other layers.
Bello,
Insights into intestinal regeneration signaling mechanisms.
2020, Pubmed,
Echinobase
Bello,
Insights into intestinal regeneration signaling mechanisms.
2020,
Pubmed
,
Echinobase
Boyko,
The Eupentacta fraudatrix transcriptome provides insights into regulation of cell transdifferentiation.
2020,
Pubmed
,
Echinobase
Brekken,
SPARC, a matricellular protein: at the crossroads of cell-matrix communication.
2001,
Pubmed
Candelaria,
Contribution of mesenterial muscle dedifferentiation to intestine regeneration in the sea cucumber Holothuria glaberrima.
2006,
Pubmed
,
Echinobase
Dolmatov,
Molecular Aspects of Regeneration Mechanisms in Holothurians.
2021,
Pubmed
,
Echinobase
Dolmatov,
Expression of Piwi, MMP, TIMP, and Sox during Gut Regeneration in Holothurian Eupentacta fraudatrix (Holothuroidea, Dendrochirotida).
2021,
Pubmed
,
Echinobase
García-Arrarás,
Cellular mechanisms of intestine regeneration in the sea cucumber, Holothuria glaberrima Selenka (Holothuroidea:Echinodermata).
1998,
Pubmed
,
Echinobase
García-Arrarás,
The mesentery as the epicenter for intestinal regeneration.
2019,
Pubmed
,
Echinobase
García-Arrarás,
Visceral regeneration in holothurians.
2001,
Pubmed
,
Echinobase
García-Arrarás,
Cell dedifferentiation and epithelial to mesenchymal transitions during intestinal regeneration in H. glaberrima.
2011,
Pubmed
,
Echinobase
Girich,
Wnt and frizzled expression during regeneration of internal organs in the holothurian Eupentacta fraudatrix.
2017,
Pubmed
,
Echinobase
Huo,
Global-warming-caused changes of temperature and oxygen alter the proteomic profile of sea cucumber Apostichopus japonicus.
2019,
Pubmed
,
Echinobase
Kretsinger,
Carp muscle calcium-binding protein. II. Structure determination and general description.
1973,
Pubmed
Latvala,
Distribution of SPARC protein (osteonectin) in normal and wounded feline cornea.
1996,
Pubmed
Li,
Identification and expression characterization of WntA during intestinal regeneration in the sea cucumber Apostichopus japonicus.
2017,
Pubmed
,
Echinobase
Marchler-Bauer,
CDD/SPARCLE: functional classification of proteins via subfamily domain architectures.
2017,
Pubmed
Mashanov,
Transdifferentiation in holothurian gut regeneration.
2005,
Pubmed
,
Echinobase
Mashanov,
Expression of Wnt9, TCTP, and Bmp1/Tll in sea cucumber visceral regeneration.
2012,
Pubmed
,
Echinobase
Mashanov,
Visceral regeneration in a sea cucumber involves extensive expression of survivin and mortalin homologs in the mesothelium.
2010,
Pubmed
,
Echinobase
McPhalen,
Calcium-binding sites in proteins: a structural perspective.
1991,
Pubmed
Miao,
Extracellular matrix remodeling and matrix metalloproteinases (ajMMP-2 like and ajMMP-16 like) characterization during intestine regeneration of sea cucumber Apostichopus japonicus.
2017,
Pubmed
,
Echinobase
Moe,
Cell healing: Calcium, repair and regeneration.
2015,
Pubmed
Nelson,
Structures of EF-hand Ca(2+)-binding proteins: diversity in the organization, packing and response to Ca2+ binding.
1998,
Pubmed
Nieves-Ríos,
The nervous system component of the mesentery of the sea cucumber Holothuria glaberrima in normal and regenerating animals.
2020,
Pubmed
,
Echinobase
Ortiz-Pineda,
Gene expression profiling of intestinal regeneration in the sea cucumber.
2009,
Pubmed
,
Echinobase
Persechini,
The EF-hand family of calcium-modulated proteins.
1989,
Pubmed
Phan,
Role of SPARC--matricellular protein in pathophysiology and tissue injury healing. Implications for gastritis and gastric ulcers.
2007,
Pubmed
Quiñones,
Extracellular matrix remodeling and metalloproteinase involvement during intestine regeneration in the sea cucumber Holothuria glaberrima.
2002,
Pubmed
,
Echinobase
Quispe-Parra,
Transcriptomic analysis of early stages of intestinal regeneration in Holothuria glaberrima.
2021,
Pubmed
,
Echinobase
Reed,
Differential expression of SPARC and thrombospondin 1 in wound repair: immunolocalization and in situ hybridization.
1993,
Pubmed
Rojas-Cartagena,
Distinct profiles of expressed sequence tags during intestinal regeneration in the sea cucumber Holothuria glaberrima.
2007,
Pubmed
,
Echinobase
Sage,
Distribution of the calcium-binding protein SPARC in tissues of embryonic and adult mice.
1989,
Pubmed
Santiago-Cardona,
Lipopolysaccharides induce intestinal serum amyloid A expression in the sea cucumber Holothuria glaberrima.
2003,
Pubmed
,
Echinobase
Sun,
Large scale gene expression profiling during intestine and body wall regeneration in the sea cucumber Apostichopus japonicus.
2011,
Pubmed
,
Echinobase
Sun,
Understanding regulation of microRNAs on intestine regeneration in the sea cucumber Apostichopus japonicus using high-throughput sequencing.
2017,
Pubmed
,
Echinobase
Sun,
Cloning and expression analysis of Wnt6 and Hox6 during intestinal regeneration in the sea cucumber Apostichopus japonicus.
2013,
Pubmed
,
Echinobase
Sun,
RNA-Seq reveals dynamic changes of gene expression in key stages of intestine regeneration in the sea cucumber Apostichopus japonicus. [corrected].
2013,
Pubmed
,
Echinobase
Tamura,
MEGA11: Molecular Evolutionary Genetics Analysis Version 11.
2021,
Pubmed
Tarnawski,
Cellular and molecular mechanisms of gastrointestinal ulcer healing.
2005,
Pubmed
Whelan,
A general empirical model of protein evolution derived from multiple protein families using a maximum-likelihood approach.
2001,
Pubmed
Yuan,
Wnt Signaling Pathway Linked to Intestinal Regeneration via Evolutionary Patterns and Gene Expression in the Sea Cucumber Apostichopus japonicus.
2019,
Pubmed
,
Echinobase
Zhang,
The sea cucumber genome provides insights into morphological evolution and visceral regeneration.
2017,
Pubmed
,
Echinobase
