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Identifying proteins that regulate fin injury is critical to our understanding of regeneration as it relates to both acute wound injury and tissue formation. We have cloned the full-length cDNA of the actinodin4 (and4) gene of Misgurnus anguillicaudatus (MaAnd4) by the RACE method (GenBank Accession No. MG385835). Quantitative RT-PCR analysis during fin regeneration indicated a sudden increase in MaAnd4 expression, with a peak at 3 days post amputation (dpa). In situ analysis showed that MaAnd4 is located in the distal blastema and cells lining the regions of actinotrichia formation at 3 dpa. The highest levels of MaAnd4 expression were observed in the adult testis as well as in the gastrulae during embryonic development. Southern blotting confirmed the existence of and4 in teleosts but not in tetrapods examined. The results show the expression of this gene in actinotrichia formation and its association with fin/limb regeneration ability in teleosts. [ABSTRACT FROM AUTHOR]

The mechanisms of sex determination and differentiation have not been elucidated in most fish species. In this study, the full-length cDNAs of DAX1 was cloned and characterized in aquaculture fish Chinese loach (Paramisgurnus dabryanus), designated as Pd-DAX1. The cDNA sequence of Pd-DAX1 was 1261 bp, including 795 bp open reading frame (ORF) encoding 264 amino acids. Pd-DAX1 shares highly identical sequence with DAX1 homologues from different species. The expression profiles of Pd-DAX1 in different developmental stages and diverse adult tissues were analyzed by quantitative real-time RT-PCR and in situ hybridization (ISH). Pd-DAX1 was continuously expressed during embryogenesis, with the extensive distribution in the development of the central nervous system. Tissue distribution analysis revealed that Pd-DAX1 expressed widely in adult tissues, with the highest expression level found in testis, moderate level in ovary, showing a sex-dimorphic expression pattern. Pd-DAX1 mainly located in spermatogonia cells, spermatocytes, primary oocytes and previtellogenic oocyte cells, implying that Pd-DAX1 may involve in gametogenesis. These preliminary findings suggest that Pd-DAX1 gene is highly conserved during vertebrate evolution and involved in a wide range of developmental processes including embryogenesis, central nervous system development and gonad development. [ABSTRACT FROM AUTHOR]

As a universal environmental contaminant, the herbicide cyhalofop-butyl is considered to have infested effects on the embryonic development of aquatic species. The present study focused on an assessment of the impacts of cyhalofop-butyl on Yellow River carp embryos. It was found that cyhalofop-butyl inhibited the hatching of the embryos, and the hatching rate decreased with higher concentrations of the herbicide. The mortality rate was increased on exposure to cyhalofop-butyl and was significantly higher in the 1.6 and 2 mg/L treatment groups over 48 h. All of the embryos of the 2 mg/L treatment group died within the 48 h post-hatching stage. And the transcription of several embryos related to apoptosis was also influenced by cyhalofop-butyl exposure. Further, cyhalofop-butyl exposure leads to a series of morphological changes (pericardial edema, tail deformation, and spine deformation) in embryos, which were consistent with significant modifications in the associated genes. These results provided a scientific basis for further studies into the effects of cyhalofop-butyl on aquatic organisms. [ABSTRACT FROM AUTHOR]

Sox7, Sox17 and Sox18 are the members of the Sry-related high-mobility group box family (SoxF) of transcription factors. SoxF factors regulate endothelial cell fate as well as development and differentiation of blood cells and lymphatic vessels. There is very less information about the functions of these genes in fish. We obtained the full-length cDNA sequence of SoxF genes including Sox7, Sox17 and Sox18 in Cyprinus carpio, where Sox7 and Sox18 had two copies. The construction of a phylogenetic tree showed that these genes were homologous to the genes in other species. Chromosome synteny analysis indicated that the gene order of Sox7 and Sox18 was highly conserved in fish. However, immense change in genomic sequences around Sox17 had taken place. Numerous putative transcription factor binding sites were identified in the 5′ flanking regions of SoxF genes which may be involved in the regulation of the nervous system, vascular epidermal differentiation and embryonic development. The expression levels of SoxF genes were highest in gastrula, and was abundantly expressed in the adult brain. We investigated the expression levels of SoxF genes in five specific parts of the brain. The expression levels of Sox7 and Sox18 were highest in the mesencephalon, while the expression level of Sox17 was highest in the epencephalon. In carp, the expression patterns of SoxF genes indicated a potential function of these genes in neurogenesis and in vascular development. These results provide new information for further studies on the potential functions of SoxF genes in carp. [ABSTRACT FROM AUTHOR]

Urodele amphibians and teleost fish are capable of nearly perfect regeneration of lost appendages. The fin constitutes an important model for studying the molecular basis of tissue regeneration. It has been known that heat shock protein 60 (Hsp60) is a multifunctional protein of the heat shock protein family. The purpose of this study is to investigate the role of hsp60 as a part of a stress response system after fin injury or in fin regeneration. We firstly cloned full-length cDNA of hsp60 from Misgurnus anguillicaudatus (designated as MaHsp60) by RACE method. The cDNA contains a 83-bp 5′UTR, a 1,728-bp open reading frame encoding 492 amino acids and a 542-bp 3′UTR (Accession No.: KF537340). The phylogenetic tree shows that the MaHsp60 fits within the hsp60 clade. Then quantitative RT-PCR detected that MaHsp60 began to increase rapidly its expression at 1 dpa and reached its peak at 2 dpa. Next, spatial distribution analysis of MaHsp60 in fins showed that MaHsp60 located mainly in the deeper layer of regenerated epidermis when MaHsp60 expressed most. After the MaHsp60 had been cloned into the pET-32a vector, SDS-PAGE analysis confirmed that the MaHsp60 protein was efficiently expressed in Escherichia coli BL21 and adjustable with the temperature. These findings have revealed that MaHsp60, a highly conserved gene during vertebrate evolution as well as related to stress response, is involved in the formation of wound epidermis which occurs as the first phase of fin regeneration after fin amputation in caudal fin regeneration. [ABSTRACT FROM AUTHOR]

To investigate the function and evolutionary origin of the SOXE subgroup, we amplified the genomic DNA of Misgurnus anguillicaudatus using a pair of degenerate primers. Using RACE, we obtained two versions of Sox8 ( MaSox8a and MaSox8b) from M. anguillicaudatus. The overall sequence identity of the deduced amino acids from the two genes was 54.38%, with only one amino acid change in the high-mobility group domain. Southern blotting and evidence from the phylogenetic tree provided further proof for the existence of two Sox8 genes at the genomic level. This is the first evidence of two distinct Sox8 genes in Cypriniformes. Semi-quantitative and real-time quantitative PCR assays showed the expression trend of the genes was opposite in early embryonic development, and both were expressed ubiquitously in several adult tissues. The similar expression patterns indicated that MaSox8a and MaSox8b have possible overlapping functions. [ABSTRACT FROM AUTHOR]