A new Agkistrodon halys venom-purified protein C activator prevents myocardial fibrosis in diabetic rats

Aim To assess the effects of protein C activator (PCA) from Agkistrodon halys snake venom on cardiac fibrosis in streptozotocin (STZ) induced diabetic rat model, and investigate the mechanisms of its action. Methods PCA was identified by one-dimensional reversed phase liquid chromatography – mass spectrometry/mass spectrometry. Male Sprague-Dawley rats (120-140 g) were randomly assigned to negative control (NC) and diabetic group. Diabetes was induced by STZ in high-fat diet fed rats. Diabetic group was subdivided into three groups: diabetic group (DM), diabetic group treated with PCA (0.5, 2, and 8 mg/kg), and diabetic group treated with metformin (5 mg/kg, positive control). NC and DM groups received the same volume of distilled water. Left ventricular mass index (LVWI) and collagen volume fraction were measured by hematoxylin and eosin and Masson staining. Transforming growth factor beta-1 (TGF-β1) and interleukin 1 beta (IL-1β) levels were determined by enzyme-linked immunosorbent assay. Results The diabetic rat model was successfully established by STZ induction and high-fat diet. Glucose level, LVWI, TGF-β1 and IL-1β level, and collagen volume fraction were significantly reduced in diabetic rats treated by PCA in a dose-dependent manner (P < 0.050), especially in the high dose (8 mg/kg) group (P < 0.010), compared to diabetes group. The high dose PCA had the same effect as metformin positive control in reducing the level of fasting blood glucose. PCA decreased the expression of MMP-2 and reduced that of TIMP-2. Conclusion Our results indicate that PCA has anti-fibrotic effects and that it may be used to treat myocardial fibrosis

Transcriptome sequencing and comparative analysis of adult ovary and testis identify potential gonadal maintenance-related genes in Mauremys reevesii with temperature-dependent sex determination

Mauremys reevesii is a classical organism with temperature-dependent sex determination (TSD). Gonad development in early life has recently received considerable attention but gonadal maintenance after sex differentiation in turtles with TSD remains a mystery. In this study, we sequenced the transcriptomes for the adult testis and ovary using RNA-seq, and 36,221 transcripts were identified. In total, 1,594 differentially expressed genes (DEGs) were identified where 756 DEGs were upregulated in the testis and 838 DEGs were upregulated in the ovary. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis suggested that the TGF-beta signaling pathway and Hedgehog signaling pathway have important roles in testis maintenance and spermatogenesis, whereas the Hippo signaling pathway and Wnt signaling pathway are likely to participate in ovary maintenance. We determined the existence of antagonistic networks containing significant specific-expressed genes and pathways related to gonadal maintenance and gametogenesis in the adult gonads of M. reevesii. The candidate gene Fibronectin type 3 and ankyrin repeat domains 1 (FANK1) might be involved with the regulation of testis spermatogenesis

Organization and variation of mitochondrial DNA control region in pleurodiran turtles

Three complete mitochondrial DNA (mtDNA) control regions (CRs) of Chelodina rugosa (Ogilby, 1890), Chelus fimbriata (Schneider, 1783), and Podocnemis unifilis (Troschel, 1848) were firstly determined using Long-PCR method and the length were 1,016 bp, 1,149 bp, and 985bp, respectively. Together with CRs of Pelomedusa subrufa (Bonnaterre, 1789) and nearly complete CR of Podocnemis expansa (Schweigger, 1812) obtained from GenBank, the structural and evolutionary characteristics of mtDNA CRs in pleurodiran turtle were analyzed in this study. We identified three functional domains (TAS, CD, and CSB domains) as well as their conservation sequences (TAS, CSB-F, and CSB-1) according to their homology to those of other turtles. Within the TAS domain, an interrupted poly-C stretch was found in C. rugosa, C. fimbriata, and P. subrufa, which also exists in the published mt DNA CRs of Chrysemys picta (Schneider, 1783), Trachemys scripta (Thunberg in Schoepff, 1792), and Trionyx triunguis (Forskål, 1775). The analysis of the origin for the poly-C sequences in TAS domain from six turtles suggested that the poly-C sequences are more related to "goose hairpin" in birds rather than CSB2 in CSB domain. In the CSB domain, CSB2 and CSB3, which were determined in CRs of Cryptodira, were absent in Pleurodira CRs, indicating the regulative mechanisms of transcription may be varied in both two suborders and the lack of CSB2 and CSB3 could be proposed as one of diagnostic characters between Pleurodira and Cryptodira at molecular level. As for CR of other cryptodiran turtles, variable number of tandem repeats (VNTRs) in the 3' end of the CRs was found in the five pleurodiran turtles. Interestingly, the long repeated motifs from each species could form stable stem-loop secondary structures, suggesting that the repeated sequences may play an important role in regulating replication of the mitochondrial genome in Pleurodiran, and the secondary structures of VNTRs may provide some potential information in phylogenetic inference

Mauremys reevesii

Raw data of <i>Mauremys reevesii</i> transcriptom

The complete mitochondrial genome of the Madagascan plowshare tortoise Astrochelys yniphora (Testudines, Testudinidae)

We sequenced and characterized the complete mitochondrial genome of the Madagascan plowshare tortoise Astrochelys yniphora (Testudines, Testudinidae). This genome was 16,527 bp and consisted of 13 protein-coding genes (PCGs), two ribosomal RNA (rRNA) genes, 22 transfer RNA (tRNA) genes, and a noncoding control region. The genome organization, gene order, and base composition were identical to the typical vertebrate. There was no an additional C in arm of tRNALys gene secondary structure. Astrochelys yniphora 945 bp control region concluded the variable number of tandem repeats (VNTRs). The complete mitogenome of A. yniphora provides the basic data to molecular systematics research of Testudinidae

Molecular cloning and expression patterns of the Vasa gene from Rana nigromaculata (Amphibia: Anura)

The Vasa protein is a member of the DEAD (Asp-Glu-Alu-Asp) box family of ATP-dependent RNA helicases. The Vasa gene is specifically expressed in germ-line cells of many metazoans and is known to play a critical role in gametogenesis and reproductive regulation. In this paper, we isolate the full length cDNA sequence of the Vasa gene from the frog Rana nigromaculata Hallowell, 1861. The open reading frame (ORF) encoding 398 amino acid residues has nine conserved motifs. According to the similarities at the amino acid sequenceythe phylogenetic analysis of Vasa gene was consistent with the evolution relationships from chordates to mammals. Furthermore, the expression pattern analysis of RnVasa mRNA, using the technique of Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR), showed a high level of transcripts in testis, ovary and kidney, whereas little to no signal was detected in other tissues, which suggests that it may play a role during gametogenesis

Molecular cloning and analysis of Myc modulator 1 (Mm-1) from Bufo gargarizans (Amphibia: Anura)

The protein of Myc modulator 1 (Mm-1) has been reported to repress the transcriptional activity of the proto-oncogene c-Myc in humans. Moreover, it was shown to be the subunit 5 of human prefoldin (PFD). So far, this gene and its homologs have been isolated and sequenced in many organisms, such as mammals and fish, but has not been sequenced for any amphibian or reptile. In order to better understand the function and evolution of Mm-1, we isolated a full-length Mm-1 cDNA (BgMm-1, GenBank accession no. EF211947) from Bufo gargarizans (Cantor, 1842) using RACE (rapid amplification of cDNA ends) methods. Mm-1 in B. gargarizans is 755 bp long, comprising an open reading frame (ORF) of 459 bp encoding 152 amino acids. The amino acid sequence had a prefoldin &#945;-like domain, partially including a typical putative leucine zipper motif. BgMm-1 showed high similarity to its homolog of Mus musculus Linnaeus, 1758 (82%) and Homo sapiens Linnaeus, 1758 MM-1 isoform a (81%) at the amino acid level. The protein secondary structure modeled with the SWISS MODEL server revealed that there were two &#945;-helices and four b-strands in BgMm-1 as its human orthologue, and both proteins belonged to the a class of PFD family. The phylogenetic relationships of Mm-1s from lower archaea to high mammals was consistent with the evolution of species, meanwhile the cluster result was consistent with the multiple alignment and the sequence identity analysis. RT-PCR (reverse transcriptase-polymerase chain reaction) analysis demonstrated that BgMm-1 expressed widely in ten tissues of adult toad. These results can be helpful for the further investigation on the evolution of Mm-1

Isolation and sequencing of the HMG domain of ten Sox genes from Odorrana schmackeri (Amphibia: Anura)

Sox (SRY-related HMG-box) genes encode a family of transcriptional regulators, which are characterized by a conserved 79-amino acid domain known as HMG-box. They play essential roles in a diverse range of processes including sex determination and the development of the central nervous system (CNS), neural crest and endoderm. In this paper, the HMG domain of ten distinct Sox gene family members (os-Sox2, os-Sox3a, os-Sox3b, os-Sox4, os-Sox11a, os-Sox11b, os-Sox14a, os-Sox14b, os-Sox21a, os-Sox21b) were isolated from both male and female Odorrana schmackeri (Boettger, 1892) using PCR, and no sexual differences were found. Molecular phylogenetic analysis of the HMG domain suggested that these ten Sox genes are members of the SoxB and SoxC groups. In addition, sequence analysis suggested that four Sox genes (os-Sox3, os-Sox11, os-Sox14, os-Sox21) were duplicated. The duplication-degeneration-complementation model should be implied to explain the evolution and diversity of the Sox gene family in O. schmackeri