Genome-wide RAD sequencing to identify a sex-specific marker in Chinese giant salamander Andrias davidianus.

BACKGROUND: Chinese giant salamander Andrias davidianus is an endangered species. The success of artificial breeding provides a useful way to protect this species. However, the method to identify the sex and mechanism of sex determination were unclear which hinder the improvement of the artificial breeding. Detection of a sex specific marker provides an effective approach to identify genetic sex and investigate the sex determination mechanism. RESULTS: We used restriction-site-associated DNA (RAD) sequencing to isolate a sex-specific genetic marker in A. davidianus to expand knowledge of the sex determination mechanism. Four male and four female specimens were subjected to RAD sequencing, which generated 934,072,989 reads containing approximately 134.4 Gb of sequences. The first round of comparison of the assembled sequence against the opposite sex raw reads revealed 19,097 female and 17,994 male unmatched sequences. Subsequently, 19,097 female sequences were subjected to a BLAST search against male genomic data, which revealed 308 sequences unmapped to the male genome. One hundred of these were randomly selected and validated by PCR in five male and five female specimens, and four putative sex-specific sequences were produced. Further validation was performed by PCR in another 24 females and 24 males, and all female individuals exhibited the expected specific bands, while the males did not. To apply the sex-specific marker, three specimens reversed from genetic female to physiological male were found in a group exposed to elevated temperature, and 13 individuals reversed from genetic male to physiological female were obtained in a 17ß-estradiol exposed group. CONCLUSION: This is the first report of a sex-specific marker in A. davidianus and may have potential for elucidation of its sex determination mechanism and, hence, its conservation.

Identification of critical sex-biased genes in Andrias davidianus by de novo transcriptome.

The Chinese giant salamander Andrias davidianus is a protected amphibian with high nutritional and economic value. Understanding its sex determination mechanism is important for improving culture techniques and sex control in breeding. However, little information on the characterization of critical genes involved in sex is available. Herein, sequencing of ovary and test produced 40,783,222 and 46,128,902 raw reads, respectively, which were jointly assembled into 80,497 unigenes. Of these, 36,609 unigenes were annotated, of which 8907 were female-biased and 10,385 were male-biased. Several sex-related pathways were observed, including the Wnt signaling pathway. After elevated temperature and estrogen exposure, neomale and neofemale specimens were identified by a female-specific marker for the first time. RT-qPCR analysis showed the expression profile of ten selected sex-biased genes to be exhibited consistently in male and neomale and in female and neofemale, with the exception of the Amh and TfIIIa genes. Results suggested that these genes may play important roles in A. davidianus sex determination and gonad development. This provides a basis for further investigation of the molecular mechanisms of sex determination in amphibians.

Characterization and expression of galectin-3 after Streptococcus agalactiae and Aeromonas hydrophila challenge in GIFT strain Nile tilapia (Oreochromis niloticus).

In mammals, Galectin-3 has been revealed to be widely expressed in immune cells and played important role in immune reactions. However, Galectin-3 is frequently less reported in teleost. In the present study, a molecular characterization and expression analysis of galectin-3 were conducted in GIFT strain Nile tilapia. The full-length cDNA is 1034 bp with 690 bp of protein coding sequences. The result of qRT-PCR showed that the mRNA of galectin-3 was widely expressed in various tissues (heart, liver, spleen, gill, kidney, brain, intestine, skin, muscle, and ovary), and the higher expression was observed in immune-related tissues (liver and spleen). The time-course expression analysis revealed that galectin-3 was significantly up-regulated in intestine (5 h, 50 h, and 7 d), liver (5 h, 50 h, and 7 d), spleen (5 and 50 h), head-kidney (5 and 50 h), gill (5 h and 7 d) after Streptococcus agalactiae challenge, and significantly up-regulated in intestine (18, 24, 36, 72, and 96 h), liver (6, 18, 24, 96 h, and 6 d), spleen (18, 24, 36, 72, and 96 h), head-kidney (6, 12, 18, 24, 36, 72, and 96 h), and gill (12, 18, 24, and 36 h) after Aeromonas hydrophila challenge. Taken together, these data suggest that galectin-3 plays a role in immune responses in Nile tilapia after bacterial challenge.

Comparative transcriptome reveal the potential adaptive evolutionary genes in Andrias davidianus.

To search the evidence of molecular evolution mechanism for aquatic and cave habitat in Andrias davidianus, the evolution analysis was carried out among several species transcriptome data. The transcriptome data of Notophthalmus viridescens, Xenopus tropicalis, Cynops pyrrhogaster, Hynobius chinensis and A. davidianus were obtained from the Genbank and reassembled except Xenopus tropicalis. The BLAST search of transcriptome data obtained 1244 single-copy orthologous genes among five species. A phylogenetic tree showed A. davidianus to have the closest relationship to H. chinensis. Fourteen positively selected genes were detected in A. davidianus and N. vridescens group and fifteen in A. davidianus and H. chinensis group. Five genes were shared in the both groups which involved in the immune system, suggesting that A. davidianus adaptation to an aquatic and cave environment required rapid evolution of the immune system compared to N. viridescens and H. chinensis.

Molecular cloning, genomic structure, polymorphism analysis and recombinant expression of a α1-antitrypsin like gene from swamp eel, Monopterus albus.

Alpha-1-antitrypsin (AAT) is a highly polymorphic glycoprotein antiprotease, involved in the regulation of human immune response. Beyond some genomic characterization and a few protein characterizations, the function of teleost AAT remains uncertain. In this study we cloned an AAT-like gene from a swamp eel liver identifying four exons and three introns, and the full-length cDNA. The elucidated swamp eel AAT amino acid sequence showed high homology with known AATs from other teleosts. The swamp eel AAT was examined both in ten healthy tissues and in four bacterially-stimulated tissues resulting in up-regulation of swamp eel AAT at different times. Swamp eel AAT transcripts were ubiquitously but unevenly expressed in ten tissues. Further, the mature peptide sequence of swamp eel AAT was subcloned and transformed into E. coli with the recombinant proteins successfully inhibiting bovine trypsin activity. Analysis of recombinant AAT showed equimolar formation of irreversible complexes with proteinases, high stability at pH 7.0-10.0 and temperatures below 55 °C. Serum AAT protein level significantly increased in response to inflammation with AAT anti-sera, and, NF-κB, apolipoprotein A1 and transferrin gene expression were dramatically decreased over 72 h post recombinant AAT injection. Lastly, examination of swamp eel AAT allelic polymorphism identified all alleles in both healthy and diseased stock except allele*g, found only in diseased stock, but without statistical difference between the distribution frequency of allele*g in the two stocks. These results are crucial to our ongoing study of the role of teleost AAT in the innate immune system.

The OXTR rs53576 impacts moral permissibility of attempted but failed harms in populations of students and prisoners.

Previous research has highlighted the roles of oxytocin in empathy and altruistic behaviors. Based on these findings, recent studies have examined the association between the oxytocin receptor gene (OXTR) and outcome-based moral judgment with sacrificial dilemmas (e.g. runaway trolley case). However, little is known about the relationships between OXTR polymorphisms and intent-based moral judgment of harms (e.g. attempted but failed harm or intentionally committed harm). This study investigated the association between the OXTR rs53576 and intent-based moral judgment in college students (N = 544) and prisoners (N = 540). Results indicated that both students and prisoners with the GG genotype of OXTR rs53576 rated attempted but failed harm as less permissible than those with the AA and AG genotypes. These findings highlight the role of the OXTR gene in intent-based moral judgment.

The interplay between parental care and OPRM1 in reward responsiveness.

The Reinforcement Sensitivity Theory (RST) explains a variety of reward-motivated behaviors as the result of the activation of biologically-based systems. Inspired by the influences of parental bonding and opioid peptide on reward system, we investigated the contributions of parental bonding and mu-opioid receptor gene (OPRM1) towards motivation systems (i.e., the BAS, BIS-anxiety, and FFFS-fear). Results indicated that (1) parental care was negatively related to FFFS-fear, but parental overprotection was positively related to both FFFS-fear and BIS-anxiety; (2) parental care significantly interacted with OPRM1 rs1799971 in reward responsiveness with diathesis-stress model. Poor parental care reduced reward responsiveness among individuals with the G allele, but not those with the AA genotype. These findings from this study demonstrate a new gene-environment interactive mechanism of the RST.

Engineering of injectable hydrogels associate with Adipose-Derived stem cells delivery for anti-cardiac hypertrophy agents.

Adipose-derived stem cells (ADSCs) treatment offers support to new methods of transporting baseline cell protein endothelial cells in alginate (A)/silk sericin (SS) lamellar-coated antioxidant system (ASS@L) to promote acute myocardial infarction. In the synthesized frames of ASS, the ratio of fixity modules, pores, the absorption and inflammation was detected at ka (65ka), 151 ± 40.12 µm, 92.8%, 43.2 ± 2.58 and 30.10 ± 2.1. In this context, ADSC-ASS@L was developed and the corresponding material was stable and physically chemical for the development of cardiac regenerative applications. ADSC-ASS@L injectable hydrogels in vitro examination demonstrated higher cell survival rates and pro-angiogenic and pro-Inflammatory expression factors, demonstrating the favorable effect of fractional ejections, fibre-areas, and low infracture vessel densities. In successful cardiac damage therapy in acute myocardial infarction the innovative ADSC injection hydrogel approach may be helpful. The approach could also be effective during coronary artery hypertrophy for successful heart damage treatment.

The rs6311 of serotonin receptor 2A (5-HT2A) gene is associated with alexithymia and mental health.

BACKGROUNDS: Alexithymia, difficulties in identifying and describing one's own feelings, is related to substantial clinical practice. Inspired by the links between serotonin functions and affective disorders, this study investigated associations of the serotonin receptor 2A (5-HT2A) gene with alexithymia and mental health. METHODS: We differentiated subjects according to two functional polymorphisms (i.e., rs6311 and rs6313) of 5-HT2A gene and scored alexithymia and mental health of college students with the Toronto Alexithymia Scale (TAS-20) and the Symptom Check List-90 (SCL-90), respectively. RESULTS: The analyses basing on sample 1 (N = 566) and sample 2 (N = 602) indicated that the G allele of rs6311 was related to higher score on the TAS-20 as compared to the AA genotype. The analysis with 467 individuals from sample 2 indicated that the rs6311 was associated with mental health, and this association was mediated by alexithymia. LIMITATIONS: The potential confounding variables such as depression and anxiety were neglected in the analyses. CONCLUSIONS: These findings demonstrate the contribution of the 5-HT2A to alexithymia, and highlight the link between alexithymia and mental health at genetic level.

The OXTR polymorphisms are not associated with attachment dimensions: A three-approach study.

Inspired by the roles of oxytocin in social behaviors, scientists have devoted considerable efforts to examine the association between the oxytocin receptor gene (OXTR) and human attachment, a personality of seek and receiving comfort from intimate figures. However, there are still a lot of controversies on the association. To clarify the relationship, this research integrated three studies: (1) A cross-sectional study indicated that the OXTR polymorphisms (i.e., rs53576 and rs2254298) were not significantly associated with attachment dimensions in a college student sample (N = 1193); (2) A three-wave study showed that the polymorphisms were not associated with the individual differences and changes of attachment dimensions in a freshmen sample (N = 657); and (3) Meta-analysis indicated that attachment dimensions were not associated with the polymorphisms of rs53576 (Anxiety: 14 samples, N = 5053; Avoidance: ten samples, N = 4273) and rs2254298 (Anxiety: ten samples, N = 3670; Avoidance: ten samples, N = 3698). Taken together, these findings provide strong evidence that the OXTR polymorphisms are not related to attachment dimensions.