Cloning, tissue distribution of desert hedgehog (dhh) gene and expression profiling during different developmental stages of Pseudopleuronectes yokohamae.

As a crucial member of the Hedgehog (Hh) protein family, desert hedgehog (dhh) plays a vital role in multiple developmental processes, cell differentiation and tissue homeostasis. However, it is unclear how it regulates development in fish. In this study, we cloned and characterized the dhh gene from Pseudopleuronectes yokohamae. The full-length cDNA of Pydhh comprises 3194 bp, with a 1386 bp open reading frame (ORF) that encodes a polypeptide of 461 amino acids with a typical HH-signal domain, Hint-N and Hint-C domains. Multiple sequence alignment revealed that the putative PyDHH protein sequence was highly conserved across species, especially in the typical domains. Phylogenetic analysis showed that the PyDHH clustered within the Pleuronectiformes. Real-time quantitative PCR showed that Pydhh was detected in fourteen different tissues in adult-female and adult-male marbled flounder, and nine different tissues in juvenile fish. During early embryonic development stages, the expression of Pydhh was revealed high levels at hatching stage of embryo development. Moreover, the relative expression of Pydhh was significantly higher in the juvenile liver than adults', and higher in the female skin than the male skin. To further investigate its location, the in situ hybridization (ISH) assay was performed, the results showed that the hybridization signal was obviously expressed in the immune organs of Pseudopleuronectes yokohamae, with weak signal expression in the other tissues. Our results suggested that Pydhh is highly conserved among species and plays a vital role in embryonic development and formation of immune related organs.

Comparison of protein differences between high- and low-quality goat and bovine parts based on iTRAQ technology.

This study was designed to investigate the differences in the proteomes of high- and low-quality goat and bovine muscles. In total, 364 and 111 differentially expressed proteins (DEPs) were identified in the high-quality/low-quality bovine and goat samples, respectively. The bioinformatic analysis demonstrated that DEPs are involved in glycolysis, the tricarboxylic acid cycle (TCA cycle) and oxidative phosphorylation. An analysis of quality-related DEPs showed that nicotinamide adenine dinucleotide dehydrogenase (NADH) and succinate dehydrogenase (SDH) could be potential biomarkers for colour, while enolase (ENO) can be used as a marker protein for tenderness qualities in goat. Lactate dehydrogenase (LDH) and 14-3-3 protein can indicate goat and bovine tenderness, while heat-shock proteins (HSPs) can also be used as tenderness marker proteins for different species. Glycerin-3-phosphate dehydrogenase (GPDH) can be used as a protein to detect fat content, and guanine ribonucleotide-binding protein (G protein) may be a flavour marker protein in bovine meat.

Quantitative phosphoproteomic analysis of caprine muscle with high and low meat quality.

During the conversion of muscle to meat, protein phosphorylation can regulate various biological processes that have important effects on meat quality. To investigate the phosphorylation pattern of protein on rigor mortis, goat longissimus thoracis and external intercostals were classified into two groups (high quality and low quality), and meat quality was evaluated according to meat quality attributes (Warner-Bratzler shear force, Color, pH and drip loss). A quantitative mass spectrometry-based phosphoproteomic study was conducted to analyze the caprine muscle at 12h postmortem applying the TiO2-SIMAC-HILIC (TiSH) phosphopeptide enrichment strategy. A total of 2125 phosphopeptides were identified from 750 phosphoproteins. Among them, 96 proteins had differed in phosphorylation levels. The majority of these proteins are involved in glucose metabolism and muscle contraction. The differential phosphorylation level of proteins (PFK, MYL2 and HSP27) in two groups may be the crucial factors of regulating muscle rigor mortis. This study provides a comprehensive view for the phosphorylation status of caprine muscle at rigor mortis, it also gives a better understanding of the regulation of protein phosphorylation on various biological processes that affect the final meat quality attributes.