Gastrodin Improves the Activity of the Ubiquitin-Proteasome System and the Autophagy-Lysosome Pathway to Degrade Mutant Huntingtin.

Gastrodin (GAS) is the main chemical component of the traditional Chinese herb Gastrodia elata (called "Tianma" in Chinese), which has been used to treat neurological conditions, including headaches, epilepsy, stroke, and memory loss. To our knowledge, it is unclear whether GAS has a therapeutic effect on Huntington's disease (HD). In the present study, we evaluated the effect of GAS on the degradation of mutant huntingtin protein (mHtt) by using PC12 cells transfected with N-terminal mHtt Q74. We found that 0.1-100 µM GAS had no effect on the survival rate of Q23 and Q74 PC12 cells after 24-48 h of incubation. The ubiquitin-proteasome system (UPS) is the main system that clears misfolded proteins in eukaryotic cells. Mutated Htt significantly upregulated total ubiquitinated protein (Ub) expression, decreased chymotrypsin-like, trypsin-like and caspase-like peptidase activity, and reduced the colocalization of the 20S proteasome with mHtt. GAS (25 µM) attenuated all of the abovementioned pathological changes, and the regulatory effect of GAS on mHtt was found to be abolished by MG132, a proteasome inhibitor. The autophagy-lysosome pathway (ALP) is another system for misfolded protein degradation. Although GAS downregulated the expression of autophagy markers (LC3II and P62), it increased the colocalization of LC3II with lysosomal associated membrane protein 1 (LAMP1), which indicates that ALP was activated. Moreover, GAS prevented mHtt-induced neuronal damage in PC12 cells. GAS has a selective effect on mHtt in Q74 PC12 cells and has no effect on Q23 and proteins encoded by other genes containing long CAGs, such as Rbm33 (10 CAG repeats) and Hcn1 (>30 CAG repeats). Furthermore, oral administration of 100 mg/kg GAS increased grip strength and attenuated mHtt aggregates in B6-hHTT130-N transgenic mice. This is a high dose (100 mg/kg GAS) when compared with experiments on HD mice with other small molecules. We will design more doses to evaluate the dose-response relationship of the inhibition effect of GAS on mHtt in our next study. In summary, GAS can promote the degradation of mHtt by activating the UPS and ALP, making it a potential therapeutic agent for HD.

Dynamic changes of miRNAs in skeletal muscle development at New Zealand rabbits.

BACKGROUND: miRNA is one of the crucial roles in the complex and dynamic network that regulates the development of skeletal muscle. The landscape of skeletal muscle miRNAs from fetus to adult in New Zealand rabbits has not been revealed yet. RESULTS: In this study, nine RNA-seq libraries of fetus, child and adult rabbits' leg muscles were constructed. A total of 278 differentially expressed miRNAs (DEmiRNAs) were identified. In the fetus vs. child group, the main functional enrichments were involved in membrane and transport. Pathway enriched terms of up-regulated DEmiRNAs were connected with the differentiation and hypertrophy of skeletal muscle, and down-regulated ones were related to muscle structure and metabolic capacity. In the child vs. adult group, functions were associated to positioning and transportation, and pathways were relevant to ECM, muscle structure and hypertrophy. Finally, ocu-miR-185-3p and ocu-miR-370-3p, which had the most target genes, were identified as hub-miRNAs in these two groups. CONCLUSIONS: In short, we summarized the highly expressed and uniquely expressed DEmiRNAs of fetus, child and adult rabbits' leg muscles. Besides, the potential functional changes of miRNAs in two consecutive stages have been explored. Among them, the ocu-miR-185-3p and ocu-miR-370-3p with the most target genes were selected as hub-miRNAs. These data improved the understanding of the regulatory molecules of meat rabbit development, and provided a novel perspective for molecular breeding of meat rabbits.

High albumen height by expression of GALNT9 and thin eggshell by decreased Ca2+ transportation caused high hatchability in Huainan partridge chicken.

Hatchability could be quite different among individuals of indigenous chicken breed which might be affected by the egg quality. In this study, hatchability was individually recorded among 800 forty-wk-old Huainan partridge chickens. The chickens were then divided into high and low hatchability groups (HH and LH group) with 50 birds in each group. Egg quality was further determined in the 2 groups. Eight birds from each group were selected for slaughtering and tissue, responsible for egg formation, collection for structure observation by staining and candidate gene expression by transcriptome analysis. The hatchability in HH was 100% and 61.18% in LH. The eggshell thickness and shell strength were significantly lower, while the albumen height and Haugh unit were significantly higher in HH group than those in LH group (P < 0.05). The magnum weight and index, and the expression of polypeptide N-acetylgalactosaminyltransferase 9 (GALNT9), which responsible for thick albumen synthesis, in HH group were also significantly higher than that of LH group (P < 0.05). Compared with the LH group, there were 702 differentially expressed genes (DEGs) in HH group, of which 402 were up-regulated and 300 were down-regulated. Candidate genes of calbindin 1 (CALB1) and solute carrier family 26 member 9 (SLC26A9), which regulate calcium signaling pathway so as to affect Ca2+ transportation, exhibited significant high and low expression, respectively, in HH group compared to those in LH group (P < 0.05). Therefore, indigenous chicken with high expression of GALNT9 in magnum to form thick albumen to provide more protein for embryo, while high CALB1 and low expression of SLC26A9 to decrease Ca2+ transportation so as to form a thinner eggshell and provide better gas exchange during embryo development.

High dietary energy decreased reproductive performance through increasing lipid deposition in Yangzhou geese at late laying stage.

Dietary metabolizable energy (ME) level could offer a well production performance through maintaining lipid homeostasis in poultry. In this study, a total of 540 geese (450 females and 90 males) at 64 wk of age with similar body weight (4,600 ± 382) were randomly divided into 5 groups with 3 replicates in each group and 30 females and 6 males (1♂:5♀) in each replicate. After 2 wk adaptation, the 5 groups were designed to provide diet with ME intakes of 9.65, 10.05, 10.70, 11.45, and 11.75 MJ/kg, respectively, according to production requirement. Body weight, egg production, hatchability, blood lipid, and fat deposition were recorded after 6 wk feeding. The expression of lipid synthesis-related genes, lipoprotein lipase (LPL) and fatty acid synthase (FASN), were determined by quantitative real-time PCR. Geese fed with high ME diet of 11.75 MJ/kg caused an increased liver and abdominal fat weight and low hatchability of set eggs. The ovarian weight and oviduct length were higher in geese fed dietary energy of 10.7 MJ/kg as compared to the 9.65 MJ/kg groups, whereas no significant difference was observed in geese fed dietary energy of 10.05 MJ/kg. Dietary energy level did not change the concentration of serum lipids at the late egg laying stage. The LPL expression exhibited linear and quadratic effect in response to dietary ME. The FASN expression showed quadratic effect and a relatively higher expression was exhibited in 10.05 and 11.45 MJ/kg than that of the 9.65 and 10.70 MJ/kg ME groups. According to the productivity, reproductive performance, and fat deposition, dietary ME of 10.13 to 10.28 MJ/kg could be suggested for breeding geese at their late laying stage.

Egg Yolk Fat Deposition Is Regulated by Diacylglycerol and Ceramide Enriched by Adipocytokine Signaling Pathway in Laying Hens.

The mechanism which regulates differential fat deposition in egg yolk from the indigenous breeds and commercial laying hens is still unclear. In this research, Chinese indigenous Huainan Partridge chickens and Nongda III commercial laying hens were used for egg collection and liver sampling. The weight of eggs and yolk were recorded. Yolk fatty acids were determined by gas chromatography-mass spectrometry. Lipid metabolites in the liver were detected by liquid chromatography-mass spectrometry. Yolk weight, yolk ratio and yolk fat ratio exhibited higher in the Huainan Partridge chicken than that of the Nongda III. Compared to the Nongda III, the content of total saturated fatty acid was lower, while the unsaturated fatty acid was higher in the yolk of the Huainan Partridge chicken. Metabolites of phosphatidylinositol and phosphatidylserine from glycerolphospholipids, and metabolites of diacylglycerol from glycerolipids showed higher enrichment in the Huainan Partridge chicken than that of the Nongda III, which promoted the activation of the adipocytokine signaling pathway. However, metabolites of phosphatidic acid, phosphatidylcholine, phosphatidylethanolamine and lysophosphatidylcholine from glycerol phospholipids, and metabolites of triacylglycerol from glycerolipids showed lower enrichment in the Huainan Partridge chicken than that of the Nongda III. The high level of yolk fat deposition in the Huainan Partridge chicken is regulated by the activation of the adipocytokine signaling pathway which can promote the accumulation of diacylglycerol and ceramide in the liver.