Molecular cloning and expression analysis of adiponectin and its receptors (AdipoR1 and AdipoR2) in the hypothalamus of the Huoyan goose during different stages of the egg-laying cycle

Multiple amino acid sequence alignment of the Huoyan goose adiponectin protein with other vertebrate species. The colour black denotes 100 % conserved sequences, and the colour grey indicates non-conservative sequences. Gaps (−) were introduced to maximize the alignment. Sequences for the alignment were obtained from GenBank (accession numbers are in brackets): Anas platyrhynchos (ADA68839.1); Ovis aries (AHV91023.1); Canis lupus familiaris (BAD15362.1); Felis catus (BAF52934.1); Gallus (AAX40986.1); Homo sapiens (NP_004788.1); Meleagris gallopavo (XP_010714799.1); Mus musculus (NP_033735.3); and Sus scrofa (ABQ95350.1). (TIFF 3483 kb

Gene Expression Profiling in the Pituitary Gland of Laying Period and Ceased Period Huoyan Geese

Huoyan goose is a Chinese local breed famous for its higher laying performance, but the problems of variety degeneration have emerged recently, especially a decrease in the number of eggs laid. In order to better understand the molecular mechanism that underlies egg laying in Huoyan geese, gene profiles in the pituitary gland of Huoyan geese taken during the laying period and ceased period were investigated using the suppression subtractive hybridization (SSH) method. Total RNA was extracted from pituitary glands of ceased period and laying period geese. The cDNA in the pituitary glands of ceased geese was subtracted from the cDNA in the pituitary glands of laying geese (forward subtraction); the reverse subtraction was also performed. After sequencing and annotation, a total of 30 and 24 up and down-regulated genes were obtained from the forward and reverse SSH libraries, respectively. These genes mostly related to biosynthetic process, cellular nitrogen compound metabolic process, transport, cell differentiation, cellular protein modification process, signal transduction, small molecule metabolic process. Furthermore, eleven genes were selected for further analyses by quantitative real-time PCR (qRT-PCR). The qRT-PCR results for the most part were consistent with the SSH results. Among these genes, Synaptotagmin-1 (SYT1) and Stathmin-2 (STMN2) were substantially over-expressed in laying period compared to ceased period. These results could serve as an important reference for elucidating the molecular mechanism of higher laying performance in Huoyan geese

Modular synthesis of glycopolymers with well-defined sugar units in the side chain via Ugi reaction and click chemistry: hetero vs. homo

For synthetic glycopolymers, multivalent carbohydrate-protein interactions have been mostly studied in homoglycopolymers so far. However, natural oligosaccharides tend to consist of various sugars and this kind of heterogeneity is used to tune affinity and selectivity towards a specific receptor. Here we report a novel method to synthesize modularized heteroglycopolymers with well-defined sugar units in the side chain via Ugi reaction and click chemistry. To verify the modularized structures, mPEG-COOH was used as the model polymer substrate to test the Ugi reaction and subsequent click chemistry, and NMR, GPC and MALDI-TOF mass spectrometry confirmed our design. The obtained heteroglycopolymers (PM-GM) and homoglycopolymers (PM-GG and PM-MM) prepared via the methods assembled in aqueous medium with a diameter of 80 nm by DLS and 50 nm by TEM, and were used to investigate their interactions with lectins. Turbidity assay, QCM-D experiments and bacterial adhesion studies all demonstrate that heteroglycopolymers have higher affinity towards specific proteins than do homoglycopolymers, as a result of the heteromultivalent effect and binding ability between different sugar moieties and lectins

Comparative proteomic analysis of pituitary glands from Huoyan geese between pre-laying and laying periods using an iTRAQ-based approach.

In this study, we performed a comprehensive evaluation of the proteomic profile of the pituitary gland of the Huoyan goose during the laying period compared to the pre-laying period using an iTRAQ-based approach. Protein samples were prepared from pituitary gland tissues of nine pre-laying period and nine laying period geese. Then the protein samples from three randomly selected geese within each period were pooled in equal amounts to generate one biological sample pool. We identified 684 differentially expressed proteins, including 418 up-regulated and 266 down-regulated proteins. GO annotation and KEGG pathway analyses of these proteins were conducted. Some of these proteins were found to be associated with hormone and neurotransmitter secretion and transport, neuropeptide signalling and GnRH signalling pathways, among others. Subsequently, the modification of the abundance of three proteins (prolactin, chromogranin-A and ITPR3) was verified using western blotting. Our results will provide a new source for mining genes and gene products related to the egg-laying performance of Huoyan geese, and may provide important information for the conservation and utilization of local goose breeds

A Versatile, Fast, and Efficient Method of Visible-Light-Induced Surface Grafting Polymerization

To overcome the problem caused by the lability of the Au–S bond, we demonstrate the first use of Mn₂(CO)₁₀ for visible-light-induced surface grafting polymerization on Au surfaces in this paper. The visible-light-induced surface grafting of poly­(<i>N</i>-isopropylacrylamide) (PNIPAAm) has the features of a “controlled” polymerization, which is characterized by a linear relationship between the thickness of the grafting layer and the monomer concentration. Ellipsometry indicated the formation of PNIPAAm films of up to ∼200 nm in thickness after only 10 min of polymerization at room temperature, demonstrating that this is a very fast process in comparison with traditional grafting polymerization techniques. Moreover, to demonstrate the potential applications of our approach, different substrates grafted by PNIPAAm and the covalent immobilization of a range of polymers on Au surfaces were also demonstrated. Considering the advantages of simplicity, efficiency, and mild reaction conditions as well as the ability of catecholic derivatives to bind to a large variety of substrates, this visible-light-induced grafting method is expected to be useful in designing functional interfaces

Transcriptome Profiling Identifies Differentially Expressed Genes in Huoyan Goose Ovaries between the Laying Period and Ceased Period

<div><p>The Huoyan goose is famous for its high egg-laying performance and is listed as a nationally protected domestic animal by the Chinese government. To elucidate the key regulatory genes involved in Huoyan goose egg laying, RNA from ovarian tissue during the ceased and laying periods was sequenced using the Illumina HiSeq 2000 sequencing platform. More than 12 million reads were produced in ceased and laying libraries that included 11,896,423 and 12,534,799 clean reads, respectively. More than 20% of the reads were matched to the reference genome, and 23% of the reads were matched to reference genes. Genes with a false discovery rate (FDR) ≤0.001 and log₂ratio ≧1 or ≤−1 were characterized as differentially expressed, and 344 up-regulated and 344 down-regulated genes were classified into functional categories. Twelve genes that are mainly involved in pathways for reproduction regulation, such as steroid hormone biosynthesis, GnRH signaling pathways, oocyte meiosis, progesterone-mediated oocyte maturation, steroid biosynthesis, calcium signaling pathways, and G-protein coupled receptor signaling pathway were selected for validation by a quantitative real-time polymerase chain reaction (qRT-PCR) analysis, the qRT-PCR results are consistent with the general expression patterns of those genes from the Illumina sequencing. These data provide comprehensive gene expression information at the transcriptional level that might increase our understanding of the Huoyan goose's reproductive biology.</p></div