Evaluation of Reference Genes for Quantitative PCR in Four Tissues from Rabbits with Hypercholesterolaemia

Abstract Rabbit with hypercholesterolaemia is an important model for studying cholesterol metabolism disease. This study aimed to evaluate the expression stability of nine reference genes for quantitative PCR (qPCR) analysis in adrenal gland, liver, spleen, and kidney tissue from rabbits with hypercholesterolaemia. In total, 30 male Harbin Large White (HLW) rabbits were fed a normal feed (n = 15) or a high cholesterol feed (n = 15) for 8 weeks to induce hypercholesterolaemia. Nine reference genes were verified by qPCR using cDNA extracted from rabbit tissue samples. For qPCR analysis, reference genes were evaluated using the RefFinder and GeNorm algorithms. Overall, seven rabbits with hypercholesterolaemia were identified based on body weight and total cholesterol measurements. Combining the results of the RefFinder and GeNorm algorithms, the most stable reference genes were hypoxanthine phosphoribosyltransferase 1 (Hprt1) and eukaryotic translation elongation factor 1 alpha 1 (Eef1a1) in the adrenal gland, β-2-microglobulin (B2m) and glyceraldehyde-3-phosphate dehydrogenase (Gapdh) in the liver, tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (Ywhaz) and Gapdh in the spleen, and peptidylprolyl isomerase (Ppia), β-actin (Actb), succinate dehydrogenase complex subunit A flavoprotein (Sdha), and B2m in the kidney. Taken together, our results confirmed that Hprt1 and Eef1a1, B2m and Gapdh, Ywhaz and Gapdh, and Ppia, Actb, Sdha, and B2m were the best reference genes for qPCR analyses in adrenal gland, liver, spleen, and kidney tissue, respectively, of rabbits with hypercholesterolaemia.

[Characteristics of fly ash particles deposition in the snowpack of Laohugou Glacier no. 12 in western Qilian Mountains, China].

This research aimed to identify and characterize individual spherical fly ash particles extracted from surface snow at Laohugou Glacier No. 12, western Qilian Mountains, China. Characterization of the spherical particles (i. e. morphology, chemical composition and genesis) was obtained by scanning electron microscopy coupled with energy dispersive X-ray spectrometer (SEM-EDX). Spherical particles and agglomerates were identified according to their morphology in nine snow samples. Prevalent particle types in all samples were granular spherical particles, hollow spherical particles, and agglomerates. The vast majority of spherical particles in our samples had mostly smooth and glossy surfaces. Individual particle analyses of elemental composition showed that particles formed in combustion were mainly composed of silicon, aluminum and trace elements. On the basis of chemical information obtained from EDX, the fly ash particles deposited in the snow could be classified into three types, which were Si-dominant particles, Fe-dominant particles, and Ti-dominant spherical particles. Backward air mass trajectory and dispersion analysis suggested that the developed urban regions of central Asia and surrounding Yumen city contributed the primary fly ash particles from industrial combustion to the study site through the high-level atmospheric circulation.

[Association analysis between SNPs of the growth hormone receptor gene and growth traits in arctic fox].

Using single-strand conformation polymorphism (PCR-SSCP) and DNA sequencing, single nucleotide polymorphisms (SNPs) of growth hormone receptor (GHR) gene were detected in an arctic fox population. Correlation analysis between GHR polymorphisms and growth traits were carried out using the appropriate model. Four SNPs, G3A in the 5'UTR, C99T in the first exon, T59C and G65A in the fifth exon were identified on the arctic fox GHR gene. The G3A and C99T polymorphisms of GHR were associated with female fox body weight (Pamp;0.05) and the T59C and G65A polymorphisms of GHR were associated with male fox body weight (Pamp;0.05) and the skin length of the female fox (Pamp;0.01). Therefore, marker assistant selection on body weight and skin length of arctic foxes using these SNPs can be applied to get big and high quality arctic foxes.

[Correlation analysis between MC3R and MC4R gene polymorphism and growth traits in pigeon.].

The SNPs in partial coding sequence of MC3R and MC4R genes were identified by polymerase chain reaction followed by single strand conformation polymorphism (PCR-SSCP) analysis and DNA sequencing in shack-Kee and Columba domestica from Harbin area. Correlation analysis between MC3R and MC4R polymorphism and growth and body composition traits was carried by the least square analysis. The genotypes of T91G mutation in MC3R gene and A903G mutation in MC4R gene proved to have significant association with body weight, carcass weight, and holo-carcass weight in shack-Kee (P<0.05). The interaction of MC3R-T91G and MC4R-A903G was discussed through combination genotype analysis. The least square analysis showed that the combined genotype had significant association with holo-carcass weight (P<0.05). Multiple comparisons revealed that BBAA genotype birds had a higher holo-carcass weight than AABB genotype birds and BBAA genotype was the beneficial genotype for the growth of body weight.

[Association analysis between SNPs of the growth hormone gene and antler production in spotted deer].

We report the effects of candidate gene growth hormone (GH) gene on antler production in the current study. Single nucleotide polymorphisms (SNPs) of the GH gene were identified and genotyped by polymerase chain reaction followed by single strand conformation polymorphism (PCR-SSCP) analysis and DNA sequencing in a deer population from the farm of Jilin Agricultural University. Correlation analysis between GH polymorphisms and antler production was carried out using the appropriate mixed model. Results showed an effect of GH gene on antler production. Deer with the SNP genotypes G-->A had a significant difference in antler production of the fifth saw (P<0.2). BB deer had a higher antler pro-duction than AA ones (P<0.2).