绝经对血液单核细胞基因表达谱的影响:基因芯片初步研究

绝经是女性一生中很重要的生理现象之一,它能增加一系列复杂免疫、神经退化、新陈代谢和心血管方面的疾病。血液单核细胞能分化成各种各样的细胞,这些细胞在组织形态发生和免疫应答方面起着很重要的作用。本研究中采用了包含大约14,500个基因探针的Affymetrix Human U133A基因芯片来研究健康的绝经前和绝经后女性外周血液单核细胞中的基因表达谱。样本之间的对比分析表明有20个基因上调,20个基因下调。其中的28个基因根据它们的生物过程如细胞繁殖、免疫应答、细胞代谢等等被分成了6个主要的GO类别;剩下的12个基因其生物学功能还没有被鉴定。研究结果支持了我们的假设:血液单核细胞的功能状态确实受到绝经的影响,而且由此带来的改变可能是由全基因组范围的基因表达谱而决定的。本研究中鉴定的一些差异表达基因有可能作为以后研究与绝经相关的系统免疫、神经退化和心血管疾病的候选基因研究。此工作是这个研究方向的第一次尝试,为将来的进一步研究奠定了基础。     

Current limitations of SNP data from the public domain for studies of complex disorders: a test for ten candidate genes for obesity and osteoporosis

Background

Public SNP databases are frequently used to choose SNPs for candidate genes in the association and linkage studies of complex disorders. However, their utility for such studies of diseases with ethnic-dependent background has never been evaluated.

Results

To estimate the accuracy and completeness of SNP public databases, we analyzed the allele frequencies of 41 SNPs in 10 candidate genes for obesity and/or osteoporosis in a large American-Caucasian sample (1,873 individuals from 405 nuclear families) by PCR-invader assay. We compared our results with those from the databases and other published studies. Of the 41 SNPs, 8 were monomorphic in our sample. Twelve were reported for the first time for Caucasians and the other 29 SNPs in our sample essentially confirmed the respective allele frequencies for Caucasians in the databases and previous studies. The comparison of our data with other ethnic groups showed significant differentiation between the three major world ethnic groups at some SNPs (Caucasians and Africans differed at 3 of the 18 shared SNPs, and Caucasians and Asians differed at 13 of the 22 shared SNPs). This genetic differentiation may have an important implication for studying the well-known ethnic differences in the prevalence of obesity and osteoporosis, and complex disorders in general.

Conclusion

A comparative analysis of the SNP data of the candidate genes obtained in the present study, as well as those retrieved from the public domain, suggests that the databases may currently have serious limitations for studying complex disorders with an ethnic-dependent background due to the incomplete and uneven representation of the candidate SNPs in the databases for the major ethnic groups. This conclusion attests to the imperative necessity of large-scale and accurate characterization of these SNPs in different ethnic groups.     

Distinct tubulin genes are differentially expressed during barley grain development

Tubulins, as major components involved in the organization of microtubules, play an important role in plant development. We describe here the expression profiles of all known α-tubulin (TUA), β-tubulin (TUB) and γ-tubulin (TUG) genes of barley ( Hordeum vulgare ), involving eight newly identified TUB sequences, five established TUA genes and one TUG gene. Macroarray and Northern blot-based expression patterns in the pericarp, endosperm and embryo were obtained over the course of the development of the grain between anthesis and maturation. These revealed that the various tubulin genes differed in their levels of expression, and to some extent were tissue specific. Two expression peaks were detected in the developing endosperm. The first and more prominent peak, at 2 days after flowering, included expression of almost all the tubulin genes. These tubulins are thought to be involved in mitoses during the formation of the syncytial endosperm. The second, less pronounced but more extended, peak included only some of the tubulin genes ( HvTUA3 , HvTUB1 and HvTUG ) and might be associated with the cell wall organization in aleurone and starchy endosperm. The HvTUA5 gene is expressed only in embryo of the developing grain and may be associated with shoot establishment. The expression profiles of the tubulin folding cofactors HvTFC A and HvTFC B as well as small G-protein HvArl2 genes were almost perfectly correlated with the global levels of tubulin mRNA, implying that they have a role in the control of the polymerization of α/β-tubulin heterodimers.     

Ca2+ channel currents and contraction in CaVbeta3-deficient ileum smooth muscle from mouse

Voltage activated L-type Ca(2+) channels are the principal Ca(2+) channels in intestinal smooth muscle cells. They comprise the ion conducting Ca(V)1 pore and the ancillary subunits alpha(2)delta and beta. Of the four Ca(V)beta subunits Ca(V)beta(3) is assumed to be the relevant Ca(V)beta protein in smooth muscle. In protein lysates isolated from mouse ileum longitudinal smooth muscle we could identify the Ca(V)1.2, Ca(V)alpha(2), Ca(V)beta(2) and Ca(V)beta(3) proteins, but not the Ca(V)beta(1) and Ca(V)beta(4) proteins. Protein levels of Ca(V)1.2, Ca(V)alpha(2) and Ca(V)beta(2) are not altered in ileum smooth muscle obtained from Ca(V)beta(3)-deficient mice indicating that there is no compensatory increase of the expression of these channel proteins. Neither the Ca(V)beta(2) nor the other Ca(V)beta proteins appear to substitute for the lacking Ca(V)beta(3). L-type Ca(2+) channel properties including current density, inactivation kinetics as well as Cd(2+)- and dihydropyridine sensitivity were identical in cells of both genotypes suggesting that they do not require the presence of a Ca(V)beta(3) protein. However, a key hallmark of the Ca(V)beta modulation of Ca(2+) current, the hyperpolarisation of channel activation is slightly but significantly reduced by 4 mV. In addition to L-type Ca(2+) currents T-type Ca(2+) currents could be recorded in the murine ileum smooth muscle cells, but T-type currents were not affected by the lack of Ca(V)beta(3). Both proteins, Ca(V)beta(2) and Ca(V)beta(3) are localized near the plasma membrane and the localization of Ca(V)beta(2) is not altered in Ca(V)beta(3) deficient cells. Spontaneous contractions and potassium and carbachol induced contractions are not significantly different between ileum longitudinal smooth muscle strips from mice of both genotypes. In summary the data show that in ileum smooth muscle cells, Ca(V)beta(3) has only subtle effects on L-type Ca(2+) currents, appears not to be required for spontaneous and potassium induced contraction but might have a function beyond being a Ca(2+) channel subunit.     

Heat shock induces oxidative stress in rotan Perccottus glenii tissues

The effects of temperature transition from 19 to 32 °C on oxidative stress indices and activities of the main antioxidant enzymes were investigated in the rotan, Perccottus glenii. Levels of lipid peroxides (LOOH), thiobarbituric acid-reactive substances (TBARS), low- (L-SH) and high-molecular mass (H-SH) thiols and activities of superoxide dismutase (SOD) and catalase were measured in rotan brain, liver and muscle over 1–12 h of high-temperature exposure followed by 3 or 24 h lower (19 °C) temperature recovery. Heat shock exposure during 1 h transiently increased 1.5–3.2-fold LOOH levels in rotan tissues with subsequent suppression of their content; however, 12 h exposure again increased LOOH levels in the brain. TBARS content were elevated by 2–3-fold during the entire heat shock exposure in the brain and liver. Levels of both products of lipid peroxidation were generally near control values during return to 19 °C. L-SH content was lowered during heat shock exposure in the brain, transiently increased after 6 h in the liver and almost disappeared after longer treatment in the muscle. Liver H-SH content slightly decreased under heat shock exposure, but was elevated after 6 h in the brain and muscle. In the latter case, L-SH level was below control values during recovery. SOD activities increased 2-fold in the liver after 6–12 h heat shock. Liver catalase activities decreased at the same conditions. Generally, a quick response to suppression of lipid peroxidation and possible involvement of its products in the up-regulation of antioxidant enzymes seem to be key adaptations to high temperature.     

Fluorescent dyes undergoing intramolecular proton transfer with improved sensitivity to surface charge in lipid bilayers.

4'-(Dialkylamino)-3-hydroxyflavones are characterized by an excited-state proton transfer reaction between two tautomeric excited states, which results in two emission bands well separated on the wavelength scale. Due to the high sensitivity of the relative intensities of the two emission bands to solvent polarity, hydrogen bonding and local electric fields, these dyes found numerous applications in biomembrane studies. In order to further improve their fluorescence characteristics, we have synthesized new dyes where the 2-phenyl group is substituted with a 2-thienyl group. In organic solvents, the new dyes exhibit red shifted absorption and dual fluorescence. Although they show lower sensitivity to solvent polarity and H-bond donor ability (acidicity) than their parent 3-hydroxyflavone dyes, they exhibit a much higher sensitivity to solvent H-bond acceptor ability (basicity). Moreover, when tested in lipid vesicles of different surface charge, the new dyes show much better resolved dual emission and higher sensitivity to the surface charge of lipid bilayers than the parent dyes. The response of the new dyes to surface charge is probably connected with the H-bond basicity of the membrane surface, which is the highest for negatively charged surfaces. As a consequence, the new dyes appear as prospective fluorophores for the development of new fluorescent probes for biomembranes.     

CCZ1, MON1 and YPT7 genes are involved in pexophagy, the Cvt pathway and non-specific macroautophagy in the methylotrophic yeast Pichia pastoris

Orthologues of Saccharomyces cerevisiae CCZ1, MON1 and YPT7 genes in the methylotrophic yeast, Pichia pastoris, have been identified. These genes encode proteins, which act as a complex, being involved in degradation of oleate-induced peroxisomes, Cvt (cytoplasm to vacuole targeting) pathway and non-specific macroautophagy in S. cerevisiae. CCZ1, MON1 and YPT7 gene orthologues are essential for multiple delivery pathways in P. pastoris. Strains with deletion of either of these genes displayed complete deficiency in pexophagy, non-specific macroautophagy and the biosynthetic Cvt pathway. The data suggest that CCZ1, MON1 and YPT7 genes are involved in degradation of both small oleate-induced and large methanol-induced peroxisomes. The data suggest conservative functions of CCZ1, MON1 and YPT7 genes among yeast species.     

Molecular Evolution of ldpA, a Gene Mediating the Circadian Input Signal in Cyanobacteria

The ldpA gene is an element of the cyanobacterial circadian system and mediates input to the clock. Using complete prokaryotic genomes from various public databases, I analyzed the structure and phylogeny of the ldpA genes. This gene belongs to the large superfamily of ferredoxins and has a HycB domain as a core element of its structure. In addition to this domain, ldpA has two conserved terminal domains that are specific to this gene and have no homologs in the databases. All three domains are under different selective constraints. The ldpA tree topology features two very distinct clades that are essentially the same as those in the previously reported trees of the sasA gene and the kaiBC operon, two other elements of the circadian system. The data on the ldpA polymorphism and evolutionary patterns give further support to the existence of two types of the system, kaiABC- and kaiBC-based, respectively. Each type has specific functional and selective constraints, which have likely been attained through highly concordant evolution of the systems components.Reviewing Editor: Dr. Rasmus Nielsen     

Adaptive response of antioxidant enzymes to catalase inhibition by aminotriazole in goldfish liver and kidney

This study was undertaken to clarify the physiological role of catalase in the maintenance of pro/antioxidant balance in goldfish tissues by inhibiting the enzyme in vivo with 3-amino 1,2,4-triazole. Intraperitoneal injection of aminotriazole (0.5 mg/g wet mass) caused a decrease in liver catalase activity by 83% after 24 h that was sustained after 168 h post-injection. In kidney catalase activity was reduced by approximately 50% and 70% at the two time points, respectively. Levels of protein carbonyls were unchanged in liver but rose by 2-fold in kidney after 168 h. Levels of thiobarbituric acid-reactive substances were elevated in both tissues after 24 h but were reversed by 168 h. Glutathione peroxidase and glutathione-S-transferase activities increased in kidney after aminotriazole treatment whereas activities of glutathione peroxidase and glutathione reductase in liver decreased after 24 h but rebounded by 168 h. Liver glucose-6-phosphate dehydrogenase activity was reduced at both time points. Activities of these three enzymes in liver correlated inversely with the levels of lipid damage products (R2=0.65-0.81) suggesting that they may have been oxidatively inactivated. Glutathione-S-transferase activity also correlated inversely with catalase (R2=0.86). Hence, the response to catalase depletion involves compensatory changes in the activities of enzymes of glutathione metabolism.