Mapping of the porcine oestrogen receptor 2 gene and association study with litter size in Iberian pigs

Genes encoding the oestrogen receptors (ESR) are considered candidate genes for prolificacy traits due to the key role these molecules play in the regulation of reproductive physiology. In this paper, we report the assignment of the pig ESR2 gene to porcine chromosome 1 by radiation hybrid mapping. Most of the ESR2 cDNA was sequenced from Iberian pig ovarian RNA samples and one A/G single nucleotide polymorphism (SNP) was found at exon 5, being associated with a Met/Val substitution at position 949. This SNP was genotyped using a PCR-RFLP (Hsp92II) protocol and its potential effect on litter size was evaluated in two Iberian pig populations. However, no statistically significant association between the ESR2 polymorphism and litter size was found.     

The effect of environmental colour on the growth,metabolism, physiology and skin pigmentation of the carnivorous freshwater catfish Lophiosilurus alexandri

The growth, physiology and skin pigmentation of pacamã Lophiosilurus alexandri juveniles were evaluated in an experiment using different tank colours (white, yellow, green, blue, brown and black) over an 80 day period. The tank colours did not cause significant differences to final body mass, total length, survival rate, carcass composition (moisture, crude protein, ash, ether extract, calcium, phosphorus, energy), or to plasma protein, triglyceride and cholesterol values. Haematocrit values, however, were highest for fish kept in white tanks (ANOVA P < 0·05), while the greatest haemoglobin levels were recorded for fish kept in blue and brown tanks (P < 0·01). The concentrations of cortisol (P < 0·001) and glucose (P < 0·01) were the most in fish in the black tanks. Tank colour affected skin pigmentation significantly, with fish in white tanks having the highest values of L* (brightness) and the lowest values in blue and black tanks. L*, however, decreased in all treatments throughout the experiment. C*_ab increased significantly over the course of the experiment in fish kept in white tanks. Similar increases of C*_ab were recorded in the other treatments but to a lesser extent. The use of black tanks during the cultivation of L. alexandri caused stress and should be avoided. Cultivation in white and yellow tanks produced individuals with a pale skin colour, while cultivation in blue and black tanks resulted in juveniles with a darker and more pigmented skin.     

Characterization of the porcine KIT ligand gene: expression analysis, genomic structure, polymorphism detection and association with coat colour traits

Kit ligand (KITLG) is the ligand for the type III receptor tyrosine kinase KIT. Studies of the KIT/KITLG pathway in a number of mammalian species have shown that it is important for the development of stem cell populations in haematopoietic tissues, germ cells in reproductive organs and the embryonic migrating melanoblasts that give rise to melanocytes. Consequently, mutations in the pathway may result in a range of defects including anaemia, sterility and de-pigmentation. The cDNA sequence of the porcine KITLG gene has been reported previously, and is an attractive candidate locus for moderating coat colour in pigs. In this paper we report the gene structure and physical mapping of the porcine gene. We also report the identification of polymorphisms in the gene, one of which was used to confirm linkage to chromosome 5. Preliminary RNA expression studies using a panel of tissues have shown that in addition to the known variant lacking exon 6, there is alternative splicing of exon 4. However, little evidence was found for the KITLG gene being linked to variation in colour in a Meishan x Large White cross.     

Cloning, sequencing and restriction fragment length polymorphism analysis of a porcine cDNA for OCT2

A full-length pig cDNA for the POU-domain protein OCT2 has been isolated and sequenced. The 478 amino acid-long reading frame in pig OCT2 is 97% identical to human OCTZA, indicating strong conservation of function for this immunoglobulin regulatory protein. To investigate the potential use of this cDNA for mapping and identifying linkage of OCT2 to economic traits, restriction fragment length polymorphism (RFLP) analysis was used to identify a TaqI polymorphism. A population of 60 unrelated animals, as well as multigenerational families, were typed for this RFLP and showed variability in several American and Chinese breeds. The Taq I polymorphism was also detected by a non-POU-domain OCT2 probe, demonstrating that this RFLP is located in the OCT2 gene. This result suggests that OCT2 is likely to be a single-copy gene in swine as seen in other mammals.     

Cloning, sequence analysis and identification of a nonsense mutation-mediated mRNA decay of porcine GSTM2 gene

The glutathione S-transferase mu 2 gene （GSTM2） encodes a GST functioning in the elimination of electrophilic compounds and the regulation of cell growth. In this study, the sequence of porcine GSTM2 gene that contains the complete sequence encoding a protein of 218 amino acids was cloned. The deduced amino acid sequence shared 76%, 78% and 76% identity with that of human, mouse and rat, respectively, mRNA expression analysis showed that the porcine GSTM2 gene was expressed at a high level in liver and testis, at a medium level in longissimus dorsi muscle, adipose tissue, spleen and lung, at a low level in kidney, and at a very low level in heart and embryo. A nonsense mutation （CGA→TGA） resulted from C27T substitution in the fifth exon to produce a premature translation termination codon was identified, and it was discovered that nonsense-mediated mRNA decay might have an effect on the regulation of porcine GSTM2 gene expression. This polymorphism was analyzed in Large White, Landrace, Meishan and Qingping pig populations using the Taq I-polymerase chain reaction-restriction fragment length polymorphism method. The result showed that allele C had a higher frequency than allele T in each population.     

Characterization of the porcine FABP5 gene and its association with the FAT1 QTL in an Iberian by Landrace cross

We have characterized and mapped the porcine fatty acid binding protein 5, epidermal (FABP5) gene. According to linkage and RH mapping, this gene is located close to the FABP4 (fatty acid binding protein 4, adipocyte) gene on swine chromosome 4. We resequenced 4.7 kb of the FABP5 gene in the parental population of an Iberian x Landrace cross (IBMAP), identifying seven SNPs arranged in two distinct FABP5 haplotypes. QTL and association analyses in the IBMAP population showed that this gene is strongly associated with fat deposition. QTL and haplotype analysis revealed that both FABP4 and FABP5 (clustered in mammals) are major candidate genes for the FAT1 QTL; the most likely position for the FAT1 QTL is between these two genes. Finally, our results suggest the presence of more than one QTL affecting fatness traits on porcine chromosome 4.     

猪雌激素受体关联受体a基因的克隆、表达及其对脂肪细胞脂质聚集的影响

雌激素受体关联受体α(Estrogen-related receptor α,ERRα)是调控机体能量代谢的关键转录调控因子,其在白色脂肪组织中的作用尚不清楚。本研究旨在通过touch down-PCR方法克隆猪ERRα基因的ORF序列;通过Western blotting和细胞免疫荧光染色法分析其在猪各组织及成熟脂肪细胞中的表达模式;利用ERRα特异性抑制剂XCT790处理原代培养的猪成熟脂肪细胞,探讨其对成熟脂肪细胞甘油三酯聚集的影响。结果显示,所克隆的猪ERRα基因ORF序列长1269bp(GenBank Accession No.FJ446485,尚未公布),编码422个氨基酸,其核苷酸和氨基酸序列与其他物种高度同源;ERRα蛋白高表达于猪白色脂肪组织(White adipose tissue,WAT)、肾脏以及心脏中,在脾脏中表达量较低;细胞免疫荧光化学染色显示,ERRα蛋白广泛分布于脂肪细胞的细胞核和胞浆中;XCT790在10μmol/L浓度时显著抑制了ERRα蛋白的表达和成熟脂肪细胞中甘油三酯的聚集。本研究将为有效调控体脂沉积提供新的靶点和理论参考。     

Genetic parameters and direct,maternal and heterosis effects on litter size in a diallel cross among three commercial varieties of Iberian pig

The Iberian pig is one of the pig breeds that has the highest meat quality. Traditionally, producers have bred one of the available varieties, exclusively, and have not used crosses between them, which has contrasted sharply with other populations of commercial pigs for which crossbreeding has been a standard procedure. The objective of this study was to perform an experiment under full diallel design among three contemporary commercial varieties of Iberian pig and estimate the additive genetic variation and the crossbreeding effects (direct, maternal and heterosis) for prolificacy. The data set comprised 18 193 records for total number born and number born alive from 3800 sows of three varieties of the Iberian breed (Retinto, Torbiscal and Entrepelado) and their reciprocal crosses (Retinto × Torbiscal, Torbiscal × Retinto, Retinto × Entrepelado, Entrepelado × Retinto, Torbiscal × Entrepelado and Entrepelado × Torbiscal), and a pedigree of 4609 individuals. The analysis was based on a multiple population repeatability model, and we developed a model comparison test that indicated the presence of direct line, maternal and heterosis effects. The results indicated the superiorities of the direct line effect of the Retinto and the maternal effect of the Entrepelado populations. All of the potential crosses produced significant heterosis, and additive genetic variation was higher in the Entrepelado than it was in the other two populations. The recommended cross for the highest yield in prolificacy is a Retinto father and an Entrepelado mother to generate a hybrid commercial sow.     

Characterization of the bioactive motif of neuregulin-1, a fibroblast-derived paracrine factor that regulates the constitutive color and the function of melanocytes in human skin

Interactions between melanocytes and neighboring cells in the skin (keratinocytes and fibroblasts) play important roles in regulating human skin color. We recently reported that neuregulin-1 (NRG1) is highly expressed in fibroblasts from Fitzpatrick type VI skin (the darkest) and at least in part determines the constitutive color of human skin. We have now characterized the bioactive motif of NRG1 that is involved in modulating melanin production in human melanocytes. We found that 8-mer motifs (PSRYLCKC and LCKCPNEF) increased melanin production but did not increase the proliferation of melanocytes; the minimum fragment that could elicit that effect was the tetrapeptide LCKC. This smaller bioactive peptide might have an advantage in clinical applications in which it modulates only pigmentation and does not stimulate melanocyte proliferation.     

生防细菌T132的鉴定及其对采后柑橘炭疽病的抑制效果

【目的】柑橘(Citri)是世界上重要的果树。由胶孢炭疽菌[Colletotrichum gloeosporioides(Penz.)]引起的柑橘炭疽病是柑橘生产的主要病害之一。为探索对采后柑橘炭疽病有效的生防措施,分离鉴定柑橘根围土壤中一株细菌T132,并研究其特性及生防效果。【方法】根据菌株T132的形态特征、生理生化特性以及16S rDNA序列对其进行鉴定;通过连续8次在人工培养基上传代培养,测定该菌株的遗传稳定性;采用柑橘果实刺伤挑战接种和拮抗菌液直接浸泡健康果实两种方法研究该菌株对柑橘炭疽病的抑菌防病效果;利用洋葱伯克霍尔德氏菌致病因子的特异性引物检测菌株T132是否为潜在的人类致病菌。【结果】菌株T132鉴定为越南伯克霍尔德氏菌(Burkholderia vietnamiensis)。连续8次在人工培养基上传代培养,菌株T132抑制胶孢炭疽病菌生长的能力没有发生明显改变。菌株T132对胶孢炭疽菌C.gloeosporioides引起的柑橘炭疽病有明显的防治作用,刺伤接种的防效为88.2%,自然发病的防效为54.9%。未检测到该拮抗菌株有人体致病相关的洋葱伯克霍尔德氏菌致病因子(BCESM)毒力基因。【结论】首次报道对柑橘采后炭疽病具有生防效果、对人类相对安全的越南伯克霍尔德氏菌生防菌株。     

猪CFL2b基因在成肌细胞中的表达及对肌球蛋白重链基因表达的影响

猪CFL2b基因主要在骨骼肌中表达,对肌肉的发育和肌纤维的形成具有一定作用。为了解猪CFL2b基因与肌纤维性状的相关性,利用定向克隆和基因转染技术获得能稳定表达猪CFL2b基因的成肌细胞株,荧光显微镜观察及Western Blotting检测CFL2b基因在成肌细胞中的表达;应用实时定量PCR技术对细胞内肌球蛋白重链基因（MyHC）的表达变化进行检测。结果显示：CFL2b基因对MyHC的表达有明显影响,其中MyHC2x基因和MyHC26基因的表达明显上调,MyHCl／slow的表达变化不明显。表明CFL2b基因与猪的肌纤维性状密切相关,推测猪CFL2b基因的高表达可能导致猪的不良肉质性状,可以考虑将CFL2b基因作为猪肉质性状的候选基因[动物学报54（6）：1014—1019,2008]。     

An effect of a missense mutation in the porcine melanocortin-4 receptor (MC4R) gene on production traits in Polish pig breeds is doubtful

A missense G-A SNP in the porcine melanocortin-4 receptor (MC4R) gene that causes an Asp-Asn substitution at position 298 of the corresponding MC4R protein is considered to be economically important, although published results on its effect are inconsistent. We analysed the association of this MC4R polymorphism with production traits in 679 gilts from two breeds, Polish Large White (PLW) and Polish Landrace (PL), as well as one synthetic line 990. The frequency of the A allele differed significantly among the breeds with frequencies of 0.76, 0.29 and 0.16 in PLW, PL and line 990 respectively. There was no evidence of an effect of this polymorphism on daily food intake, backfat thickness or abdominal fat. The A allele was correlated with higher test daily gains and lower levels of intramuscular fat in PL, and increased levels of intramuscular fat in PLW.     

Comparison of three different farrowing systems: skin lesions and behaviour of sows with special regard to nursing behaviour in a group housing system for lactating sows

While group housing (GH) is mandatory in the European Union for the greater part of pregnancy, single housing in farrowing crates (FCs) during lactation that restrict sows in most of their natural behaviour patterns is still practised on a large scale. Research is urgently needed to develop alternative farrowing systems that improve sows’ welfare. Therefore, sows in three different farrowing systems – pens with FC, loose housing (LH) pens and GH for six sows – were compared regarding the level of skin injuries and their active and resting behaviour. A skin injury score was assessed for 15 body parts of 102 sows in six batches on 3 days (days 1, 14 and 34). In total, the active and resting behaviour of 77 sows in six batches was examined on 3 days (days 18, 25 and 32) between 0700 h and 1900 h by means of a scan sampling method. The suckling behaviour and the level of cross-suckling were analysed in GH by means of direct observation in four batches during three 4-h sampling periods (days 17, 24 and 31). No significant differences were found in total skin injuries when the sows entered the systems (day 1), but GH sows showed significantly higher total skin injuries compared to FC and LH sows in the middle (day 14) and at the end (day 34) of the lactation period. A significant difference between FC and LH sows was never seen. Differences were found for the proportion of different body postures between the three systems. The odds for lying in lateral recumbency versus standing and sitting versus standing were significantly higher for FC and LH sows compared to GH sows. Additionally, sows were significantly more likely to be standing as opposed to lying in lateral recumbency as the lactation period progressed. Cross-suckling was a frequent behaviour in GH, seen in 35.0% of all successful suckling bouts. However, only an average of 0.56 piglets per successful suckling bout was observed cross-suckling, suggesting only a few piglets were engaged in cross-suckling. In conclusion, the skin injury score was only moderately increased in GH compared to FC and LH and comparable to pregnant group-housed sows, both free farrowing systems seemed to be an environmental enrichment for lactating sows and good management cannot prevent the occurrence of cross-suckling in a GH system, but can probably reduce it.     

cDNA macroarray analysis of gene expression changes in rat brain after a single administration of a 2-aminoadamantane derivative

The Atlas Rat cDNA Expression Array (BD Biosciences, United States) has been used to analyze changes in the expression of 588 genes in rat brain cells in response to a single administration of Ladasten, a 2-aminoadamantane derivative that has psychostimulating and anxiolytic effects. The analysis of hybridization on macroarrays, confirmed by the results of real-time quantitative RT-PCR, has demonstrated that Ladasten alters the expression of 12 genes in the rat brain. The GAT3 and CARBH genes are presumed to be pharmacologically important targets of Ladasten. The changes in their activity explain the mechanisms of the anxiolytic and mood-stabilizing effects of the drug. Ladasten has been shown to induce the genes whose products are involved in various signal pathways (APC, Rb, PKCIP, and PMCA), as well as the genes of cytoskeletal proteins (Tub1 and actin), synaptic proteins (SynIA&IB and PLP), and enzymes (Gapdh and NSE). The proteins encoded by these genes are presumably involved in compensatory and/or neuroplastic adaptation to the effects of Ladasten.__________Translated from Molekulyarnaya Biologiya, Vol. 39, No. 2, 2005, pp. 276–285.Original Russian Text Copyright © 2005 by Vakhitova, Yamidanov, Vakhitov, Seredenin.     

QTL and association analysis for skin and fibre pigmentation in sheep provides evidence of a major causative mutation and epistatic effects

The pursuits of white features and white fleeces free of pigmented fibre have been important selection objectives for many sheep breeds. The cause and inheritance of non‐white colour patterns in sheep has been studied since the early 19th century. Discovery of genetic causes, especially those which predispose pigmentation in white sheep, may lead to more accurate selection tools for improved apparel wool. This article describes an extended QTL study for 13 skin and fibre pigmentation traits in sheep. A total of 19 highly significant, 10 significant and seven suggestive QTL were identified in a QTL mapping experiment using an Awassi × Merino × Merino backcross sheep population. All QTL on chromosome 2 exceeded a LOD score of greater than 4 (range 4.4–30.1), giving very strong support for a major gene for pigmentation on this chromosome. Evidence of epistatic interactions was found for QTL for four traits on chromosomes 2 and 19. The ovine TYRP1 gene on OAR 2 was sequenced as a strong positional candidate gene. A highly significant association (P < 0.01) of grandparental haplotypes across nine segregating SNP/microsatellite markers including one non‐synonymous SNP with pigmentation traits could be shown. Up to 47% of the observed variation in pigmentation was accounted for by models using TYRP1 haplotypes and 83% for models with interactions between two QTL probabilities, offering scope for marker‐assisted selection for these traits.     

The effect of low-intensity laser irradiation in the blue, green, and red spectral ranges on healing of experimental skin wounds in rats

The effects of low-intensity laser irradiation in the red (632.8 nm), green (532 nm), and blue (441.2 nm) spectral ranges on wound healing has been studied in rats. The effect of the traditionally used red laser irradiation has been compared with the effect caused by laser irradiation in other spectral ranges, aiming to support the provisional hypothesis that a similar healing effect could be achieved at lower doses of wound irradiation by lasers emitting in the blue and green spectral ranges. The following parameters have been used to characterize healing of the experimental wounds: the functional activity of phagocytes in the wound exudate, which was determined from luminol-dependent chemiluminescence, the phagocyte number; the wound exudates’ antioxidant activity; and the rate of healing, which was determined as the change of the wound surface area. It was found that in all cases the laser irradiation accelerated the healing of wounds. Exposure to red laser irradiation at the dose of 1.5 J/cm²), and to blue or green laser irradiation at a dose of 0.75 J/cm² shortened the time of the wound healing from 22 to 17 and 19 days, respectively. The functional activity of leukocytes in irradiated groups increased by day 5 after surgery, whereas in the control group it decreased. The superoxide dismutase activity increased in all experimental groups by day 5 after surgery. Laser irradiation in the red spectral range at a dose of 1.5 J/cm² resulted in a larger increase in superoxide dismutase activity, as compared to that found after exposure to laser irradiation in the blue and green spectral ranges at a dose of 0.75 J/cm².