共查询到19条相似文献，搜索用时 156 毫秒
1.
DNA甲基化参与调节动物配子发生和胚胎发育过程,TET基因负责DNA主动去甲基化,在基因印迹去除和细胞全能性获得中起重要作用。为了解海洋贝类配子发生和胚胎发育过程中DNA去甲基化如何发生,本研究以虾夷扇贝为研究对象,鉴定了其TET基因,并分析了该基因在性腺和胚胎幼虫发育中的表达变化。结果表明,虾夷扇贝基因组中含有1个TET基因(PyTET),该基因长39127 bp,包含10个外显子,编码1592个氨基酸,其蛋白具有完整的2OGFeDO超家族的加氧酶结构域。在性腺发育过程中,PyTET基因表达峰值出现在休止期精巢和增殖期卵巢,原位杂交结果显示其在精原细胞、精母细胞中均有表达,在卵母细胞中的表达明显高于卵原细胞;在早期发育过程中,其峰值出现在囊胚期。以上结果提示虾夷扇贝配子发生和胚胎发育过程中均发生了DNA主动去甲基化,这将有助于系统了解表观调控在贝类发育中的作用。 相似文献
2.
为探究亮氨酸和精氨酸作为信号分子参与调控mTOR信号通路对大菱鲆(Scophthalmus maximus)生长的作用,本研究以大菱鲆幼鱼(初始体质量(13.50±0.19) g)为实验对象,设置50%蛋白水平饲料作为阳性对照组,45%蛋白水平饲料作为阴性对照组,设置两个实验组,分别为在阴性对照组中添加1%亮氨酸的实验组和添加1%精氨酸的实验组,饲养周期为56 d,测量大菱鲆的生长性能和饲料利用水平。研究表明,添加亮氨酸、精氨酸均能一定程度提高因饲料蛋白水平不足导致的大菱鲆的特定生长率、蛋白质效率和增重率下降。添加1%亮氨酸和1%精氨酸均能在摄食后有效增强大菱鲆肌肉与肝脏mTOR信号通路中的雷帕霉素靶蛋白(mTOR)、核糖体蛋白S6和真核起始因子4E结合蛋白1(4E-BP1)的磷酸化。添加1%精氨酸还能够提高大菱鲆肠道淀粉酶和脂肪酶活性,显著提高血清过氧化氢酶和溶菌酶水平并增加血清总抗氧化能力。研究结果表明,在低蛋白饲料中添加1%亮氨酸、1%精氨酸能够激活大菱鲆mTOR信号通路,有效提高其生长性能和蛋白质效率;此外,添加1%精氨酸还能够提高大菱鲆的消化酶活性、增强非特异性免疫反应。 相似文献
3.
酪蛋白激酶1 (CK1)是一种重要的蛋白激酶家族,其在DNA损伤应答和修复、细胞增殖和凋亡、胚胎发育和稳态等重要的生物学过程中都具有复杂多样的调控作用。为了理解CK1基因家族在双壳贝类中的特征、进化及生物学功能,实验采用比较基因组学及生物信息学的方法对双壳贝类CK1基因家族进行了鉴定分析,通过虾夷扇贝高温应激实验,研究了CK1基因家族在高温应激时的表达规律。结果显示,在虾夷扇贝、栉孔扇贝、长牡蛎与侏儒蛤中均存在CK1α,CK1αlike,CK1δ,CK1γ3,并发现CK1ε,CK1γ1,CK1γ2在双壳贝类中发生了丢失。时空表达分析发现,双壳贝类CK1基因均在胚胎发育早期集中表达,并以多细胞/囊胚期为界呈现出母源/合子特异性表达模式。CK1为关键基因的基因共表达模块显著富集在错配修复、核苷酸剪切修复等相关通路上,暗示了CK1基因在双壳贝类胚胎发育过程中参与调控DNA损伤修复过程,从而维持早期胚胎发育时的基因组稳定性。双壳贝类CK1基因在成体中呈现组织特异的表达模式,主要在鳃和雄性性腺中具有相对较高的表达量。虾夷扇贝受到高温应激后,其鳃中PyCK1α, PyCK1αlike和PyCK1δ... 相似文献
4.
为研究mTOR信号通路在凡纳滨对虾(Litopenaeus vannamei)氨基酸代谢中的调控作用,本研究利用RACE技术首次克隆获得了凡纳滨对虾肌肉组织中eif4e2和eif4e1a基因的全长c DNA序列。eif4e2基因c DNA序列全长1 069 bp,开放阅读框699 bp,编码232个氨基酸;eif4e1a基因c DNA序列全长3579bp,开放阅读框627bp,编码208个氨基酸。氨基酸序列比对和进化分析均证实eIF4E2和eIF4E1A为目前已知的eIF4E家族蛋白的同源蛋白。利用实时荧光定量PCR的方法研究了eif4e2和eif4e1a基因在凡纳滨对虾不同组织中的表达差异以及注射雷帕霉素或氨基酸后肌肉中eif4e2和eif4e1a基因表达量的变化情况,探讨了eif4e2和eif4e1a基因在细胞生长中的调控作用。结果表明,凡纳滨对虾eif4e2和eif4e1a基因在眼柄、肝胰脏、肠道、胃、鳃丝和肌肉等组织中均有表达;其中eif4e2基因在肌肉中的表达量显著高于其他组织(P0.05),eif4e1a基因在肠道和肌肉中都有较高表达量,且在肠道中的表达量显著高于其在肌肉中的表达量(P0.05);注射雷帕霉素后2 h内肌肉中eif4e2和eif4e1a基因表达量都出现显著下降(P0.05);肌肉中eif4e1a基因的表达量在单独注射亮氨酸或精氨酸4 h内均未出现变化,但在同时注射亮氨酸和精氨酸后表达量显著增加(P0.05);肌肉中eif4e2基因在注射亮氨酸、精氨酸及亮氨酸加精氨酸后表达量都明显提高(P0.05),且同时注射亮氨酸和精氨酸后,基因表达变量明显比单独注射亮氨酸或精氨酸后变化量大。本研究从动物分子营养学角度出发,克隆了凡纳滨对虾mTOR信号通路中eif4e2和eif4e1a两个基因,并证明其能够通过mTOR信号通路接收不同氨基酸信号来调控细胞生长,对于深入了解对虾的生长和代谢调控机制、饲料配方的优化以及建立合理的养殖管理模式都具有重要的意义。 相似文献
5.
热刺激对栉孔扇贝免疫功能和热休克蛋白表达的影响 总被引：1，自引：0，他引：1
《中国海洋大学学报(自然科学版)》2017,(8)
为探究热刺激对栉孔扇贝(Chlamysfarreri)免疫功能的影响,以及热休克蛋白在应对急性高温胁迫时的作用。本研究对一龄栉孔扇贝进行28℃热刺激,采用流式细胞术于0、1、2、4、8h测定血细胞的吞噬率和活性氧ROS含量;以鳗弧菌和溶壁微球菌为底物测定血清抗菌、溶菌活力;用彗星实验测定DNA损伤;用AO/EB(吖啶橙/溴化乙啶)荧光染色法检测细胞凋亡;用荧光实时定量PCR检测HSP70和HSP90mRNA相对表达量。研究表明:热刺激使血细胞中活性氧含量、吞噬活性以及血淋巴抗菌、溶菌活力受到抑制,总体表现为下降趋势;DNA损伤和细胞凋亡受热刺激诱导显著上升,具有明显的时间-效应关系;HSP70、HSP90mRNA表达量显著上调,反应迅速,且HSP70受热刺激诱导更显著。研究结果表明,热刺激导致栉孔扇贝血细胞发生细胞凋亡、加剧DNA损伤,造成免疫力下降,而机体通过大量表达热休克蛋白HSP70、HSP90保护细胞和组织免受损伤。本研究为探究夏季易发扇贝大规模死亡原因提供了理论依据。 相似文献
6.
阳光紫外辐射对水生动物个体水平如生长、发育和存活等方面产生不利影响.从DNA损伤、蛋白质损伤、呼吸爆发和细胞凋亡等方面较系统地分析了影响机制,另外,还从逃避紫外辐射、光保护物质、DNA修复、体内的抗氧化系统等方面分析了水生动物对阳光紫外辐射的响应机制.最后提出应当重点开展原位实验、长期效应、间接影响、群落及生态系统和响应机制等工作. 相似文献
7.
利用120#燃料油分散液和消油剂处理的乳化液测定海胆急性毒性和慢性DNA损伤及全基因组甲基化水平变化,确定消油剂处理的120#燃料油对海胆的毒性影响。结果表明:消油剂单独使用对海胆的影响微乎其微,经消油剂处理的燃料油乳化液中的多环芳烃质量浓度明显高于分散液中的质量浓度;乳化液和分散液对海胆都有明显的毒性作用,其96h半数效应浓度(EC50)值分别为12.03g/L和29.15g/L;DNA损伤研究表明,乳化液处理的海胆肠细胞彗星拖尾情况比同质量浓度分散液中拖尾情况更加严重;全基因组甲基化水平比分散液中更低,表明乳化液的急性毒性更大,所造成的DNA损伤更加严重。结果提示消油剂的使用会增大燃料油对海胆的毒性效应。 相似文献
8.
克隆得到了合浦珠母贝(Pinctada fucata)PF-CREB3L2蛋白的c DNA序列,其c DNA全长1 983 bp,其中开放阅读框长度为1 728 bp,编码的蛋白含有575个氨基酸残基。组织表达分布实验发现,其在合浦珠母贝内脏囊中表达量最高,在外套膜和鳃中也有大量表达,推测其广泛参与合浦珠母贝的贝壳形成等生理活动。贝壳损伤修复实验中发现,在合浦珠母贝贝壳受到损伤后,PF-CREB3L2和基质蛋白的表达量会迅速上升,且PF-CREB3L2的峰值出现更早,推测其通过影响基质蛋白转录,参与合浦珠母贝生物矿化的调控过程。PF-CREB3L2与合浦珠母贝基质蛋白表达相关性的分析发现,PF-CREB3L2与Prisilkin39、KRMP的表达量呈现显著的正相关,说明其可能特异性地调控某些基质蛋白的转录。 相似文献
9.
全基因组DNA甲基化作为重要的表观遗传学现象,其在生物体诸多生理生化过程中起了重要作用。研究表明全基因组DNA甲基化在基因的表达调控、维持胚胎正常发育、染色体结构稳定等方面发挥了重要作用。DNA甲基化也成为当今的研究热点之一,目前在脊椎动物领域研究相对较多,但是对无脊椎动物的甲基化规律的研究比较匮乏,因此本研究以棘皮动物门中仿刺参作为研究对象,采用最新的全基因组DNA甲基化检测方法 MethylRAD-Seq技术探究了仿刺参三种组织的甲基化状况,得到了甲基化标签数量、甲基化位点分布以及甲基化标签密度等信息;此外通过整合仿刺参甲基化文库数据和表达谱数据发现基因甲基化水平与表达水平之间存在弱的正相关性,这暗示了无脊椎动物基因组DNA甲基化可能起到促进基因表达的作用。 相似文献
10.
应用MSAP技术研究扇贝全基因组DNA甲基化水平 总被引：1，自引：0，他引：1
DNA甲基化作为真核生物基因组重要的表观遗传学修饰,对生物体基因的表达有重要的调控作用。为获得扇贝基因组DNA甲基化修饰水平及模式等表观遗传学信息,以栉孔扇贝(Chlamys farreri)、海湾扇贝(Argopecten irradians)、虾夷扇贝(Mizuhopecten yessoensis)以及本课题组培育的"海大金贝"为材料,建立了甲基化敏感扩增多态性方法(Methylation-sensitive amplified polymorphism,MSAP)的反应体系,利用该方法对其基因组DNA CCGG区域的甲基化水平进行分析。结果表明,本研究筛选得到的引物组合可用于贝类DNA甲基化的研究,在栉孔扇贝、海湾扇贝、普通虾夷扇贝和"海大金贝"的甲基化比例分别为32.08%、25.99%、32.88%和34.97%。几种扇贝基因组CCGG序列中,胞嘧啶的全甲基化率要高于半甲基化率,推测扇贝基因组中主要的甲基化模式是CpG型。通过对"海大金贝"和普通虾夷扇贝闭壳肌甲基化谱进行比较,筛查得到46个差异位点,这些位点可能参与"海大金贝"闭壳肌积累类胡萝卜素的调控。 相似文献
11.
Comet assays to assess DNA damage and repair in grass shrimp embryos exposed to phototoxicants 总被引：4，自引：0，他引：4
Exposure of grass shrimp (Palaemonetes pugio) embryos to four compounds (anthracene, pyrene, alpha-terthienyl, methylene blue) along with solar exposure resulted in extensive DNA strand damage using the comet assay. DNA tail moments of embryos exposed to these chemicals in the dark ranged from 1.8 to 4.3, while exposure to chemicals and solar resulted in tail moments of 14.3-15.3. Reduction of DNA tail moments when solar exposed embryos were transferred to the dark, suggested DNA repair systems were active. The comet assay can be used to follow both DNA damage and repair following exposure to phototoxic chemicals. 相似文献
12.
以大弹涂鱼为研究对象,应用单细胞凝胶电泳技术并结合外周血基因组DNA琼脂糖凝胶电泳,研究了镉胁迫对大弹涂鱼外周血细胞的遗传损伤。结果表明:(1)镉对大弹涂鱼外周血细胞是遗传毒性而非细胞毒性,并且产生的遗传损伤存在显著(P0.01或P0.05)的剂量效应,染毒1h的大弹涂鱼的全长和尾长与浓度的相关方程分别是y=29.592x-10.576(r2=0.9786),y=28.417x-14.859(r2=0.9857),染毒2h的大弹涂鱼的全长和尾长与浓度的相关方程分别是y=32.044x-5.0235(r2=0.941),y=29.911x-9.539(r2=0.9635)。(2)DNA的损伤程度和污染胁迫之间存在着时间效应。以上结果表明,DNA损伤可作为大弹涂鱼受镉胁迫时的生物指标。 相似文献
13.
Hartl MG Coughlan BM Sheehan D Mothersill C van Pelt FN O'Reilly SJ Heffron JJ O'Halloran J O'Brien NM 《Marine environmental research》2004,57(4):295-310
We explore the use of the clam Tapes semidecussatus Reeves 1864 as an indicator for the presence of potentially genotoxic substances in estuarine sediments. The limitations associated with the interpretation of Comet assay data (expressed as % DNA in tail) in terms of clam reproductive state, size (age) and thermal exposure history following laboratory acclimation are discussed. Hatchery-reared clams, subjected to ambient temperature fluctuations during growth, were exposed in vivo under laboratory conditions for three weeks to sediment samples collected from a polluted site and a "clean" reference site. The DNA damage observed in haemocytes, gill and digestive gland cells was significantly higher in animals exposed to contaminated sediment compared to those exposed to sediment from the reference site. The extent of DNA damage recorded was not correlated with size (age). Spawning was not observed during the experiment. Nevertheless, clams with well-developed gonads showed a statistically higher degree of DNA damage in gill and digestive gland cells- but not haemocytes, demonstrating an increased sensitivity to potential genotoxic compounds, possibly caused by impaired DNA repair capacity due to reproductive activity. Furthermore, the degree of DNA damage in clams exposed to contaminated sediments was higher in autumn and winter compared to spring and summer, suggesting an effect of seasonal priming. 相似文献
14.
Grass shrimp embryos develop in egg sacs (stages 1-10) attached to the female for 14-20 days after which they 'hatch' from the egg sacs into a swimming zoea stage (stage 11). Until they emerge from the egg sacs, embryos depend on lipids and lipovitellin stored within the egg. The percent of embryos which hatch after exposure to toxicants relative to controls was the basis of an embryo development assay. Exposure of embryos to chromium(III) chloride, sodium chromate, mercuric chloride, and 2-methyl-1,2-naphthoquinone (MNQ) resulted in a reduced hatching rate. In addition to effects on embryo development, DNA strand damage tests were carried out on contaminant-exposed embryos, using the single-cell electrophoresis method often referred to as comet assay. Development of stage 4 embryos was more affected by MNQ exposure than stage 7 embryos. The hatching rates of stages 4 and 7 embryos exposed to MNQ (172 micrograms/l) were 0 and 90%, respectively. DNA strand damage, measured as DNA tail moments, were 3.4 and 4.4, respectively. Thus, exposure of an early embryo stage to MNQ prevented full embryo development while development of later embryo stages was not affected. It may be that the DNA repair systems are more efficient in later embryo stages than in early stages and thus DNA damaged in the early stages affects development. 相似文献
15.
16.
采用单细胞凝胶电泳实验研究了不同浓度的Cu2+(10、50、100、500、1000、2500、5000μg/L)和不同粒径(0.1、0.2、0.3、0.5、0.7、1.0、2.0μm)、不同浓度(1、10、15、20、25 mg/mL)的聚苯乙烯微塑料对人单核细胞白血病(Human Acute Monocytic Leukemia,THP-1)细胞的DNA损伤效应,结果表明:单一污染时,Cu2+(≥50μg/L)对THP-1细胞的DNA损伤极显著(p<0.01)。而且,DNA损伤值随Cu2+浓度的增大而增加。2.0μm粒径的聚苯乙烯微塑料(10 mg/mL)对THP-1细胞产生明显的DNA损伤效应。复合污染作用下对THP-1细胞的DNA损伤效应,相比单一污染明显增强,而且DNA损伤值比单一污染的理论加和值高。本研究表明Cu2+和聚苯乙烯微塑料均对DNA产生损伤效应,显示一定的基因毒性,二者复合对损伤具有交互增强的作用。 相似文献
17.
J P Shaw A T Large J K Chipman D R Livingstone L D Peters 《Marine environmental research》2000,50(1-5):405-409
Mytilus edulis digestive gland microsomes were prepared from indigenous populations sampled from a clean reference site (Port Quin) and an urban-industrial contaminated site (Blackpool) in the UK. Samples were collected in March/April, May, August and December 1998. Western blot analysis was performed using polyclonal antibodies to fish CYP1A and rat CYP2E using partially purified M. edulis CYP as a positive control, to aid identification. CYP1A- and CYP2E-immunopositive protein levels showed different site-specific seasonal variation with higher levels of CYP2E determined in May (P < 0.05). At both sites, lower levels of CYP1A-immunopositive protein but not CYP2E-immunopositive protein were observed in the samples collected in December (P < 0.05). This correlated with lower levels of nuclear DNA damage (Comet assay expressed as per cent tail DNA) observed in December compared to August (P < 0.05). 相似文献
18.
Shaw JP Large AT Livingstone DR Doyotte A Renger J Chipman JK Peters LD 《Marine environmental research》2002,54(3-5):505-509
Mytilus edulis were collected from a reference site (Port Quin) and an urban/industrial contaminated site (New Brighton) in the UK during June 1999. Levels of PCBs (sigma7 congeners) and CB-138 were determined to be, respectively, 21 fold and 16 fold higher in the mussel digestive glands from New Brighton. Levels of CYPIA-immunopositive protein were 1.5 fold higher (P < 0.05) at the polluted site but the levels of DNA strand breaks were 1.3 fold higher (P<0.05) at the reference site. Mussels from Port Quin were placed in cages at both sites and both transplanted and indigenous populations sampled in September (13 weeks). Mussels transplanted from the reference site to the industrial site, reported elevated levels of CYP1A-immunopositive protein (1.4 fold; P < 0.05) and higher levels of DNA damage (1.2 fold; P < 0.05) compared to caged populations at the reference site and a PCB loading similar to the populations from the polluted site. Moreover, transplanted mussels had DNA damage 1.8 fold greater (P < 0.05) than indigenous mussels at the transplant site. These changes were small but significant when compared to the observed temporal changes in the indigenous populations. 相似文献
19.
Effects of genotoxic compounds on DNA and development of early and late grass shrimp embryo stages 总被引：1，自引：0，他引：1
Early and late developmental stages of grass shrimp embryos were exposed to different concentrations of two genotoxicants, 2-methyl-1,4-naphthoquinone (MNQ) and 4-nitroquinoline-N-oxide (NQO). DNA strand breaks were assessed by the comet assay while embryo development effects were determined by % of embryos hatching. Early embryo stage embryos were significantly more sensitive to genotoxicants than late stages. For example, all stage 4 embryos failed to hatch at 1 microM NQO while 95% of stage 8 hatched at this concentration. High DNA tail moments, which are a measure of the number of DNA strand breaks, were found in late stage embryos exposed to genotoxicants. Early stage embryo development was effected by low concentrations of genotoxicants but no changes were observed in DNA tail moments. We suggest that high DNA moments in late embryo stages reflect high DNA repair activity, while early stages may lack a fully developed DNA repair system. 相似文献