The generation of mature and healthy oocytes is the most critical event in the entire female reproductive process,and the mechanisms regulating this process remain to be studied.Here,we demonstrate that Smith-like(LSM...The generation of mature and healthy oocytes is the most critical event in the entire female reproductive process,and the mechanisms regulating this process remain to be studied.Here,we demonstrate that Smith-like(LSM)family member 14B(LSM14B)regulates oocyte maturation,and the loss of LSM14B in mouse ovaries leads to abnormal oocyte MII arrest and female infertility.Next,we find the aberrant transcriptional activation,indicated by abnormal non-surrounded nucleolus and surrounded nucleolus oocyte proportions,and abnormal chromosome assembly and segregation in Lsm14b-deficient mouse oocytes.The global transcriptome analysis suggests that many transcripts involved in cytoplasmic processing body(P-body)function are altered in Lsm14b-deficient mouse oocytes.Deletion of Lsm14b results in the expression and/or localization changes of P-body components(such as LSM14A,DCP1A,and 4E-T).Notably,DDX6,a key component of the P-body,is downregulated and accumulates in the nuclei in Lsm14b-deficient mouse oocytes.Taken together,our data suggest that LSM14B links mouse oocyte maturation to female fertility through the regulation of the P-body.展开更多
To modulate gene expression in research studies or in potential clinical therapies,transfection of exogenous nucleic acids including plasmid DNA and small interference RNA(siRNA)are generally performed.However,the cel...To modulate gene expression in research studies or in potential clinical therapies,transfection of exogenous nucleic acids including plasmid DNA and small interference RNA(siRNA)are generally performed.However,the cellular processing and the fate of these nucleic acids remain elusive.By investigating the cellular behavior of transfected nucleic acids using confocal imaging,here we show that when siRNA was cotransfected into cultured cells with other nucleic acids,including single-stranded RNA oligonucleotides,single and double-stranded DNA oligonucleotides,as well as long double-stranded plasmid DNA,they all aggregate in the same cytoplasmic granules.Interestingly,the amount of siRNA aggregating in granules was found not to correlate with the gene silencing activity,suggesting that assembly of cytoplasmic granules triggered by siRNA transfection may be separable from the siRNA silencing event.Our results argue against the claim that the siRNAaggregating granules are the functional site of RNA interference(RNAi).Taken together,our studies suggest that,independent of their types or forms,extraneously transfected nucleic acids are processed through a common cytoplasmic pathway and trigger the formation of a new type of cytoplasmic granules“transfection granules”.展开更多
人脱帽酶Dcp1a(human m RNA decapping enzyme 1a)是m RNA降解过程中的重要成员之一,构建其真核表达载体,并对其表达和细胞内定位进行分析,可为进一步研究Dcp1a的功能提供基础。本研究以He La c DNA文库为模板,采用PCR方法扩增Dcp1a c ...人脱帽酶Dcp1a(human m RNA decapping enzyme 1a)是m RNA降解过程中的重要成员之一,构建其真核表达载体,并对其表达和细胞内定位进行分析,可为进一步研究Dcp1a的功能提供基础。本研究以He La c DNA文库为模板,采用PCR方法扩增Dcp1a c DNA全长序列并克隆到pm Cherry-N1载体。转化大肠杆菌DH5α后,挑取阳性克隆提取质粒,分别进行Xho I/Sal I双酶切及测序鉴定。将重组质粒用Vigo Fect转染试剂转染He La细胞,随后用RT-PCR和Western-blot检测Dcp1a在He La细胞中的表达,并采用激光共聚焦显微镜观察Dcp1a的亚细胞定位情况。通过DNA测序分析证实目的基因Dcp1a的序列完全正确,人Dcp1a基因真核表达载体pm Cherry-N1-Dcp1a构建成功,并在He La细胞中获得表达;激光共聚焦显微镜观察显示Dcp1a蛋白定位在细胞质中,而且He La细胞中过表达Dcp1a在胞质中明显形成了P小体(processing body,P-body)。实验结果为进一步探讨该基因的功能及其与P小体的相关性奠定了基础。展开更多
In this paper, the concepts of the ith L;-mixed affine surface area and Lp-polar curvature images are introduced, some new inequalities connecting these new notions with Lρ-centroid bodies and ρ-Blaschke bodies are ...In this paper, the concepts of the ith L;-mixed affine surface area and Lp-polar curvature images are introduced, some new inequalities connecting these new notions with Lρ-centroid bodies and ρ-Blaschke bodies are showed. Moreover, a Blaschke-Santalo type inequality for Lρ-mixed affine surface area is established. Our results also imply the similar to the inequalities for Marcus-Lopes, Bergstrom and Ky Fan.展开更多
基金supported by the National Key R&D Program of China(2021YFC2700201 to X.W.)the National Natural Science Foundation of China(32070831,32270897 to X.W.).
文摘The generation of mature and healthy oocytes is the most critical event in the entire female reproductive process,and the mechanisms regulating this process remain to be studied.Here,we demonstrate that Smith-like(LSM)family member 14B(LSM14B)regulates oocyte maturation,and the loss of LSM14B in mouse ovaries leads to abnormal oocyte MII arrest and female infertility.Next,we find the aberrant transcriptional activation,indicated by abnormal non-surrounded nucleolus and surrounded nucleolus oocyte proportions,and abnormal chromosome assembly and segregation in Lsm14b-deficient mouse oocytes.The global transcriptome analysis suggests that many transcripts involved in cytoplasmic processing body(P-body)function are altered in Lsm14b-deficient mouse oocytes.Deletion of Lsm14b results in the expression and/or localization changes of P-body components(such as LSM14A,DCP1A,and 4E-T).Notably,DDX6,a key component of the P-body,is downregulated and accumulates in the nuclei in Lsm14b-deficient mouse oocytes.Taken together,our data suggest that LSM14B links mouse oocyte maturation to female fertility through the regulation of the P-body.
文摘To modulate gene expression in research studies or in potential clinical therapies,transfection of exogenous nucleic acids including plasmid DNA and small interference RNA(siRNA)are generally performed.However,the cellular processing and the fate of these nucleic acids remain elusive.By investigating the cellular behavior of transfected nucleic acids using confocal imaging,here we show that when siRNA was cotransfected into cultured cells with other nucleic acids,including single-stranded RNA oligonucleotides,single and double-stranded DNA oligonucleotides,as well as long double-stranded plasmid DNA,they all aggregate in the same cytoplasmic granules.Interestingly,the amount of siRNA aggregating in granules was found not to correlate with the gene silencing activity,suggesting that assembly of cytoplasmic granules triggered by siRNA transfection may be separable from the siRNA silencing event.Our results argue against the claim that the siRNAaggregating granules are the functional site of RNA interference(RNAi).Taken together,our studies suggest that,independent of their types or forms,extraneously transfected nucleic acids are processed through a common cytoplasmic pathway and trigger the formation of a new type of cytoplasmic granules“transfection granules”.
基金Supported by the NNSF of China(11161024)Supported by the NSF of Jiangxi Province (2010GZC0115)
文摘In this paper, the concepts of the ith L;-mixed affine surface area and Lp-polar curvature images are introduced, some new inequalities connecting these new notions with Lρ-centroid bodies and ρ-Blaschke bodies are showed. Moreover, a Blaschke-Santalo type inequality for Lρ-mixed affine surface area is established. Our results also imply the similar to the inequalities for Marcus-Lopes, Bergstrom and Ky Fan.
