Cold Spring Harbor protocols最新文献_第3页

Membrane Protein Fractionation and Analysis through Western Blot in Aedes aegypti Malpighian Tubules. 埃及伊蚊马氏管的膜蛋白分馏和 Western 印迹分析。

Cold Spring Harbor protocols Pub Date : 2024-12-02 DOI: 10.1101/pdb.prot108283

Britney Picinic, Jean-Paul V Paluzzi

引用次数: 0

Mapping Transcript Cell-Specific Localization and Protein Subcellular Localization in the Adult Mosquito Aedes aegypti. 绘制埃及伊蚊成蚊转录本细胞特异性定位和蛋白质亚细胞定位图。

Cold Spring Harbor protocols Pub Date : 2024-12-02 DOI: 10.1101/pdb.top107698

Farwa Sajadi, Salwa Afifi, Britney Picinic, Jean-Paul V Paluzzi

{"title":"Mapping Transcript Cell-Specific Localization and Protein Subcellular Localization in the Adult Mosquito Aedes aegypti.","authors":"Farwa Sajadi, Salwa Afifi, Britney Picinic, Jean-Paul V Paluzzi","doi":"10.1101/pdb.top107698","DOIUrl":"10.1101/pdb.top107698","url":null,"abstract":"This introduction reviews techniques used to examine the distribution and expression of gene transcripts and proteins in a variety of tissues/organs in the medically important global disease vector mosquito, Aedes aegypti Specifically, these methods allow the detection of cell-specific transcript expression by fluorescent in situ hybridization; facilitate immunohistochemical mapping of a protein of interest in whole-mount small tissue/organ samples; examine the subcellular localization of proteins, such as membrane transporters, through sectioning of paraffin-embedded tissue/organ samples; and finally, enable the efficient separation of cytosolic and membrane proteins for western blot analysis without the need for specialized equipment (e.g., ultracentrifuge) in the mosquito Ae. aegypti Such techniques are useful to help answer fundamental questions in mosquito scientific research including (but not limited to) the identification of specific cells in an organ responsible for expressing a receptor of particular interest and necessary for eliciting a response to exogenous signals, including hormones. Moreover, changes in the subcellular localization of specific targets of interest can be assessed both qualitatively and quantitatively, providing insight into transient or long-term physiologically relevant regulation necessary for activity under experimental treatments or varied internal (e.g., development) or external (e.g., environmental stress) factors that might be normally experienced by the organism.","PeriodicalId":10496,"journal":{"name":"Cold Spring Harbor protocols","volume":" ","pages":"pdb.top107698"},"PeriodicalIF":0.0,"publicationDate":"2024-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138795556","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Examining Apical or Basolateral Protein Localization in Aedes aegypti Tissues: Cross-Section Immunohistochemistry. 检查埃及伊蚊组织中的顶端或基底侧蛋白定位：横切面免疫组化。

Cold Spring Harbor protocols Pub Date : 2024-12-02 DOI: 10.1101/pdb.prot108282

Farwa Sajadi, Britney Picinic, Salwa Afifi, Jean-Paul V Paluzzi

引用次数: 0

Use of DeTerm for Automated Drosophila Dendrite Arbor Terminal Counts. 使用 DeTerm 自动进行果蝇树突顶端计数。

Cold Spring Harbor protocols Pub Date : 2024-12-02 DOI: 10.1101/pdb.prot108151

Minami Kimura, Jason Y Tann, Oliver R Wilkes, Fangke Xu, Henrik Skibbe, Adrian W Moore

引用次数: 0

Examining Protein Localization in Aedes aegypti Cells, Tissues, and Organs: Whole-Mount Immunohistochemistry. 检查埃及伊蚊细胞、组织和器官中的蛋白质定位：整装免疫组化。

Cold Spring Harbor protocols Pub Date : 2024-12-02 DOI: 10.1101/pdb.prot108281

Salwa Afifi, Farwa Sajadi, Jean-Paul V Paluzzi

{"title":"Examining Protein Localization in Aedes aegypti Cells, Tissues, and Organs: Whole-Mount Immunohistochemistry.","authors":"Salwa Afifi, Farwa Sajadi, Jean-Paul V Paluzzi","doi":"10.1101/pdb.prot108281","DOIUrl":"10.1101/pdb.prot108281","url":null,"abstract":"Immunohistochemistry (IHC) is a powerful technique used for visualizing cellular components and determining the presence and/or location of proteins or other macromolecules in tissue samples. The classical IHC process involves the detection of epitopes using a highly specific primary antibody. This is followed by a secondary antibody that is coupled to a reporter molecule or fluorophore and capable of binding to the primary antibody and allowing for protein immunodetection. Although IHC does not routinely provide quantitative results compared to an enzyme-linked immunoassay or western blotting, it permits the localization of the proteins in intact tissues. This protocol describes an IHC assay for whole-body Aedes aegypti mosquito tissues that is used to detect small proteins, specifically neuropeptide hormones. This method is useful for protein detection in whole-mount preparations; however, cross-section IHC is recommended to determine if a protein is localized in the apical versus basolateral membrane of tissues/organs or to visualize immunological distribution in larger, more complex preparations.","PeriodicalId":10496,"journal":{"name":"Cold Spring Harbor protocols","volume":" ","pages":"pdb.prot108281"},"PeriodicalIF":0.0,"publicationDate":"2024-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138795544","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Study of Dendrite Differentiation Using Drosophila Dendritic Arborization Neurons. 利用果蝇树突分化神经元研究树突分化

Cold Spring Harbor protocols Pub Date : 2024-12-02 DOI: 10.1101/pdb.top108146

Jason Y Tann, Fangke Xu, Minami Kimura, Oliver R Wilkes, Li-Foong Yoong, Henrik Skibbe, Adrian W Moore

引用次数: 0

Mounting of Embryos, Larvae, and Pupae for Live Drosophila Dendritic Arborization Neuron Imaging. 为果蝇树突分化神经元活体成像安装胚胎、幼虫和蛹。

Cold Spring Harbor protocols Pub Date : 2024-12-02 DOI: 10.1101/pdb.prot108149

Fangke Xu, Jason Y Tann, Oliver R Wilkes, Minami Kimura, Li-Foong Yoong, Adrian W Moore

引用次数: 0

Mosaic Analysis with a Repressible Cell Marker (MARCM) Clone Generation in Drosophila Dendritic Arborization Neurons. 果蝇树突分化神经元可抑制细胞标记（MARCM）克隆生成的镶嵌分析。

Cold Spring Harbor protocols Pub Date : 2024-12-02 DOI: 10.1101/pdb.prot108147

Jason Y Tann, Fangke Xu, Minami Kimura, Adrian W Moore

{"title":"Mosaic Analysis with a Repressible Cell Marker (MARCM) Clone Generation in Drosophila Dendritic Arborization Neurons.","authors":"Jason Y Tann, Fangke Xu, Minami Kimura, Adrian W Moore","doi":"10.1101/pdb.prot108147","DOIUrl":"10.1101/pdb.prot108147","url":null,"abstract":"Mosaic analysis with a repressible cell marker (MARCM) is used in Drosophila research to create labeled homozygous mutant clones of cells in an otherwise heterozygous fly. It allows the study of the effect of embryonically lethal genes and the determination of cell autonomy for a mutant phenotype. When used in dendritic arborization (da) neurons with a fluorescent protein targeted to the plasma membrane, MARCM allows the identification of homozygous mutant neurons and clear imaging of the dendrite arbor in both live and fixed preparations. Previous protocols that outlined experimental procedures to create MARCM clones in da neurons used a heat shock promoter to drive Flippase (FLP) expression; such an approach requires laborious embryo collection and heat shock steps, and it creates clones in other tissues besides the da neurons. The updated protocol described here outlines the use of FLP expression driven by a sensory organ precursor promoter (SOP-FLP); it requires no embryo collection or manipulation steps and creates clones exclusively in the peripheral sensory neuron lineage.","PeriodicalId":10496,"journal":{"name":"Cold Spring Harbor protocols","volume":" ","pages":"pdb.prot108147"},"PeriodicalIF":0.0,"publicationDate":"2024-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139039617","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Culture of Larval and Pupal Drosophila Dendritic Arborization Neurons. 幼虫和蛹果蝇树突分化神经元的培养

Cold Spring Harbor protocols Pub Date : 2024-12-02 DOI: 10.1101/pdb.prot108150

Jason Y Tann, Fangke Xu, Li-Foong Yoong, Adrian W Moore

引用次数: 0

Optimized Methods for Applying and Assessing Heat, Drought, and Nutrient Stress of Maize Seedlings in Controlled Environment Experiments. 在受控环境实验中应用和评估玉米幼苗热、旱和养分胁迫的优化方法。

Cold Spring Harbor protocols Pub Date : 2024-11-18 DOI: 10.1101/pdb.top108467

Alejandra Quiñones, Leonardo W Lima, Katherine M Murphy, Anna L Casto, Malia A Gehan, Cory D Hirsch

{"title":"Optimized Methods for Applying and Assessing Heat, Drought, and Nutrient Stress of Maize Seedlings in Controlled Environment Experiments.","authors":"Alejandra Quiñones, Leonardo W Lima, Katherine M Murphy, Anna L Casto, Malia A Gehan, Cory D Hirsch","doi":"10.1101/pdb.top108467","DOIUrl":"10.1101/pdb.top108467","url":null,"abstract":"Maize (Zea mays), also known as corn, is an important crop that plays a crucial role in global agriculture. The economic uses of maize are numerous, including for food, feed, fiber, and fuel. It has had a significant historical importance in research as well, with important discoveries made in maize regarding plant domestication, transposons, heterosis, genomics, and epigenetics. Unfortunately, environmental stresses cause substantial yield loss to maize crops each year. Yield losses are predicted to increase in future climate scenarios, posing a threat to food security and other sectors of the global economy. Developing efficient methods to study maize abiotic stress responses is a crucial step toward a more resilient and productive agricultural system. This review describes the importance of and methods for studying the effects of heat, drought, and nutrient deficiency on early developmental stages of maize grown in controlled environments. Studying the early effects of environmental stressors in controlled environments allows researchers to work with a variety of environmental conditions with low environmental variance, which can inform future field-based research. We highlight the current knowledge of physiological responses of maize to heat, drought, and nutrient stress; remaining knowledge gaps and challenges; and information on how standardized protocols can address these issues.","PeriodicalId":10496,"journal":{"name":"Cold Spring Harbor protocols","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142459804","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0