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> Anatomy > Cell Component > Nuclear Envelope

Nuclear Envelope

The Nuclear Envelope is the double-membrane structure that surrounds the nucleus of eukaryotic cells.
It acts as a barrier, controlling the exchange of materials between the nucleus and cytoplasm.
The envelope is composed of an outer nuclear membrane, an inner nuclear membrane, and nuclear pore complexes that facilitate selective transport.
It plays a crucial role in gene expression, chromatin organization, and cell signaling.
Researchers can unlock the secrets of the Nuclear Envelope using PubCompare.ai, an AI-driven platform that helps optimize research protocols by comparing the latest literature, preprints, and patents.
Tis platform enables researchers to streamline their process and achieve better results in their studies of this essential cellular structure.

Most cited protocols related to «Nuclear Envelope»

1
Xenopus Neural Crest Migration Assay
Xenopus neural crest was labelled with nuclear-RFP/membrane-GFP or membrane-RFP/nuclear-GFP. In vitro analysis of neural crest migration was performed using Xenopus neural crest cultured on fibronectin-coated plates. For in vivo studies we used Xenopus embryos grafted with labelled neural crest or zebrafish transgenic lines embryos that express cytoplasm or membrane-GFP under the neural crest promoter sox10. Time-lapse was carried out using DIC or fluorescent/confocal microscopy. FRET analysis was performed as described in14 (link). For full methods, see Supplementary Material.
Carmona-Fontaine C., Matthews H.K., Kuriyama S., Moreno M., Dunn G.A., Parsons M., Stern C.D, & Mayor R. (2008). Contact Inhibition of Locomotion in vivo controls neural crest directional migration. Nature, 456(7224), 957-961.
Publication 2008
Animals, Transgenic Cytoplasm Embryo Fibronectins Fluorescence Resonance Energy Transfer Microscopy, Confocal Neural Crest Nuclear Envelope SOX10 Transcription Factor Tissue, Membrane Xenopus laevis Zebrafish
2
Xenopus Neural Crest Migration Assay
Xenopus neural crest was labelled with nuclear-RFP/membrane-GFP or membrane-RFP/nuclear-GFP. In vitro analysis of neural crest migration was performed using Xenopus neural crest cultured on fibronectin-coated plates. For in vivo studies we used Xenopus embryos grafted with labelled neural crest or zebrafish transgenic lines embryos that express cytoplasm or membrane-GFP under the neural crest promoter sox10. Time-lapse was carried out using DIC or fluorescent/confocal microscopy. FRET analysis was performed as described in14 (link). For full methods, see Supplementary Material.
Carmona-Fontaine C., Matthews H.K., Kuriyama S., Moreno M., Dunn G.A., Parsons M., Stern C.D, & Mayor R. (2008). Contact Inhibition of Locomotion in vivo controls neural crest directional migration. Nature, 456(7224), 957-961.
Publication 2008
Animals, Transgenic Cytoplasm Embryo Fibronectins Fluorescence Resonance Energy Transfer Microscopy, Confocal Neural Crest Nuclear Envelope SOX10 Transcription Factor Tissue, Membrane Xenopus laevis Zebrafish
3
3D Deep Learning for Subcellular Localization
Supplementary Table 1 outlines the data used to train and evaluate the models based on 3D live cell z-stacks, including train-test data splits. All multi-channel z-stacks were obtained from a database of images produced by the Allen Institute for Cell Science’s microscopy pipeline (see http://www.allencell.org). For each of the 11 hiPSC cell lines, we randomly selected z-stacks from the database and paired the transmitted light channel with the EGFP/RFP channel to train and evaluate models (Fig. 1c) to predict the localization of the tagged subcellular structure. The transmitted light channel modality was bright-field for all but the DIC-to-nuclear envelope model. For the DNA model data, we randomly selected 50 z-stacks from the combined pool all bright-field-based z-stacks and paired the transmitted light channel with the Hoechst channel. The training set for the DNA+ model was further expanded to 540 z-stacks with additional images from the Allen Institute for Cell Science’s database. Note that while a CellMask channel was available for all z-stacks, we did not use this channel because the CAAX-membrane cell line provided higher quality images for training cell membrane models. A single z-stack time series of wild-type hiPSCs was used only for evaluation (Fig. 1e).
For experiments testing the effects of number of training images on model performance (Supplementary Fig. 3), we supplemented each model’s training set with additional z-stacks from the database. Z-stacks of HEK-293 cells were used to train and evaluate DNA models whereas all z-stacks of cardiomyocytes and of HT-1080 cells were used only for evaluation (Supplementary Fig. 5). The 2D DNA model (Supplementary Fig. 4) used the same data as the DNA+ model.
All z-stacks were converted to floating-point and were resized via cubic interpolation such that each voxel corresponded to 0.29 μm × 0.29 μm × 0.29 μm, and resulting images were 244 px × 366 px for 100x-objective images or 304 px × 496 px for 63x-objective images in Y and X respectively and between 50 and 75 pixels in Z. Pixel intensities of all input and target images were z-scored on a per-image basis to normalize any systematic differences in illumination intensity.
Ounkomol C., Seshamani S., Maleckar M.M., Collman F, & Johnson G.R. (2018). Label-free prediction of three-dimensional fluorescence images from transmitted light microscopy. Nature methods, 15(11), 917-920.
Publication 2018
Cell Lines Cells Cuboid Bone HEK293 Cells Human Induced Pluripotent Stem Cells Light Microscopy Myocytes, Cardiac Nuclear Envelope Plasma Membrane Tissue, Membrane
4
Sequential Detergent Extraction for Polysome Fractionation
This method takes advantage of the relatively high cholesterol content of the plasma membrane, as compared to other cellular membranes. Digitonin is a ß-sterol binding detergent that selectively solubilizes the plasma membrane, leaving the ER- and nuclear membranes intact. Hence, sequential treatment with digitonin followed by a more lytic detergent, such as an NP-40/DOC cocktail, yields cytosolic- and membrane-bound polysome fractions, respectively (schematically illustrated in Fig. 1A). The various steps of the sequential detergent extraction procedure have been validated by immunofluorescene microscopy, where it can be seen that disruption of the plasma membrane with digitonin results in the release of (depolymerized) tubulin, without effect on the ER, the actin cytoskeleton, or the intermediate filament network (Fig. 1 B). Following addition of the ER lysis buffer, the ER fraction is recovered in a soluble fraction and the nuclei, actin cytoskeleton, and intermediate filament network remain (Fig. 1B). Companion immunoblot analyses of marker protein distributions show that the cytosolic proteins GAPDH and tubulin are present in the cytosol fraction, as expected, and the ER-membrane proteins, TRAPα and ER-lumenal protein, GRP94 are present in the ER fraction (Fig. 2 A). The NP-40 insoluble material consists primarily of nuclear and cytoskeletal elements, as evidenced by the marker proteins histone H3 and actin, respectively (Fig. 2 A). Similarly, Northern blot analysis of the mRNA composition of the cytosol and membrane fractions show that the cytosol fraction is enriched for mRNAs encoding histone (H3F3A) and GAPDH, whereas the membrane fraction is enriched in mRNAs encoding ER resident proteins, such as GRP94 and calreticulin (Fig. 2 B).
The method described below is for cells grown in monolayer. However, the protocol can be easily adapted for non-adherent cells by performing permeabilization, wash and lysis in suspension and pelleting cells at 3000 × g for 5 minutes between the different steps. The volumes of reagents mentioned in the following protocol are scaled to extract polysomes from 10 million cells.
Jagannathan S., Nwosu C, & Nicchitta C.V. (2011). Analyzing Subcellular mRNA Localization via Cell Fractionation. Methods in molecular biology (Clifton, N.J.), 714, 301-321.
Publication 2011
Actins Buffers Calreticulin Cell Nucleus Cells Cytoskeleton Cytosol Detergents Digitonin GAPDH protein, human Gastrin-Secreting Cells GRP94 Histone H3 Histones Hypercholesterolemia Immunoblotting Intermediate Filaments Membrane Proteins Microfilaments Microscopy Nonidet P-40 Northern Blotting Nuclear Envelope Pets Plasma Membrane Polyribosomes Proteins RNA, Messenger Sterols Tissue, Membrane Tubulin Vision
5
3D Deep Learning for Subcellular Localization
Supplementary Table 1 outlines the data used to train and evaluate the models based on 3D live cell z-stacks, including train-test data splits. All multi-channel z-stacks were obtained from a database of images produced by the Allen Institute for Cell Science’s microscopy pipeline (see http://www.allencell.org). For each of the 11 hiPSC cell lines, we randomly selected z-stacks from the database and paired the transmitted light channel with the EGFP/RFP channel to train and evaluate models (Fig. 1c) to predict the localization of the tagged subcellular structure. The transmitted light channel modality was bright-field for all but the DIC-to-nuclear envelope model. For the DNA model data, we randomly selected 50 z-stacks from the combined pool all bright-field-based z-stacks and paired the transmitted light channel with the Hoechst channel. The training set for the DNA+ model was further expanded to 540 z-stacks with additional images from the Allen Institute for Cell Science’s database. Note that while a CellMask channel was available for all z-stacks, we did not use this channel because the CAAX-membrane cell line provided higher quality images for training cell membrane models. A single z-stack time series of wild-type hiPSCs was used only for evaluation (Fig. 1e).
For experiments testing the effects of number of training images on model performance (Supplementary Fig. 3), we supplemented each model’s training set with additional z-stacks from the database. Z-stacks of HEK-293 cells were used to train and evaluate DNA models whereas all z-stacks of cardiomyocytes and of HT-1080 cells were used only for evaluation (Supplementary Fig. 5). The 2D DNA model (Supplementary Fig. 4) used the same data as the DNA+ model.
All z-stacks were converted to floating-point and were resized via cubic interpolation such that each voxel corresponded to 0.29 μm × 0.29 μm × 0.29 μm, and resulting images were 244 px × 366 px for 100x-objective images or 304 px × 496 px for 63x-objective images in Y and X respectively and between 50 and 75 pixels in Z. Pixel intensities of all input and target images were z-scored on a per-image basis to normalize any systematic differences in illumination intensity.
Ounkomol C., Seshamani S., Maleckar M.M., Collman F, & Johnson G.R. (2018). Label-free prediction of three-dimensional fluorescence images from transmitted light microscopy. Nature methods, 15(11), 917-920.
Publication 2018
Cell Lines Cells Cuboid Bone HEK293 Cells Human Induced Pluripotent Stem Cells Light Microscopy Myocytes, Cardiac Nuclear Envelope Plasma Membrane Tissue, Membrane

Most recents protocols related to «Nuclear Envelope»

1
Visualizing Nuclear Envelope Elongation
To visualize the nuclear envelope, we followed the same procedures described previously (Maitra et al. 2022 (link)). Briefly, when the nucleus is near-spherical, the ratio of the long to short nuclear axes is 1. However, as the nucleus starts to expand, the ratio increases, signifying the elongation of the nuclear envelope (Fig. 6).
Hammer S.E, & Polymenis M. (2023). One-carbon metabolic enzymes are regulated during cell division and make distinct contributions to the metabolome and cell cycle progression in Saccharomyces cerevisiae. G3: Genes|Genomes|Genetics, 13(3), jkad005.
Full text: Click here
Publication 2023
Cell Nucleus Epistropheus Nuclear Envelope
2
Annotating Mitotic Trait Combinations
All four analyzed traits were coded as binary traits (presence “1” or absence “0”). While the sexual reproduction may be either present or absence, the mitotic traits have to be seen a little different. An absence for the closed nuclear division means that the nuclear envelope is open or semi-open during mitosis. For the intranuclear spindles, an absence corresponds to extranuclear spindles and the absence of orthomitosis (axial symmetry) stands for pleuromitosis (bilateral symmetry). The annotation of traits is based on literature (supplementary table S1, Supplementary Material online). Not every species in our data set is annotated for every trait in literature. We therefore applied a majority rule for each group in question. If there is data on the exact ancestral state of a trait, we annotated it to be present in the whole group. In cases where only one representative of a group is annotated in literature, this annotation was suspected to be present in the whole group. Groups with different annotations for different members were annotated by majority rule. No cases with a 50:50 distribution were found in our data set. Two species in our data set are annotated with incompatible mitotic combinations (closed orthomitosis with extranuclear spindle): Chlamydomonas reinhardtii and Volvox carteri, both members of the taxon Chlorophyceae. Although the combination of traits itself is incompatible, the majority rule resulted in this combination for the group.
Bremer N., Tria F.D., Skejo J, & Martin W.F. (2023). The Ancestral Mitotic State: Closed Orthomitosis With Intranuclear Spindles in the Syncytial Last Eukaryotic Common Ancestor. Genome Biology and Evolution, 15(3), evad016.
Publication 2023
Chlamydomonas reinhardtii Chlorophyceae Mitosis Nuclear Division Nuclear Envelope Reproduction Vision Volvox
3
Quantifying Chromosome Pairing Dynamics
Leptotene/zygotene nuclei staining positively for S8 pSUN-1 were randomly selected from high-resolution Z-stacks. The region of the inner nuclear envelope where chromosomes cluster during pairing (i.e., the region corresponding to adjacent or overlapping DAPI and NPC staining) was selected as a region of interest (ROI). A line-plot of raw pixel intensities for S8 pSUN-1 and GRAS-1::GFP signals along the ROI was generated using the Multichannel Plot Profile plugin for Fiji ImageJ [70 (link)]. Pixel intensity values for each ROI were used to calculate the Pearson correlation coefficient as a measure of normalized covariance of the signals. Coefficients calculated from 45 leptotene/zygotene nuclei across 9 gonads were then averaged.
Martinez-Garcia M., Naharro P.R., Skinner M.W., Baran K.A., Lascarez-Lagunas L.I., Nadarajan S., Shin N., Silva-García C.G., Saito T.T., Beese-Sims S., Diaz-Pacheco B.N., Berson E., Castañer A.B., Pacheco S., Martinez-Perez E., Jordan P.W, & Colaiácovo M.P. (2023). GRAS-1 is a novel regulator of early meiotic chromosome dynamics in C. elegans. PLOS Genetics, 19(2), e1010666.
Full text: Click here
Publication 2023
Cell Nucleus Chromosomes DAPI Glucocorticoid-Remediable Aldosteronism Gonads Nuclear Envelope
4
Cytoplasmic and Nuclear Protein Extraction
The cell pellet was resuspended in five pellet volumes of extraction buffer A (cytoplasmic extraction buffer: Combine 20 mm Tris, pH 7.6, 0.1 mm EDTA, 2 mm MgCl2, 0.5 mm NaF, 0.5 m Na3VO4. To yield ready‐to‐use extraction buffer, protease inhibitors were supplemented by adding 10 µL of 100× protease inhibitor and 10 µL of 100 mm PMSF to 980 µL of extraction buffer A shortly before use. The cells were incubated for 15 min on ice to induce hypotonic swelling of cells as a preparative step for subsequent cell lysis. Nonidet P‐40 was added to obtain a final concentration of 1% and was mixed gently by vortexing or inverting the tube. This induced cell membrane disruption and the release of cytoplasmic proteins while keeping nuclear membranes intact. Broken cells were homogenized by gently pipetting up and down three times.4 °C, 500 × g for 5 min. ≈80% of the supernatant was aspirated and transferred it to a new 1.5‐mL microcentrifuge tube (cytoplasmic extract). The residual 20% was thoroughly discarded, 10–15 pellet volumes buffer (as above) was added to the pellet of crude nuclei. The nuclei were gently resuspended by pipetting up and down and further purifying the crude nuclear preparation. The nuclei were pelleted by centrifugation at 4 °C and 500 × g for 3 min and the supernatant was roughly discarded. The rest was the nuclear fraction.
Li Y., Fu Y., Zhang Y., Duan B., Zhao Y., Shang M., Cheng Y., Zhang K., Yu Q, & Wang T. (2023). Nuclear Fructose‐1,6‐Bisphosphate Inhibits Tumor Growth and Sensitizes Chemotherapy by Targeting HMGB1. Advanced Science, 10(7), 2203528.
Full text: Click here
Publication 2023
Buffers Cell Nucleus Cells Centrifugation Cytoplasm Edetic Acid Magnesium Chloride Nonidet P-40 Nuclear Envelope Plasma Membrane Protease Inhibitors Proteins Tromethamine
5
Karyosome Defects in Oocytes
The workflow in the second round of the screen is exactly the same as described above, except that 20–29 oocytes were examined per candidate gene. One hundred six genes showed karyosome abnormalities in 25% or more of all examined oocytes in the second round. These 106 genes were considered to show reproducible karyosome defects and therefore studied further. In all 106 genes with karyosome defects, there were two major types of karyosome defects: chromatin attachment to the nuclear envelope and chromatin distortion in the nucleus.
Nieken K.J., O’Brien K., McDonnell A., Zhaunova L, & Ohkura H. (2023). A large-scale RNAi screen reveals that mitochondrial function is important for meiotic chromosome organization in oocytes. Chromosoma, 132(1), 1-18.
Full text: Click here
Publication 2023
Cell Nucleus Chromatin Congenital Abnormality congenital defects Genes Nuclear Envelope Oocytes

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More about "Nuclear Envelope"

The nuclear envelope, also known as the nuclear membrane, is a crucial cellular structure that surrounds the nucleus of eukaryotic cells.
It acts as a selective barrier, controlling the exchange of materials between the nucleus and the cytoplasm.
This double-membrane structure is composed of an outer nuclear membrane, an inner nuclear membrane, and nuclear pore complexes that facilitate the transport of molecules.
The nuclear envelope plays a vital role in various cellular processes, including gene expression, chromatin organization, and cell signaling.
Researchers can unlock the secrets of this essential structure using advanced tools and techniques.
For example, the SimpleChIP Enzymatic Chromatin IP Kit can be used to study chromatin-associated proteins, while the Subcellular Protein Fractionation Kit for Cultured Cells can help isolate and analyze the nuclear envelope proteins.
Imaging techniques, such as the Eclipse Ti microscope, can provide high-resolution visualizations of the nuclear envelope and its dynamics.
Fluorescent dyes like Hoechst 33342 can be used to stain the nuclear envelope and track its movements.
Computational tools, such as MATLAB, can be employed to analyze and model the complex interactions within the nuclear envelope.
To ensure the integrity of their experiments, researchers may utilize protease inhibitor cocktails, such as the Complete protease inhibitor cocktail, to protect proteins from degradation.
Fluorescent labeling with Alexa Fluor 488 can also be used to visualize and track specific proteins within the nuclear envelope.
By leveraging these advanced tools and techniques, researchers can gain a deeper understanding of the nuclear envelope and unlock its secrets.
The AI-driven platform PubCompare.ai can help optimize research protocols by comparing the latest literature, preprints, and patents, enabling researchers to streamline their process and achieve better results in their studies of this essential cellular structure.