Cell-Specific Markers for the Identification of Retinal Cells and Subcellular Organelles by Immunofluorescence Microscopy

2019 ◽  
pp. 293-310 ◽  
Author(s):  
Laurie L. Molday ◽  
Christiana L. Cheng ◽  
Robert S. Molday
Keyword(s):  
Immunofluorescence Microscopy ◽  
Retinal Cells ◽  
Subcellular Organelles

scholarly journals Interferon action: nucleolar and nucleoplasmic localization of the interferon-inducible 72-kD protein that is encoded by the Ifi 204 gene from the gene 200 cluster.

1992 ◽  
Vol 116 (6) ◽  
pp. 1333-1341 ◽  
Author(s):  
D Choubey ◽  
P Lengyel
Keyword(s):  
Ribosomal Rna ◽  
Protein Function ◽  
Immunofluorescence Microscopy ◽  
Polyclonal Antibodies ◽  
Sequence Similarity ◽  
Chromosome 1 ◽  
Cdna Clones ◽  
Subcellular Organelles ◽  
Indirect Immunofluorescence Microscopy ◽  
Alpha Spectrin

The interferons are cytokines with antiviral, cell growth regulatory, and immunomodulatory activities. These activities are mediated by the proteins induced by the interferons. Earlier we described a gene cluster (the 200 cluster) consisting of at least six adjacent, interferon-activatable genes located next to the erythroid alpha-spectrin locus on murine chromosome 1. The genes of the cluster arose by repeated gene duplication and they specify proteins with pronounced sequence similarity. We have now raised polyclonal antibodies against a segment from one of these proteins (the 204 protein of 72 kD). Using these, we established that the 204 protein is a phosphoprotein whose level in cells from various murine lines can be increased up to 75-fold upon treatment with alpha interferon. Experiments involving fractionation of cell lysates and indirect immunofluorescence microscopy of control and interferon-treated cells revealed that the 204 protein is nucleolar and nucleoplasmic. This conclusion was confirmed by co-localization with B23, a known nucleolar protein. The 204 protein is the first interferon-induced protein found to be located in the nucleoli, the subcellular organelles of ribosomal RNA production and ribosome assembly. It remains to be seen whether the 204 protein affects any of these processes. Studies on 204 protein function should be facilitated by the availability of complete cDNA clones and the finding of cell lines in which the expression of this protein is impaired.

The application of video-enhanced constrast/differential interference constrast microscopy in conjunction with whole mount electron microscopy to the study of organelle transport

1988 ◽  
Vol 46 ◽  
pp. 66-67
Author(s):  
J. H. Hayden
Keyword(s):  
Electron Microscopy ◽  
Rana Pipiens ◽  
Silver Film ◽  
Immunofluorescence Microscopy ◽  
Organelle Transport ◽  
Linear Element ◽  
Whole Mount ◽  
Carbon Coated ◽  
Linear Elements ◽  
Dic Microscopy

In a previous study, Allen video-enhanced constrast/differential interference constrast (AVEC-DIC) microscopy was used in conjunction with immunofluorescence microscopy to demonstrate that organelles and vesicle move in either direction along linear elements composed of microtubules. However, this study was limited in that the number of microtubules making up a linear element could not be determined. To overcome this limitation, we have used AVEC-DIC microscopy in conjunction with whole mount electron microscopy.Keratocytes from Rana pipiens were grown on glass coverslips as described elsewhere. Gold London Finder grids were Formvar- and carbon coated, and sterilized by exposure to ultraviolet light. It is important to select a Formvar film that gives a grey reflection when it is floated on water. A silver film is too thick and will detract from the image in the light microscope.

Assemblies of D-peptides for Targeting Cell Nucleolus

2019 ◽  
Author(s):  
Huaimin Wang ◽  
Zhaoqianqi Feng ◽  
Weiyi Tan ◽  
Bing Xu
Keyword(s):  
Particle Morphology ◽  
Side Chain ◽  
Mechanism Study ◽  
Structural Analogue ◽  
Hydrophobic Residues ◽  
Subcellular Organelles ◽  
First Case ◽  
New Strategy ◽  
Peptide Derivative ◽  
Peptide Nanoparticles

<p>Selectively targeting cell nucleolus remains a challenge. Here we report the first case that D-peptides form membraneless molecular condensates with RNA for targeting cell nucleolus. A D-peptide derivative, enriched with lysine and hydrophobic residues, self-assembles to form nanoparticles, which enter cells through clathrin dependent endocytosis and mainly accumulate at cell nucleolus. Structural analogue of the D-peptide reveals that particle morphology of the assemblies, which depends on the side chain modification, favors the cellular uptake. Contrasting to those of the D-peptide, the assemblies of the corresponding L-enantiomer largely localize in cell lysosomes. Preliminary mechanism study suggests that the D-peptide nanoparticles interact with RNA to form membraneless condensates in the nucleolus, which further induces DNA damage and results in cell death. This work illustrates a new strategy for rationally designing supramolecular assemblies of D-peptides for targeting subcellular organelles.</p>

The Effect of Claudin-5 Overexpression on the Interactions of Claudin-1 and -2 and Barrier Function in Retinal Cells

2014 ◽  
Vol 14 (9) ◽  
pp. 1226-1237 ◽  
Author(s):  
R. Tian ◽  
Y. Luo ◽  
Q. Liu ◽  
M. Cai ◽  
J. Li ◽  
...  
Keyword(s):  
Barrier Function ◽  
Retinal Cells

Antiproliferative Effects of Ocimum basilicum Methanolic Extract and Fractions, Oleanolic Acid and 3-epi-Ursolic Acid

2020 ◽  
Vol 6 (2) ◽  
pp. 134-146 ◽  
Author(s):  
Kehkashan Arshad Qamar ◽  
Ahsana Dar Farooq ◽  
Bina S. Siddiqui ◽  
Nurul Kabir ◽  
Sabira Begum
Keyword(s):  
Ursolic Acid ◽  
Oleanolic Acid ◽  
Mitotic Spindle ◽  
Immunofluorescence Microscopy ◽  
Methanolic Extract ◽  
Folk Medicine ◽  
Ocimum Basilicum ◽  
Antiproliferative Effects ◽  
Ancient Times ◽  
Mcf 7

Aims: The aim of the current study was to identify active compound(s) responsible for the antiproliferative effects of O. basilicum and explore their underlying mechanism/s. Background: Plants have been the source of medicines for the treatment of various diseases since ancient times. Ocimum basilicum (Sweet Basil, Bobai Tulsi) has been used in the folk medicine for the treatment of human liver, spleen and stomach cancers. Background: Plants have been the source of medicines for the treatment of various diseases since ancient times. Ocimum basilicum (Sweet Basil, Bobai Tulsi) has been used in the folk medicine for the treatment of human liver, spleen and stomach cancers. Objective: To emphasize the importance of O. basilicum as a potential novel non-toxic alternative to the conventional anticancer therapy. Method: O. basilicum (aerial parts) methanolic extract and fractions were screened against HT-144, MCF-7, NCI-H460 and SF-268 human cancer cell lines using sulforhodamine B assay. The more active Petroleum Ether Insoluble (PEI) fraction was fractionated into six sub-fractions (OB-1 to OB-6). Four pure compounds (3-O-methyl ursolic acid, oleanolic acid, 3-epi-ursolic acid and ursolic acid) were isolated from the more potent sub-fraction OB- 6. Triple channel immunofluorescence microscopy was employed to observe the effects of methanolic extract, PEI fraction, sub-fractions OB-5 and OB-6, 3-epi-ursolic acid and oleanolic acid on the cytoskeleton and nuclei of MCF-7 cells. Result: The methanolic extract and the PEI fraction exhibited selectively greater growth inhibition against MCF-7 cell line (TGI: 56 and 36.2 µg/ml, respectively). By using triple channel immunofluorescence microscopy, it was observed that the methanolic extract, PEI fraction, sub-fraction OB-5 and 3-epi-ursolic acid induced irregular mitotic spindle formation and slowing of mitotic progression in MCF-7 cells while sub-fraction OB-6 induced mitotic arrest in the prophase stage. F-actin aggregation was also visible in PEI fraction, subfraction OB-5 and 3-epi-ursolic acid treated MCF-7 cells. Conclusion: These results emphasize the importance of O. basilicum as a potential novel non-toxic alternative to the conventional anticancer therapy and suggest that it inhibits the growth of MCF-7 cancer cells via multiple mechanisms such as interaction with the microtubules and mitotic spindle apparatus, and F-actin aggregation.

Enhanced Multiplexing of Immunofluorescence Microscopy Using a Long‐Stokes‐Shift Fluorophore

2021 ◽  
Vol 1 (8) ◽  
Author(s):  
Sydney J. Reitz ◽  
Andrew D. Sauerbeck ◽  
Terrance T. Kummer
Keyword(s):  
Immunofluorescence Microscopy ◽  
Stokes Shift

scholarly journals Long-term effects of human induced pluripotent stem cell-derived retinal cell transplantation in Pde6b knockout rats

2021 ◽  
Author(s):  
Jee Myung Yang ◽  
Sunho Chung ◽  
KyungA Yun ◽  
Bora Kim ◽  
Seongjun So ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cell Transplantation ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Histological Evaluation ◽  
Long Term Effects ◽  
Retinal Cells ◽  
Rpe Cells ◽  
Induced Pluripotent

AbstractRetinal degenerative disorders, including age-related macular degeneration and retinitis pigmentosa (RP), are characterized by the irreversible loss of photoreceptor cells and retinal pigment epithelial (RPE) cells; however, the long-term effect of implanting both human induced pluripotent stem cell (hiPSC)-derived RPE and photoreceptor for retinal regeneration has not yet been investigated. In this study, we evaluated the long-term effects of hiPSC-derived RPE and photoreceptor cell transplantation in Pde6b knockout rats to study RP; cells were injected into the subretinal space of the right eyes of rats before the appearance of signs of retinal degeneration at 2–3 weeks of age. Ten months after transplantation, we evaluated the cells using fundus photography, optical coherence tomography, and histological evaluation, and no abnormal cell proliferation was observed. A relatively large number of transplanted cells persisted during the first 4 months; subsequently, the number of these cells decreased gradually. Notably, immunohistochemical analysis revealed that the hiPSC-derived retinal cells showed characteristics of both RPE cells and photoreceptors of human origin after transplantation. Functional analysis of vision by scotopic electroretinogram revealed significant preservation of vision after transplantation. Our study suggests that the transplantation of hiPSC-derived retinal cells, including RPE cells and photoreceptors, has a potential therapeutic effect against irreversible retinal degenerative diseases.

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