Non-isotopic electron microscope in situ hybridization for studying the functional sub-compartmentalization of the cell nucleus

Puvion‐Dutilleul, Francine; Puvion, Edmond

doi:10.1007/bf02473202

Cited by 18 publications

(13 citation statements)

References 79 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…32. Electron microscopy in situ hybridization was performed over ultrathin sections from Lowicryl K4M embedded samples (37). Immunocytochemical detection of digoxigenin was performed with antidigoxigenin antibody conjugated to gold particles that were 10 nm in diameter (Boehringer Mannheim).…”

Section: Methodsmentioning

confidence: 99%

Isolation and expression of a Pax-6 gene in the regenerating and intact Planarian Dugesia(G)tigrina

Callaerts

Munoz-Marmol

Glardon

et al. 1999

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

The Pax-6 gene encodes a transcription factor containing both a paired and a homeodomain and is highly conserved among Metazoa. In both vertebrates and invertebrates, Pax-6 is required for eye morphogenesis, development of parts of the central nervous system, and, in some phyla, for the development of olfactory sense organs. Ectopic expression of Pax-6 from insects, mammals, cephalopods, and ascidians induces ectopic eyes in Drosophila, suggesting that Pax-6 may be a universal master control gene for eye morphogenesis. Platyhelminthes are an ancient phylum, originating from the base of spiralian protostomes, that bear primitive eyes, consisting of a group of rhabdomeric photoreceptor cells enclosed in a cup of pigment cells. The analysis of Pax-6 and its expression pattern should provide insights into the ancestral function of Pax-6 in eye morphogenesis. We have identified the Pax-6 gene of the planarian Dugesia(G)tigrina (Platyhelminthes; Turbellaria; Tricladida). This gene shares significant sequence identity and conserved genomic organization with Pax-6 proteins from other phyla. Phylogenetic analysis indicates that it clusters with the other Pax-6 genes, but in the most basal position. DtPax-6 is expressed as a single transcript in both regenerating and fully grown eyes, and electron microscopy studies show strong expression in the perykarion of both photoreceptor and pigment cells. Very low levels of expression also are detectable in other body regions. Because a bona fide Pax-6 homolog so far has not been detected in diploblastic animals, we speculate that Pax-6 may be typical for triploblasts and that the appearance of additional Pax genes may have coincided with increasingly complex body plans.

show abstract

Section: Methodsmentioning

confidence: 99%

Isolation and expression of a Pax-6 gene in the regenerating and intact Planarian Dugesia(G)tigrina

Callaerts

Munoz-Marmol

Glardon

et al. 1999

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…Herpes simplex virus type 1 (HSV-1) Us11 protein is a true late gene product packaged within the virion and is delivered into cells after infection [l-41. It exhibits a nucleo-cytoplasmic localization at early times of infection, and later accumulates in the nucleoli, but is still present in the cytoplasm [5,6]. In addition to these characteristics, Us1 1 protein is an RNA-binding protein and even associates with a specific nucleotide sequence localized at the 3'-end of a truncated UL34 mRNA [7].…”

Section: Introductionmentioning

confidence: 99%

Phosphorylation of herpes simplex virus type 1 Us11 protein is independent of viral genome expression

et al. 1995

View full text Add to dashboard Cite

The Us11 protein is a true late gene product of herpes simplex virus type 1 (HSV-1), whose exact function is unknown but which exhibits RNA-binding properties and which is phosphorylated on serine residues. In order to determine whether the Us11 protein is phosphorylated by cellular kinase(s) or by virally encoded kinase(s), the Us11 gene has been cloned and transiently expressed in HeLa cells. In addition, HeLa-derived cell lines have been selected for their ability to express Us11 protein constitutively. 32P-Labeling and analysis by two-dimensional electrophoresis of transiently and constitutively expressed Us11 protein demonstrated that, indeed, multiple phosphorylation of the protein occurs in absence of HSV-1 genome expression, indicating that the protein behaves as a natural substrate for cellular kinase(s). In addition, a sequence heterogeneity of the Us11 protein, due to a difference in the number of SPREPR repeats, has been characterized between different strains of HSV-1.

show abstract

“…Historically, utilization of AuNPs in the electron microscopy-based ISH could date back to the late 1980s. 35 However, as a labeling material for mRNA detection AuNP did not gain much popularity until its optical properties were fully discovered. Upon irradiating a sub-200 nm AuNP with a visible wavelength of light, the free electrons in gold atom would oscillate with the incident photons, called localized surface plasmon resonance (LSPR).…”

Section: Plasmonic Ishmentioning

confidence: 99%

Beyond quantification: in situ analysis of transcriptome and pre‐mRNA alternative splicing at the nanoscale

Cui

Liu

Irudayaraj

2016

WIREs Nanomed Nanobiotechnol

View full text Add to dashboard Cite

In situ analysis offers a venue for dissecting the complex transcriptome in its natural context to tap into cellular processes that could explain the phenotypic physiology and pathology yet to be understood. Over the past decades, enormous progress has been made to improve the resolution, sensitivity, and specificity of single-cell technologies. The continued efforts in RNA research not only facilitates mechanistic studies of molecular biology but also provides state-of-the-art strategies for diagnostic purposes. The implementation of novel bio-imaging platforms has yielded valuable information for inspecting gene expression, mapping regulatory networks, and classifying cell types. In this article, we discuss the merits and technical challenges in single-molecule in situ RNA profiling. Advanced in situ hybridization methodologies developed for a variety of detection modalities are reviewed. Considering the fact that in mammalian cells the number of protein products immensely exceeds that of the actual coding genes due to pre-mRNA alternative splicing, tools capable of elucidating this process in intact cells are highlighted. To conclude, we point out future directions for in situ transcriptome analysis and expect a plethora of opportunities and discoveries in this field. WIREs Nanomed Nanobiotechnol 2017, 9:e1443. doi: 10.1002/wnan.1443 For further resources related to this article, please visit the WIREs website.

show abstract

Non-isotopic electron microscope in situ hybridization for studying the functional sub-compartmentalization of the cell nucleus

Cited by 18 publications

References 79 publications

Isolation and expression of a Pax-6 gene in the regenerating and intact Planarian Dugesia(G)tigrina

Isolation and expression of a Pax-6 gene in the regenerating and intact Planarian Dugesia(G)tigrina

Phosphorylation of herpes simplex virus type 1 Us11 protein is independent of viral genome expression

Beyond quantification: in situ analysis of transcriptome and pre‐mRNA alternative splicing at the nanoscale

Contact Info

Product

Resources

About