scholarly journals A novel immunofluorescence detection method for renal cell-type specific in situ cytokine production by confocal microscopy

MethodsX ◽  
2020 ◽  
Vol 7 ◽  
pp. 100935
Author(s):  
Sun-sang J. Sung ◽  
Shu Man Fu
Keyword(s):  
Confocal Microscopy ◽  
Renal Cell ◽  
Cytokine Production ◽  
Detection Method ◽  
Cell Type ◽  
Cell Type Specific

scholarly journals Transcriptome-based network analysis reveals renal cell type-specific dysregulation of hypoxia-associated transcripts

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Natallia Shved ◽  
Gregor Warsow ◽  
Felix Eichinger ◽  
David Hoogewijs ◽  
Simone Brandt ◽  
...  
Keyword(s):  
Network Analysis ◽  
Renal Cell ◽  
Cell Type ◽  
Cell Type Specific

scholarly journals Control of Cell Type Proportioning in Dictyostelium discoideum by Differentiation-Inducing Factor as Determined by In Situ Hybridization

2004 ◽  
Vol 3 (5) ◽  
pp. 1241-1248 ◽  
Author(s):  
Toshinari Maruo ◽  
Haruyo Sakamoto ◽  
Negin Iranfar ◽  
Danny Fuller ◽  
Takahiro Morio ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Dictyostelium Discoideum ◽  
Wild Type ◽  
Cell Type ◽  
Type Conversion ◽  
New Genes ◽  
A Cell ◽  
Cell Type Specific ◽  
Cell Type Conversion

ABSTRACT We have determined the proportions of the prespore and prestalk regions in Dictyostelium discoideum slugs by in situ hybridization with a large number of prespore- and prestalk-specific genes. Microarrays were used to discover genes expressed in a cell type-specific manner. Fifty-four prespore-specific genes were verified by in situ hybridization, including 18 that had been previously shown to be cell type specific. The 36 new genes more than doubles the number of available prespore markers. At the slug stage, the prespore genes hybridized to cells uniformly in the posterior 80% of wild-type slugs but hybridized to the posterior 90% of slugs lacking the secreted alkylphenone differentiation-inducing factor 1 (DIF-1). There was a compensatory twofold decrease in prestalk cells in DIF-less slugs. Removal of prespore cells resulted in cell type conversion in both wild-type and DIF-less anterior fragments. Thus, DIF-1 appears to act in concert with other processes to establish cell type proportions.

scholarly journals Spatially patterned excitatory neuron subtypes and circuits within the claustrum

2021 ◽  
Author(s):  
Sarah R Erwin ◽  
Brianna N Bristow ◽  
Kaitlin E Sullivan ◽  
Brian Marriott ◽  
Lihua Wang ◽  
...  
Keyword(s):  
Neural Circuits ◽  
Spatial Arrangement ◽  
Retrograde Tracing ◽  
Excitatory Neuron ◽  
Cell Type ◽  
Narrow Region ◽  
Single Cell Rna Sequencing ◽  
Cell Type Specific ◽  
Functional Complexity

The claustrum is a functionally and structurally complex brain region, whose very spatial extent remains debated. Histochemical-based approaches typically treat the claustrum as a relatively narrow region that primarily projects to the neocortex, whereas circuit-based approaches suggest a broader region embedding neocortical and other neural circuits. Here, we took a bottom up, cell-type-specific approach to complement and possibly unite these seemingly disparate conclusions. Using single-cell RNA-sequencing, we found that the claustrum is comprised of two excitatory neuron subtypes that are differentiable from the surrounding cortex. Multicolor retrograde tracing in conjunction with 12-channel multiplexed in situ hybridization revealed a core-shell spatial arrangement of these subtypes, as well as differential projection targets. Thus, the claustrum is comprised of excitatory neuron subtypes with distinct molecular and circuit properties, whose spatial patterns reflect the narrower and broader claustral extents debated in previous research. This subtype-specific heterogeneity likely shapes the functional complexity of the claustrum.

scholarly journals The landscape of S100B+ and HLA-DR+ dendritic cell subsets in tonsils at the single cell level via high-parameter mapping

2018 ◽  
Author(s):  
Maddalena Maria Bolognesi ◽  
Francesca Maria Bosisio ◽  
Marco Manzoni ◽  
Denis Schapiro ◽  
Riccardo Tagliabue ◽  
...  
Keyword(s):  
Cellular Interactions ◽  
Cell Type ◽  
Cell Level ◽  
Parameter Mapping ◽  
Hla Dr ◽  
Monocytes And Macrophages ◽  
Cell Type Specific ◽  
Dendritic Cell Subsets

SUMMARYDendritic cells (DC) (classic, plasmacytoid, inflammatory) are an intense focus of interest because of their role in inflammation, autoimmunity, vaccination and cancer. We present a tissue-based classification of human DC subsets in tonsils with a high-parameter (>40 markers) immunofluorescent approach, cell type-specific image segmentation and the use of bioinformatics platforms. Through this deep phenotypic and spatial examination, classic cDC1, cDC2, pDC subsets have been further refined and a novel subset of DC co-expressing IRF4 and IRF8 identified. Based on unique tissue locations within the tonsil, and close interactions with T cells (cDC1) or B cells (cDC2), DC subsets can be further subdivided by correlative phenotypic changes associated with these interactions. In addition, monocytes and macrophages expressing HLA-DR or S100AB are identified and localized in the tissue. This study thus provides a whole tissue in situ catalog of human DC subsets and their cellular interactions within spatially defined niches.

scholarly journals Induction of Golli-MBP Expression in CNS Macrophages During Acute LPS-Induced CNS Inflammation and Experimental Autoimmune Encephalomyelitis (EAE)

2007 ◽  
Vol 7 ◽  
pp. 112-120 ◽  
Author(s):  
Tracey L. Papenfuss ◽  
J. Cameron Thrash ◽  
Patricia E. Danielson ◽  
Pamela E. Foye ◽  
Brian S. Hllbrush ◽  
...  
Keyword(s):  
Experimental Autoimmune Encephalomyelitis ◽  
Cell Type ◽  
Autoimmune Encephalomyelitis ◽  
Cns Inflammation ◽  
Cell Type Specific ◽  
Candidate Molecule ◽  
Experimental Autoimmune

Microglia are the tissue macrophages of the CNS. Microglial activation coupled with macrophage infiltration is a common feature of many classic neurodegenerative disorders. The absence of cell-type specific markers has confounded and complicated the analysis of cell-type specific contributions toward the onset, progression, and remission of neurodegeneration. Molecular screens comparing gene expression in cultured microglia and macrophages identified Golli-myelin basic protein (MBP) as a candidate molecule enriched in peripheral macrophages.In situhybridization analysis of LPS/IFNg and experimental autoimmune encephalomyelitis (EAE)–induced CNS inflammation revealed that only a subset of CNS macrophages express Golli-MBP. Interestingly, the location and morphology of Golli-MBP+ CNS macrophages differs between these two models of CNS inflammation. These data demonstrate the difficulties of extendingin vitroobservations toin vivobiology and concretely illustrate the complex heterogeneity of macrophage activation states present in region- and stage-specific phases of CNS inflammation. Taken altogether, these are consistent with the emerging picture that the phenotype of CNS macrophages is actively defined by their molecular interactions with the CNS microenvironment.

scholarly journals Combination of in situ hybridization and immunocytochemistry to detect messenger RNAs in identified CNS neurons and glia in tissue culture.

1991 ◽  
Vol 39 (7) ◽  
pp. 891-898 ◽  
Author(s):  
P A Trimmer ◽  
L L Phillips ◽  
O Steward
Keyword(s):  
In Situ Hybridization ◽  
Cdna Probe ◽  
Cell Types ◽  
Intermediate Filament Protein ◽  
Messenger Rnas ◽  
Cell Type ◽  
Protein Subunit ◽  
The Central Nervous System ◽  
Cell Type Specific

We have developed a technique in which immunofluorescence is combined with in situ hybridization using cDNA and RNA probes to assess the expression and distribution of messenger RNAs (mRNA) by neurons and neuroglia in tissue cultures of the rat dentate gyrus. The probes used in this study include a cDNA probe for ribosomal RNA (rRNA) and an RNA probe (cRNA) for glial fibrillary acidic protein (GEAP), an intermediate filament protein subunit expressed by astrocytes in the central nervous system. Both ubiquitous (tubulin) and cell type-specific (MAP-2 and GEAP) antibodies were used to identify neurons and neuroglia in culture. Using this procedure, the mRNA for rRNA was found in the cell bodies and large processes of MAP-2-positive neurons and throughout the cytoplasm of GEAP-positive flat astrocytes. In process-bearing astrocytes, GEAP mRNA is concentrated in the cell body, although some hybridization also occurred in astrocyte cell processes. With this combined in situ hybridization-immunofluorescence technique, the expression and distribution of an mRNA can be examined in different immunocytochemically identified cell types under identical culture and hybridization conditions. It is also possible to determine if there is a differential subcellular distribution of an mRNA in a single cell and if the distribution of the mRNA reflects the distribution of the protein itself. Finally, this technique can be utilized to verify the specificity of probes for cell type-specific mRNAs and to determine appropriate hybridization conditions to produce a specific signal.

scholarly journals Identifying gene locus associations with promyelocytic leukemia nuclear bodies using immuno-TRAP

2013 ◽  
Vol 201 (2) ◽  
pp. 325-335 ◽  
Author(s):  
Reagan W. Ching ◽  
Kashif Ahmed ◽  
Paul C. Boutros ◽  
Linda Z. Penn ◽  
David P. Bazett-Jones
Keyword(s):  
Gene Locus ◽  
Immunofluorescence Microscopy ◽  
Physiological State ◽  
Tp53 Gene ◽  
Promyelocytic Leukemia ◽  
Nuclear Bodies ◽  
Cell Type ◽  
Cell Type Specific ◽  
Promyelocytic Leukemia Nuclear Bodies

Important insights into nuclear function would arise if gene loci physically interacting with particular subnuclear domains could be readily identified. Immunofluorescence microscopy combined with fluorescence in situ hybridization (immuno-FISH), the method that would typically be used in such a study, is limited by spatial resolution and requires prior assumptions for selecting genes to probe. Our new technique, immuno-TRAP, overcomes these limitations. Using promyelocytic leukemia nuclear bodies (PML NBs) as a model, we used immuno-TRAP to determine if specific genes localize within molecular dimensions with these bodies. Although we confirmed a TP53 gene–PML NB association, immuno-TRAP allowed us to uncover novel locus-PML NB associations, including the ABCA7 and TFF1 loci and, most surprisingly, the PML locus itself. These associations were cell type specific and reflected the cell’s physiological state. Combined with microarrays or deep sequencing, immuno-TRAP provides powerful opportunities for identifying gene locus associations with potentially any nuclear subcompartment.

Sign in / Sign up

Export Citation Format

Share Document