The Origins and Functions of Tissue-Resident Macrophages in Kidney Development

Munro, David A. D.; Hughes, Jeremy

doi:10.3389/fphys.2017.00837

Cited by 101 publications

(92 citation statements)

References 124 publications

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“…As expected, Actb was uniformly expressed in kidney and heart while Scgb1a1 and Sftpc were undetectable (Supplementary Figure 2 A, B). In both tissues, Lyz2 was expressed by a few scattered cells, which presumably correspond to macrophages (37, 38) (Supplementary Figure 2). In a subset of kidney tubular structures, we detected Napsa (Supplementary Figure 2 A), which agrees with the previously described immunohistochemical detection of the marker in renal proximal tubule cells (39).…”

Section: Resultsmentioning

confidence: 99%

SCRINSHOT, a spatial method for single-cell resolution mapping of cell states in tissue sections

Sountoulidis¹,

Liontos²,

Nguyen³

et al. 2020

Preprint

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16 17 ǂ Corresponding authors: Christos Samakovlis (christos.samakovlis@scilifelab.se), Alexandros 18 Sountoulidis (alexandros.sountoulidis@scilifelab.se) 19 20 Keywords: SCRINSHOT, padlock-probe, multiplex RNA FISH, spatial cell-type map, lung 21 22 2 23 Abstract 24 Changes in cell identities and positions underlie tissue development and disease progression. 25 Although, single-cell mRNA sequencing (scRNA-Seq) methods rapidly generate extensive lists of 26 cell-states, spatially resolved single-cell mapping presents a challenging task. We developed 27 SCRINSHOT (Single Cell Resolution IN Situ Hybridization On Tissues), a sensitive, multiplex 28 RNA mapping approach. Direct hybridization of padlock probes on mRNA is followed by 29 circularization with SplintR ligase and rolling circle amplification (RCA) of the hybridized padlock 30 probes. Sequential detection of RCA-products using fluorophore-labeled oligonucleotides profiles 31 thousands of cells in tissue sections. We evaluated SCRINSHOT specificity and sensitivity on 32 murine and human organs. SCRINSHOT quantification of marker gene expression shows high 33 correlation with published scRNA-Seq data over a broad range of gene expression levels. We 34 demonstrate the utility of SCRISHOT by mapping the locations of abundant and rare cell types 35 along the murine airways. The amenability, multiplexity and quantitative qualities of SCRINSHOT 36 facilitate single cell mRNA profiling of cell-state alterations in tissues under a variety of native and 37 experimental conditions. 38 39 40 41 42 43 44 45 65addressed by the sequential fluorescence in situ hybridization (seqFISH) (14, 15) and the 66 multiplexed error-robust FISH (MERFISH) (4). These approaches utilize sequential rounds of 67 hybridization of FISH probes or barcode-based primary probes to detect multiple RNA species. 68The outstanding throughput of these methods makes them strong candidates for generation of 69 spatial transcriptome maps in tissues. However, since the principle of these techniques is similar 70 to smFISH, they require large number of gene-specific probes, confocal or super-resolution 4 71 microscopy to deconvolve the signals and complicated algorithms for both probe design and 72 analysis. Nevertheless, the low signal-to-noise ratio still remains a major technical challenge of 73 these methods, especially for tissue sections with strong auto-fluorescence from structural 74 extracellular matrix components like collagen and elastin (16). New strategies for signal 75 amplifications, such as branched-DNA amplification (RNAScope) (17) and hybridization chain 76 reaction (18, 19), have been recently combined with sophisticated probe design (AmpFISH) (20) 77 to increase sensitivity and specificity of smFISH. 78 Padlock probes have been successfully used to detect RNA species (21). They are linear DNA 79 molecules, with complementary arms to the target mRNA sequence and a common "backbone". 80 Upon hybridization with the target sequence, they can be ligated, creating circular single-stranded 81...

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Section: Resultsmentioning

confidence: 99%

SCRINSHOT, a spatial method for single-cell resolution mapping of cell states in tissue sections

Sountoulidis¹,

Liontos²,

Nguyen³

et al. 2020

Preprint

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“…The vast majority of our knowledge on human DCs comes from work on human peripheral blood. However, it is becoming increasingly clear that the function of DCs and many immune cells differs depending on their anatomical location (14)(15)(16)(17)(18)(19)(20)(21), which highlights the importance of studying immune cells at the site where they exert their function.…”

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confidence: 99%

Human Blood and Tonsil Plasmacytoid Dendritic Cells Display Similar Gene Expression Profiles but Exhibit Differential Type I IFN Responses to Influenza A Virus Infection

Vangeti

Gertow

et al. 2019

The Journal of Immunology

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Influenza A virus (IAV) infection constitutes an annual health burden across the globe. Plasmacytoid dendritic cells (PDCs) are central in antiviral defense because of their superior capacity to produce type I IFNs in response to viruses. Dendritic cells (DCs) differ depending on their anatomical location. However, only limited host-pathogen data are available from the initial site of infection in humans. In this study, we investigated how human tonsil PDCs, likely exposed to virus because of their location, responded to IAV infection compared with peripheral blood PDCs. In tonsils, unlike in blood, PDCs are the most frequent DC subset. Both tonsil and blood PDCs expressed several genes necessary for pathogen recognition and immune response, generally in a similar pattern. MxA, a protein that renders cells resistant to IAV infection, was detected in both tonsil and blood PDCs. However, despite steady-state MxA expression and contrary to previous reports, at high IAV concentrations (typically cytopathic to other immune cells), both tonsil and blood PDCs supported IAV infection. IAV exposure resulted in PDC maturation by upregulation of CD86 expression and IFN-α secretion. Interestingly, blood PDCs secreted 10-fold more IFN-α in response to IAV compared with tonsil PDCs. Tonsil PDCs also had a dampened cytokine response to purified TLR ligands compared with blood PDCs. Our findings suggest that tonsil PDCs may be less responsive to IAV than blood PDCs, highlighting the importance of studying immune cells at their proposed site of function.

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“…Tissue resident macrophages also play critical roles in tissue remodeling during development; the macrophages are involved in killing and removing interdigital cells in the mouse paw (Hopkinson-Woolley et al, 1994), developing the mouse eye hyaloid vessels and pupillary membrane (Bishop et al, 2016), bridging blood vessel anastomoses in the zebrafish embryo (Fantin et al, 2010), and branching morphogenesis in the murine kidney (Munro and Hughes, 2017). On the basis of the observation that the hemogenic activity of the endocardium in the heart tube coincides with the developmental remodeling of cardiac tissue, the authors hypothesized that these events might be intrinsically linked.…”

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confidence: 99%