Range of SHH signaling in adrenal gland is limited by membrane contact to cells with primary cilia

Mateska, Ivona; Nanda, Kareena; Dye, Natalie A.; Alexaki, Vasileia Ismini; Eaton, Suzanne

doi:10.1083/jcb.201910087

Cited by 13 publications

(13 citation statements)

References 93 publications

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“…Vimentin-deficient SW13/cl.2 adrenocortical carcinoma cells, used a negative control, displayed short structures positive for ACE2 and acetylated tubulin or ARL13B (Suppl. Figure 6 ), consistent with the imperfect cilia formation of this cell type 59 . Furthermore, these structures lacked vimentin staining, confirming the specificity of vimentin detection at primary cilia shown in Fig.…”

Section: Resultssupporting

confidence: 81%

Cell surface detection of vimentin, ACE2 and SARS-CoV-2 Spike proteins reveals selective colocalization at primary cilia

Lalioti

González-Sanz

Lois-Bermejo

et al. 2022

Sci Rep

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The SARS-CoV-2 Spike protein mediates docking of the virus onto cells prior to viral invasion. Several cellular receptors facilitate SARS-CoV-2 Spike docking at the cell surface, of which ACE2 plays a key role in many cell types. The intermediate filament protein vimentin has been reported to be present at the surface of certain cells and act as a co-receptor for several viruses; furthermore, its potential involvement in interactions with Spike proteins has been proposed. Nevertheless, the potential colocalization of vimentin with Spike and its receptors on the cell surface has not been explored. Here we have assessed the binding of Spike protein constructs to several cell types. Incubation of cells with tagged Spike S or Spike S1 subunit led to discrete dotted patterns at the cell surface, which consistently colocalized with endogenous ACE2, but sparsely with a lipid raft marker. Vimentin immunoreactivity mostly appeared as spots or patches unevenly distributed at the surface of diverse cell types. Of note, vimentin could also be detected in extracellular particles and in the cytoplasm underlying areas of compromised plasma membrane. Interestingly, although overall colocalization of vimentin-positive spots with ACE2 or Spike was moderate, a selective enrichment of the three proteins was detected at elongated structures, positive for acetylated tubulin and ARL13B. These structures, consistent with primary cilia, concentrated Spike binding at the top of the cells. Our results suggest that a vimentin-Spike interaction could occur at selective locations of the cell surface, including ciliated structures, which can act as platforms for SARS-CoV-2 docking.

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Section: Resultssupporting

confidence: 81%

Cell surface detection of vimentin, ACE2 and SARS-CoV-2 Spike proteins reveals selective colocalization at primary cilia

Lalioti

González-Sanz

Lois-Bermejo

et al. 2022

Sci Rep

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“…As a negative control for these observations we studied vimentin-deficient SW13/cl.2 adrenocortical carcinoma cells. Both primary cells and carcinoma cell lines from the cortex of the adrenal gland are defective in primary cilia formation, in relation with low levels of acetylated tubulin and marginal expression of ARL13B [57]. Consistent with this, SW13/cl.2 cells displayed scarce and short structures positive for ACE2 and acetylated tubulin or ARL13B, and did not show vimentin staining (Suppl.…”

Section: Detection Of Ace2 and Vimentin On The Surface Of Cellsmentioning

confidence: 54%

“…As a negative control for these observations we studied vimentin-deficient SW13/cl.2 adrenocortical carcinoma cells. Although cells and from the cortex of the adrenal gland display imperfect primary cilia formation in relation with low levels of acetylated tubulin and ARL13B [57], we were able to spot some short structures positive for ACE2 and acetylated tubulin or ARL13B in SW13/cl.2 cells, consistent with short cilia (Suppl. Fig.…”

Section: Detection Of Ace2 and Vimentin On The Surface Of Cellsmentioning

confidence: 56%

Immunolocalization studies of vimentin and ACE2 on the surface of cells exposed to SARS-CoV-2 Spike proteins

Lalioti

González-Sanz

Lois-Bermejo

et al. 2021

Preprint

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The Spike protein from SARS-CoV-2 mediates docking of the virus onto cells and contributes to viral invasion. Several cellular receptors are involved in SARS-CoV-2 Spike docking at the cell surface, including ACE2 and neuropilin. The intermediate filament protein vimentin has been reported to be present at the surface of certain cells and act as a co-receptor for several viruses; furthermore, its potential involvement in interactions with Spike proteins has been proposed. Here we have explored the binding of Spike protein constructs to several cell types using low-temperature immunofluorescence approaches in live cells, to minimize internalization. Incubation of cells with tagged Spike S or Spike S1 subunit led to discrete dotted patterns at the cell surface, which showed scarce colocalization with a lipid raft marker, but consistent coincidence with ACE2. Under our conditions, vimentin immunoreactivity appeared as spots or patches unevenly distributed at the surface of diverse cell types. Remarkably, several observations including potential antibody internalization and adherence to cells of vimentin-positive structures present in the extracellular medium exposed the complexity of vimentin cell surface immunoreactivity, which requires careful assessment. Notably, overall colocalization of Spike and vimentin signals markedly varied with the cell type and the immunodetection sequence. In turn, vimentin-positive spots moderately colocalized with ACE2; however, a particular enrichment was detected at elongated structures positive for acetylated tubulin, consistent with primary cilia, which also showed Spike binding. Thus, these results suggest that vimentin-ACE2 interaction could occur at selective locations near the cell surface, including ciliated structures, which can act as platforms for SARS-CoV-2 docking.

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“…Indeed, at the time of floor-plate induction, junctions between neural tube cells loosen, changing the restrictive nature of this barrier (Aaku-Saraste et al, 1996). Shh might therefore move between cells by diffusion, although recent work in cell culture (Li et al, 2018) and the adrenal gland (Mateska et al, 2020) suggests that membrane contact with ciliated receiving cells, perhaps by the extension of thin cytonemes through the space between cells (Sanders et al, 2013), is necessary for Shh transport.…”

Section: Ligand Source and Its Relationship To Cilium Location In Epitheliamentioning

confidence: 99%

Hedgehog signaling and the primary cilium: implications for spatial and temporal constraints on signaling

Stearns

2021

Development

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The mechanisms of vertebrate Hedgehog signaling are linked to the biology of the primary cilium, an antenna-like organelle that projects from the surface of most vertebrate cell types. Although the advantages of restricting signal transduction to cilia are often noted, the constraints imposed are less frequently considered, and yet they are central to how Hedgehog signaling operates in developing tissues. In this Review, we synthesize current understanding of Hedgehog signal transduction, ligand secretion and transport, and cilia dynamics to explore the temporal and spatial constraints imposed by the primary cilium on Hedgehog signaling in vivo.

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Range of SHH signaling in adrenal gland is limited by membrane contact to cells with primary cilia

Cited by 13 publications

References 93 publications

Cell surface detection of vimentin, ACE2 and SARS-CoV-2 Spike proteins reveals selective colocalization at primary cilia

Cell surface detection of vimentin, ACE2 and SARS-CoV-2 Spike proteins reveals selective colocalization at primary cilia

Immunolocalization studies of vimentin and ACE2 on the surface of cells exposed to SARS-CoV-2 Spike proteins

Hedgehog signaling and the primary cilium: implications for spatial and temporal constraints on signaling

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