GenEPi: Piezo1-based fluorescent reporter for visualizing mechanical stimuli with high spatiotemporal resolution

Yaganoglu, Sine; Helassa, Nordine; Bm, Gaub; Welling, Maaike; Shi, Jing; Müller, DJ; Török, Katalin; Pantazis, Periklis

doi:10.1101/702423

Cited by 6 publications

(5 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Next, we evaluated Piezo1 localization and activity in both resting and dividing cells transfected with a genetically encoded fluorescent Ca 2+ reporter tagged to Piezo1 (GenEPI) ( 25 ). Resting cells showed plasma membrane GenEPI signal reinforcement (Fig.…”

Section: Resultsmentioning

confidence: 99%

The mechanosensitive Piezo1 channel controls endosome trafficking for an efficient cytokinetic abscission

et al. 2021

View full text Add to dashboard Cite

show abstract

Section: Resultsmentioning

confidence: 99%

The mechanosensitive Piezo1 channel controls endosome trafficking for an efficient cytokinetic abscission

et al. 2021

View full text Add to dashboard Cite

show abstract

“…Furthermore, other mechanosensor channels, such as Trpv4, show enrichment in the AV region 50 . Therefore, it would be interesting to generate fluorescent fusion lines of the mechanosensors essential for cardiac valve formation in order to track their trafficking within endocardial cells and study the signals required for their activation 51 .…”

Section: Discussionmentioning

confidence: 99%

Multiple pkd and piezo gene family members are required for atrioventricular valve formation

et al. 2023

View full text Add to dashboard Cite

Cardiac valves ensure unidirectional blood flow through the heart, and altering their function can result in heart failure. Flow sensing via wall shear stress and wall stretching through the action of mechanosensors can modulate cardiac valve formation. However, the identity and precise role of the key mechanosensors and their effectors remain mostly unknown. Here, we genetically dissect the role of Pkd1a and other mechanosensors in atrioventricular (AV) valve formation in zebrafish and identify a role for several pkd and piezo gene family members in this process. We show that Pkd1a, together with Pkd2, Pkd1l1, and Piezo2a, promotes AV valve elongation and cardiac morphogenesis. Mechanistically, Pkd1a, Pkd2, and Pkd1l1 all repress the expression of klf2a and klf2b, transcription factor genes implicated in AV valve development. Furthermore, we find that the calcium-dependent protein kinase Camk2g is required downstream of Pkd function to repress klf2a expression. Altogether, these data identify, and dissect the role of, several mechanosensors required for AV valve formation, thereby broadening our understanding of cardiac valvulogenesis.

show abstract

“…Alternatively, assuming that local intracellular calcium concentration is the highest at the vicinity of the inner mouth of the P2X permeation pathway, using GCaMPs in which point mutations are introduced in EF hand and RS20 peptide in order to lower the affinity for calcium (Helassa et al, 2016) should reduce nonspecific activation of PG6. Such a strategy was successfully used in the case of GenEPi, a Piezo1-based fluorescent reporter for visualizing mechanical stimuli (Yaganoglu et al, 2019).…”

Section: Discussionmentioning

confidence: 99%

P2X-GCaMPs as versatile tools for imaging extracellular ATP signaling

Ollivier

Beudez²,

Linck

et al. 2020

Preprint

View full text Add to dashboard Cite

Adenosine 5' triphosphate (ATP) is an extracellular signaling molecule involved in numerous physiological and pathological processes. Yet, in situ characterization of the spatiotemporal dynamic of extracellular ATP is still challenging due to the lack of sensor with appropriate specificity, sensitivity and kinetics. Here we report the development of biosensors based on the fusion of cation permeable ATP receptors (P2X) to genetically encoded calcium sensors (GECI). By combining the features of P2X receptors with the high signal to noise ratio of GECIs, we generated ultrasensitive green and red fluorescent sniffers that detect nanomolar ATP concentrations in situ and also enable the tracking of P2X receptor activity. We provide the proof of concept that these sensors can dynamically track ATP release evoked by neuronal depolarization or by extracellular hypotonicity. Targeting these P2X-based biosensors to diverse cell types should advance our knowledge of extracellular ATP dynamics in vivo.coupled P2Y receptors (von Kügelgen and Harden, 2011), mainly P2Y1, P2Y2 and P2Y11, although these receptors display higher affinity for other endogenous nucleotide such as ADP or UTP (von Kügelgen, 2019). Yet, activation of P2Y receptors by ATP seems to have important physiological functions, particularly in the central nervous system (A. Weisman et al., 2012). Pharmacology of P2Y receptors is considerably more developed than that of P2X receptors, particularly for P2Y receptors that are sensitive to ATP (Jacobson and Müller, 2016). However, because these proteins are coupled to intracellular signaling pathways, studying the dynamic of their activation in integrated preparation mostly relies on indirect readthrough.Studying extracellular ATP signaling in multicellular preparations and in vivo is complex. First, in vertebrate virtually all cells can release ATP through various mechanisms such as classical vesicular release, lysosomal exocytosis, transmembrane channels or cell lysis (Lohman et al., 2012;Praetorius and Leipziger, 2009;Rassendren and Audinat, 2016). Signals triggering ATP release remain poorly characterized, although there is clear evidence that cells constitutively release ATP (Lazarowski et al., 2011;Sivaramakrishnan et al., 2012) and that evoked release can be triggered in defined physiological and pathological contexts (Lazarowski, 2008). A second level of complexity comes from the short half-life of extracellular ATP. Due to the ubiquitous expression of membrane-bound and soluble ectonucleotidases, extracellular ATP is within seconds to minutes degraded in ADP and ultimately adenosine, which both also act as signaling molecules (Deaglio and Robson, 2011;Kukulski et al., 2011).Various methods have been developed to measure extracellular ATP (Wu and Li, 2020). The most common is the luciferase-luciferin assay which allows bioluminescent quantification of ATP in solution. A genetically encoded version of this assay was developed (Pellegatti et al., 2005), however because of its low quantic yield, this approach h...

show abstract

GenEPi: Piezo1-based fluorescent reporter for visualizing mechanical stimuli with high spatiotemporal resolution

Cited by 6 publications

References 42 publications

The mechanosensitive Piezo1 channel controls endosome trafficking for an efficient cytokinetic abscission

The mechanosensitive Piezo1 channel controls endosome trafficking for an efficient cytokinetic abscission

Multiple pkd and piezo gene family members are required for atrioventricular valve formation

P2X-GCaMPs as versatile tools for imaging extracellular ATP signaling

Contact Info

Product

Resources

About