FocalPlane features… preLights recording

In our FocalPlane features… preLights webinar, Felipe Del Valle Batalla and Vibha Singh, two members of the preLights team, presented live preLights highlighting preprinted research from Carmen Rodilla-Ramírez, Francesca Bottanelli and colleagues on the link between T-cell protrusions and CAR signalling, and from Eva Mejía-Ramírez, Carolina Florian and colleagues on the role of RhoA in stem cell ageing. Following the preLight of their work, Carmen and Eva presented research talks. Each talk was followed by a short Q&A, and we wrapped up with a general discussion on preprinting research.

You can view the recording below and view the answers to the questions that we didn’t have time to address during the webinar. You can read Vibha’s written preLight on ‘Turning back the clock on stem cells‘ over on the preLights website.

Questions for Carmen:

Q: Great talk! Did you have any problems with phototoxicity?
A: Addressing phototoxicity is particularly important for STED imaging. When we image with STED, we use far-red lasers, that are less energetic and have lower associated phototoxicity. Nevertheless, we make sure that we do not image repeatedly the same cell and we acquire a single frame. The only case in which we imaged twice the same area using STED was for the Lck imaging, and we space the acquisition time frames enough to let the cell
recover.

Q: Thank you this is excellent! The proteins clearly localise to the protrusions. Are the ZAP etc phosphorylating also at that initial contact, or do they get phosphorylated when the full IS contact happens?
A: Yes, they are phosphorylated already at these early contacts. We can confidently assume this because very clear LAT clustering occurs at the protrusions. As ZAP-70 is the kinase responsible for LAT recruitment, we can conclude that ZAP-70 has been phosphorylated and has then phosphorylated LAT. LAT clustering further suggests that downstream proteins (e.g., Sos1, Grb2) are recruited.

Q: Do you read preprints and published manuscripts ‘differently’; or, on the other side, do you notice any difference in quality between what they readnin preprint and in press?
A: For me, peer review usually increases the trust in the results (for me as a PhD student). However, I cannot say that I see an overall quality difference between preprints and published manuscripts, as this strongly depends on which specific manuscript/preprint I am reading.

Questions for Eva:

Q: Have you tried Heterochronic parabiosis between the young and aged mice.
A: No, we have not. We are focused on investigating specific targets in molecular mechanisms of aging in haematopoietic stem cells. Haematopoietic stem cells rejuvenation contributes to improve the life span of aged mice as we have demonstrated previously (10.1038/s41536-022-00275-y). We are interested in the BM residing stem cells function rather than on the circulating, with the purpose of improving the regenerative capacity and differentiation of aged stem cells. We understand that by doing parabiosis experiments we are not going to see the function of stem cells in the bone marrow.

Q: You mentioned that this was your first preprint, why did you decide to go for it for this research?
A: We chose to go for the preprint to give more visibility to our work while we were performing the revision on peer reviewed journals. We understood that our results were of high relevance and novelty since we demonstrate for the first time that nuclear mechanics have a consequence in vivo in haematopoietic stem cells, and, moreover, we have identified a targetable mechanism of ageing.

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