Cool microscopy following AMF formation and collapse on plant root for several days live imaging

Such a cool method visualization of spatial temporal patterns

See their paper on AMslide:
onlinelibrary.wiley.com/doi/full/10....

#KNVMmycology

Q2) We haven't tried. I know that there are some methods to visualize cytoplasmic dynamics using single-GFP tracking. But just by intensity, I don't know, maybe combine with FRAP?. @robertarkowitz.bsky.social do you think is possible?

Hi Darren! Thanks for your interest :)
To answer your questions: Q1) I haven't look into it in detail, but check this very interesting work pubmed.ncbi.nlm.nih.gov/37740366/ in yeast; not immobilised but confined and under pressure, from @delarueresearch.bsky.social

Ready :) bsky.app/profile/anto...

PS: I am missing many funding organizations and colleagues in this network, mainly in Europe, I hope they will join soon!

If you would like to listen to an AI-generated summary of this work, it can be found here: sciencecast.org/casts/rvmtna... - Automatically done after submission of our work to
@biorxivpreprint.bsky.social, really cool use of technology! The AI summary is surprisingly good! (11/11)

and of course the funding organizations for making it possible! @cnrs.bsky.social, Inserm, Université Cote d'Azur, Université Toulouse, UMassChan, Charite Berlin @hhmi.bsky.social, MSCActions, ERC_Research, FRM, Laas-CNRS and Institute Biology Valrose (10/11)

This work wouldn’t be possible without the incredible collaboration of wonderful colleagues.Thanks to Charles Puerner, Emily Plumb, Louis Chevalier, Johannes Elferich, Ludwig Sinn, Niko Grigorieff, Markus Ralser, @delarueresearch.bsky.social, Martine Bassilana and
@robertarkowitz.bsky.social (9/11)

In summary, our results reveal a substantial fluidization of the cytoplasm during morphogenesis, a key process for fungal virulence, via a reduction in ribosome concentration and that importantly inhibition of ribosome biogenesis triggers filamentous growth (8/11)

Depletion of a protein critical for ribosome biogenesis resulted in a reduction of rRNA levels and an increase in GEM Deff. Strikingly this also resulted in the induction of the yeast-to-filament transition (both morphologically and of hyphal specific genes) w/o serum. (7/11)

Mass spectrometry revealed that ribosomal protein levels drop during filamentation. To directly visualize this, we carried out Cryo-EM+2D template matching and observed that filamentous cells have a lower ribosome concentration, which was proportional to their length. (6/11)

🧮 Mathematical modeling (both theoretical predictive equations and simulations) recapitulate our observations, that is the ~2-fold increase in Deff as cell volume increases. This supports our initial hypothesis: there is a substantial dilution of a cytoplasmic crowder. (5/11)

We found GEM Deff increases 2x during filamentation, indicating that the cytoplasm becomes more fluid and likely less crowded. Fluidity correlates very nicely with filament length/volume, suggesting a substantial dilution of molecular crowders e.g. ribosomes. (4/11)

The cytoplasm is highly crowded, and this affects a range of biological functions. We tracked how crowded the cytoplasm is during morphogenesis by using genetically encoded GEMs. Lower GEM effective diffusion (Deff) = higher crowding. This is the cytoplasm at 30 fps 🎥 (3/11)

Using a passive fluorescent microrheological probe (GEMs), we reveal how cytoplasmic fluidity changes during yeast-to-filament morphogenesis, a key process for fungal virulence. (2/11)