Direct investigation of viscosity of an atypical inner membrane of Bacillus spores: A molecular rotor/FLIM study

Pauline Loison 1 Neveen Hosny 2 Patrick Gervais 1 Dominique Champion 3 Marina Kuimova 2 Jean-Marie Perrier-Cornet 1
1 PMB - Procédés Microbiologiques et Biotechnologiques
PAM - Procédés Alimentaires et Microbiologiques, PAM - Procédés Alimentaires et Microbiologiques [Dijon]
2 Department of Chemistry [Imperial College London]
3 PAPC - Procédés Alimentaires et Physico-Chimie
PAM - Procédés Alimentaires et Microbiologiques
Abstract : We utilize the fluorescent molecular rotor Bodipy-C12 to investigate the viscoelastic properties of hydrophobic layers of bacterial spores Bacillus subtilis. The molecular rotor shows a marked increase in fluorescence lifetime, from 0.3 to 4ns, upon viscosity increase from 1 to 1500cP and can be incorporated into the hydrophobic layers within the spores from dormant state through to germination. We use fluorescence lifetime imaging microscopy to visualize the viscosity inside different compartments of the bacterial spore in order to investigate the inner membrane and relate its compaction to the extreme resistance observed during exposure of spores to toxic chemicals. We demonstrate that the bacterial spores possess an inner membrane that is characterized by a very high viscosity, exceeding 1000cP, where the lipid bilayer is likely in a gel state. We also show that this membrane evolves during germination to reach a viscosity value close to that of a vegetative cell membrane, ca. 600cP. The present study demonstrates quantitative imaging of the microscopic viscosity in hydrophobic layers of bacterial spores Bacillus subtilis and shows the potential for further investigation of spore membranes under environmental stress.
Keywords : Bacillus subtilis spores Fluorescence Lifetime Imaging (FLIM) Lipid membranes Microviscosity Molecular rotors
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Journal articles
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Submitted on : Tuesday, October 1, 2019 - 2:40:46 PM
Last modification on : Thursday, October 24, 2019 - 4:32:05 PM

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Pauline Loison, Neveen Hosny, Patrick Gervais, Dominique Champion, Marina Kuimova, et al.. Direct investigation of viscosity of an atypical inner membrane of Bacillus spores: A molecular rotor/FLIM study. Biochimica et Biophysica Acta:Biomembranes, Elsevier, 2013, 1828 (11), pp.2436-2443. ⟨10.1016/j.bbamem.2013.06.028⟩. ⟨hal-02302386⟩

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