High resolution microscopy reveals the nuclear shape of budding yeast during cell cycle and in various biological states.
How spatial organization of the genome depends on nuclear shape is unknown, mostly because accurate nuclear size and shape measurement is technically challenging. In large cell populations of the yeast Saccharomyces cerevisiae, we assessed the geometry (size and shape) of nuclei in three dimensions with a resolution of 30 nm. We improved an automated fluorescence localization method by implementing a post-acquisition correction of the spherical microscopic aberration along the z-axis, to detect the three dimensional (3D) positions of nuclear pore complexes (NPCs) in the nuclear envelope. Here, we used a method called NucQuant to accurately estimate the geometry of nuclei in 3D throughout the cell cycle. To increase the robustness of the statistics, we aggregated thousands of detected NPCs from a cell population in a single representation using the nucleolus or the spindle pole body (SPB) as references to align nuclei along the same axis. We could detect asymmetric changes of the nucleus associated with modification of nucleolar size. Stereotypical modification of the nucleus toward the nucleolus further confirmed the asymmetric properties of the nuclear envelope.
Laboratoire de Biologie Moléculaire Eucaryote, Centre de Biologie Intégrative (CBI), Université de Toulouse, CNRS, UPS, Toulouse 31000, France.
Renjie
0000-0002-2460-2175
R
Wang
Laboratoire de Biologie Moléculaire Eucaryote, Centre de Biologie Intégrative (CBI), Université de Toulouse, CNRS, UPS, Toulouse 31000, France.
Alain
A
Kamgoue
Laboratoire de Biologie Moléculaire Eucaryote, Centre de Biologie Intégrative (CBI), Université de Toulouse, CNRS, UPS, Toulouse 31000, France.
Christophe
C
Normand
Laboratoire de Biologie Moléculaire Eucaryote, Centre de Biologie Intégrative (CBI), Université de Toulouse, CNRS, UPS, Toulouse 31000, France.
Isabelle
I
Léger-Silvestre
Laboratoire de Biologie Cellulaire et Moléculaire du Contrôle de la Prolifération, Centre de Biologie Intégrative (CBI), Université de Toulouse, CNRS, UPS, Toulouse 31000, France.
Thomas
T
Mangeat
Laboratoire de Biologie Moléculaire Eucaryote, Centre de Biologie Intégrative (CBI), Université de Toulouse, CNRS, UPS, Toulouse 31000, France olivier.gadal@ibcg.biotoul.fr.
Olivier
0000-0001-9421-0831
O
Gadal
D002244:Carbon / Q000494:pharmacology; D002453:Cell Cycle / Q000187:drug effects; D053144:Cell Nucleus Shape / Q000187:drug effects; D059585:G1 Phase Cell Cycle Checkpoints / Q000187:drug effects; D021621:Imaging, Three-Dimensional; D007399:Interphase / Q000187:drug effects; D018613:Microscopy, Confocal / Q000379:methods*; D009685:Nuclear Envelope / Q000187:drug effects / Q000378:metabolism; D028861:Nuclear Pore Complex Proteins / Q000378:metabolism; D004718:Saccharomycetales / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism
D028861:Nuclear Pore Complex Proteins; D002244:Carbon
Localization microscopy; Nuclear geometry; Nuclear pore complex; Super resolution microscopy