title: Matching Patterns of Gene Expression to Mechanical Stiffness at Cell Resolution through Quantitative Tandem Epifluorescence and Nanoindentation.
abstract: Cell differentiation has been associated with changes in mechanical stiffness in single-cell systems, yet it is unknown whether this association remains true in a multicellular context, particularly in developing tissues. In order to address such questions, we have developed a methodology, termed quantitative tandem epifluorescence and nanoindentation, wherein we sequentially determine cellular genetic identity with confocal microscopy and mechanical properties with atomic force microscopy. We have applied this approach to examine cellular stiffness at the shoot apices of Arabidopsis (Arabidopsis thaliana) plants carrying a fluorescent reporter for the CLAVATA3 (CLV3) gene, which encodes a secreted glycopeptide involved in the regulation of the centrally located stem cell zone in inflorescence and floral meristems. We found that these CLV3-expressing cells are characterized by an enhanced stiffness. Additionally, by tracking cells in young flowers before and after the onset of GREEN FLUORESCENT PROTEIN expression, we observed that an increase in stiffness coincides with this onset. This work illustrates how quantitative tandem epifluorescence and nanoindentation can reveal the spatial and temporal dynamics of both gene expression and cell mechanics at the shoot apex and, by extension, in the epidermis of any thick tissue.
authors.affiliation: Reproduction et Développement des Plantes, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique (P.M., V.M., C.C., F.R., O.H., P.D., A.B.), and Laboratoire Joliot-Curie, Centre National de la Recherche Scientifique (P.M., V.M., O.H., P.D., A.B.), Ecole Normal Supérieure de Lyon, Université de Lyon, 69364 Lyon cedex 07, France.
authors.forename: Pascale
authors.initials: P
authors.lastname: Milani
authors.affiliation: Reproduction et Développement des Plantes, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique (P.M., V.M., C.C., F.R., O.H., P.D., A.B.), and Laboratoire Joliot-Curie, Centre National de la Recherche Scientifique (P.M., V.M., O.H., P.D., A.B.), Ecole Normal Supérieure de Lyon, Université de Lyon, 69364 Lyon cedex 07, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Mirabet
authors.affiliation: Reproduction et Développement des Plantes, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique (P.M., V.M., C.C., F.R., O.H., P.D., A.B.), and Laboratoire Joliot-Curie, Centre National de la Recherche Scientifique (P.M., V.M., O.H., P.D., A.B.), Ecole Normal Supérieure de Lyon, Université de Lyon, 69364 Lyon cedex 07, France.
authors.forename: Coralie
authors.initials: C
authors.lastname: Cellier
authors.affiliation: Reproduction et Développement des Plantes, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique (P.M., V.M., C.C., F.R., O.H., P.D., A.B.), and Laboratoire Joliot-Curie, Centre National de la Recherche Scientifique (P.M., V.M., O.H., P.D., A.B.), Ecole Normal Supérieure de Lyon, Université de Lyon, 69364 Lyon cedex 07, France.
authors.forename: Frédérique
authors.initials: F
authors.lastname: Rozier
authors.affiliation: Reproduction et Développement des Plantes, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique (P.M., V.M., C.C., F.R., O.H., P.D., A.B.), and Laboratoire Joliot-Curie, Centre National de la Recherche Scientifique (P.M., V.M., O.H., P.D., A.B.), Ecole Normal Supérieure de Lyon, Université de Lyon, 69364 Lyon cedex 07, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Hamant
authors.affiliation: Reproduction et Développement des Plantes, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique (P.M., V.M., C.C., F.R., O.H., P.D., A.B.), and Laboratoire Joliot-Curie, Centre National de la Recherche Scientifique (P.M., V.M., O.H., P.D., A.B.), Ecole Normal Supérieure de Lyon, Université de Lyon, 69364 Lyon cedex 07, France pradeep.das@ens-lyon.fr arezki.boudaoud@ens-lyon.fr.
authors.forename: Pradeep
authors.initials: P
authors.lastname: Das
authors.affiliation: Reproduction et Développement des Plantes, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique (P.M., V.M., C.C., F.R., O.H., P.D., A.B.), and Laboratoire Joliot-Curie, Centre National de la Recherche Scientifique (P.M., V.M., O.H., P.D., A.B.), Ecole Normal Supérieure de Lyon, Université de Lyon, 69364 Lyon cedex 07, France pradeep.das@ens-lyon.fr arezki.boudaoud@ens-lyon.fr.
authors.forename: Arezki
authors.initials: A
authors.lastname: Boudaoud