Actin Colocalization with Metaphase Chromosomes of the Second Meiosis in Ovulated Mouse Oocytes
Actin distribution in the area of meiosis II metaphase plate. (a–c) Monomeric actin distribution in oocytes stained with Alexa Fluor 488-DNase-I for G-actin and TO-PRO-3 for DNA; (a) actin fluorescence after Alexa Fluor 488-DNase-I labeling; (b) DNA staining by TO-PRO-3; (c) merged image, arrow—metaphase plate, bar −20 μm. (d–f) Double staining for F-actin with TRITC-phalloidin and for DNA with TO-PRO-3; (d) actin stained with TRITC-phalloidin; (e) chromosomes stained with DNA probe TO-PRO-3; (f) merged image, bar −8 μm. (g–i) Double staining for actin by antibody against fragment of N-terminal domain of actin and for DNA by TO-PRO-3; (g) fluorescence of antibody-labeled actin; (h) DNA staining by TO-PRO-3; (i) merged image, bar–24 μm. (j-k) Antibody against fragment of N-terminal domain of actin recognizes microfilaments near metaphase plate; (j) fluorescent staining with antibody, arrows—actin filaments; (k) double staining for actin by antibody and for DNA by TO-PRO-3, arrow—metaphase plate, bar −8 μm. (m-n) After latrunculin B treatment antibody against N-terminus cannot detect actin filaments near chromosomes; (m) fluorescent staining with antibody; (n) double staining for actin by antibody and for DNA by TO-PRO-3, bar −10 μm. (l and o) Disruption of cortical microfilament layer in oocytes after latrunculin B treatment; (l) F-actin stained with TRITC- phalloidin, bar −24 μm; (o) actin fluorescence after labeling by antibody against fragment of N-terminal domain, bar −24 μm.