Supplemental Movies

Images of sperm obtained after incubation in media of low, medium, or high osmolality, or with cytochalasin, are shown in 3D to display F-actin structure and staining patterns throughout the sperm. The images obtained were deconvolved (using the deconvolution module in softWoRx) and reconstructed into 3D volumes. The movies shown are 3D representations of the specimens and are not time-lapse videos. Note that the contrast in videos was optimized to demonstrate the F-actin cytoskeletal architecture along the sides of the sperm head, which stained quite intensely

Files in this Data Supplement:

• Supplemental Movie Fig6B3D4 - "Sperm incubated in medium-osmolality media containing 20 μM CB display the brightly stained F-actin structures along the sperm head similar to that seen in sperm after incubation in control and high-osmolality conditions. This movie shows the same sperm depicted in Fig. 6B and 6D, in a 3D projection."
• Supplemental Movie Fig3C3D2 - "Sperm subjected to low-osmolality conditions exhibited a diminished distribution of F-actin stain throughout the sperm. This movie shows the same sperm depicted in Fig. 3 (C, D)."
• Supplemental Movie Fig7A3D5 - "Sperm incubated in control medium exhibited F-actin in the flagellum that is organized in a discrete, punctate pattern. This movie shows the same sperm depicted in Fig. 7A and 7E, in a 3D projection."
• Supplemental Movie Fig3E3D3 - "Subjection to high-osmolality conditions induced the formation of areas of F-actin structure (also referred to as bands) in the apical portion of the sperm head above the PA region. Areas of brightly stained F-actin cytoskeletal structures along the head and in the midpiece are also apparent. This movie shows the same sperm depicted in Fig. 3 (E, F)."
• Supplemental Movie Fig3a3D1 - "Sperm incubated in medium-osmolality media exhibited areas of brightly stained F-actin structure along the head, neck and in the mid and proximal pieces of the tail. The sperm shown (same image as in Fig. 3, A and B) was stained with phalloidin-FITC and was imaged using a Delta Vision System, Olympus IX70 microscope and cooled CCD camera. Data were collected as z sections and were deconvolved (using the deconvolution module in softWoRx) and reconstructed into 3D volumes. This movie contains a 3D projection that was rotated 180° for a full view of the sample, displayed at 15 frames/sec. Note that the contrast in videos was optimized to demonstrate the F-actin cytoskeletal architecture along the sides of the sperm head, which stained quite intensely, therefore the head staining patterns are not as apparent as in the 2D images."