|
|
|
|
|
|
|
||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | ||||
| ||||||||||||||||||||
Files in this Data Supplement:
Movie 1. Proximal streaming of mitochondria (green) and PGL-1::mRFP-1 particles (red) around the loop (right) and into enlarging oocytes (bottom). Total time, 30 minutes; 2 frames/minute. At about 20 minutes, an ovulation event leads to brief, reversed streaming. Oocyte cell membranes and outer margins of the gonad are marked in white. The region shown corresponds to Fig. 1H-L. Note that the focal plane differs from the one shown in Fig. 1H-L.
Movie 2. An injected mineral oil drop (arrowhead) moves along the path of proximal streaming. The same gonad is represented in Fig. 2. Arrowhead, injected oil drop. Oocyte cell membranes and outer margins of the gonad are marked with dark-gray lines. Total time, 30 minutes; 1 frame per minute.
Movie 3. Movement of cytoplasmic PGL-1::GFP particles (green) and injected red fluorescent microspheres along the core. This low magnification movie shows most of the one gonad arm except for the distal third. Microspheres were injected into the mid-pachytene area, where most of them remain stationary throughout the movie. Some microspheres start moving in the late pachytene area, and around the loop (right) into oocytes. The oocyte at the lower left was ovulated just before the sequence shown, and a second oocyte is ovulated during the movie. Perinuclear P granules labeled with PGL-1::GFP are visible throughout the pachytene region, and cytoplasmic PGL-1::GFP particles travel along the core at about the same speed as microspheres. Note the perinuclear P granule detaching from a young oocyte nucleus (top right) and traveling through the loop into an older oocyte. Total time, 56 minutes; 2 frames/minute.
Movie 4. Proximal streaming persists after treatment with tba-2 (α-tubulin) feeding RNAi for 48 hours. Oocyte cell membranes and outer margins of the gonad are marked with dark-gray lines; broken lines indicate poorly visible membranes on highly abnormal oocytes. In the pachytene area, nuclei are inappropriately present in the core and move proximally along with streaming DIC particles. Total time, 5 minutes; 4 frames/minute.
Movie 5. Proximal streaming persists after treatment with dhc-1 (dynein heavy chain) RNAi. This worm had been injected with dhc-1 double-stranded RNA and the movie started 28 hours post-injection. Red, PGL-1::mRFP-1; green, mitochondria; white lines, outer margins of the gonad and some oocyte membranes that are clearly visible. Mitochondria and pachytene nuclei move proximally along the core and into abnormally shaped and positioned oocytes. Total time, 15 minutes.
Movie 6. Actin distribution in enlarging oocytes. This movie shows a confocal z-stack through a phalloidin-stained gonad fixed with method II. Proximal is to the left. Distance between two focal planes (z-step size) is 0.8 μm. Oriented filaments extend through the ring channels. Some of these filaments appear to be connected to the outer plasma membrane of the core (top).
Movie 7. Actin distribution in the pachytene (top), loop (right) and enlarging oocytes (bottom) at lower magnification. This movie shows a confocal z-stack through a phalloidin-stained gonad fixed with method II. Distance between two focal planes (z-step size) is 1.0 μm.
Movie 8. Normal proximal streaming in a gonad extruded in culture medium containing 10 mM EGTA (to inhibit sheath cell contractions). Oocyte cell membranes and outer margins of the gonad are marked with dark-gray lines. For corresponding particle tracks, see Fig. 4F. Total time, 5 minutes; 4 frames/minute.
Movie 9. Inhibition of myosin light chain kinase activity results in abnormal, reversed cytoplasmic streaming. Extruded gonads were incubated in 83 μM ML-7, and EGTA. In this gonad, cytoplasm exits the enlarging oocytes (streaming is reversed), while the oocyte membranes appear to stay intact. Oocyte cell membranes and outer margins of the gonad are marked with dark-gray lines. Total time, 7.5 minutes; 4 frames/minute.
Movie 10. Blocking the area between late pachytene area and loop results in slow reversed or random movement in the pachytene area (upper left), but normal proximal movement of particles in the loop (right). Note that some particles exit small oocytes in the loop area and then join streaming in the core. For corresponding particle tracks and summary diagram, see Fig. 5E,F. Oocyte cell membranes and outer margins of the gonad are marked with dark-gray lines. Injected oil drop, light-gray outline. Total time, 10 minutes; 4 frames/minute.
Movie 11. Blocking all enlarging oocytes with a large injected oil drop inhibits streaming throughout the gonad. Oocyte cell membranes and outer margins of the gonad are marked with dark-gray lines. Injected oil drop, light-gray outline. Total time, 5 minutes; 4 frames/minute.
Movie 12. Mechanically damaged oocytes lose the ability to receive cytoplasm. An oocyte was damaged with a microneedle and injected with a small oil droplet (light-gray circle, white arrowhead) as a marker. Cytoplasm streams out of the damaged oocyte and into neighboring oocytes on both sides, while normal proximal streaming continues in the loop. Particle movements are summarized by gray arrows in the final frame. Also see Fig. 5H,I for particle tracks and summary diagram. Oocyte cell membranes and outer margins of the gonad are marked with dark-gray lines. Total time, 10 minutes; 2 frames/minute.
| ||||||||||||||||||||
