DEV00733 Movies

Files in this Data Supplement:

• Movie 1 - DIC videomicrograph of circulation in the tail of a 3 dpf zebrafish embryo (real time, 30 frames/second). Anterior is towards the left, dorsal is upwards. The functional vessels present at this stage are the anteroposteriorly aligned dorsal aorta (DA) and posterior cardinal vein (PCV), and dorsoventrally aligned intersegmental arteries (ISA) and intersegmental veins (ISV)
• Movie 2 - Timelapse multiphoton confocal images of vessel dynamics in fli-EGFP transgenic zebrafish. Initial emergence of primary angiogenic sprouts from the dorsal aorta. Filopodial activity precedes synchronous emergence of sprouts. Anterior is towards the left, dorsal is upwards. Images were collected beginning at ~18 somite stage. Forty-one time points were collected with 5 minutes between time points (~3.5 hours total time). Movie frame rate is 6 frames/second.
• Movie 3 - Timelapse multiphoton confocal images of vessel dynamics in fli-EGFP transgenic zebrafish. Emergence and elongation of primary angiogenic sprouts. Anterior is towards the left, dorsal is up. Images were collected beginning at ~20 somite stage. Ninety-five time points were collected, with 5 minutes between time points (~8 hours total time). Movie frame rate is 30 frames/second.
• Movie 4 - Timelapse multiphoton confocal images of vessel dynamics in fli-EGFP transgenic zebrafish. Highly dynamic behavior of an elongating primary angiogenic sprout, and formation of a dorsal branch. Anterior is towards the left, dorsal is upwards. Images were collected beginning at ~1.2 days post-fertilization. Seventy-five time points were collected, with 1.6 minutes between time points (~2 hours total time). Movie frame rate is 10 frames/second.
• Movie 5 - Timelapse multiphoton confocal images of vessel dynamics in fli-EGFP transgenic zebrafish. Elongation of a primary angiogenic sprout and formation of a dorsal branch. Anterior is towards the left, dorsal is upwards. Images were collected beginning at ~1.2 days post-fertilization. Forty-one time points were collected with 5 minutes between time points (~3.5 hours total time). Movie frame rate is 6 frames/second.
• Movie 6 - Timelapse multiphoton confocal images of vessel dynamics in fli-EGFP transgenic zebrafish. Emergence and fate of a secondary angiogenic sprout from the posterior cardinal vein. After initial extension and retraction of a number of filopodia on the posterior cardinal vein, a ‘definitive’ secondary sprout emerges and extends dorsally. This secondary sprout makes contact with the primary angiogenic segment, and after an extended period of time fuses with it and lumenizes. This results in formation of an intersegmental vein at this position. The connection to the dorsal aorta regresses, although this is not visualized well here. The arrow indicates the relevant position (there are shifts in this position because of movement of the animal during time-lapse collection). Anterior is towards the left, dorsal is upwards. Images were collected beginning at ~1.5 days post-fertilization. Two-hundred and two time points were collected with 6.6 minutes between time points (~22 hours total time). Movie frame rate is 15 frames/second.
• Movie 7 - Timelapse multiphoton confocal images of vessel dynamics in fli-EGFP transgenic zebrafish. Dynamics of a secondary intersegmental vessel sprout that does not connect to the primary intersegmental vessel. Numerous filopodial extensions can be seen extending from this sprout as it tracks dorsally in close apposition to the adjacent primary vessel. Although it appears that some of these filopodia contact the primary vessel, an effective connection between these vessels is not made. The vessel at this position therefore becomes an intersegmental artery. Images were collected beginning at ~1.6 days post-fertilization. One-hundred and eighty-six time points were collected with 5 minutes between time points (~15.5 hours total time). Movie frame rate is 15 frames/second.
• Movie 8 - Timelapse multiphoton confocal images of vessel dynamics in fli-EGFP transgenic zebrafish. Regression of the ventral primary connection to the dorsal aorta occurs in a vessel functioning robustly as an intersegmental vein. A thin vestigial vascular segment connects a large patent intersegmental vein to the dorsal aorta. This segment is initially partially patent, but gradually loses all patency, continues to thin and eventually entirely regresses, disconnecting from the dorsal aorta and collapsing into the intersegmental vein. At the ‘jump’ in the movie, there is ~1 hour gap in the time-lapse sequence. Images were collected beginning at ~1.6 days post-fertilization. One-hundred and eighty-six time points were collected with 5 minutes between time points (~15.5 hours total time). Movie frame rate is 15 frames/second.
• Movie 9 - Timelapse multiphoton confocal images of vessel dynamics in fli-EGFP transgenic zebrafish. Dynamics of a secondary angiogenic sprout that serves as a ventral root for the parachordal vessel system. A secondary sprout can be seen extending dorsally and rostrally past the primary vessel (which has become an intersegmental artery). As this secondary sprout reaches the horizontal myoseptum, it branches rostrally and caudally along the myoseptum. Images were collected beginning at ~1.8 days post-fertilization. Forty-eight time points were collected with 10 minutes between time points (~8 hours total time). Movie frame rate is 6 frames/second.
• Movie 10 - DIC videomicrograph of circulation through a ‘dual connected’ intersegmental in the trunk of a 2.5 dpf zebrafish embryo (this is the vessel shown in Fig. 7B). Blood flow is shunted ventrally from the dorsal aorta to posterior cardinal vein (red arrow). There is no flow through dorsal portions of the vessel (the blood cells observed moving ventralwards in the more dorsal parts of the trunk at this position are actually moving through a different vessel on the opposite side of the trunk). Anterior is towards the left, dorsal is upwards.
• Movie 11 - DIC videomicrograph of circulation through two separate ‘dual connected’ intersegmentals in the tail of a 2.5 dpf zebrafish embryo (these are the vessels shown in Figures 7C,D). Blood flow is shunted ventrally from the dorsal aorta to posterior cardinal vein (red arrows). There is no flow through dorsal regions of either vessel. Anterior is towards the left, dorsal is upwards.