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Development of blastomere clones in the Ilyanassa embryo: transformation of the spiralian blastula into the larval body plan

Abstract

Spiralian embryogenesis is deeply conserved and seems to have been in place in the last common ancestor of the large assemblage of protostome phyla known as the Lophotrochozoa. While the blastula fate maps of several spiralian embryos have been determined, little is known about the events that link the early embryo and the larva. For all cells in the Ilyanassa blastula, we determined the clonal morphology at four time points between the blastula and veliger stages. We found that ectomesoderm comes mostly from 3a and 3b, but also from 2c and 2b. We also observed the ingression and early proliferation of 3a- and 3b-derived ectomesoderm. We found cells in the 2b clone that marked the anterior edge of the blastopore and later the mouth and cells in the 3c/3d clones that marked the posterior edges of these structures. This demonstrates directly that the mouth forms in the same location as the blastopore. In the development of the shell field, we observed dramatic cell migration events that invert the positions of the 2b and 2d clones that contribute to the shell. Using time-lapse imaging, we followed and described the cleavage pattern of the conserved endomesodermal blast cell, 4d, up to 4d + 45 h, when there were 52 cells in the clone. Our results show the growth and movement of clones derived from cells of the spiralian blastula as they transform into the trochophore-like and veliger stages. They have implications for the evolution of the shell in gastropods, the origins of mesoderm in spiralians, and the evolution of mouth formation in metazoans.

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Acknowledgments

We thank Morgan Goulding, Pamela Agbu and Adam Johnson for thoughtful comments on the manuscript. This work was supported by a grant by the N.S.F to J.D.L. (IOB-1146782).

Author information

Affiliations

Authors

Corresponding author

Correspondence to J. David Lambert.

Additional information

Communicated by: David A Weisblat

Electronic supplementary material

Below is the link to the electronic supplementary material.

427_2014_474_MOESM1_ESM.mpg

Stack of confocal images of 1a clone at veliger larva stage. Anterior view. (MPG 1,374 kb)

427_2014_474_MOESM2_ESM.mpg

Stack of confocal images of 1c clone at veliger larva stage. Anterior view. (MPG 1,622 kb)

427_2014_474_MOESM3_ESM.mpg

Stack of confocal images of 1a clone at two day-old stage. Dorsal view. (MPG 2,012 kb)

427_2014_474_MOESM4_ESM.mpg

Stack of confocal images of 1c clone at two day-old stage. Dorsal view. (MPG 1,674 kb)

427_2014_474_MOESM5_ESM.mpg

Stack of confocal images of 1a clone at three day-old stage. Dorsal view. (MPG 1,326 kb)

427_2014_474_MOESM6_ESM.mpg

Stack of confocal images of 1c clone at three day-old stage. Dorsal view. (MPG 2,016 kb)

427_2014_474_MOESM7_ESM.mpg

Stack of confocal images of 1a clone at four day-old stage. Dorsal view. (MPG 1,732 kb)

427_2014_474_MOESM8_ESM.mpg

Stack of confocal images of 1c clone at four day-old stage. Dorsal view. (MPG 2,158 kb)

427_2014_474_MOESM9_ESM.mpg

Stack of confocal images of 1b clone at veliger larva stage. Anterior view. (MPG 766 kb)

427_2014_474_MOESM10_ESM.mpg

Stack of confocal images of 1b clone at two day-old stage. Dorsal view. (MPG 1,634 kb)

427_2014_474_MOESM11_ESM.mpg

Stack of confocal images of 1b clone at two day-old stage. Ventral view. (MPG 1,580 kb)

427_2014_474_MOESM12_ESM.mpg

Stack of confocal images of 1b clone at three day-old stage. Dorsal view. (MPG 1,530 kb)

427_2014_474_MOESM13_ESM.mpg

Stack of confocal images of 1b clone at four day-old stage. Dorsal view. (MPG 1,244 kb)

427_2014_474_MOESM14_ESM.mpg

Stack of confocal images of 1d clone at veliger larva stage. Anterior view. (MPG 870 kb)

427_2014_474_MOESM15_ESM.mpg

Stack of confocal images of 1d clone at two day-old stage. Dorsal view. (MPG 1,102 kb)

427_2014_474_MOESM16_ESM.mpg

Stack of confocal images of 1d clone at three day-old stage. Dorsal view. (MPG 1,134 kb)

427_2014_474_MOESM17_ESM.mpg

Stack of confocal images of 1d clone at four day-old stage. Ventral view. (MPG 2,316 kb)

427_2014_474_MOESM18_ESM.mpg

Stack of confocal images of 2a clone at veliger larva stage. Ventral view. (MPG 1,594 kb)

427_2014_474_MOESM19_ESM.mpg

Stack of confocal images of 2c clone at veliger larva stage. Ventral view. (MPG 1,316 kb)

427_2014_474_MOESM20_ESM.mpeg

Stack of confocal images of 2c clone at veliger larva stage. Left view. (MPEG 1,109 kb)

427_2014_474_MOESM21_ESM.mpg

Stack of confocal images of 2a clone at two day-old stage. Posterior ventral view. (MPG 2,036 kb)

427_2014_474_MOESM22_ESM.mpg

Stack of confocal images of 2c clone at two day-old stage. Posterior ventral view. (MPG 2,156 kb)

427_2014_474_MOESM23_ESM.mpg

Stack of confocal images of 2a clone at three day-old stage. Ventral view. (MPG 1,358 kb)

427_2014_474_MOESM24_ESM.mpg

Stack of confocal images of 2c clone at three day-old stage. Ventral view. (MPG 2,020 kb)

427_2014_474_MOESM25_ESM.mpg

Stack of confocal images of 2a clone at four day-old stage. Ventral view. (MPG 924 kb)

427_2014_474_MOESM26_ESM.mpg

Stack of confocal images of 2c clone at four day-old stage. Ventral view. (MPG 992 kb)

427_2014_474_MOESM27_ESM.mpg

Stack of confocal images of 2c clone at four day-old stage. Dorsal view. (MPG 1,220 kb)

427_2014_474_MOESM28_ESM.mpg

Stack of confocal images of 2b clone at veliger larva stage. Anterior view. (MPG 1,566 kb)

427_2014_474_MOESM29_ESM.mpg

Stack of confocal images of 2b clone at two day-old stage. Ventral view. (MPG 1,344 kb)

427_2014_474_MOESM30_ESM.mpg

Stack of confocal images of 2b clone at three day-old stage. Dorsal view. (MPG 1,794 kb)

427_2014_474_MOESM31_ESM.mpg

Stack of confocal images of 2b clone at three day-old stage. Ventral view. (MPG 1,390 kb)

427_2014_474_MOESM33_ESM.mpg

Stack of confocal images of 2b clone at four day-old stage. Dorsal view. (MPG 1,154 kb)

427_2014_474_MOESM34_ESM.mpg

Stack of confocal images of 2b clone at four day-old stage. Ventral view. (MPG 1,930 kb)

427_2014_474_MOESM35_ESM.mpg

Stack of confocal images of 2d clone at veliger larva stage. Anterior view. (MPG 908 kb)

427_2014_474_MOESM36_ESM.mpg

Stack of confocal images of 2d clone at two day-old stage. Posterior dorsal view. (MPG 1,224 kb)

427_2014_474_MOESM37_ESM.mpg

Stack of confocal images of 2d clone at three day-old stage. Ventral view. (MPG 2,096 kb)

427_2014_474_MOESM38_ESM.mpg

Stack of confocal images of 2d clone at four day-old stage. Ventral view. (MPG 1,186 kb)

427_2014_474_MOESM39_ESM.mpg

Stack of confocal images of 3a clone at veliger larva stage. Anterior view. (MPG 3,396 kb)

427_2014_474_MOESM40_ESM.mpg

Stack of confocal images of 3b clone at veliger larva stage. Anterior view. (MPG 1,968 kb)

427_2014_474_MOESM41_ESM.mpg

Stack of confocal images of 3a clone at two day-old stage. Ventral view. (MPG 606 kb)

427_2014_474_MOESM42_ESM.mpg

Stack of confocal images of 3b clone at two day-old stage. Ventral view. (MPG 432 kb)

427_2014_474_MOESM44_ESM.mpeg

Stack of confocal images of 3a clone at three day-old stage. Ventral view. Red: rhodamine dextran; cyan blue: phalloidin; Blue: DAPI. (MPEG 1,109 kb)

427_2014_474_MOESM45_ESM.mpg

Stack of confocal images of 3b clone at three day-old stage. Ventral view. (MPG 1,128 kb)

427_2014_474_MOESM46_ESM.mpg

Stack of confocal images of 3a clone at four day-old stage. Ventral view. Red: rhodamine dextran; cyan blue: phalloidin; Blue: DAPI. (MPG 2,132 kb)

427_2014_474_MOESM47_ESM.mpg

Stack of confocal images of 3b clone at four day-old stage. Ventral view. (MPG 892 kb)

427_2014_474_MOESM48_ESM.mpg

Stack of confocal images of 3c clone at veliger larva stage. Anterior view. (MPG 1,046 kb)

427_2014_474_MOESM49_ESM.mpg

Stack of confocal images of 3d clone at veliger larva stage. Anterior view. (MPG 746 kb)

427_2014_474_MOESM50_ESM.mpeg

Stack of confocal images of 3c clone at two day-old stage. Posterior ventral view. (MPEG 869 kb)

427_2014_474_MOESM51_ESM.mpg

Stack of confocal images of 3d clone at two day-old stage. Posterior ventral view. (MPG 1,350 kb)

427_2014_474_MOESM52_ESM.mpg

Stack of confocal images of 3c clone at three day-old stage. Ventral view. (MPG 3,512 kb)

427_2014_474_MOESM53_ESM.mpg

Stack of confocal images of 3d clone at three day-old stage. Ventral view. (MPG 1,558 kb)

427_2014_474_MOESM54_ESM.mpg

Stack of confocal images of 3c clone at four day-old stage. Ventral view. (MPG 1,120 kb)

427_2014_474_MOESM55_ESM.mpg

Stack of confocal images of 3d clone at four day-old stage. Ventral view. (MPG 2,798 kb)

427_2014_474_MOESM56_ESM.mpeg

Stack of confocal images of 3c clone at veliger larva stage. Right view. (MPEG 1,019 kb)

427_2014_474_MOESM57_ESM.mpg

Stack of confocal images of 4A clone at two day-old stage. Ventral view. (MPG 1,058 kb)

427_2014_474_MOESM58_ESM.mpg

Stack of confocal images of 4B clone at two day-old stage. Ventral view. (MPG 1,356 kb)

427_2014_474_MOESM59_ESM.mpg

Stack of confocal images of 4C clone at two day-old stage. Ventral view. (MPG 1,306 kb)

427_2014_474_MOESM60_ESM.mpg

Stack of confocal images of 4A clone at three day-old stage. Ventral view. (MPG 962 kb)

427_2014_474_MOESM61_ESM.mpg

Stack of confocal images of 4B clone at three day-old stage. Ventral view. (MPG 1,870 kb)

427_2014_474_MOESM62_ESM.mpg

Stack of confocal images of 4C clone at three day-old stage. Ventral view. (MPG 1,646 kb)

427_2014_474_MOESM63_ESM.mpg

Stack of confocal images of 4A clone at four day-old stage. Right view. (MPG 1,462 kb)

427_2014_474_MOESM64_ESM.mpg

Stack of confocal images of 4B clone at four day-old stage. Right view. (MPG 1,094 kb)

427_2014_474_MOESM65_ESM.mpg

Stack of confocal images of 4C clone at four day-old stage. Right view. (MPG 1,500 kb)

427_2014_474_MOESM66_ESM.mpg

Stack of confocal images of 4A clone at veliger larva stage. Left view. (MPG 6,466 kb)

427_2014_474_MOESM67_ESM.mpg

Stack of confocal images of 4B clone at veliger larva stage. Left view. (MPG 1,268 kb)

427_2014_474_MOESM68_ESM.mpg

Stack of confocal images of 4C clone at veliger larva stage. Left view. (MPG 1,576 kb)

427_2014_474_MOESM69_ESM.mpg

Stack of confocal images of 4a clone at two day-old stage. Posterior ventral view. (MPG 1,246 kb)

427_2014_474_MOESM70_ESM.mpg

Stack of confocal images of 4b clone at two day-old stage. Posterior ventral view. (MPG 1,204 kb)

427_2014_474_MOESM71_ESM.mpg

Stack of confocal images of 4c clone at two day-old stage. Posterior ventral view. (MPG 1,124 kb)

427_2014_474_MOESM72_ESM.mpg

Stack of confocal images of 4a clone at three day-old stage. Ventral view. (MPG 1,434 kb)

427_2014_474_MOESM73_ESM.mpg

Stack of confocal images of 4b clone at three day-old stage. Ventral view. (MPG 2,014 kb)

427_2014_474_MOESM74_ESM.mpg

Stack of confocal images of 4c clone at three day-old stage. Ventral view. (MPG 1,654 kb)

427_2014_474_MOESM75_ESM.mpg

Stack of confocal images of 4a clone at four day-old stage. Ventral view. (MPG 884 kb)

427_2014_474_MOESM76_ESM.mpg

Stack of confocal images of 4b clone at four day-old stage. Dorsal view. (MPG 1,668 kb)

427_2014_474_MOESM77_ESM.mpg

Stack of confocal images of 4c clone at four day-old stage. Ventral view. (MPG 1,058 kb)

427_2014_474_MOESM78_ESM.mpg

Stack of confocal images of 4a clone at veliger larva stage. Right view. (MPG 1,044 kb)

427_2014_474_MOESM79_ESM.mpg

Stack of confocal images of 4b clone at veliger larva stage. Right view. (MPG 942 kb)

427_2014_474_MOESM80_ESM.mpg

Stack of confocal images of 4c clone at veliger larva stage. Left view. (MPG 2,228 kb)

427_2014_474_MOESM81_ESM.mpg

Stack of confocal images of 4d clone at two day-old stage. Posterior dorsal view. (MPG 1,968 kb)

427_2014_474_MOESM82_ESM.mpg

Stack of confocal images of 4d clone at three day-old stage. Ventral view. (MPG 2,226 kb)

427_2014_474_MOESM83_ESM.mpg

Stack of confocal images of 4d clone at four day-old stage. Ventral view. (MPG 4,234 kb)

427_2014_474_MOESM84_ESM.mpg

Stack of confocal images of 4d clone at veliger larva stage. Dorsal view. (MPG 2,800 kb)

Time-lapse movie of cell divisions in the 4d lineages. This time-lapsed movie was recorded at 4d + 26.5 h when each side of the lineage contains 12 cells. Chromatin is visualized with H2B-eGFP. Dorsal view. (MPG 5,794 kb)

427_2014_474_MOESM88_ESM.mpeg

Stack of confocal images of 2a and 3a double labeling at two day-old stage. Ventral view. Red: Rhodamine dextran (3a); Green: Oregon green dextran (2a); cyan blue: phalloidin; Blue: DAPI. (MPEG 3,374 kb)

427_2014_474_MOESM89_ESM.mpeg

Stack of confocal images of 2a and 3a double labeling at three day-old stage. Ventral view. Red: Rhodamine dextran (3a); Green: Oregon green dextran (2a); cyan blue: phalloidin; Blue: DAPI. (MPEG 1,198 kb)

427_2014_474_MOESM90_ESM.mpeg

Stack of confocal images of 2b and 3a double labeling at two day-old stage. Ventral view. Red: Rhodamine dextran (3a); Green: Oregon green dextran (2b); cyan blue: phalloidin; Blue: DAPI. (MPEG 923 kb)

427_2014_474_MOESM91_ESM.mpeg

Stack of confocal images of 2b and 3a double labeling at three day-old stage. Ventral view. Red: Rhodamine dextran (3a); Green: Oregon green dextran (2b); cyan blue: phalloidin; Blue: DAPI. (MPEG 1,241 kb)

Movie S1

Stack of confocal images of 1a clone at veliger larva stage. Anterior view. (MPG 1,374 kb)

Movie S2

Stack of confocal images of 1c clone at veliger larva stage. Anterior view. (MPG 1,622 kb)

Movie S3

Stack of confocal images of 1a clone at two day-old stage. Dorsal view. (MPG 2,012 kb)

Movie S4

Stack of confocal images of 1c clone at two day-old stage. Dorsal view. (MPG 1,674 kb)

Movie S5

Stack of confocal images of 1a clone at three day-old stage. Dorsal view. (MPG 1,326 kb)

Movie S6

Stack of confocal images of 1c clone at three day-old stage. Dorsal view. (MPG 2,016 kb)

Movie S7

Stack of confocal images of 1a clone at four day-old stage. Dorsal view. (MPG 1,732 kb)

Movie S8

Stack of confocal images of 1c clone at four day-old stage. Dorsal view. (MPG 2,158 kb)

Movie S9

Stack of confocal images of 1b clone at veliger larva stage. Anterior view. (MPG 766 kb)

Movie S10

Stack of confocal images of 1b clone at two day-old stage. Dorsal view. (MPG 1,634 kb)

Movie S11

Stack of confocal images of 1b clone at two day-old stage. Ventral view. (MPG 1,580 kb)

Movie S12

Stack of confocal images of 1b clone at three day-old stage. Dorsal view. (MPG 1,530 kb)

Movie S13

Stack of confocal images of 1b clone at four day-old stage. Dorsal view. (MPG 1,244 kb)

Movie S14

Stack of confocal images of 1d clone at veliger larva stage. Anterior view. (MPG 870 kb)

Movie S15

Stack of confocal images of 1d clone at two day-old stage. Dorsal view. (MPG 1,102 kb)

Movie S16

Stack of confocal images of 1d clone at three day-old stage. Dorsal view. (MPG 1,134 kb)

Movie S17

Stack of confocal images of 1d clone at four day-old stage. Ventral view. (MPG 2,316 kb)

Movie S18

Stack of confocal images of 2a clone at veliger larva stage. Ventral view. (MPG 1,594 kb)

Movie S19

Stack of confocal images of 2c clone at veliger larva stage. Ventral view. (MPG 1,316 kb)

Movie S20

Stack of confocal images of 2c clone at veliger larva stage. Left view. (MPEG 1,109 kb)

Movie S21

Stack of confocal images of 2a clone at two day-old stage. Posterior ventral view. (MPG 2,036 kb)

Movie S22

Stack of confocal images of 2c clone at two day-old stage. Posterior ventral view. (MPG 2,156 kb)

Movie S23

Stack of confocal images of 2a clone at three day-old stage. Ventral view. (MPG 1,358 kb)

Movie S24

Stack of confocal images of 2c clone at three day-old stage. Ventral view. (MPG 2,020 kb)

Movie S25

Stack of confocal images of 2a clone at four day-old stage. Ventral view. (MPG 924 kb)

Movie S26

Stack of confocal images of 2c clone at four day-old stage. Ventral view. (MPG 992 kb)

Movie S27

Stack of confocal images of 2c clone at four day-old stage. Dorsal view. (MPG 1,220 kb)

Movie S28

Stack of confocal images of 2b clone at veliger larva stage. Anterior view. (MPG 1,566 kb)

Movie S29

Stack of confocal images of 2b clone at two day-old stage. Ventral view. (MPG 1,344 kb)

Movie S30

Stack of confocal images of 2b clone at three day-old stage. Dorsal view. (MPG 1,794 kb)

Movie S31

Stack of confocal images of 2b clone at three day-old stage. Ventral view. (MPG 1,390 kb)

Fig. S32

The development of the 2b clone. (AD) Ventral view. At two days-old, the future mouth cells are positioned on the ventral midline and connected to the band of cells. During the transition from two day-old to three day-old, they disconnected from the band of cells and remain on the ventral midline. (EH) Dorsal view. At two day-old, the arms of the band of cells are apart from each other on the dorsal side. Transitioning from two day-old to three day-old, the arms of the band of cells extend and join at the dorsal side. In a three day-old embryo, the cells that are part of the shell field bud off from the joined arms. Green: Oregon Green dextran; Blue: DAPI. Dotted line: blastopore; asterisk: developing mouth. (DOCX 322 kb)

Movie S33

Stack of confocal images of 2b clone at four day-old stage. Dorsal view. (MPG 1,154 kb)

Movie S34

Stack of confocal images of 2b clone at four day-old stage. Ventral view. (MPG 1,930 kb)

Movie S35

Stack of confocal images of 2d clone at veliger larva stage. Anterior view. (MPG 908 kb)

Movie S36

Stack of confocal images of 2d clone at two day-old stage. Posterior dorsal view. (MPG 1,224 kb)

Movie S37

Stack of confocal images of 2d clone at three day-old stage. Ventral view. (MPG 2,096 kb)

Movie S38

Stack of confocal images of 2d clone at four day-old stage. Ventral view. (MPG 1,186 kb)

Movie S39

Stack of confocal images of 3a clone at veliger larva stage. Anterior view. (MPG 3,396 kb)

Movie S40

Stack of confocal images of 3b clone at veliger larva stage. Anterior view. (MPG 1,968 kb)

Movie S41

Stack of confocal images of 3a clone at two day-old stage. Ventral view. (MPG 606 kb)

Movie S42

Stack of confocal images of 3b clone at two day-old stage. Ventral view. (MPG 432 kb)

Fig. S43

The mesodermal population derived from the ectomesodermal clone, 3a. (AB) Ventral view of two day-old embryo. (A) Projection of stacks of mesodermal layers in the two day-old embryo. The mesodermal progenitor cells derived from the 3a clone is shown. (B) Projection of a whole mount embryo showing the clonal distribution of the 3a cell. (CD) Ventral view of three day-old embryo. (C) Projection of stacks of mesodermal layers in the three day-old embryo. The mesodermal population has divided and extended to the posterior region. Meanwhile, the esophageal cell sheet (ectodermal) has started internalizing through the future mouth opening. (D) Projection of a whole mount three day-old embryo showing the 3a lineage. Green is lineage tracer. Blue is DAPI staining of nuclei. Red is phalloidin staining of filamentous actin. (DOCX 239 kb)

Movie S44

Stack of confocal images of 3a clone at three day-old stage. Ventral view. Red: rhodamine dextran; cyan blue: phalloidin; Blue: DAPI. (MPEG 1,109 kb)

Movie S45

Stack of confocal images of 3b clone at three day-old stage. Ventral view. (MPG 1,128 kb)

Movie S46

Stack of confocal images of 3a clone at four day-old stage. Ventral view. Red: rhodamine dextran; cyan blue: phalloidin; Blue: DAPI. (MPG 2,132 kb)

Movie S47

Stack of confocal images of 3b clone at four day-old stage. Ventral view. (MPG 892 kb)

Movie S48

Stack of confocal images of 3c clone at veliger larva stage. Anterior view. (MPG 1,046 kb)

Movie S49

Stack of confocal images of 3d clone at veliger larva stage. Anterior view. (MPG 746 kb)

Movie S50

Stack of confocal images of 3c clone at two day-old stage. Posterior ventral view. (MPEG 869 kb)

Movie S51

Stack of confocal images of 3d clone at two day-old stage. Posterior ventral view. (MPG 1,350 kb)

Movie S52

Stack of confocal images of 3c clone at three day-old stage. Ventral view. (MPG 3,512 kb)

Movie S53

Stack of confocal images of 3d clone at three day-old stage. Ventral view. (MPG 1,558 kb)

Movie S54

Stack of confocal images of 3c clone at four day-old stage. Ventral view. (MPG 1,120 kb)

Movie S55

Stack of confocal images of 3d clone at four day-old stage. Ventral view. (MPG 2,798 kb)

Movie S56

Stack of confocal images of 3c clone at veliger larva stage. Right view. (MPEG 1,019 kb)

Movie S57

Stack of confocal images of 4A clone at two day-old stage. Ventral view. (MPG 1,058 kb)

Movie S58

Stack of confocal images of 4B clone at two day-old stage. Ventral view. (MPG 1,356 kb)

Movie S59

Stack of confocal images of 4C clone at two day-old stage. Ventral view. (MPG 1,306 kb)

Movie S60

Stack of confocal images of 4A clone at three day-old stage. Ventral view. (MPG 962 kb)

Movie S61

Stack of confocal images of 4B clone at three day-old stage. Ventral view. (MPG 1,870 kb)

Movie S62

Stack of confocal images of 4C clone at three day-old stage. Ventral view. (MPG 1,646 kb)

Movie S63

Stack of confocal images of 4A clone at four day-old stage. Right view. (MPG 1,462 kb)

Movie S64

Stack of confocal images of 4B clone at four day-old stage. Right view. (MPG 1,094 kb)

Movie S65

Stack of confocal images of 4C clone at four day-old stage. Right view. (MPG 1,500 kb)

Movie S66

Stack of confocal images of 4A clone at veliger larva stage. Left view. (MPG 6,466 kb)

Movie S67

Stack of confocal images of 4B clone at veliger larva stage. Left view. (MPG 1,268 kb)

Movie S68

Stack of confocal images of 4C clone at veliger larva stage. Left view. (MPG 1,576 kb)

Movie S69

Stack of confocal images of 4a clone at two day-old stage. Posterior ventral view. (MPG 1,246 kb)

Movie S70

Stack of confocal images of 4b clone at two day-old stage. Posterior ventral view. (MPG 1,204 kb)

Movie S71

Stack of confocal images of 4c clone at two day-old stage. Posterior ventral view. (MPG 1,124 kb)

Movie S72

Stack of confocal images of 4a clone at three day-old stage. Ventral view. (MPG 1,434 kb)

Movie S73

Stack of confocal images of 4b clone at three day-old stage. Ventral view. (MPG 2,014 kb)

Movie S74

Stack of confocal images of 4c clone at three day-old stage. Ventral view. (MPG 1,654 kb)

Movie S75

Stack of confocal images of 4a clone at four day-old stage. Ventral view. (MPG 884 kb)

Movie S76

Stack of confocal images of 4b clone at four day-old stage. Dorsal view. (MPG 1,668 kb)

Movie S77

Stack of confocal images of 4c clone at four day-old stage. Ventral view. (MPG 1,058 kb)

Movie S78

Stack of confocal images of 4a clone at veliger larva stage. Right view. (MPG 1,044 kb)

Movie S79

Stack of confocal images of 4b clone at veliger larva stage. Right view. (MPG 942 kb)

Movie S80

Stack of confocal images of 4c clone at veliger larva stage. Left view. (MPG 2,228 kb)

Movie S81

Stack of confocal images of 4d clone at two day-old stage. Posterior dorsal view. (MPG 1,968 kb)

Movie S82

Stack of confocal images of 4d clone at three day-old stage. Ventral view. (MPG 2,226 kb)

Movie S83

Stack of confocal images of 4d clone at four day-old stage. Ventral view. (MPG 4,234 kb)

Movie S84

Stack of confocal images of 4d clone at veliger larva stage. Dorsal view. (MPG 2,800 kb)

Fig. S85

4d blast cell lineage. The reference lineage is the most extended lineage obtained and thus, is used as reference for comparison to other lineages examined. Lineages after the 4d + 24 h mark are derived from the live-time imaging result of four different embryos including the reference embryo. Left axis is time after the birth of 4d. Color code: long orange line (t = 4d + 24 h mark); black lines (reference); red, blue and green lines each represents division happened in three other individual lineages examined. (DOCX 98 kb)

Fig. S87

The development of the foregut: mouth and esophagus. The positions of the 2nd quartet cells (2a and 2b) and 3rd quartet cells (3a) in relation to the blastopore and the developing mouth. (AB) Double labeling of the 2a (green) and 3a (red) clones in two day-old and three day-old embryos. (CD) Double labeling of the 2b (green) and 3a (red) clones in two day-old and three day-old embryos. Green: Oregon Green dextran; Red: Rhodamine dextran; Blue: DAPI. Dotted line: blastopore/developing mouth. (DOCX 219 kb)

Movie S88

Stack of confocal images of 2a and 3a double labeling at two day-old stage. Ventral view. Red: Rhodamine dextran (3a); Green: Oregon green dextran (2a); cyan blue: phalloidin; Blue: DAPI. (MPEG 3,374 kb)

Movie S89

Stack of confocal images of 2a and 3a double labeling at three day-old stage. Ventral view. Red: Rhodamine dextran (3a); Green: Oregon green dextran (2a); cyan blue: phalloidin; Blue: DAPI. (MPEG 1,198 kb)

Movie S90

Stack of confocal images of 2b and 3a double labeling at two day-old stage. Ventral view. Red: Rhodamine dextran (3a); Green: Oregon green dextran (2b); cyan blue: phalloidin; Blue: DAPI. (MPEG 923 kb)

Movie S91

Stack of confocal images of 2b and 3a double labeling at three day-old stage. Ventral view. Red: Rhodamine dextran (3a); Green: Oregon green dextran (2b); cyan blue: phalloidin; Blue: DAPI. (MPEG 1,241 kb)

Fig. S92

Shell field invagination in early trochophores. (AD) Individual labeling of all 2nd quartet micromeres. (A) 2a clone contributes to upper left part of the shell field. (B) 2c clone contributes to upper right part of the shell field. (C) Higher magnification of the 2b clone contribution to the upper middle part of the shell gland. (D) 2d clone gives rise to a portion of the ventral part of the shell gland that is similar in size to the 2a and 2c contributions. Green: Oregon Green dextran; Blue: DAPI. (DOCX 220 kb)

Fig. S93

Summary of the early cell lineages and the contributions derived from fate mapping. Abbreviation: L, Left; R, Right. (DOCX 340 kb)

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Chan, X.Y., Lambert, J.D. Development of blastomere clones in the Ilyanassa embryo: transformation of the spiralian blastula into the larval body plan. Dev Genes Evol 224, 159–174 (2014). https://doi.org/10.1007/s00427-014-0474-z

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Keywords

  • Mollusc
  • Spiralian
  • Fate map
  • Lineage tracing