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367
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Abitua, P.B., Gainous, T.B., Kaczmarczyk, A.N., Winchell, C.J., Hudson, C., Kamata, K., Nakagawa,
M., Tsuda, M., Kusakabe, T.G., Levine, M., (2015) The pre-vertebrate origins of neurogenic
placodes. Nature 524: 462-465. https://doi.org/10.1038/nature14657
Abitua, P.B., Wagner, E., Navarrete, I.A., Levine, M., (2012) Identification of a rudimentary neural
crest in a non-vertebrate chordate. Nature 492: 104-107. https://doi.org/10.1038/nature11589
Ahrens, K., Schlosser, G., (2005) Tissues and signals involved in the induction of placodal Six1
expression in Xenopus laevis. Developmental Biology 288: 40-59.
https://doi.org/10.1016/j.ydbio.2005.07.022
Brugmann, S.A., Moody, S.A., (2005) Induction and specification of the vertebrate ectodermal
placodes: precursors of the cranial sensory organs. Biology of the cell / under the auspices of the
European Cell Biology Organization 97: 303-319. https://doi.org/10.1042/BC20040515
Brugmann, S.A., Pandur, P.D., Kenyon, K.L., Pignoni, F., Moody, S.A., (2004) Six1 promotes a
placodal fate within the lateral neurogenic ectoderm by functioning as both a transcriptional
activator and repressor. Development 131: 5871-5881. https://doi.org/10.1242/dev.01516
Cao, C., Lemaire, L.A., Wang, W., Yoon, P.H., Choi, Y.A., Parsons, L.R., Matese, J.C., Wang, W.,
Levine, M., Chen, K., (2019) Comprehensive single-cell transcriptome lineages of a protovertebrate. Nature 571: 349-354. https://doi.org/10.1038/s41586-019-1385-y
Delsuc, F., Brinkmann, H., Chourrout, D., Philippe, H., (2006) Tunicates and not cephalochordates are
the closest living relatives of vertebrates. Nature 439: 965-968. https://doi.org/10.1038/nature04336
Esterberg, R., Fritz, A., (2009) dlx3b/4b are required for the formation of the preplacodal region and
otic placode through local modulation of BMP activity. Developmental Biology 325: 189-199.
https://doi.org/10.1016/j.ydbio.2008.10.017
Gans, C., Northcutt, R.G., (1983) Neural Crest and the Origin of Vertebrates - a New Head. Science
220: 268-273. https://doi.org/DOI 10.1126/science.220.4594.268
Glavic, A., Honore, S.M., Feijoo, C.G., Bastidas, F., Allende, M.L., Mayor, R., (2004) Role of BMP
signaling and the homeoprotein iroquois in the specification of the cranial placodal field.
Developmental Biology 272: 89-103. https://doi.org/10.1016/j.ydbio.2004.04.020
Graham, A., Shimeld, S.M., (2013) The origin and evolution of the ectodermal placodes. Journal of
anatomy 222: 32-40. https://doi.org/10.1111/j.1469-7580.2012.01506.x
Hino, K., Satou, Y., Yagi, K., Satoh, N., (2003) A genomewide survey of developmentally relevant
genes in Ciona intestinalis. VI. Genes for Wnt, TGFbeta, Hedgehog and JAK/STAT signaling
pathways. Dev Genes Evol 213: 264-272. https://doi.org/10.1007/s00427-003-0318-8
Horie, R., Hazbun, A., Chen, K., Cao, C., Levine, M., Horie, T., (2018) Shared evolutionary origin of
vertebrate neural crest and cranial placodes. Nature 560: 228-232. https://doi.org/10.1038/s41586018-0385-7
Hudson, C., Yasuo, H., (2005) Patterning across the ascidian neural plate by lateral Nodal signalling
sources. Development 132: 1199-1210. https://doi.org/10.1242/dev.01688
Ikeda, T., Matsuoka, T., Satou, Y., (2013) A time delay gene circuit is required for palp formation in the
ascidian embryo. Development 140: 4703-4708. https://doi.org/10.1242/dev.100339
Ikeda, T., Satou, Y., (2017) Differential temporal control of Foxa.a and Zic-r.b specifies brain versus
notochord fate in the ascidian embryo. Development 144: 38-43.
https://doi.org/10.1242/dev.142174
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395
396
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398
399
400
401
402
403
404
405
406
407
408
409
410
411
412
Imai, K.S., Daido, Y., Kusakabe, T.G., Satou, Y., (2012) Cis-acting transcriptional repression
establishes a sharp boundary in chordate embryos. Science 337: 964-967.
https://doi.org/10.1126/science.1222488
Imai, K.S., Hino, K., Yagi, K., Satoh, N., Satou, Y., (2004) Gene expression profiles of transcription
factors and signaling molecules in the ascidian embryo: towards a comprehensive understanding of
gene networks. Development 131: 4047-4058. https://doi.org/10.1242/dev.01270
Imai, K.S., Levine, M., Satoh, N., Satou, Y., (2006) Regulatory blueprint for a chordate embryo.
Science 312: 1183-1187. https://doi.org/10.1126/science.1123404
Kwon, H.J., Bhat, N., Sweet, E.M., Cornell, R.A., Riley, B.B., (2010) Identification of Early
Requirements for Preplacodal Ectoderm and Sensory Organ Development. PLoS genetics 6.
https://doi.org/ARTN e1001133
10.1371/journal.pgen.1001133
Litsiou, A., Hanson, S., Streit, A., (2005) A balance of FGF, BMP and WNT signalling positions the
future placode territory in the head. Development 132: 4051-4062.
https://doi.org/10.1242/dev.01964
Liu, B., Satou, Y., (2019) Foxg specifies sensory neurons in the anterior neural plate border of the
ascidian embryo. Nat Commun 10: 4911. https://doi.org/10.1038/s41467-019-12839-6
Liu, B., Satou, Y., (2020) The genetic program to specify ectodermal cells in ascidian embryos. Dev
Growth Differ 62: 301-310. https://doi.org/10.1111/dgd.12660
Manni, L., Agnoletto, A., Zaniolo, G., Burighel, P., (2005) Stomodeal and neurohypophysial placodes
in Ciona intestinalis: insights into the origin of the pituitary gland. Journal of experimental zoology.
Part B, Molecular and developmental evolution 304: 324-339. https://doi.org/10.1002/jez.b.21039
Manni, L., Lane, N.J., Joly, J.S., Gasparini, F., Tiozzo, S., Caicci, F., Zaniolo, G., Burighel, P., (2004)
Neurogenic and non-neurogenic placodes in ascidians. Journal of experimental zoology. Part B,
Molecular and developmental evolution 302: 483-504. https://doi.org/10.1002/jez.b.21013
Mazet, F., Hutt, J.A., Milloz, J., Millard, J., Graham, A., Shimeld, S.M., (2005) Molecular evidence
from Ciona intestinalis for the evolutionary origin of vertebrate sensory placodes. Dev Biol 282:
494-508. https://doi.org/10.1016/j.ydbio.2005.02.021
Oda-Ishii, I., Kubo, A., Kari, W., Suzuki, N., Rothbacher, U., Satou, Y., (2016) A maternal system
initiating the zygotic developmental program through combinatorial repression in the ascidian
embryo. PLoS genetics 12: e1006045. https://doi.org/10.1371/journal.pgen.1006045
Ohta, N., Satou, Y., (2013) Multiple signaling pathways coordinate to induce a threshold response in a
chordate embryo. PLoS genetics 9: e1003818. https://doi.org/10.1371/journal.pgen.1003818
Pasini, A., Amiel, A., Rothbacher, U., Roure, A., Lemaire, P., Darras, S., (2006) Formation of the
ascidian epidermal sensory neurons: insights into the origin of the chordate peripheral nervous
system. PLoS Biol 4: e225. https://doi.org/10.1371/journal.pbio.0040225
Putnam, N.H., Butts, T., Ferrier, D.E., Furlong, R.F., Hellsten, U., Kawashima, T., Robinson-Rechavi,
M., Shoguchi, E., Terry, A., Yu, J.K., Benito-Gutierrez, E.L., Dubchak, I., Garcia-Fernandez, J.,
Gibson-Brown, J.J., Grigoriev, I.V., Horton, A.C., de Jong, P.J., Jurka, J., Kapitonov, V.V., Kohara,
Y., Kuroki, Y., Lindquist, E., Lucas, S., Osoegawa, K., Pennacchio, L.A., Salamov, A.A., Satou, Y.,
Sauka-Spengler, T., Schmutz, J., Shin, I.T., Toyoda, A., Bronner-Fraser, M., Fujiyama, A., Holland,
L.Z., Holland, P.W., Satoh, N., Rokhsar, D.S., (2008) The amphioxus genome and the evolution of
the chordate karyotype. Nature 453: 1064-1071. https://doi.org/10.1038/nature06967
Satou, Y., Kawashima, T., Shoguchi, E., Nakayama, A., Satoh, N., (2005) An integrated database of the
17
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ascidian, Ciona intestinalis: Towards functional genomics. Zool Sci 22: 837-843.
https://doi.org/10.2108/zsj.22.837
Satou, Y., Nakamura, R., Yu, D., Yoshida, R., Hamada, M., Fujie, M., Hisata, K., Takeda, H., Satoh, N.,
(2019) A nearly complete genome of Ciona intestinalis type A (C. robusta) reveals the contribution
of inversion to chromosomal evolution in the genus Ciona. Genome biology and evolution 11:
3144-3157. https://doi.org/10.1093/gbe/evz228
Satou, Y., Tokuoka, M., Oda-Ishii, I., Tokuhiro, S., Ishida, T., Liu, B., Iwamura, Y., (2022) A Manually
Curated Gene Model Set for an Ascidian, Ciona robusta (Ciona intestinalis Type A). Zool Sci 39:
253-260. https://doi.org/10.2108/zs210102
Schlosser, G., (2014) Early embryonic specification of vertebrate cranial placodes. Wires Dev Biol 3:
349-363. https://doi.org/10.1002/wdev.142
Schneider, C.A., Rasband, W.S., Eliceiri, K.W., (2012) NIH Image to ImageJ: 25 years of image
analysis. Nature methods 9: 671-675. https://doi.org/10.1038/nmeth.2089
Singh, S., Groves, A.K., (2016) The molecular basis of craniofacial placode development. Wiley
Interdiscip Rev Dev Biol 5: 363-376. https://doi.org/10.1002/wdev.226
Steventon, B., Mayor, R., Streit, A., (2014) Neural crest and placode interaction during the
development of the cranial sensory system. Dev Biol 389: 28-38.
https://doi.org/10.1016/j.ydbio.2014.01.021
Stolfi, A., Ryan, K., Meinertzhagen, I.A., Christiaen, L., (2015a) Migratory neuronal progenitors arise
from the neural plate borders in tunicates. Nature 527: 371-374.
https://doi.org/10.1038/nature15758
Stolfi, A., Sasakura, Y., Chalopin, D., Satou, Y., Christiaen, L., Dantec, C., Endo, T., Naville, M.,
Nishida, H., Swalla, B.J., Volff, J.N., Voskoboynik, A., Dauga, D., Lemaire, P., (2015b) Guidelines
for the nomenclature of genetic elements in tunicate genomes. Genesis 53: 1-14.
https://doi.org/10.1002/dvg.22822
Tokuoka, M., Maeda, K., Kobayashi, K., Mochizuki, A., Satou, Y., (2021) The gene regulatory system
for specifying germ layers in early embryos of the simple chordate. Sci Adv 7: eabf8210.
https://doi.org/10.1126/sciadv.abf8210
Tribulo, C., Aybar, M.J., Nguyen, V.H., Mullins, M.C., Mayor, R., (2003) Regulation of Msx genes by
a Bmp gradient is essential for neural crest specification. Development 130: 6441-6452.
https://doi.org/10.1242/dev.00878
Wagner, E., Levine, M., (2012) FGF signaling establishes the anterior border of the Ciona neural tube.
Development 139: 2351-2359. https://doi.org/10.1242/dev.078485
Wagner, E., Stolfi, A., Choi, Y.G., Levine, M., (2014) Islet is a key determinant of ascidian palp
morphogenesis. Development 141: 3084-3092. https://doi.org/10.1242/dev.110684
Waki, K., Imai, K.S., Satou, Y., (2015) Genetic pathways for differentiation of the peripheral nervous
system in ascidians. Nat Commun 6: 8719. https://doi.org/10.1038/ncomms9719
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Figure 1. BMP signaling is activated in ANB cells. (A) Schematic illustration of the anterior
border of the neural plate in ascidian late gastrula and early neurula embryos. Because
embryos are bilaterally symmetrical, names of individual cells are indicated only in the left
half, and are prefixed with IDs shown next to the illustration, e.g., the left upper cell of the
late gastrula embryo is called a9.36. Vertical bars indicate sister cell relationships. (B, C)
Immunostaining of (B) a late gastrula embryo and (C) an early neurula embryo to detect
phosphorylated Smad1/5/9 (pSmad1/5/9). ANB cells were marked by Foxc>GFP expression,
detected with an anti-GFP antibody. Note that GFP was detected only on the right side
because of mosaic incorporation. Right images are overlaid in pseudocolor. The brightness
and contrast of these photographs were adjusted linearly. ANB cells are enclosed by broken
lines. Note that strong signals are observed in the anterior medial ANB cells and relatively
weak signals are also observed in the remaining ANB cells. Dorsal views are shown. The
scale bar in (B) represents 50 μm.
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Figure 2. Admp and BMP5/6/7/8 are expressed in ANB cells. (A-G) In situ hybridization
for (A,C) Admp (magenta) expression at (A) the late gastrula stage and (C) early neurula
stage, and for (E, G) Bmp5/6/7/8 (magenta) expression at (E) the early neurula stage and (G)
the middle neurula stage. Foxc in (A) marks ANB cells (green). Foxg in (C) and (E) marks the
most anterior and posterior rows of the ANB region (green). Six1/2 marks the most posterior
row of the ANB (green), although this gene is also expressed in the two flanking cells on both
sides of the ANB. The results shown in (A), (C), and (E) are illustrated in (B), (D), and (F),
respectively. Dorsal views are shown. (H, I) Optical slices of pSmad1/5/9 immunostaining of
(H) an unperturbed control early neurula embryo and (I) an early neurula embryo injected
with the Admp MO (green). Nuclei are stained with DAPI (gray). ANB cells are marked by
arrowheads. Photographs are pseudocolored, and lateral views are shown. (J) Quantification
of fluorescence intensities of signals for pSmad1/5/9 in the ANB region of control
unperturbed embryos and Admp morphants. Each dot represents relative signal intensities of a
line of four medial ANB cells, and bars represent median values. Relative intensities were
calculated by dividing sums of pSmad1/5/9 signals of four ANB cells with sums of DAPI
signals of the same cells. As we used tyramide signal amplification, signal levels might not be
amplified linearly. However, a Wilcoxon’s rank sum test indicated that signal levels are
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significantly different between the control and experimental specimens. The scale bar in (A)
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represents 50 μm.
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Figure 3. Chordin and Noggin are expressed in cells next to the ANB cells. (A-D) In situ
hybridization for (A) Foxc (green) and Chordin (magenta), (B) Foxc (green) and Noggin
(magenta) at late gastrula stage, and for (C) Foxg (green) and Chordin (magenta), and (D)
Foxg (green) and Noggin (magenta) at the early neurula stage. Nuclei were stained with DAPI
(gray). Photographs are pseudocolored Z-projected image stacks. Dorsal views are shown.
Note that the anterior row of the ANB is not visible in (D). (E) Optical slices of pSmad1/5/9
immunostaining of an unperturbed control early neurula embryo and early neurula embryos
injected with Foxc>Chordin or Foxc>Noggin. ANB cells are marked with arrowheads.
Lateral views are shown. The scale bar in (A) represents 50 μm. (F) Quantification of
fluorescence intensities of signals for pSmad1/5/9 in the ANB region of unperturbed control
embryos and embryos injected with Foxc>Chordin or Foxc>Noggin. Each dot represents
relative signal intensities of a line of four medial ANB cells, and bars represent median
values. Relative intensities were calculated by dividing sums of pSmad1/5/9 signals of four
ANB cells with sums of DAPI signals of the same cells. As we used tyramide signal
amplification, signal levels might not be amplified linearly. However, Wilcoxon’s rank sum
tests indicated that signal levels are significantly different between the control and
experimental specimens.
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Figure 4. Suppression of BMP signaling activity affects expression of Foxg and Six1/2.
(A) A dorsal view of in situ hybridization for Foxg in an embryo in which Foxc>Noggin was
introduced by electroporation. Expression of Foxg in the posterior row (white arrowhead) was
lost, while expression in the anterior row was not affected in this embryo (black arrowhead).
(B) Percentages of embryos that normally expressed Foxg, or lost Foxg expression. (C) A
dorsal view of in situ hybridization for Six1/2 in an embryo in which Foxc>Noggin was
introduced by electroporation. Expression of Six1/2 in the ANB region was lost or greatly
reduced (white arrowheads). (D) Percentages of embryos that normally expressed or lost
Six1/2 expression in the posterior row of the ANB region of normal and embryos
electroporated with Foxc>Noggin. (E) Percentages of embryos that normally expressed or
lost Six1/2 expression in one or more ANB cells in embryos injected with Foxc>Noggin at
different concentrations. (F) Percentages of embryos that ectopically expressed Six1/2 in
embryos injected with Foxc>Noggin at different concentrations. (G) A dorsal view of an
embryo in which Foxc>Noggin was injected at 20 ng/μL, expressed Six1/2 ectopically
(arrowheads). The scale bar in (A) represents 50 μm.
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Figure 5. BMP signaling activity is required for proper palp formation. (A) Morphology
of the trunk of larvae developed from (A) a control unperturbed egg and (B) an egg
electroporated with the Foxc>Noggin construct. Two of three palp protrusions are visible in
this larva. We examined 16 control larvae and 24 experimental larvae, and all of them
exhibited phenotypes represented by these photographs. (C-F) In situ hybridization for Foxg
in (C) a control unperturbed embryo, (D) an embryo electroporated with the Foxc>Noggin
construct, (E) a control DMSO-treated embryo, and (F) an embryo treated with 50 μM
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dorsomorphin at the middle tailbud stage. We examined 45 embryos electroporated with
Foxc>Noggin, and 101 embryos treated with dorsomorphin. Among them, 93% and 75% of
embryos exhibited the phenotypes represented in (D) and (F), respectively. (G, H) Double
fluorescence in situ hybridization for Foxg (green) and Zf220 (magenta) in (G) a control
embryo, (H) an embryo treated with 50 μM dorsomorphin. In (H), the experimental embryo
lost Zf220 in the central region, and ectopically expressed Foxg. The expression pattern of
Foxg and Zf220 in the most anterior row of normal embryos are depicted on the right of (G).
These most-anterior-row cells are daughters of a10.72 and a10.80 as illustrated. Note that
Zf220 is also expressed in the second row (Liu and Satou, 2019), which are not depicted.
Photographs from C to H are anterior views, and the dorsal side of the trunk is down. (I)
Percentages of embryos that normally expressed Zf220 (black), lost Zf220 expression in the
central cells (gray), and lost it in both of the central and flanking cells (white) in the anterior
row of unperturbed control embryos and embryos treated with dorsomorphin. We examined
Zf220 expression by non-fluorescence in situ hybridization in addition to double fluorescence
in situ hybridization shown in (G) and (H). The scale bars in (A), (C), and (G) represent 50
μm.
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