Oocytes of a single individual of Ciona intestinalis (Roscoff, France) were fertilized by a mixture of sperm of two individuals and development was followed microscopically. Embryos were snapfrozen in liquid nitrogen at the appropriate stage and kept at -80°C. Samples at stage 0, 8, 12, 15, 21, 26 were collected from the same fertilzation, while sample from stage 11 was independently collected from a different individual. Total RNA was prepared with the RNA II kit of Magerey Nagel and the quality was checked with the Bioanalyzer, all samples had a RIN value of higher than 9. Strand specific libraries were constructed and the sequencing (50PE) was performed with Illumina HiSeq 2000 technology (BGI, Hong Kong, China). Reads were aligned onto C.robusta genome and considered to reflect the expression of the C.robusta orthologous genes.

Conversion to Aniseed format of data from Yutaka Satou's gene expression profiles of transcription factors and signalling molecule ISH screen (Development. 2004 131:4047-58).

Original annotation :
Gene name :nodal.
Expression pattern : No signal was detected

Conversion to Aniseed format of data from Yutaka Satou's gene expression profiles of transcription factors and signalling molecule ISH screen (Development. 2004 131:4047-58).

Original annotation :
Gene name :nodal.
Expression pattern : No distinct zygotic signal was detected

Animal views with anterior to the top of a 32-cell stage embryo stained with Nodal. Anterior ectoderm is marked in white, posterior ectoderm in yellow and gene expression in blue. Expression was observed in the b6.5 cell pair.

Animal view of a 32-cell stage embryo stained with Nodal.
Expression was observed in b6.5 cell pair.

Animal and vegetal views of a 32-cell stage wild type embryo probed for nodal expression.

Conversion to Aniseed format of data from Yutaka Satou's gene expression profiles of transcription factors and signalling molecule ISH screen (Development. 2004 131:4047-58).

Original annotation :
Gene name :nodal.
Expression pattern : A6.1, A6.3, B6.1, b6.5

Animal and vegetal views of a late 32-cell stage embryo. Embryos showed Ci-Nodal expression in A6.1, A6.3 and B6.1 cells of the vegetal hemisphere and also in b6.5 cells of the animal hemisphere.

Vegetal and animal views of a late 32-cell stage embryo showing Nodal expression in endoderm/endomesoderm blastomeres and in b6.5 cell pair.

Vegetal view of a 64-cell wild type stage embryo probed with Nodal expression.

Original annotation :
Gene name :nodal.
Expression pattern : b7.9, b7.10

Conversion to Aniseed format of data from Yutaka Satou's gene expression profiles of transcription factors and signalling molecule ISH screen (Development. 2004 131:4047-58).

Original annotation :
Gene name :nodal.
Expression pattern : b7.9, b7.10

Animal view with anterior to the top of a 64-cell stage embryo stained with Nodal. Expression was observed in the b7.9 and b7.10 cell pairs.

Vegetal and lateral views of a 110-cell stage wild type embryo probed for nodal expression.

Vegetal view of 110-cell stage wild type embryo probed for Nodal expression.

Original annotation :
Gene name :nodal.
Expression pattern : b8.17, b8.18, b8.19, b8.20

Conversion to Aniseed format of data from Yutaka Satou's gene expression profiles of transcription factors and signalling molecule ISH screen (Development. 2004 131:4047-58).

Original annotation :
Gene name :nodal.
Expression pattern : No distinct zygotic signal was detected

Dorsal view of a WT Ciona robusta late gastrula embryo electroporated with Nodal b-line>H2B::Venus (red) and Neurog(Full lenght)>H2B::CFP (blue).
Venus staining is observed in b-line tail epidermis.
Original annotation : Late gastrula embryo (stage 13) electroporated with full-length Neurog (blue) and Nodal b-line (red) reporter constructs. Reporter co-expression is seen in b9.36 descendants on either side of the neural plate. Scale bar, 25 μm

Conversion to Aniseed format of data from Yutaka Satou's gene expression profiles of transcription factors and signalling molecule ISH screen (Development. 2004 131:4047-58).

Original annotation :
Gene name :nodal.
Expression pattern : No distinct zygotic signal was detected

Dorsal view of an early tailbud embryo probed for ci-Nodal expression.

Original annotation:
Left - right asymmetric expression of Ci-Nodal became detectable by WISH in the left epidermis at the early tail bud stage (Fig. 1A; Yoshida and Saiga, 2008).

Dorsal view of an early tailbud stage probed for Nodal expression. The embryo developed within the chorion

Original annotation :
Ci-Nodal is expressed in the left epidermis.

Dorsal view of an early tailbud stage probed for Nodal expression.

Aniseed Comment : update of annotation of (Imai et al.,2004), based on (Imai et al.,2006).

Conversion to Aniseed format of data from Yutaka Satou's gene expression profiles of transcription factors and signalling molecule ISH screen (Development. 2004 131:4047-58).

Original annotation :
Gene name :nodal.
Expression pattern : epidermis

Lateral and dorsal views of early tailbud stage embryos showing reporter expression of the region upstream of nodal in tail epidermis, nerve cord and in part of head epidermis.

Lateral view of a full embryo (left pannel) and the tail (right pannel) of a WT Ciona robusta early tailbud embryo co-electroporated with Neurog b-line>unc-76::VenusYFP (green) and Nodal b8.18>H2B::mCherry (magenta). mCherru signalling is observed in the posterior dorsal midline tail epidermis.
Original annotation : Embryos co-electroporated with Neurog b-line>unc-76::VenusYFP (green) and Nodal b8.18>H2B::mCherry (magenta). Right panels are enlarged views of the images on the left. Dashed lines indicate displacement from clonally related epidermal cells (indicated by colour-coded brackets). Scale bars, 25 μm.

Dorsal view of a mid tailbud embryo probed for ci-Nodal expression.
Original annotation:
Left - right asymmetric expression of Ci-Nodal became detectable by WISH in the left epidermis at the early tail bud stage (Fig. 1A; Yoshida and Saiga, 2008). This expression continued until the mid tail bud stage (Fig. 1B).

Dorsal view of a mid tailbud embryo (10 hpf) probed for ci-Nodal (red) and Ci-Rx (green) expression (focus on the sensory vesicle region).
Image of DAPI staining is additionally merged (blue). Dorsal views are shown with anterior to the top. Dashed lines indicate the midline.

Original annotation:
Ci-Rx is expressed bilaterally in the SV until 10 hpf and then down regulated on the left side by 11 hpf when the expression of Ci-Nodal starts to be detectable in the left SV. At 12 hpf, Ci-Nodal expression in the left SV expands, while the expression of Ci-Rx is no longer detectable on the left side of the SV.

Dorsal view of a mid tailbud embryo (11 hpf) probed for ci-Nodal (red) and Ci-Rx (green) expression (focus on the sensory vesicle region).
Image of DAPI staining is additionally merged (blue). Dorsal views are shown with anterior to the top. Dashed lines indicate the midline.

Original annotation:
Ci-Rx is expressed bilaterally in the SV until 10 hpf and then down regulated on the left side by 11 hpf when the expression of Ci-Nodal starts to be detectable in the left SV. At 12 hpf, Ci-Nodal expression in the left SV expands, while the expression of Ci-Rx is no longer detectable on the left side of the SV.

Aniseed Comment : update of annotation of (Imai et al.,2004), based on (Imai et al.,2006).

Conversion to Aniseed format of data from Yutaka Satou's gene expression profiles of transcription factors and signalling molecule ISH screen (Development. 2004 131:4047-58).

Original annotation :
Gene name :nodal.
Expression pattern : nervous system, epidermis

Lateral view of a full embryo (left pannel) and the tail (right pannel) of a WT Ciona robusta mid tailbud embryo co-electroporated with Neurog b-line>unc-76::VenusYFP (green) and Nodal b8.18>H2B::mCherry (magenta).
mCherry signalling is observed in the dorsal midline tail epidermis including the anterior bipolar tail neuro (aBTN) precursor.
Original annotation : Embryos co-electroporated with Neurog b-line>unc-76::VenusYFP (green) and Nodal b8.18>H2B::mCherry (magenta). Right panels are enlarged views of the images on the left. Dashed lines indicate displacement from clonally related epidermal cells (indicated by colour-coded brackets). Scale bars, 25 μm.

Lateral view of a full embryo (left pannel) and the tail (right pannel) of a WT Ciona robusta mid tailbud embryo co-electroporated with Neurog b-line>unc-76::VenusYFP (green) and Nodal b8.18>H2B::mCherry (magenta).
mCherry signalling is observed in the posterior dorsal midline tail epidermis and in the anterior bipolar tail neuron (pink arrowhead) but not in the posterior bipolar tail neuron (white arrowhead).
Right panels are enlarged views of the images on the left. Dashed lines indicate displacement from clonally related epidermal cells (indicated by colour-coded brackets). Scale bars, 25 μm.

Dorsal view of a late tailbud embryo probed for ci-Nodal expression.

Original annotation:
As development proceeded, Ci-Nodal expression in the left epidermis became down regulated. At the late tail bud stage, expression of Ci-Nodal was detected on the left side of the SV and on the left side of the posterior endoderm.

Dorsal view of a late tailbud embryo (12 hpf) probed for ci-Nodal (red) and Ci-Rx (green) expression (focus on the sensory vesicle region).
Image of DAPI staining is additionally merged (blue). Dorsal views are shown with anterior to the top. Dashed lines indicate the midline.

Original annotation:
Ci-Rx is expressed bilaterally in the SV until 10 hpf and then down regulated on the left side by 11 hpf when the expression of Ci-Nodal starts to be detectable in the left SV. At 12 hpf, Ci-Nodal expression in the left SV expands, while the expression of Ci-Rx is no longer detectable on the left side of the SV.

Dorsal view of a late tailbud embryo probed for ci-Nodal expression.

Aniseed Comment : update of annotation of (Imai et al.,2004), based on (Imai et al.,2006).

View of larvae showing reporter expression of the region upstream of nodal in tail epidermis, nerve cord and in part of head epidermis.