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.

Expression of Ci-opsin3 in Ciona intestinalis embryo. Ci-opsin3 was not detected in gastrula embryos by whole-mount in situ hybridization using a digoxigenin-labeled RNA probe.

Expression of Ci-opsin3 in Ciona intestinalis embryos. Ci-opsin3 was detected at the neural plate stage by whole-mount in situ hybridization using a digoxigenin-labeled RNA probe.

Original annotation: the Ci-opsin3 mRNA first appeared in anterior brain progenitor cells at the neural plate stage.

Aniseed annotation: precursors of the anterior brain are localized in row III and row IV in the neural plate.

Expression of Ci-opsin3 in Ciona intestinalis embryos. Ci-opsin3 was detected in early neurula embryos by whole-mount in situ hybridization using a digoxigenin-labeled RNA probe.

Original annotation: the expression extended posteriorly within the central nervous system at the early neurula stage.

Expression of Ci-opsin3 in C. intestinalis embryos. Ci-opsin3 was detected in mid neurula embryos by whole-mount in situ hybridization using a digoxigenin-labeled RNA probe.

Original annotation:at the mid-neurula stage, Ci-opsin3 transcripts were abundantly present in the developing brain and weaker signals were also found in cells at the posterior neural fold, which would eventually form the dorsal part of the nerve cord (roof plate).

Lateral (A') and dorsal (A'') views of an early tailbud stage wild type embryo probed for Ci-opsin3 (cilv025d01) expression.

Original annotation:
Expression was detected in the whole brain.

Expression of Ci-opsin3 in Ciona intestinalis embryos. Ci-opsin3 was detected in tailbud embryos by whole-mount in situ hybridization using a digoxigenin-labeled RNA probe.

Original annotation: at the tailbud stage, Ci-opsin3 was expressed trongly in the brain and faintly in the dorsal midline of the tail nerve cord.

Lateral view of a mid tailbud stage wild type embryo probed for Ci-opsin3 expression.

Original annotation:
Expression was detected in the whole brain.

Lateral (C') and dorsal (C'') views of a late tailbud stage wild type embryo probed for Ci-opsin3 expression.

Original annotation:
Expression was detected in the whole brain.

Dorsal view of a late tailbud stage wild type embryo probed for Ci-opsin3 expression.

Dorsal view of a late tailbud stage wild type embryo probed for Ci-opsin3 expression. This picture shows expression in the photosensory organs.

Conversion to aniseed format of DBTGR database entry from Kusakabe team.

Original annotation:
Brain vesicle.

Tadpole wild type embryo probed for a retinal G protein-coupled receptor opsin 3.

Expression of Ci-opsin3 in Ciona intestinalis embryo. Ci-opsin3 was detected in tadpole larvae by whole-mount in situ hybridization using a digoxigenin-labeled RNA probe.

Original annotation: in the larva, Ci-opsin3 is expressed in the entire region of brain vesicle containing the pigment cup cells of the ocellus and the pigment cell of the otolith.

Lateral view of a tadpole larva. Spatial localization of transcripts of Ci-opsin3 was visualized by whole-mount in situ hybridization.