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.

Assay platform	Illumina HiSeq 2000
Layout	Paired-end
Stranded	Yes
Read length	49
Normalized data types	FPKM RLE
Wild type - Replica 1
	Territories	whole embryo -
	Biomaterial(s)	C. robusta formely Ciona int. type A, Stage 0 (Unfertilized egg)
		C. robusta formely Ciona int. type A, Stage 8 (64-cell)
		C. robusta formely Ciona int. type A, Stage 11 (early gastrula)
		C. robusta formely Ciona int. type A, Stage 12 (mid gastrula)
		C. robusta formely Ciona int. type A, Stage 15 (mid neurula)
		C. robusta formely Ciona int. type A, Stage 21 (mid tailbud I)
		C. robusta formely Ciona int. type A, Stage 26 (hatching larva)
Wild type - Replica 2
	Territories	whole embryo -
	Biomaterial(s)	C. robusta formely Ciona int. type A, Stage 0 (Unfertilized egg)
		C. robusta formely Ciona int. type A, Stage 8 (64-cell)
		C. robusta formely Ciona int. type A, Stage 11 (early gastrula)
		C. robusta formely Ciona int. type A, Stage 12 (mid gastrula)
		C. robusta formely Ciona int. type A, Stage 15 (mid neurula)
		C. robusta formely Ciona int. type A, Stage 21 (mid tailbud I)
		C. robusta formely Ciona int. type A, Stage 26 (hatching larva)

Dorsal view of neurula embryo probed for Ci-ChAT expression.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	No expression -

Dorsal view of neurula embryo probed for Ci-vAChTP expression.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	No expression -

Dorsal view of an early tailbud wild type embryo probed for ChAT expression.
Expression on the left-hand side (L) or right-hand side (R) is indicated at the top and bottom of each panel, respectively.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	lateral VG ependymal precursors -

Dorsal and Lateral view of a early Tailbud stage wild type embryo for Ci-ChAT expression. Expression is seen in the lateral cells of the visceral ganglion.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	lateral VG ependymal precursors -

Dorsal view of an early tailbud embryo in which b5.3 cell was ablated at 16-cell stage and probed for ChAT expression. Expression on the left-hand side (L) or right-hand side (R) is indicated at the top and bottom of each panel, respectively. The ablated side is indicated by arrowheads.

Orignal annotation :
Expression of low level of ChAT on the ablated side may be due to the recovery of lateral Nodal expression that is seen occasionally following ablation (Hudson and Yasuo, 2006).

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	No expression -

Lateral view of an early tailbud ermbryo probed for Ci-ChAT expression. A the early tailbud stage, Ci-Chat was detected in part of the caudal neural tube (nt, neurual tube; nc, notochord).

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	tail nerve cord -

Dorsal views of early tailbud stage embryo expressing vAChTP. The upper panel shows WISH signals, the middle DAPI signals and the lower the merged images.
Arrowheads indicate nuclei of the cells that express vAChTP. Open arrows indicate the constrictions in the row of nuclei of the lateral cells. Red dots indicate nuclei of the landmark cells.

Original annotation:
Ci-vAChTP and Ci-Lhx3 are co-expressed in the three landmark cells on each side.
NB: All genes described in figure 4 are expressed exclusively in the lateral cell rows.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	lateral TNC ependymal precursors - lateral VG ependymal precursors -

Dorsal view of an early tailbud wild type embryo probed for vAChTP expression.
Numbers indicate the number of embryos showing expression over the total number of embryos analysed. 'r' indicates severely reduced expression or expression on only one side of the embryo.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	lateral TNC ependymal precursors - lateral VG ependymal precursors -

lateral view of an early tailbud embryo probed for Ci-vAChTP expression. At the early tailbud stage, Ci-vAChTP was first detected in the part of the caudal neural tube (nt, neural tube; nc, notochord).

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	visceral ganglion -

Two-color fluorescence WISH was performed at the early tailbud stage. vAChTP expression is shown in green, Hox1 in red.

Original annotation:
Ci-vAChTP is expressed in the visceral ganglion and in part of tail nerve cord. The anterior expression domain of vAChTP is located in the gap between the two expression domains of Ci-Hox1, and the posterior domain of Ci-vAChTP is located postero-lateraly in relation to the second domain of Ci-Hox1.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	tail nerve cord - visceral ganglion -

Dorsal view of a mid tailbud embryo in which A7.4, a7.10 and notochord cells were ablated at 64-cell stage and probed for vAChTP expression. Numbers indicate the number of embryos showing expression over the total number of embryos analysed. 'r' indicates severely reduced expression or expression on only one side of the embryo.

Original annotation:
Expression of vAChTP was detected in N+FP ablated embryos by conventional in situ hybridisation.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	lateral VG ependymal precursors -

Lateral view of a middle tailbud embryo probed for Ci-ChAT expression.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	lateral VG ependymal precursors -

Dorsal views of mid tailbud stage embryo expressing vAChTP. The upper panel shows WISH signals, the middle DAPI signals and the lower the merged images.
Arrowheads indicate nuclei of the cells that express vAChTP. Open arrows indicate the constrictions in the row of nuclei of the lateral cells. Red dots indicate nuclei of the landmark cells.

Original annotation:
Ci-vAChTP is expressed in the M2, M3 and M4 cells.
NB: All genes described in figure 4 are expressed exclusively in the lateral cell rows.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	lateral TNC ependymal precursors - lateral VG ependymal precursors -

Dorsal view of an early tailbud wild type embryo probed for vAChTP expression.
Expression on the left-hand side (L) or right-hand side (R) is indicated at the top and bottom of each panel, respectively.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	lateral TNC ependymal precursors - lateral VG ependymal precursors -

Antero-dorsal and lateral views of a middle tailbud embryo probed for Ci-vAChTP expression. The positive region divided in a double line along the neural tube of the tailbud.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	lateral VG ependymal precursors -

Two-color fluorescence WISH was performed at the mid tailbud stage. vACHTP expression is shown in red, Fgf8/17/18 in green.
Ci-vACHTP is expressed the in visceral ganglion.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	tail nerve cord - visceral ganglion -

Two-color fluorescence WISH was performed at the mid tailbud stage. vACHTP expression is shown in green.

Original annotation:
Ci-vACHTP is expressed in part of visceral ganglion and in part of tail nerve cord. Ci-vACHTP is expressed in the gap between the two Ci-Hox1 domains as at the early stage and in the domain just ventrolateral to the second domain of Ci-Hox1.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	tail nerve cord - visceral ganglion -

The central nervous system of a mid tailbud wild type embryo co-electroporated with FGF8/17/18>RFP (in red) and AchTP>GFP (in green).

Original annotation:
AchTP was detected in A11.118, A11.117 and A10.57 cell pairs (visceral ganglion and tail nerve cord).

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	tail nerve cord - visceral ganglion -

Dorsal view of an early tailbud embryo in which b5.3 cell was ablated at 16-cell stage and probed for vAChTP expression.
Expression on the left-hand side (L) or right-hand side (R) is indicated at the top and bottom of each panel, respectively. The ablated side is indicated by arrowheads.

Orignal annotation :
Expression of low level of vAChTP on the ablated side may be due to the recovery of lateral Nodal expression that is seen occasionally following ablation (Hudson and Yasuo, 2006).

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	No expression -

Lateral view of a tailbud embryo probed for ChAT expression. ChAT is expressed in the latteral cells of the visceral ganglion derived from A9.30 and A9.29 A-neural plate cells. (ChAT corresponds to KH2012:KH.C1.498.v2.A.SL1-2 transcript)

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	visceral ganglion -

Lateral view of a tailbud embryo probed for vChATP expression. vAChTP is expressed in the latteral cells of the visceral ganglion derived from A9.30 and A9.29 A-neural plate cells.

(vChATP corresponds to KH2012:KH.C1.498.v1.A.SL1-1 transcript)

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	visceral ganglion -

Dorsal view of a late tailbud wild type embryo probed for ChAT expression.
Numbers indicate the number of embryos showing expression over the total number of embryos analysed. 'r' indicates severely reduced expression or expression on only one side of the embryo.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	anterior sensory vesicle - lateral VG ependymal cells -

Dorsal view of a late tailbud embryo in which notochord cells were ablated at 64-cell stage and probed for ChAT expression.
An inset showing the control expression of the corresponding batch of embryos is presented.
Numbers indicate the number of embryos showing expression over the total number of embryos analysed. 'r' indicates severely reduced expression or expression on only one side of the embryo.

Original annotation:
ChAT expression was not affected following ablation of the four anterior notochord precursors.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	lateral VG ependymal cells -

Dorsal view of a late tailbud embryo in which A7.4 cells were ablated at 64-cell stage and probed ChAT expression.
Numbers indicate the number of embryos showing expression over the total number of embryos analysed. 'r' indicates severely reduced expression or expression on only one side of the embryo.

Original annotation:
Ablation of these cells had no effect on ChAT expression.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	lateral VG ependymal cells -

Dorsal view of a late tailbud embryo in which A7.4 and a7.10 cells were ablated at 64-cell stage and probed for ChAT expression.
An inset showing the control expression of the corresponding batch of embryos is presented.
Numbers indicate the number of embryos showing expression over the total number of embryos analysed. 'r' indicates severely reduced expression or expression on only one side of the embryo.

Original annotation:
ChAT expression still persisted after the removal of a7.10 together with A7.4.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	lateral VG ependymal cells - tail nerve cord -

Dorsal view of a late tailbud embryo in which A7.4, a7.10 and notochord cells were ablated at 64-cell stage and probed for ChAT expression.
An inset showing the control expression of the corresponding batch of embryos is presented.
Numbers indicate the number of embryos showing expression over the total number of embryos analysed. 'r' indicates severely reduced expression or expression on only one side of the embryo.

Original annotation:
Expression of ChAT was detected in N+FP ablated embryos by conventional in situ hybridisation.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	lateral VG ependymal cells -

Lateral view of a late tailbud probed for ChAT expression. ChAT is expressed in the latteral cells of the visceral ganglion derived from A9.30 and A9.29 A-neural plate cells.

(ChAT corresponds to KH2012:KH.C1.498.v2.A.SL1-2 transcript)

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	lateral VG ependymal cells -

Lateral view of a late tailbud probed for vAChTP expression. vAChTP is expressed in the latteral cells of the visceral ganglion derived from A9.30 and A9.29 A-neural plate cells.
(vChATP corresponds to KH2012:KH.C1.498.v1.A.SL1-1 transcript)

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	lateral VG ependymal cells -

Late tailbud embryo probed for Ci-vAChTP expression (blue), Ci-Pax2/5/8 expression (green) and Ci-En expression (red).

Original annotation:
According to the gene expression map (Ikuta and Saiga, 2007), expression of Ci-vAChTP was detected in A11.117, A11.118 and A10.57 cell pairs.

Aniseed annotation:
These cells have several fates corresponding to lateral visceral ganglion and lateral tail nerve cord.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	lateral TNC ependymal cells - lateral VG ependymal cells -

Dorsal views of late tailbud stage embryo expressing vAChTP. The upper panel shows WISH signals, the middle DAPI signals and the lower the merged images.
Arrowheads indicate nuclei of the cells that express vAChTP. Open arrows indicate the constrictions in the row of nuclei of the lateral cells. Red dots indicate nuclei of the landmark cells.

Original annotation:
Ci-vAChTP is expressed exclusively in the four landmarks cells.
NB: All genes described in figure 4 are expressed exclusively in the lateral cell rows.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	lateral TNC ependymal cells - lateral VG ependymal cells -

Two-color fluorescence WISH was performed at the late tailbud stage. vAChTP expression is shown in green, Hox1 in red.

Original annotation:
vAChTP is expressed in the visceral ganglion. At this stage, vAChTP is expressed in two distinct domains aligned along the A-P axis. While the posterior domain is located in the gap between the two domains of Ci-Hox1 continuously, the anterior domain lies the middle of the first domain of Ci-Hox1.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	tail nerve cord - visceral ganglion -

Two-color fluorescence WISH was performed at the late tailbud stage. vAChTP expression is shown in red, Dmbx in green.

Original annotation:
Ci-vAChTP is expressed in the visceral ganglion. The small anterior domain of Ci-vAChTP coincides with the Ci-Dmbx expression domain.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	tail nerve cord - visceral ganglion -

Two-color fluorescence WISH was performed at the late tailbud stage. vAChTP expression is shown in green, Lhx3 in red.

Original annotation:
Ci-vAChTP is expressed in the visceral ganglion. Expression of Ci-Lhx3 in the posterior CNS is detected in two domains with a narrow gap between them. The first domain of Ci-Lhx3 does not correspond to that of Ci-vAChTP.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	tail nerve cord - visceral ganglion -

Two-color fluorescence WISH was performed at the late tailbud stage. vAChTP expression is shown in red, Fgf8/17/18 in green.
Ci-vAChTP is expressed in the visceral ganglion and in the neck.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	neck - tail nerve cord - visceral ganglion -

The central nervous system of a late tailbud wild type embryo co-electroporated with Fgf8/17/18>RFP (in red) and AchTP>GFP (in green).
Arrowheads indicate motoneurons.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	tail nerve cord - visceral ganglion -

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

Original annotation:
Brain vesicle; visceral ganglion.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	anterior sensory vesicle - posterior sensory vesicle - visceral ganglion -

Lateral (A, B) and dorsal (C, D) views of a larvae probed for Ci-ChAT expression.
(B, D) High magnification views of the black rectangles in (A) and (C), respectively. The dotted lines in (B) and (D) indicate the boundary between the trunk and tail.

Original Annotation : Ci-ChAT is expressed mainly in the ventral region of the visceral ganglion. The white arrowheads (A, B) indicate expression in the dorsal region of the visceral ganglion. The black arrowhead in (C) indicates expression in the sensory vesicle.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	posterior sensory vesicle - visceral ganglion -

Dorsal and lateral views of larvae probed for Ci-ChAT. The positive region divided in a double line in the visceral ganglion of the larva (sv, sensory vesicle; vg, visceral ganglion; oc, ocellus; ot, otolith).

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	visceral ganglion -

Larvae wild type probed for Ci-vAChTP (a cholinergic neuron marker) expression (in red) and Ci-vGAT (a GABAergic neuron marker) expression (in green). The white arrowhead shows positive signal in the dorsal visceral ganglion.

Original annotation:
In uninjected control larvae, expression of Ci-vAChTP was detected in the sensory vesicle and the ventral visceral ganglion.

Aniseed annotation:
Expression in the tail nerve cord?

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	posterior sensory vesicle - tail nerve cord - visceral ganglion -

Late tailbud embryo probed for Ci-vAChTP expression (blue), Ci-Lhx3 expression (green) and Ci-SoxB1 expression (red).

Original annotation:
According to the gene expression map (Ikuta and Saiga, 2007), expression of Ci-vAChTP in A11.117, A11.118, A12.239 and A10.57 cell pairs.

Aniseed annotation:
These cells have several fates corresponding to lateral visceral ganglion and lateral tail nerve cord.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	lateral TNC ependymal cells - visceral ganglion -

Lateral view of a wild type tadpole embryo stained for Ci-vAChTP mRNA by in situ hybridization. Staining is observed in the dorsocaudal epidermal neurons (en), the visceral ganglion (vg), and in the otolith (ot) and ocellus (oc) cells.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	neck - ocellus - otolith - tail nerve cord - visceral ganglion -

Right side of a larvae probed for Ci-vAChTP expression. Cells positive for Ci-vAChTP numbered at least two on the ventral side of the visceral ganglion (sv, sensory vesicle; vg, visceral ganglion; oc, ocellus; ot, otolith).

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	neck - visceral ganglion -

Larvae probed for GFP reporter expression of the promoter region of Ci-vAChTP (named dCi-TPup in the paper).

Original Annotation :
Pictures 1 and 3 : left lateral views, anterior is to the left.
Pictures 2 and 4 : Right lateral views, anterior is to the right.
Picture 5 : Right dorsal view, anterior is to the right.

Pictures 1, 2, 5 : White arrowheads indicate the axon terminals of the cells labeled with asterisks. Some of the labeled cells have an axon with terminal endplates that seem to be neuromuscular junctions (white rectangles). Dotted lines indicate the approximate boundary of the anterior and posterior parts of the visceral ganglion.

Pictures 3, 4 : The white arrowheads indicate three GFP-labeled cells, the white arrows indicate axonal fibers from these cells. The dotted line indicates the trunk-tail boundary.

Picture 6 : The green cells in (A) and (B) are GFP-labeled cells, with expression driven by the Ci-vAChTP promoter. The red cells are LacZ-labeled cells with GAD reporter expression.

The red staining in the pics 1, 2, 3, 4 and 5 is due to the UA301 detection. UA301 is a nervous system marker.

In the posterior sensory vesicle, GFP expression driven by the dCi-TPup/GFP transgene was observed. Some extended posteriorly and ended in the middle part of the visceral ganglion.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	posterior sensory vesicle - tail nerve cord - visceral ganglion -

Larvae probed for GFP reporter expression of the promoter region of Ci-vAChTP (named dCi-TPup in the paper).

Original Annotation :

Pictures 1 and 3: Left lateral views, anterior is to the left.
Pictures 2 and 4 : Right lateral views, anterior is to the right.
Picture 5 (E) : left dorsal view
Picture 6 (J) : right dorsal view

Picture 1 and 2 : White arrowheads indicate the axon terminals of cells with an asterisk. The open arrowheads indicate three GFP-labeled cells around the ocellus. The white arrows indicate the boundary between the trunk and tail. OC, ocellus, OT, otolith.

Picture 3, 4, 5 and 6 : Arabic numerals indicate the conventional numbers of the GFP-labeled cells. Some additional cells also are evident (white arrowheads). Arrows indicate axonal fibers from No. 1 cells. The open arrowhead indicates the terminal end of the axon of the No. 2 cell. Dotted lines indicate the approximate boundaries of the anterior and posterior parts of the visceral ganglion.

Picture 7 : GFP labeled cells in the caudal nerve cord extend short processes posteriorly (white arrows).

In the posterior sensory vesicle, GFP expression driven by the dCi-TPup/GFP transgene was observed. Some extended posteriorly and ended in the middle part of the Visceral Ganglion.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	posterior sensory vesicle - tail nerve cord - visceral ganglion -

Expression GFP in larvae that developed from fertilized eggs electroporated with pCi-vAChTP-EGFP construct.

Original annotation :
Picture a: the GFP expression recapitulated the endogenous expression patterns of Ci-vAChTP. GFP fluorescence was observed in the brain vesicle (bv), visceral ganglion (vg), and dorsocaudal epidermal neurons (en) on the dorsal midline of the tail.

Picture b: in a larva showing GFP expression only in the visceral ganglion (vg), connections between the brain vesicle and visceral ganglion were not observed, while the motor axons (arrows) extending from the visceral ganglion were heavily labeled.

Picture c: in a larva showing GFP expression only in the visceral ganglion (vg), connections between the brain vesicle and visceral ganglion were not observed, while the motor axons (arrows) extending from the visceral ganglion were heavily labeled.

Picture d: a GFP-positive dorsocaudal epidermal neuron on the dorsal midline of the tail showed an axon (arrow) extending anteriorly along the nerve cord.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	neck - ocellus - otolith - visceral ganglion -

Lateral view of a WT Ciona robusta larva electroporated with VAChT>GFP reporter construct.
Original comment: Visualization of the motor ganglion (MG) by VACHT>GFP cholinergic reporter. The MG contains five bilaterally symmetrical pairs of cholinergic neurons that project their axons along the muscle bands in the tail.

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)	visceral ganglion -

Ciona intestinalis juveniles probed for Chat expression.
Cholinergic neurons are marked with Chat RNA probe, localized in cerebral ganglion (cg) as well as nerve fibers (arrowheads).
Original comment: ISH assay detected Chat expression throughout the cerebral ganglion and on nerve roots. Expression decreased in strength as nerves became thinner.
Scale bars: 100 μm

Stained molecule	KH2012:KH.C1.498 (CHAT; CPT2; CRAT)
Stained region(s)