Dataset DOI: 10.5061/dryad.80gb5mm5j

Description of the data and file structure

We addressed neural cell origins during development of the Clytia hemisphaerica planula larva. From our results we propose that two distinct neurogenesis pathways operate during Clytia embryogenesis, one involving ectodermal delamination, and one generating many nematoblasts and some neurons from i-cell-like precursors.

This dataset includes images and image stacks acquired in the course of studies of neurogenesis during larva formation in the hydrozoan jellyfish .

They are organised into 6 folders, containing images aquired by different kinds of microscopy following different experimental approaches, as presented in Ruggiero et al, Development 2026

Complete lists of the all image files are also provided in the excel file. list_of_images_.xlsx

Folder 1-ISH  (In situ Hybridisation images)-by Sandra Chevalier Antonella Ruggiero, Anna Ferraioli and Carine Barreau  

JPG mages of fixed Clytia hemisphaerica embryos and larvae following whole-mount colorimetric in situ hybridization detecting mRNAs for a panel of genes chosen for their selective expression in different neural cell types or their precursors. Images were aquired on Zeiss A2 and Olympus BX51 microscopes, and then converted to jpg formal.Files names each include tags as follows for Gene abbreviations, Experimental conditions (where applicable) and Developmental stages.

 

Gene abbreviations: (individual references provided in Ruggiero et al Development 2026)

Bra1 =Brachyury1

FoxQ2A =Fox Q2A

WegA1 = WegA1

Wnt3= Wnt3

ELAV= ELAV2

GLWamide=  GLWamide precursor PP2

Nematocilin= Nematocilin

RFamide = RFamide precursor PP5

Hlh6 =Hlh6

Neurogenin= Neurogenin

M14peptidase = M14 peptidase

Minicollagen34a = Minicollagen 3/4a

Mos3 =Mos3

Nanos = Nanos1

Piwi=Piwi1

Znf845=Znf845

Sox10=Sox10  (SoxB2)

 

Conditions:

cont = control, untreated

PKF = treated with the Wnt pathway inhibitor PKF118-310 at 0.8-1.3 μM

PKF1= treated with the Wnt pathway inhibitor PKF118-310 at 1.0μM

PKF0_6  = treated with the Wnt pathway inhibitor PKF118-310 at 0.6 µM

MoWnt = developed from an egg injected with a morpholino antisense oligonucleotide targeting Wnt3

 

Stages :

EG =Early Gastrula

MG= Mid Gastrula

P1  = Parenchymula

P2  =  Planula 2 days post fertilisation

P3  =  Planula 3 days post fertilisation

 

The subfolders contain images resulting from the experiments documented in the corresponding figures from Ruggiero et al 2006, Development

 

Subfolder figS2 

Effects of PKF treatment on polarised gene expression

File names : Gene abbreviation_condition_stage.jpg

 

Bra1_cont_P1.jpg

Bra1_cont_P2.jpg

Bra1_PKF_P2.jpg

FoxQ2a_cont_P1.jpg

FoxQ2a_cont_P2.jpg

FoxQ2a_PKF_P2.jpg

WegA1_cont_P1.jpg

WegA1_cont_P2.jpg

WegA1_PKF_P2.jpg

Wnt3_cont_P1.jpg

Wnt3_cont_P2.jpg

Wnt3_PKF_P2.jpg

 

Subfolder figS4

Effects of low-dose PKF treatment on neural gene expression

File names all : Gene abbreviation_stageCondition.jpg   b is added to distinsh files of samples with  the same genes, stages and conditions

ELAV_P2cont.jpg

ELAV_P2PKF0_6.jpg

ELAV_P2PKF1

GLWamide_P2cont

GLWamide_P2PKF0_6

GLWamide_P2PKF1

Nematocilin_P2cont

Nematocilin_P2PKF0_6

Nematocilin_P2PKF0_6b

Nematocilin_P2PKF1

RFamide_P2cont

RFamide_P2PKF0_6

RFamide_P2PKF1

 

Subfolder fig1

Neural gene expression across stages

File names : Gene abbreviation_stage.jpg

 

ELAV_EG.jpg

ELAV_P1.jpg

ELAV_P2.jpg

ELAV_P3.jpg

GLWamide_EG.jpg

GLWamide_P1.jpg

GLWamide_P2.jpg

GLWamide_P3.jpg

            Hlh6_EG.jpg

Hlh6_P1.jpg

Hlh6_P2.jpg

Hlh6_P3.jpg

M14peptidase_P1.jpg

M14peptidase_P2.jpg

M14peptidase_P3.jpg

Minicollagen34a_P1.jpg

Minicollagen34a_P2.jpg

Minicollagen34a_P3.jpg

Mos3_EG.jpg

Mos3_P1.jpg

Mos3_P2.jpg

Mos3_P3.jpg

Nanos_EG.jpg

Nanos_P1.jpg

Nanos_P2.jpg

Nanos_P3.jpg

Nematocilin_P1.jpg

Nematocilin_P2.jpg

Nematocilin_P3.jpg

Neurogenin_EG.jpg

Neurogenin_P1.jpg

Neurogenin_P2.jpg

Neurogenin_P3.jpg

Piwi_EG.jpg

Piwi_P1.jpg

Piwi_P2.jpg

Piwi_P3.jpg

RFamide_EG.jpg

RFamide_P1.jpg

RFamide_P2.jpg

RFamide_P3.jpg

Znf845_EG

Znf845_P1.jpg

Znf845_P2.jpg

Znf845_P3.jpg 

 

Subfolder fig3 ISH

Neurogenic gene expression in Mid-gastrula embryos

File names : Gene abbreviation_stage_Focal plane.jpg

 

ELAV_MG_1.jpg

ELAV_MG_2.jpg

Hlh6_MG_1.jpg

Hlh6_MG_2.jpg

Sox10_MG_1.jpg

Sox10_MG_2.jpg

 

Subfolder fig6

Effects of PKF treatment on neural gene expression

File names all : Gene abbreviation_condition_stage.jpg   To distinsh files of samples with  the same genes, stages and conditions , B is added to the condition, or a, b or c to the stage.

 

            ELAV_cont_P1.jpg

ELAV_cont_P2.jpg

ELAV_PKF_P1.jpg

ELAV_PKF_P2.jpg

GLWa_cont_P2.jpg

GLWa_MoWnt_P2.jpg

GLWamide_contB_P2.jpg

GLWamide_PKF_P2.jpg

HlH6_cont_P1.jpg

HlH6_PKF_P1.jpg

Minicollagen34a_cont_P1.jpg

Minicollagen34a_cont_P2.jpg

Minicollagen34a_contB_P1.jpg

Minicollagen34a_contB_P2.jpg

Minicollagen34a_MoWnt_P1.jpg

Minicollagen34a_MoWnt_P2.jpg

Minicollagen34a_PKF_P1.jpg

Minicollagen34a_PKF_P2.jpg

Mos3_cont_EG.jpg

Mos3_cont_P1.jpg

Mos3_cont_P2.jpg

Mos3_contB_P1.jpg

Mos3_MoWnt_EG.jpg

Mos3_MoWnt_P1.jpg

Mos3_MoWnt_P1a.jpg

Mos3_MoWnt_P1b.jpg

Mos3_MoWnt_P2.jpg

Mos3_MoWnt_P2a

Mos3_MoWnt_P2b

Mos3_PKF_P1.jpg

Nanos1_cont_EG.jpg

Nanos1_cont_P1.jpg

Nanos1_cont_P2.jpg

Nanos1_contB_P1.jpg

Nanos1_contB_P2.jpg

Nanos1_MoWnt_EG.jpg

Nanos1_MoWnt_P1.jpg

Nanos1_MoWnt_P2

Nanos1_MoWnt_P2a

Nanos1_MoWnt_P2b

Nanos1_PKF_P1

Nanos1_PKF_P2

Nematocilin_cont_P2

Nematocilin_contB_P2

Nematocilin_MoWnt_P2

Nematocillin_MoWnt_P2a

Nematocilin_MoWnt_P2b

Nematocilin_PKF_P2.jpg

Neurogenin_cont_P1.jpg

Neurogenin_PKF_P1.jpg

RFamide_cont_P2.jpg

RFamide_contB_P2.jpg

RFamide_MoWnt_P2.jpg

RFamide_PKF_P2.jpg

RFamide_PKF_P2b.jpg

Znf845_cont_EG.jpg

Znf845_cont_P1.jpg

Znf845_cont_P2.jpg

Znf845_contB_P1.jpg

Znf845_contB_P2.jpg

Znf845_MoWnt_EG.jpg

Znf845_MoWnt_P1.jpg

Znf845_MoWnt_P1a.jpg

Znf845_MoWnt_P1b.jpg

Znf845_MoWnt_P1c.jpg

Znf845_MoWnt_P2.jpg

Znf845_MoWnt_P2a.jpg

Znf845_MoWnt_P2b.jpg

Znf845_PKF_P1.jpg

Znf845_PKF_P2.jpg

 

Folder 2-Fluorescence ISH by Antonella Ruggiero, Evelyn Houliston (.tif stacks)

 

Confocal image stacks of Clytia planulae fixed 2 days post fertilization and processed for fluorescence in situ hybridization. Stacks were  aquired on a Leica SP5 confocal microscope.

In each case images from three channels are combined as follows

Piwi1 mRNA detection (green) , Znf854 mRNA detection (magenta);  Hoechst staining of nuclei (blue)

or

Znf845 mRNA detection (magenta) and Minicollagen3/4a mRNA detection (yellow). .

 

File names : Planula First gene abbreviation Second gene abbreviation StackID.tif

 

Where Gene abbreviations are

Piwi = Piwi1

Znf = Znf854

Minicol= Minicollagen3/4a

 

PlanulaPiwiZnf2-6.tif

PlanulaPiwiZnf2-6b.tif

PlanulaPiwiZnf2-a.tif

PlanulaPiwiZnf2-hm.tif

PlanulaPiwiZnf2-1.tif

PlanulaZnfMinicol2-5hm.tif

PlanulaZnfMinicol2-6.tif

 

 

Folder 3-HCR by Sandra Chevalier (.tif stacks)

Confocal image stacks of Clytia mid-Gastrula stage embryos fixed and processed for Hybridisation Chain Reaction detections of  Sox10 and Hlh6 mRNAs. Stacks were aquired on a Leica Stellaris confocal microscope, and three channels combined using Fiji..

Images from three channels detected :  Sox10 mRNA (yellow) ; Hlh6 mRNA  (magenta) mRNAs; Hoechst staining of DNA (cyan).

File names : Gastrula EmbryoID First gene Second gene -overlay-.tif

Gastrula2-Hlh6Sox10.tif

Gastrula5-Hlh6Sox10.tif 

 

Folder 4-Gastrulation morphology by Evelyn Houliston, Sandra Chevalier (.tif stacks)

Confocal image stacks of Clytia gastrulae fixed and stained with rhodamine-phalloidin (magenta) for F-actin  and Hoechst (cyan) for DNA.  Stacks were  aquired on a Leica SP8 confocal microscope and the two channels combined using Fiji.

File names : StageEmbryoID- PhalloidinHoechst.tif

Stages :

EG =Early Gastrula

MG= Mid Gastrula

EG1-PhalloidinHoechst.tif

EG2-PhalloidinHoechst.tif

EG3-PhalloidinHoechst.tif

MG1-PhalloidinHoechst.tif

MG2-PhalloidinHoechst.tif

 

Folder 5-Gastrula Biscection experiments by Carine Barreau, Evelyn Houliston, Sandra Chevalier (.tif stacks)

 

Confocal images of Planulae stained with Hoechst for DNA (blue) ) Alexa488-Phalloidin (yellow;  Endogenous GFP is also detected in this channel), anti-FMRFamide (magenta) and anti-LWamide (green) antibodies

Some of these developed from gastrula stage embryos that were manually bisected at the gastrula stage into oral and aboral fragments.

 

File names : Date_ConditionEmbryoID-stain.tif

 

Conditions:

cont = control derived from uncut gastrula

oral = derived from oral gastrula fragment

aboral = derived from aboral gastrula fragment

 

Stains:

Phal = Phalloidin

Ho= Hoechst

GLW- =anti-LWamide

RF = anti-FMRFamide

 

020621-cont5-GLW-RF.tif

020621-cont5-PhalHo.tif

020621-oral12-GLW-RF.tif

020621-oral12-PhalHo.tif

020621-oral17-GLW-RF.tif

020621-oral17-PhalHo.tif

020621-aboral13-GLW-RF.tif

020621-aboral13-PhalHo.tif

020621-aboral23-GLW-RF.tif

020621-aboral23-PhalHo.tif

 

Folder 6-Lineage tracing by Sandra Chevalier ,Carine Barreau, Evelyn Houliston, Sandra Chevalier

Lineage tracing of gastrula regions to the planula stage by photoconversion of Dendra-2 from green to red. The Dendra2 protein was expressed from mRNA injected into the egg.

Confocal .tif  image  or stacks were aquired for each channel on a Leica Stellaris confocal microscope and colour-combined using in FIJI :

Photoconverted Dendra2 (magenta) , Phalloidin-Alexa488/endogenous GFP (green) , Hoechst staining of nuclei (cyan), anti-LWamide staining or anti-RFamide staining (yellow).

File names : experimentID_Condition-AntibodyEmbryoID-imageID-overlay or separate channel-overlay version (optional).tif

 

Conditions:

Oral  = oral patch of photoconverted Dendra2

Aboral=  Aboral patch of photoconverted Dendra2

 

Antibodies:

LWa =anti LWamide

RFa = anti-RFamide

 

ex4-Oral-RFa3-10-separate.tif

ex4-Oral-RFa3-10-overlay.tif

ex8-Oral-RFa4-2-separate.tif

ex8-Oral-RFa4-2-overlay.tif

ex8-Oral-RFa4-4-separate.tif

ex8-Oral-RFa4-4-overlay.tif

ex5-Aboral-LWa11-1-overlay.tif

ex5-Aboral-LWa10-4-overlay-1.tif

ex5-Aboral-LWa10-4-overlay-2.tif