DOI: 10.5061/dryad.9s4mw6mt5
title: Data from: Opposing roles for Bmp signalling during the development of electrosensory lateral line organs

description: The lateral line system enables fishes and aquatic-stage amphibians to
 detect local water movement via mechanosensory hair cells in neuromasts,
 and many species to detect weak electric fields via electroreceptors
 (modified hair cells) in ampullary organs. Both neuromasts and ampullary
 organs develop from lateral line placodes, but the molecular mechanisms
 underpinning ampullary organ formation are understudied relative to
 neuromasts. This is because the ancestral lineages of zebrafish (teleosts)
 and Xenopus (frogs) independently lost electroreception. We identified
 Bmp5 as a promising candidate via differential RNA-seq in an
 electroreceptive ray-finned fish, the Mississippi paddlefish (Polyodon
 spathula; Modrell et al., 2017, eLife 6: e24197). In an experimentally
 tractable relative, the sterlet sturgeon (Acipenser ruthenus), we found
 that Bmp5 and four other Bmp pathway genes are expressed in the developing
 lateral line, and that Bmp signalling is active. Furthermore,
 CRISPR/Cas9-mediated mutagenesis targeting Bmp5 in G0-injected sterlet
 embryos resulted in fewer ampullary organs. Conversely, when Bmp
 signalling was inhibited by DMH1 treatment shortly before the formation of
 ampullary organ primordia, supernumerary ampullary organs developed. These
 data suggest that Bmp5 promotes ampullary organ development, whereas Bmp
 signalling via another ligand(s) prevents their overproduction. Taken
 together, this demonstrates opposing roles for Bmp signalling during
 ampullary organ formation.
description: # Data from: Opposing roles for Bmp signalling during the development of
 electrosensory lateral line organs Dataset DOI:
 [https://doi.org/10.5061/dryad.9s4mw6mt5](https://doi.org/10.5061/dryad.9s4mw6mt5) ## Description of the data and file structure In this dataset you will find images of sterlet sturgeon (*Acipenser ruthenus*) larvae underlying the data reported in **Supplementary File 1**, **Supplementary File 2** and **Supplementary File 3** of Campbell AS, Minařík M, Franěk R, Vazačová M, Havelka M, Gela D, Pšenička M, Baker CVH (2025) *eLife* 14, e99798 (DOI: 10.7554/eLife.99798) The dataset contains four main folders (zip archives). The first two folders (**Suppl_File_1-Bmp5** and **Suppl_File_1-Tyr)** correspond to the data in Supplementary File 1 of the paper: Breakdown by single-guide (sg)RNA mix of CRISPR/Cas9 experiments targeting *Bmp5* or *tyrosinase (Tyr, control)*. *Note:* *Tyr* crispant data are shared between this study and Minařík et al. (2024, preprint: DOI: [https://doi.org/10.1101/2023.04.15.537030](https://doi.org/10.1101/2023.04.15.537030)).  The third folder (**Suppl_File_2)** corresponds to Supplementary File 2 of the paper: Breakdown by individual crispant of CRISPR/Cas9 experiments targeting *Bmp5* or *tyrosinase* (*Tyr*, control). The fourth folder (**Suppl_File_3)** corresponds to Supplementary File 3 of the paper: Breakdown of ampullary organ counting data for DMH1 experiments (to block Bmp signalling) and older wild-type larvae. Images are of sterlet sturgeon larvae after chromogenic *in situ* hybridization (ISH) and in some cases after post-ISH immunostaining for Sox2. Images were focus-stacked using HeliconFocus software (HeliconSoft). Individual focus slices are not included here. For phenotype assessment and publication, the brightness and contrast of some images were adjusted, but these further changes were not applied to the image files presented here. All methods are fully described in the Materials and Methods section of the associated publication. **FILES** **File: Suppl_File_1-Bmp5.zip** **Description:** Images in this folder (zip archive) underlie the *Bmp5* crispant data reported in **Supplementary File 1** of the paper [Breakdown by single-guide (sg)RNA mix of CRISPR/Cas9 experiments targeting *Bmp5* or *tyrosinase (control)]*. Image files are organised into subfolders based on the sgRNA mix used and the ISH marker analysed (*Cacna1d* or *Kcnab3*). Within each subfolder, images of *Bmp5* crispants without a phenotype are included in a separate "no_phenotype" subfolder, where relevant. *Note:* Most of the non-phenotypic *Bmp5* crispants analysed with *Cacna1d* from Supplementary File 1 of the paper were recorded as "no phenotype" but not imaged. *Filenames in Suppl_File_1-Bmp5* Filenames include the CRISPR/Cas9 target gene (Bmp5), sgRNA mix (sgRxxx) and a unique 2-digit code (see section "Abbreviations in filenames" below). Filenames include the ISH marker used (Cacna1a or Kcnab3) and end in a 6-digit date in yy.mm.dd format (e.g., 21.03.08), corresponding to the date the ISH protocol was started. Filenames starting with AOxx\_ (e.g., AO07_, AO12_, AO22_) indicate the images used for the ampullary organ counting data reported for individual *Bmp5* crispants in Supplementary File 2 of the paper (these images are also provided, with the same filenames, in the **Suppl_File_2** folder, where the files are ordered by AOxx code to match the organisation of Supplementary File 2 of the paper). *Abbreviations in filenames:* Ar, *Acipenser ruthenus*; Ar45, *Acipenser ruthenus* stage 45; sgRxx_NPyy_, sgRNA mix plus 2-digit code for non-phenotypic crispant (e.g., sgR14_NP03 corresponds to sgRNA mix 1+4, non-phenotypic crispant 3); sgRxx_YPyy_, sgRNA mix plus 2-digit code for phenotypic crispant (e.g., sgR14_YP19 corresponds to sgRNA mix 1+4, phenotypic crispant 19). **File: Suppl_File_1-Tyr.zip** **Description:** Images in this folder (zip archive) underlie the *tyrosinase* (*Tyr*) crispant data reported in **Supplementary File 1** of the paper [Breakdown by single-guide (sg)RNA mix of CRISPR/Cas9 experiments targeting *Bmp5* or *tyrosinase* (control)]. *Note: Tyr* crispant data are shared between this study and Minařík et al. (2024, preprint: DOI: [https://doi.org/10.1101/2023.04.15.537030](https://doi.org/10.1101/2023.04.15.537030)).  Image files are organised into **AO number** (AO, ampullary organ) and **Pigmentation** subfolders (matching the organisation of Supplementary File 1 of the paper). In the **AO number** subfolder, images are further organised into subfolders based on the sgRNA mix used and the ISH marker analysed (*Cacna1d* or *Kcnab3*). Filenames starting with AOxx\_ (e.g., AO07_, AO12_, AO22_) indicate the images used for the ampullary organ counting data reported for individual *Tyr* crispants in Supplementary File 2 of the paper (these images are also provided, with the same filenames, in the **Suppl_File_2** folder, where the files are ordered by AOxx code to match the organisation of Supplementary File 2 of the paper). In the **Pigmentation** subfolder, images are further organised into subfolders based on the sgRNA mix used; each sgRNA-mix subfolder includes a "no_phenotype" subfolder for images of *Tyr* crispants without a phenotype (i.e., no effect on melanin pigmentation). Images of melanin pigmentation were not taken for 31 non-phenotypic Tyr crispants from Supplementary File 1 of the paper (all 24 analysed larvae for *Tyr* sgRNA 5+6, and 9 out of 15 non-phenotypic larvae for *Tyr* sgRNA 7+8). *Filenames in Suppl_File_1-Tyr:* Filenames include the CRISPR/Cas9 target gene (Tyr), sgRNA mix (sgRxxx) and a unique 2-digit or 3-digit code (see section "Abbreviations in filenames" below). Filenames include, where relevant, the ISH marker used (Cacna1d or Kcnab3) and "a-Sox2" if post-ISH immunostaining was performed with the anti-Sox2 antibody. Filenames end in a 6-digit date in yy.mm.dd format, with or without periods (e.g., 21.03.08 or 211020). This corresponds to the date the ISH protocol was started or, for images of melanin pigmentation, the date of the imaging. *Abbreviations in filenames:* Ar, *Acipenser ruthenus*; Ar45, *Acipenser ruthenus* stage 45; sgRxxx_yyy_, sgRNA mix plus 3-digit crispant code (e.g., sgR12_097\_ corresponds to sgRNA mix 1+2, crispant 97); sgRxx_NPyy_, sgRNA mix plus 2-digit code for non-phenotypic crispant (e.g., sgR14_NP03\_ corresponds to sgRNA mix 1+4, non-phenotypic crispant 3); sgRxx_YPyy_, sgRNA mix plus 2-digit code for phenotypic crispant (e.g., sgR14_YP19\_ corresponds to sgRNA mix 1+4, phenotypic crispant 19); wmD, wholemount dorsal view; wmL, wholemount left-side lateral view; wmR, wholemount right-side lateral view; wmV, wholemount ventral view. **File: Suppl_File_2.zip** **Description:** Images in this folder (zip archive) underlie the data reported in **Supplementary File 2** of the paper [Breakdown by individual crispant of CRISPR/Cas9 experiments targeting *Bmp5* or *tyrosinase* (control)]. Image files are organised into two main subfolders based on the target gene: **Bmp5 crispants** or **Tyr control crispants**. Within each of these subfolders, all filenames start with AOxx\_ (for example, AO07_, AO12_, AO22_), indicating that these images were used for the ampullary organ counting data reported for the individual *Bmp5* crispants or *Tyr* control crispants in Supplementary File 2 of the paper. These image files are also included, with the same filenames, in the relevant folders in **Suppl_File_1-Bmp5** and **Suppl_File_1-Tyr** (in which the image files are organised by sgRNA mix and marker used). *Filenames in Suppl_File_2* Filenames include the CRISPR/Cas9 target gene (Bmp5 or Tyr), sgRNA mix (sgRxxx) and a unique 2-digit or 3-digit code (see section "Abbreviations in filenames" below). Filenames include the ISH marker used (Cacna1d or Kcnab3), plus "a-Sox2" if post-ISH immunostaining was performed with the anti-Sox2 antibody. Filenames end in a 6-digit date in yy.mm.dd format, with or without periods (e.g., 21.02.22 or 210702), corresponding to the date the ISH protocol was started. *Abbreviations in filenames:* Ar, *Acipenser ruthenus*; Ar45, *Acipenser ruthenus* stage 45; sgRxxx_yyy_, sgRNA mix plus 3-digit crispant code (e.g., sgR12_097\_ corresponds to sgRNA mix 1+2, crispant 97); sgRxx_NPyy_, sgRNA mix plus 2-digit code for non-phenotypic crispant (e.g., sgR14_NP03\_ corresponds to sgRNA mix 1+4, non-phenotypic crispant 3); sgRxx_YPyy_, sgRNA mix plus 2-digit code for phenotypic crispant (e.g., sgR14_YP19\_ corresponds to sgRNA mix 1+4, phenotypic crispant 19); wmD, wholemount dorsal view; wmL, wholemount left-side lateral view; wmR, wholemount right-side lateral view; wmV, wholemount ventral view. **File: Suppl_File_3.zip** **Description:** Images in this folder (zip archive) underlie the data reported in **Supplementary File 3** of the paper [Breakdown of ampullary organ counting data for DMH1 experiments (to block Bmp signalling) and older wild-type larvae]. Image files are organised into three main subfolders, corresponding to the different categories in Supplementary File 3 of the paper: **DMH1-treated**, **DMSO_Controls** and **Older_WT_Larvae**. *Filenames in Suppl_File_3* Filenames start with a 2-digit code for the individual larva, corresponding to the numbers in Supplementary File 3 of the paper. Filenames include the ISH marker used (Cacna1d or Kcnab3), plus "Sox2" if post-ISH immunostaining was performed with the anti-Sox2 antibody. Filenames end in a 6-digit date in yy.mm.dd format (e.g., 22.04.05), corresponding to the date the ISH protocol was started. For the older wild-type (WT) larvae, the filename includes the length of the larva in cm (2.0 cm or 2.8 cm). *Abbreviations in filenames:* Ar, *Acipenser ruthenus*; S45, stage 45