SSBD:database

289-Fujimoto-MitralCell/Fig4H_RhoABC-KO_RhoA_3

Author reviewed

Z-series images of RhoABC-knockout and RhoA-rescued mouse brain at P6 in which mitral cells were labed by tdTomato

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Project

289-Fujimoto-MitralCell

Dataset ID

Fig4H_RhoABC-KO_RhoA_3

Kind

Image data

Title

Z-series images of RhoABC-knockout and RhoA-rescued mouse brain at P6 in which mitral cells were labed by tdTomato

Description

Z-series images of RhoA, RhoB, and RhoC knockout mouse brain at P6 in which mitral cells were labed by tdTomato. RhoA, B, and C were knocked out by using CRISPR-Cas9 and in utero electroporation. In this experoment, CRISPR- resistant-pCAG-RhoA was also introduced by in utero electroporation to test effect of the compensatory exprssion of RhoA.

Folder Name

Fig4H_RhoABC-KO_RhoA_3

File Format

.lif

File Size

785MB

File Separation Method

3 series

Figure Number

Fig4H

Dimensions

1024x1024x248x2x1

X Scale

0.568 micrometer

Y Scale

0.569 micrometer

Z Scale

5 micrometer

T Scale

Channel

2 channel

Quantitative Data

Paper(s)

Biosample

Description

A fixed and cleared olfactory bulb of P6 mouse in which RhoA, RhoB, and RhoC were knocked out by CRISPR-Cas9. In addition, RhoA expression was rescued by introduction of RhoA plasmid. tdTomato was expressed in mitral/tufted cells.

Organism

Mus musculus (NCBITaxon:10090)

Strain

ICR (https://www.jslc.co.jp/pdf/mouse/002_ICR2013.pdf)

Cell

mitral cell (CL:1001502)

Cell line

MeSH

Neurons (D009474)

Microscopy, Confocal (D018613)

Olfactory Bulb (D009830)

Neuronal Plasticity (D009473)

Cell Body (D066146)

Brain (D001921)

Dendrites (D003712)

Intrinsic Variable

pCAX-hCas9 and the gRNA for RhoA, RhoB, and RhoC were introduced to knockout RhoA, RhoB, and RhoC by CRISPR-Cas9 method.

Extrinsic Variable

By using in utero electroporation, the authors electroporated a pCAFNF-tdTomato and pCAG-FLPo at E12. CRISPR- resistant-pCAG-RhoA were also introduced.

Ontology

Anatomical Entity

UBERON

olfactory bulb brain (UBERON:0000955)

Biological Process

neuron development neuron remodeling (GO:0016322)

Cellular Component

cell body (GO:0044297)

dendrite (GO:0030425)

Molecular Function

MeSH Anatomy

Neurons (D009474)

Microscopy, Confocal (D018613)

Olfactory Bulb (D009830)

Neuronal Plasticity (D009473)

Cell Body (D066146)

Brain (D001921)

Dendrites (D003712)

MeSH BP

Neurons (D009474)

Microscopy, Confocal (D018613)

Olfactory Bulb (D009830)

Neuronal Plasticity (D009473)

Cell Body (D066146)

Brain (D001921)

Dendrites (D003712)

MeSH CC

Neurons (D009474)

Microscopy, Confocal (D018613)

Olfactory Bulb (D009830)

Neuronal Plasticity (D009473)

Cell Body (D066146)

Brain (D001921)

Dendrites (D003712)

MeSH MF

Neurons (D009474)

Microscopy, Confocal (D018613)

Olfactory Bulb (D009830)

Neuronal Plasticity (D009473)

Cell Body (D066146)

Brain (D001921)

Dendrites (D003712)

Genetics

Gene

Protein / Probe

Method / Genotype

transfection electroporation

Gene

Rhoa

Protein / Probe

Method / Genotype

gene knock out electroporation

Gene

Rhob

Protein / Probe

Method / Genotype

gene knock out electroporation

Gene

Rhoc

Protein / Probe

Method / Genotype

gene knock out electroporation

Gene

Protein / Probe

Transforming protein RhoA

Method / Genotype

transfection electroporation

Treatments

Reagent / Compound

Concentration

Fold Dilution

Imaging Method

confocal microscopy (FBbi:00000251)

Detection Method

Visualization Method

tdTomatoFP 4',6-diamidino-2-phenylindole (DAPI)

Illumination Method

Source of Contrast

distribution of a specific protein

Imaged Parameter

fluorescence emission

Sample Preparation Method

formaldehyde fixed tissue vibratome-sectioned tissue tissue clearing

MeSH

Neurons (D009474)

Microscopy, Confocal (D018613)

Olfactory Bulb (D009830)

Neuronal Plasticity (D009473)

Cell Body (D066146)

Brain (D001921)

Dendrites (D003712)

Instrument

Body

Leica DMi8

Module

Light Source

Detector

Objective

Leica HC PL APO 20x/0,75 IMM CORR CS2

Filter Set

Dichroic