Expression Database

GUDMAP:21893

Gene:	Cited1, Cbp/p300-interacting transactivator with Glu/Asp-rich carboxy-terminal domain 1
Stage:	Theiler Stage 23
Tissue:	urinary system
Species:	Mus musculus
Assay Type:	TG mouse marker strain

Images

	Wholemount nuc-TagRFP-T detection in 15.5dpc Cited1nuc-TagRFP-T/+ UGS. Nuc-TagRFP-T fluorescence was visible in the cap mesenchyme cells of the nephrogenic zone in 15.5dpc kidneys and the male reproductive system. Nuc-TagRFPT expression was limited to the Cited1 domain.
	TagRFP-T fluorescent cells apposing the ureteric bud, co-localize with cells detectable with TagRFP-T antibody. Cited1nuc-TagRFP-T 15.5 dpc kidneys were probed with anti-TagRFP-T and anti-Cytokeratin 5 antibodies. Co-localization of native TagRFPT fluorescent cells and cells positive for anti-TagRFP-T antibody are observable in the cap mesenchyme at the tips of the ureteric buds (Cytokeratin positive).
	TagRFP-T fluorescent cells in the cap mesenchyme co-localize with cells detectable with anti-Cited 1 antibody. Cited1nuc-TagRFP-T 15.5 dpc kidneys were probed with anti-Cited1 and anti-Cytokeratin 5 antibodies. Co-localization of native TagRFP-T fluorescent cells and cells positive for anti-Cited1 antibody are observable in the cap mesenchyme at the tips of the ureteric buds (Cytokeratin positive).
	TagRFP-T fluorescent cells in the cap mesenchyme co-localize with cells detectable with anti-Six2 antibody. Cited1nucTagRFP-T 15.5 dpc kidneys were probed with anti-Six2 and anti-Cytokeratin 5 antibodies. Co-localization of native TagRFP-T fluorescent cells and cells positive for anti-Six2 antibody are observable in the cap mesenchyme at the tips of the ureteric buds (Cytokeratin positive) but not in the Six2 positive pre-tubular aggregates.
	TagRFP-T expressing cells in the cap mesenchyme co-localize with a subpopulation of cells expressing GFP in Cited1nuc-TagRFP-T; Six2TGC/+ 15.5 dpc kidneys. UGSs were probed with anti-TagRFP-T, anti-GFP and anti-Cytokeratin 5 antibodies. Co-localization of TagRFP-T and GFP positive cells were observable in the cap mesenchyme at the tips of the ureteric buds (Cytokeratin positive) but not in the Six2 positive pre-tubular aggregates.
	TagRFP-T expressing cells in the cap mesenchyme co-localize with a subpopulation of cells expressing GFP in Cited1nucTagRFP-T; Six2TGC/+ 15.5 dpc kidneys. UGSs were probed with anti-TagRFP-T, anti-GFP and anti-Cytokeratin 5 antibodies. Co-localization of TagRFP-T and GFP positive cells were observable in the cap mesenchyme at the tips of the ureteric buds (Cytokeratin positive) but not in the Six2 positive population of cells undergoing mesenchyme to epithelial transition.

Results Notes

Submitter findings: Analysis confirms the expression of nuc-TagRFP-T under the regulation of Cited1 in cap mesenchyme cells apposing the ureteric buds in 15.5 dpc kidneys and in the male reproductive system. At 15.5dpc nuc-TagRFP-T co-localizes with Six2 expressing cells at the tips of the ureteric buds but not in the mesenchymal population of pre-tubular aggregate cells. Analysis of the expression in the male reproductive system was not included in this characterization.

Mouse marker strain entries are unique GUDMAP entries that can contain data for multiple probes at multiple stages of development (more info).

Submitters

Principal Investigator(s):	Andrew P McMahon, Keck School of Medicine, Los Angeles, CA, USA, amcmahon@med.usc.edu
Contributors:	Andrew P McMahon, M Todd Valerius, Jinjin Guo, Jill McMahon
Submitted By:	Jill McMahon, Keck School of Medicine, Los Angeles, CA, USA
Batch ID:	555
Submission ID:	21893

Expression Mapping

Expression Strengths Key:

	Present (unspecified strength)
	Present (strong)
	Present (moderate)
	Present (weak)
	Uncertain
	Not Detected

Expression Patterns Key:

	Homogeneous
	Graded
	Regional
	Spotted
	Ubiquitous

	Restricted
	Single cell

Nerve Density:

Relative to Total:

	High
	Medium
	Low

Relative to P0/Adult:

	Increase, large
	Increase, small
	Decrease, large
	Decrease, small

	Contains note

View annotated components as a list

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Specimen

Stage:

TS23

Other Staging System:

15.5 dpc

Tissue:

urinary system

Strain:

mixed background

Genotype:

Non-wild type Allele:
Gene		Cited1
MGI ID		MGI:4940389
MGI Symbol or lab name		Tg(Cited1-TagRFP)26Amc
Type		Transgenic, (Reporter)
Allele First Chromatid		Tg(Cited1-TagRFP)26Amc
Allele Second Chromatid		wild type
Notes:		Mouse strain is available from the Jackson Lab, see here: http://jaxmice.jax.org/strain/015853.html

Sex:

both

Specimen Preparation:

mouse marker strain

Fixation Method:

unspecified

Experiment Notes:

Immunohistochemistry was performed to examine whether TagRFP-T was expressed in the expected Cited1 domain. Nuc-TagRFP-T expression was examined by probing Cited1nuc-TagRFP-T/+ 15.5dpc embryos with rabbit-anti-Cited1, rabbit-anti-TagRFP-T and rabbit-anti-Six2 antibodies and comparing these positive cell populations to native nuc- TagRFP-T expression. Co-localization of native nuc-TagRFP-T and cells positive for the Cited1 and TagRFP-T antibodies were observed in the cap mesenchyme cells surrounding the tips of the ureteric buds (Fig 3 and 4). The population of nuc-TagRFP-T expressing cells observable by native fluorescence is apparently contained within the domain visualized with anti-Six2 antibodies (Fig 5). To clarify the extent of the overlapping domains Cited1nuc-TagRFP-T/+ male mice were mated to Six2TGC/+ females. The Six2-TGC mouse strain is a BAC transgenic line expressing GFP:Cre in the Six2 domain. Cited1nuc-TagRFP-T/+;; Six2TGC/+ 15.5dpc embryos were probed with rabbit-anti-TagRFP-T and chicken-anti-GFP antibodies. Anti-TagRFP-T and anti-GFP positive cells co-localize in cap mesenchyme cells surrounding the tips of the ureteric bud, but not with GFP positive cells delineating the Six2 population in pretubular aggregates undergoing mesenchyme to epithelial transition (Fig 5 and 6). Whole UGSs were fixed in 4% paraformaldehyde at 4ºC 2 hours, washed 3 times in PBS, equilibrated in 30% sucrose overnight and then embedded in OCT and flash frozen on dry-ice. The UGSs were sectioned at 16μm and probed with: rabbit anti- TagRFP-T/mouse-anti-Cytokeratin, rabbit-anti-Cited1/mouse-anti-Cytokeratin, rabbitanti- Six2/mouse-anti-Cytokeratin, rabbit-anti-Six2/mouse-anti-Cytokeratin and rabbitanti- TagRFP-T/ Chicken-anti-GFP/mouse-anti-Cytokeratin respectively, incubated overnight at 4ºC and detected with secondary antibodies.

Transgene Visualisation

Reporter:	RFP
Direct method for visualising reporter:	Fluorescence

Acknowledgements

Comprehensive PDFs containing allele verification and characterisation can be downloaded from the GUDMAP mouse strains page (http://www.gudmap.org/Resources/MouseStrains/Mouse_Strains_Summary.html). GUDMAP:21893 entry compiled from these PDFs by GUDMAP Editorial Office.