Expression of molecular markers in subpopulations of mouse celiac ganglion neurons using small molecule FISH (RNAScope)
Study Purpose: Sympathetic regulation of peripheral organ function is achieved through the coordinated actions of secretomotor, vasomotor and erector sympathetic neurons, in a manner that is highly conserved across species. Functionally distinct classes of sympathetic neurons have been defined by their firing properties and the expression of different combinations of neurotransmitters and neuropeptides. The identification of unique molecular markers for these populations would open the door to the application of genetic tools to parse the actions of secretomotor, vasomotor and erector sympathetic neurons. We identified unique molecular markers that can distinguish between subpopulations of stellate ganglion (SG) neurons in the mouse. Here we investigated whether the expression patterns of these markers are conserved in the celiac ganglion (CG) by multiplex single molecule fluorescent in situ hybridization (smFISH) on SCG cryosections.
Data Collection: Data consists of 36 images (CZI format) of combinations of RNAScope probes marking distinct populations of CG neurons in Npy-GFP mice using a Zeiss LSM 710 inverted confocal microscope.
Primary Conclusion: Gene expression patterns detected with smFISH in the CG were different than those observations in the SG.
Experimental Design: Mouse ganglia were dissected, fixed in 4% PFA in 0.1M PB overnight at 4°C, washed with Phosphate Buffer Saline (PBS) at 4°C for at least 1 h and encased in Optimal Cutting Temperature compound (Tissue-Tek) then cut into 10 μm cryo-sections on slides, and stored at -80°C. RNAscope single-molecule fluorescent in situ hybridization (smFISH) multiplex version was performed as instructed by Advanced Cell Diagnostics (ACDBio, Document Number 320293-USM) without modification. Expression patterns were visualized by confocal microscopy.
Completeness: This dataset is a part of a larger study, " Expression of molecular markers for SG neuronal subpopulations in 3 sympathetic ganglia."
Subjects & Samples: Male (n=3) and female (n=3) adult mice (RRID:IMSR_JAX:006417) were used in this study.
Primary vs derivative data: Primary data is organized by the subject ID folders and sample ID subfolder. Each sample subfolder contains confocal images (CZI format). Image data in the derivative folder (JPEG2000 and OME-TIFF) was derived from primary images (.CZI).
Important Notes: A complete list of molecular probes is shown in the associated protocol https://www.protocols.io/view/expression-of-molecular-markers-in-subpopulations-b62mrgc6/materials, and is available as a pdf file in the protocol folder in this dataset.
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