Study purpose: Sympathetic innervation is critical for cold-induced changes in blood flow, oxidative capacity, and activity of brown adipose tissue (BAT). In addition to regulating BAT function, cold is also reported to stimulate branching of sympathetic fibers within the BAT parenchyma. Due to the high degree of variability in the pattern of sympathetic innervation between animals and between different lobes of BAT within a single animal, it is critical to assess these endpoints in an intact depot (as opposed to "representative sections").
Data collection: We used the iDISCO technique to stain and visualize peripheral axons (defined by Pgp9.5 and beta 3 tubulin-immunoreactivity) and sympathetic axons (defined by tyrosine hydroxylase-immunoreactivity). Data consist of 8 stacked images (ND2 or TIF format) of intact BAT depots from a Zeiss 710 microscope.
Primary conclusion: None stated
Experimental Design: Brown adipose tissue was excised from surrounding tissue and immunohistochemistry was completed using Pgp9.5, beta 3 tubulin, and tyrosine hydroxylase antibodies, using the iDISCO technique.
Subjects & Samples: Juvenile male mice C57BL/6J, between 1-4 weeks old (n = 8). Brown adipose tissue samples were taken from the interscapular region of each mouse. Tyrosine hydroxylase, Pgp9.5 and beta 3 tubulin immunoreactivity was examined in the 3 week old mice Tyrosine hydroxylase immunoreactivity was examined in the 1 week old mouse Beta 3 tubulin immunoreactivity was examined in the 4 week old mouse
Primary vs derivative data: The primary and derivative folders consist of folders divided by mouse age and specimen. The primary folder contains nd2 formatted images, while the derivative folder includes images (tif and jpx) and videos (mp4) files. Folders contain data from mouse brown adipose tissues (BAT), and a trunk with BAT attached labeled with 3 different axonal markers (TH, Pgp9.5, B3tub) using iDISCO method and confocal or lightsheet microscopy.
Important notes: (1) The manifest file within the derived folder is very helpful in understanding all files in this folder. (2) Fiji software can be used to open nd2 files (https://imagej.net/Fiji/Downloads) > Open Fiji > Plugins (top of screen) > Bio-Formats > Bio-Formats Importer > Select the file.
Open Questions: (1)Protocol: The protocol is lacking in information regarding how animal was anesthetized, where the brown adipose tissue was removed from and which secondary antibodies were used.
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