Characterization of adeno associated virus serotypes 4 weeks after pancreas injection in mice

Study Purpose: To identify viral vectors capable of gene delivery to specific pancreatic nerves we characterize and assess multiple adeno-associated virus (6, 8, and 9) serotypes, viral titers, and delivery routes.

Data Collection: Adeno-associated virus, expressing mCherry fluorophore under synapsin promoter (AAV-hSyn-mCherry) was injected in the parenchyma of the mouse pancreas. Different serotypes (6, 8, 9) were tested. The coeliacs were harvested four weeks afterward. Tissue clearing and immunohistochemistry staining were applied to identify AAV infected neurons. A confocal microscope was used for image acquisition. Analysis and quantification were performed with IMARIS software.

Primary Conclusion: Viral serotypes have been optimized to deliver to pancreatic nerves. Pancreatic innervation was mapped using viral and non-viral traces with inputs from parasympathetic and somatosensory ganglia.

Curator's Notes

Experimental Design: Animals were divided into three groups based on adeno-associated virus serotypes: AAV serotype 6, AAV serotype 8, and AAV serotype 9 . AAV was injected in the parenchyma of the pancreas and the coeliac ganglia was harvested four weeks after injection. The iDISCO procedure was used to visualize mCherry via confocal microscopy.

Completeness: This is complete.

Subjects & Samples: This study used adult C57BL/6J male mice (n=18) between 12 - 14 weeks old.

Primary vs derivative data: The primary folder consists of 2 images per sample (czi and ims formats). The derivative folder contains 3 files: csv formatted file with the quantification results of the raw data, pzfz formatted graphics of the quantification results and a tiff formatted representative image.

Important Notes: (1) There is a doc contains additional metadata and a summary image of the results (2) Special file types: (a) to open ims images- use Fiji (https://imagej.net/Fiji/Downloads) > Open Fiji > File (top of screen) > Open > Select the file. (b) to open pzfz file - use GraphPad Prism