human synapsin promoter  (Addgene inc)

Journal: bioRxiv

Article Title: A neural entero-pancreatic pathway that regulates insulin secretion and glucose tolerance

doi: 10.1101/2025.08.14.670343

Figure Lengend Snippet: a , Schematic of the entero-pancreatic neural circuit and retrograde tracing strategy. AAVrg-hSyn-mCherry was injected intraductally into the pancreas of ChAT–GFP mice. b , An in toto preparation of the duodenum and pancreas illustrating their anatomical continuity at the level of the common bile duct. Tissues were immunostained for βIII-tubulin (magenta), ChAT–GFP (cyan) and viral mCherry (yellow). c–e , Flat-mount preparations showing traced neurons in myenteric but not submucosal ganglia. Immunostaining for nNOS (magenta), ChAT–GFP (cyan) and mCherry (yellow). c, c′ , Retrogradely labeled neurons in the duodenal myenteric plexus d , Submucosal ganglia lack labeled neurons. e , Distribution of traced neurons along the myenteric plexus, distances from the pylorus indicated. f–h , Quantification of neuronal phenotypes (3–15 randomly selected areas per region, 100–1000 neurons per area, n = 3 mice). f , Percentage of total neurons positive for ChAT, nNOS and the tracer AAV PHP.S . g , Percentages of AAV PHP.S + neurons of the total neuronal population in peripheral ganglia known to innervate the pancreas: myenteric plexus (2–4 cm from pylorus), nodose ganglion (NG), thoracic dorsal root ganglia (DRG; T9–T11) and coeliac ganglia. h , Neurochemical identity of traced neurons in myenteric, pancreatic and nodose ganglia: ChAT+ (cyan), nNOS+ (magenta), ChAT+/nNOS+ (pink) or double-negative (grey).

Article Snippet: For retrograde tracing studies, we injected the recombinant adeno-associated virus (rAAV) expressing reporter mCherry under neuronal human synapsin promoter (rAAV-hSyn-mCherry, AddGene 114472) into the pancreatic duct of ChAT-GFP transgenic mice ( ).

Techniques: Retrograde Tracing, Injection, Immunostaining, Labeling

Journal: bioRxiv

Article Title: A neural entero-pancreatic pathway that regulates insulin secretion and glucose tolerance

doi: 10.1101/2025.08.14.670343

Figure Lengend Snippet: a , Schematic of the entero-pancreatic neural axis and anterograde viral tracing strategy. AAVphp.s-hSyn-mCherry was injected submuscularly into the stomach antrum and duodenum of wild-type mice. b , Duodenal flat mount showing a viral injection site (mCherry, yellow). c–i , Representative pancreatic sections immunostained for βIII-tubulin (cyan), VIP (magenta), mCherry (yellow) and DAPI (grey). c , Low-magnification view of a section containing three endocrine islets. d–f , High-magnification view of a representative islet (highlighted in c) with numerous traced varicosities in the endocrine parenchyma and fewer in a neighboring ganglion; some varicosities are VIP-positive ( e ). g–i , Islet with few traced varicosities in the endocrine parenchyma but more in the adjacent ganglion. j , Quantification of traced varicosities in endocrine islets as in d and g . Varicosities were present in 8 of 39 analyzed islets ( n = 3 mice).

Article Snippet: For retrograde tracing studies, we injected the recombinant adeno-associated virus (rAAV) expressing reporter mCherry under neuronal human synapsin promoter (rAAV-hSyn-mCherry, AddGene 114472) into the pancreatic duct of ChAT-GFP transgenic mice ( ).

Techniques: Injection

Journal: bioRxiv

Article Title: A neural entero-pancreatic pathway that regulates insulin secretion and glucose tolerance

doi: 10.1101/2025.08.14.670343

Figure Lengend Snippet: a, Schematic of the experimental design where entero-pancreatic neurons were labeled with GFP using the INTACT technique followed by snRNA-seq. To achieve population-specific nuclear labeling, floxed Sun1-GFP transgenic mice received intraductal pancreatic injection of AAVrg-hSyn-Cre. b, Reference UMAP of 111 GFP+ enteric nuclei from mouse duodenum. c, Representative images of the duodenal flat mounts depicting Sun-1-GFP+ nuclei in nNos+ myenteric neurons. Immunostaining for bIII-Tubulin (cyan), nNos (magenta), vAChT (white), DAPI (blue), and endogenous Sun1-GFP (yellow). d, Transcriptomic signature of pancreas-projecting enteric neurons reported in a series of dot plots for genes associated with major groups of neurotransmitters and neuronal receptors. Dot size reflects the fraction of nuclei expressing the gene, dot color indicates the mean expression level in expressing nuclei. Some of the gene names were swapped with the names of the protein they encode, for ease of identification (red). For a comparison of the transcriptional profiles of entero-pancreatic neurons with all enteric neurons from Drokholyanski et al. , see Extended Data Fig.3.

Article Snippet: For retrograde tracing studies, we injected the recombinant adeno-associated virus (rAAV) expressing reporter mCherry under neuronal human synapsin promoter (rAAV-hSyn-mCherry, AddGene 114472) into the pancreatic duct of ChAT-GFP transgenic mice ( ).

Techniques: Labeling, Transgenic Assay, Injection, Immunostaining, Expressing, Comparison