Core C facility provides state of the art molecular biological and morphological support to investigators and their staff as well as new investigators collecting data for future directions of the PPG. The molecular component has developed RT-PCR protocols for enzymatically dispersed, isolated smooth muscle and ICCs with green reporters to identify transcript expression within these specific cell types within the tunica muscularis of several animal species including humans. The Core continually performs RT-PCR to verify the identity of cell populations within the tunica muscularis that have been purified by fluorescent-activated cell sorting (FACS;conducted by Core B). These analyses have been extended to include real-time PCR analysis of specific transcript expression in these cell populations. The Core provides routine genotyping of transgenic animals for all applicable projects, designs and tests primers for RT-PCR and constructs vectors for proposed experiments. The Core also continues to provide day to day maintenance of genomic clones, cDNAs and cultures for molecular biological investigations. The core provides DNA sequence analysis of clones and amplification products and provides support with mammalian cell lines expressing various project specific cDNAs for several ion channels and receptors. The morphology component of the Core continues to provide expertise in the areas of conventional light microscopy, phase contrast microscopy, fluorescence microscopy, laser scanning confocal microscopy, digital imaging, transmission electron microscopy, in-situ hybridization, immunohistochemistry and immunocytochemistry, to support individual projects that have the need to utilize these techniques. This component holds a high standard and is continually developing novel approaches to determine cellular numbers and volumes within the tunica muscularis. New protocols and alogrithims in combination with confocal microscopy and deconvolution have set new standards for the quantitiative analysis of cells types within the gut wall including enteric nerves and ICC populations. Appropriate quantatitive structural and ultrascructural analysis of cells will provide valuable information of changes which occur in different cell types within the in gut wall in health and disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Program Projects (P01)
Project #
5P01DK041315-25
Application #
8469498
Study Section
Special Emphasis Panel (ZDK1-GRB-9)
Project Start
Project End
Budget Start
2013-05-01
Budget End
2014-04-30
Support Year
25
Fiscal Year
2013
Total Cost
$214,393
Indirect Cost
$61,799
Name
University of Nevada Reno
Department
Type
DUNS #
146515460
City
Reno
State
NV
Country
United States
Zip Code
89557
Sanders, Kenton M; Salter, Anna K; Hennig, Grant W et al. (2014) Responses to enteric motor neurons in the gastric fundus of mice with reduced intramuscular interstitial cells of cajal. J Neurogastroenterol Motil 20:171-84
Zheng, Haifeng; Park, Kyung Sik; Koh, Sang Don et al. (2014) Expression and function of a T-type Ca2+ conductance in interstitial cells of Cajal of the murine small intestine. Am J Physiol Cell Physiol 306:C705-13
McCann, Conor J; Hwang, Sung-Jin; Hennig, Grant W et al. (2014) Bone Marrow Derived Kit-positive Cells Colonize the Gut but Fail to Restore Pacemaker Function in Intestines Lacking Interstitial Cells of Cajal. J Neurogastroenterol Motil 20:326-37
Durnin, Leonie; Hwang, Sung Jin; Kurahashi, Masaaki et al. (2014) Uridine adenosine tetraphosphate is a novel neurogenic P2Y1 receptor activator in the gut. Proc Natl Acad Sci U S A 111:15821-6
Okamoto, T; Barton, M J; Hennig, G W et al. (2014) Extensive projections of myenteric serotonergic neurons suggest they comprise the central processing unit in the colon. Neurogastroenterol Motil 26:556-70
Drumm, Bernard T; Koh, Sang Don; Andersson, Karl-Erik et al. (2014) Calcium signalling in Cajal-like interstitial cells of the lower urinary tract. Nat Rev Urol 11:555-64
Mutafova-Yambolieva, Violeta N; Durnin, Leonie (2014) The purinergic neurotransmitter revisited: a single substance or multiple players? Pharmacol Ther 144:162-91
Sanders, Kenton M; Ward, Sean M; Koh, Sang Don (2014) Interstitial cells: regulators of smooth muscle function. Physiol Rev 94:859-907
Kurahashi, Masaaki; Nakano, Yasuko; Peri, Lauren E et al. (2013) A novel population of subepithelial platelet-derived growth factor receptor *-positive cells in the mouse and human colon. Am J Physiol Gastrointest Liver Physiol 304:G823-34
Keef, K D; Saxton, S N; McDowall, R A et al. (2013) Functional role of vasoactive intestinal polypeptide in inhibitory motor innervation in the mouse internal anal sphincter. J Physiol 591:1489-506

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