Confocal Microscopy Image Analysis of Pancreatic beta-Cells K-ATP Channel Proteins in Congenital Hyperinsulinism (CHI)

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dc.contributorSistema FMUSP-HC: Faculdade de Medicina da Universidade de São Paulo (FMUSP) e Hospital das Clínicas da FMUSP
dc.contributor.authorLOVISOLO, S. M.
dc.contributor.authorGARIPPO, A. L.
dc.contributor.authorGUEDES, F.
dc.contributor.authorZERBINI, M. C.
dc.date.accessioned2013-10-11T21:25:47Z
dc.date.available2013-10-11T21:25:47Z
dc.date.issued2012
dc.description.abstractBackground: CHI is a life-threatening disorder of glucose metabolism of neonates characterized by serum insulin levels unresponsive to blood glucose concentrations. Pancreatic ß-cells regulates insulin secretion through ATP sensitive potassium channels (KATP channel) formed by sulfonylurea receptor 1 (SUR1) and potassium inward rectifying 6.2 (Kir 6.2) proteins. The ß-cell K ATP channel dependent CHI is classically associated with loss-of-function mutations of these subunits genes. In a previous study [1], no mutations in the Kir6.2 gene and in the 33-37 exons hot spot region of the SUR1 gene were identified in these patients. The aim of this study is to investigate if these subunits were present in the ß-cells of CHI patients. Design: Pancreatic surgical paraffin tissue from 7 neonates (3F/4M, 4-13 mo) with CHI and 8 autopsy pancreatic control tissue (5F/3M, 4-11 mo) were included in the study. Confocal microscopy double-staining immunofluorescence (insulin/SUR1 and insulin/Kir6.2) was performed in order to detect each protein specifically in the ß-cells All sections were analyzed under a Zeiss 510 Meta confocal laser scanning microscope (63x). At least 10 different endocrine islets were captured to evaluate the expression of either Kir6.2 or SUR1 (green-488nm) and insulin (red-633nm). Co-expression of insulin/SUR1 and insulin/Kir6.2 was analysed visually (green x red→yellow) and through correlation Pearson’s coefficient(PC) that estimates the goodness of rate association of the two fluorochromes. PC was compared among cases and controls using a nonparametric method (Mann-Whitney). Results: Visual observation clearly revealed the presence of Kir6.2 and SUR1 in a granular cytoplasmic pattern in the ß-cells of cases and controls. Nevertheless, overlap of insulin/Kir6.2 and insulin/SUR1 seemed to be more constant and uniform in controls than in CHI cases, confirmed by the analysis through PC showing a statistically significant decrease in Kir6.2 (P= 0.0084) and SUR1 (P= 0.041) in the ß-cells of CHI patients. Conclusions: This is the first demonstration of K ATP channels subunits Kir6.2 and SUR1 in the pancreatic ß-cells of CHI patients using an in situ method. Our results show that both subunits were present in the ß-cells, but are under-expressed in CHI patients compared to controls, adding a new information to this rare condition with a complex pathogenesis.
dc.description.conferencedateMAR 17-23, 2012
dc.description.conferencelocalVancouver, CANADA
dc.description.conferencename101st Annual Meeting of the United-States-and-Canadian-Academy-of-Pathology (USCAP)
dc.description.indexMEDLINE
dc.identifier.citationLABORATORY INVESTIGATION, v.92, suppl.1, p.470A-470A, 2012
dc.identifier.issn0023-6837
dc.identifier.urihttps://observatorio.fm.usp.br/handle/OPI/3028
dc.language.isoeng
dc.publisherNATURE PUBLISHING GROUP
dc.relation.ispartofLaboratory Investigation
dc.rightsrestrictedAccess
dc.rights.holderCopyright NATURE PUBLISHING GROUP
dc.subject.wosMedicine, Research & Experimental
dc.subject.wosPathology
dc.titleConfocal Microscopy Image Analysis of Pancreatic beta-Cells K-ATP Channel Proteins in Congenital Hyperinsulinism (CHI)
dc.typeconferenceObject
dc.type.categorymeeting abstract
dc.type.versionpublishedVersion
dspace.entity.typePublication
hcfmusp.contributor.author-fmusphcANA LUCIA GARIPPO
hcfmusp.contributor.author-fmusphcFERNANDA GUEDES LUIZ
hcfmusp.contributor.author-fmusphcMARIA CLAUDIA NOGUEIRA ZERBINI
hcfmusp.description.beginpage470A
hcfmusp.description.endpage470A
hcfmusp.description.issuesuppl 1
hcfmusp.description.volume92
hcfmusp.origemWOS
hcfmusp.origem.wosWOS:000299799902469
hcfmusp.publisher.cityNEW YORK
hcfmusp.publisher.countryUSA
hcfmusp.relation.referenceLovisolo SM, 2010, PEDIATR DEVEL PATHOL, V13, P375, DOI 10.2350/08-12-0578.1
relation.isAuthorOfPublication90f6a3d4-6c85-41f0-8738-f8fe993ae35b
relation.isAuthorOfPublicationc33153a4-0e6b-4895-b087-b65dbae733e1
relation.isAuthorOfPublication71cbe6cf-ec3c-4df8-bc56-ff122fd165f6
relation.isAuthorOfPublication.latestForDiscovery90f6a3d4-6c85-41f0-8738-f8fe993ae35b
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