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Imaging NF‐κB activity in a murine model of early stage diabetes
Author(s) -
Taghian Toloo,
Metelev Valeriy G.,
Zhang Surong,
Bogdanov Alexei A.
Publication year - 2020
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.201801147r
Subject(s) - nf κb , diabetes mellitus , stage (stratigraphy) , cancer research , medicine , endocrinology , inflammation , biology , paleontology
Early pro‐inflammatory signaling in the endocrine pancreas involves activation of NF‐κB, which is believed to be important for determining the ultimate fate of β‐cells and hence progression of type 1 diabetes (T1D). Thus, early non‐invasive detection of NF‐κB in pancreatic islets may serve as a potential strategy for monitoring early changes in pancreatic endocrine cells eventually leading to T1D. We investigated the feasibility of optical imaging of NF‐κB transcription factor activation induced by low‐dose streptozocin (LD‐STZ) treatment in the immunocompetent SKH1 mouse model of early stage diabetes. In this model, we showed that the levels of NF‐κB may be visualized and measured by fluorescence intensity of specific near‐infrared (NIR) fluorophore‐labeled oligodeoxyribonucleotide duplex (ODND) probes. In addition, NF‐κB activation following LD‐STZ treatment was validated using immunofluorescence and transgenic animals expressing NF‐κB inducible imaging reporter. We showed that LD‐STZ‐treated SKH1 mice had significantly higher (2‐3 times, P < .01) specific NIR FI in the nuclei and cytoplasm of islets cells than in non‐treated control mice and this finding was corroborated by immunoblotting and electrophoretic mobility shift assays. Finally, using semi‐quantitative confocal analysis of non‐fixed pancreatic islet microscopy we demonstrated that ODND probes may be used to distinguish between the islets with high levels of NF‐κB transcription factor and control islet cells.