Combined micro-osmotic pump infusion and intracerebroventricular injection to study FGF1 signaling pathways in the mouse brain
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Intracerebroventricular (icv) injection of fibroblast growth factor 1 (FGF1) elicits remission of diabetic hyperglycemia in rodent models of type 2 diabetes. Here, we present an optimized protocol to study the intracellular signaling pathways underlying the FGF1-induced sustained glucose lowering in the mouse brain. This protocol combines icv injection of FGF1 and osmotic mini-pump infusion of U0126, an inhibitor of MAPK/ERK signaling. We describe the surgical procedure and verification of U0126 inhibition of FGF1-stimulated hypothalamic MAPK/ERK signaling via western blot. For complete details on the use and execution of this protocol, please refer to Brown et al. (2021).
| Originalsprog | Engelsk |
|---|---|
| Artikelnummer | 101329 |
| Tidsskrift | STAR Protocols |
| Vol/bind | 3 |
| Udgave nummer | 2 |
| Antal sider | 16 |
| ISSN | 2666-1667 |
| DOI | |
| Status | Udgivet - 2022 |
Bibliografisk note
Funding Information:
J.M.B. was supported by National Heart, Lung, and Blood Institute T32 training grant HL-007312 and the Diabetes Research Center Samuel and Althea Stroum Endowed Graduate Fellowship at the University of Washington. This work was supported by NIH-NIDDK grants K08DK114474 (J.M.S.), R03 DK128383 (J.M.S.), R01DK101997 (M.W.S.), R01DK089056 (G.J.M.), R01DK124238 (G.J.M.), and R01DK083042 (G.J.M. and M.W.S.), and an American Diabetes Association Innovative Basic Science Award (ADA; G.J.M.). This work was also supported by the NIH-NIDDK?funded Nutrition Obesity Research Center (NORC; P30DK035816) and the Diabetes Research Center (DRC; P30DK017047) at the University of Washington. Additional funding to support these studies was provided to J.M.S. by the Department of Defense (DoD W81XWH2110635) and UW Royalty Research Fund (RRF; A139339). Funding was also provided to M.W.S. by Novo Nordisk (CMS-431104) and to J.M.B and M.A.B. by the Novo Nordisk Foundation Center for Basic Metabolic Research, which is an independent research center at the University of Copenhagen partially funded by an unrestricted donation from the Novo Nordisk Foundation (NNF10CC1016515). J.B. B.P. and J.S. performed the experiments. J.B. B.P. N.A. G.M. M.S. and J.S. designed the experiments. J.B. B.P. N.A. M.S. and J.S. wrote and revised the manuscript. All authors read, revised, and approved the manuscript. The authors declare no competing interests.
Funding Information:
J.M.B. was supported by National Heart, Lung, and Blood Institute T32 training grant HL-007312 and the Diabetes Research Center Samuel and Althea Stroum Endowed Graduate Fellowship at the University of Washington . This work was supported by NIH - NIDDK grants K08DK114474 (J.M.S.), R03 DK128383 (J.M.S.), R01DK101997 (M.W.S.), R01DK089056 (G.J.M.), R01DK124238 (G.J.M.), and R01DK083042 (G.J.M. and M.W.S.), and an American Diabetes Association Innovative Basic Science Award (ADA; G.J.M.). This work was also supported by the NIH-NIDDK–funded Nutrition Obesity Research Center (NORC; P30DK035816 ) and the Diabetes Research Center (DRC; P30DK017047 ) at the University of Washington. Additional funding to support these studies was provided to J.M.S. by the Department of Defense (DoD W81XWH2110635 ) and UW Royalty Research Fund (RRF; A139339 ). Funding was also provided to M.W.S. by Novo Nordisk (CMS- 431104 ) and to J.M.B and M.A.B. by the Novo Nordisk Foundation Center for Basic Metabolic Research , which is an independent research center at the University of Copenhagen partially funded by an unrestricted donation from the Novo Nordisk Foundation ( NNF10CC1016515 ).
Publisher Copyright:
© 2022 The Author(s)
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