PMID- 30567483 OWN - NLM STAT- MEDLINE DCOM- 20191203 LR - 20200309 IS - 1524-4636 (Electronic) IS - 1079-5642 (Print) IS - 1079-5642 (Linking) VI - 39 IP - 2 DP - 2019 Feb TI - Perivascular Adipocytes Store Norepinephrine by Vesicular Transport. PG - 188-199 LID - 10.1161/ATVBAHA.118.311720 [doi] AB - Objective- Perivascular adipose tissue (PVAT) contains an independent adrenergic system that can take up, metabolize, release, and potentially synthesize the vasoactive catecholamine norepinephrine. Norepinephrine has been detected in PVAT, but the mechanism of its protection within this tissue is unknown. Here, we investigate whether PVAT adipocytes can store norepinephrine using VMAT (vesicular monoamine transporter). Approach and Results- High-performance liquid chromatography identified norepinephrine in normal male Sprague Dawley rat aortic, superior mesenteric artery, and mesenteric resistance vessel PVATs, and retroperitoneal fat. Real-time polymerase chain reaction revealed VMAT1 and VMAT2 mRNA expression in the adipocytes and stromal vascular fraction of mesenteric resistance vessel PVAT. Immunofluorescence demonstrated the presence of VMAT1 and VMAT2, and the colocalization of VMAT2 with norepinephrine, in the cytoplasm of adipocytes in mesenteric resistance vessel PVAT. A protocol was developed to capture real-time uptake of Mini 202-a functional and fluorescent VMAT probe-in live rat PVAT adipocytes. Mini 202 was taken up by freshly isolated and differentiated adipocytes from mesenteric resistance vessel PVAT and adipocytes from thoracic aortic and superior mesenteric artery PVATs. In adipocytes freshly isolated from mesenteric resistance vessel PVAT, addition of rose bengal (VMAT inhibitor), nisoxetine (norepinephrine transporter inhibitor), or corticosterone (organic cation 3 transporter inhibitor) significantly reduced Mini 202 signal. Immunofluorescence supports that neither VMAT1 nor VMAT2 is present in retroperitoneal adipocytes, suggesting that PVAT adipocytes may be unique in storing norepinephrine. Conclusions- This study supports a novel function of PVAT adipocytes in storing amines in a VMAT-dependent manner. It provides a foundation for future studies exploring the purpose and mechanisms of norepinephrine storage by PVAT in normal physiology and obesity-related hypertension. FAU - Ahmad, Maleeha F AU - Ahmad MF AD - From the Department of Pharmacology and Toxicology (M.F.A., D.F., E.D., J.T., A.I., R.B., S.W.W.), Michigan State University, East Lansing. FAU - Ferland, David AU - Ferland D AD - From the Department of Pharmacology and Toxicology (M.F.A., D.F., E.D., J.T., A.I., R.B., S.W.W.), Michigan State University, East Lansing. FAU - Ayala-Lopez, Nadia AU - Ayala-Lopez N AD - Department of Laboratory Medicine, Yale University, New Haven, CT (N.A.-L.). FAU - Contreras, G Andres AU - Contreras GA AD - Department of Large Animal Clinical Sciences (G.A.C., K.T., A.J.M.), Michigan State University, East Lansing. FAU - Darios, Emma AU - Darios E AD - From the Department of Pharmacology and Toxicology (M.F.A., D.F., E.D., J.T., A.I., R.B., S.W.W.), Michigan State University, East Lansing. FAU - Thompson, Janice AU - Thompson J AD - From the Department of Pharmacology and Toxicology (M.F.A., D.F., E.D., J.T., A.I., R.B., S.W.W.), Michigan State University, East Lansing. FAU - Ismail, Alexander AU - Ismail A AD - From the Department of Pharmacology and Toxicology (M.F.A., D.F., E.D., J.T., A.I., R.B., S.W.W.), Michigan State University, East Lansing. FAU - Thelen, Kyan AU - Thelen K AD - Department of Large Animal Clinical Sciences (G.A.C., K.T., A.J.M.), Michigan State University, East Lansing. FAU - Moeser, Adam J AU - Moeser AJ AD - Department of Large Animal Clinical Sciences (G.A.C., K.T., A.J.M.), Michigan State University, East Lansing. FAU - Burnett, Robert AU - Burnett R AD - From the Department of Pharmacology and Toxicology (M.F.A., D.F., E.D., J.T., A.I., R.B., S.W.W.), Michigan State University, East Lansing. FAU - Anantharam, Arun AU - Anantharam A AD - Department of Pharmacology, University of Michigan Medical School, Ann Arbor (A.A.). FAU - Watts, Stephanie W AU - Watts SW AD - From the Department of Pharmacology and Toxicology (M.F.A., D.F., E.D., J.T., A.I., R.B., S.W.W.), Michigan State University, East Lansing. LA - eng GR - F31 HL128035/HL/NHLBI NIH HHS/United States GR - R01 HD072968/HD/NICHD NIH HHS/United States GR - T32 GM092715/GM/NIGMS NIH HHS/United States GR - F31 HL143937/HL/NHLBI NIH HHS/United States GR - P01 HL070687/HL/NHLBI NIH HHS/United States GR - T32 HL007974/HL/NHLBI NIH HHS/United States GR - R01 GM111997/GM/NIGMS NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't PL - United States TA - Arterioscler Thromb Vasc Biol JT - Arteriosclerosis, thrombosis, and vascular biology JID - 9505803 RN - 0 (Slc18a1 protein, rat) RN - 0 (Slc18a2 protein, rat) RN - 0 (Vesicular Monoamine Transport Proteins) RN - X4W3ENH1CV (Norepinephrine) SB - IM MH - Adipocytes/*metabolism MH - Animals MH - Biological Transport MH - Chromaffin Cells/metabolism MH - Female MH - Male MH - Mesenteric Arteries/metabolism MH - Norepinephrine/*metabolism MH - Rats MH - Rats, Sprague-Dawley MH - Vesicular Monoamine Transport Proteins/*physiology PMC - PMC6344267 MID - NIHMS1515962 OTO - NOTNLM OT - adipocytes OT - adipose tissue OT - catecholamines OT - norepinephrine OT - obesity EDAT- 2018/12/21 06:00 MHDA- 2019/12/04 06:00 PMCR- 2020/02/01 CRDT- 2018/12/21 06:00 PHST- 2018/12/21 06:00 [pubmed] PHST- 2019/12/04 06:00 [medline] PHST- 2018/12/21 06:00 [entrez] PHST- 2020/02/01 00:00 [pmc-release] AID - 10.1161/ATVBAHA.118.311720 [doi] PST - ppublish SO - Arterioscler Thromb Vasc Biol. 2019 Feb;39(2):188-199. doi: 10.1161/ATVBAHA.118.311720.