The secretory pathway is the major pathway for the extracellular release of proteins. In nucleated eukaryotic cells, the pathway is organized into a series of subcompartments with protein synthesis and folding being initiated in the endoplasmic reticulum and the downstream Golgi apparatus acting as a hub for protein modification, in particular, glycosylation, and sorting to various destinations including storage granules/vesicles, plasma membrane, and lysosomes. The laboratory concentrates on two examples of secretory pathway organization. Most efforts focus on the contributed of regulated a-granule secretion in platelets to the cessation of bleeding. In response to wounding and the creation of a hole in a vein or artery, platelets differentiate to become adherent and form a platelet aggregate that fills the hole and stops bleeding. This overall process is referred to as thrombus formation. We wish to understand the mechanism of a-granule secretion in both a detailed in vitro model and within the functional context of the thrombus. To do this, the laboratory is using structural readouts from light and electron microscopy and the tools of mouse genetics to probe granule release mechanisms both in vitro and in vivo. Portions of this work is done collaboratively with other laboratories with the mouse knockouts in SNARE proteins being contributed by the Whiteheart laboratory at the University of Kentucky and puncture wound examples being contributed by the Stalker/Brass laboratory at the University of Pennsylvania.
The other side of the laboratory concentrates on revealing the molecular links between mutations in Rab33b, a small GTPase that localizes to the medial, middle portion of the Golgi apparatus, and skeletal defects in humans. Mutations in Rab33b in humans cause severe skeletal defects and pre-mature death. As the Golgi apparatus is so important to biochemical processes such as glycosylation, we hypothesize that the Rab33b defect altered glycosylation and hence defects in secreted proteins that are essential to normal bone formation. Initial experiments indicate that significant changes in glycosylation in Rab33b mutant fibroblasts and a RNAi-knockdown HeLa cell model. The use of cell culture models enables the application of all the techniques of molecular cell biology to the research problem while the development of a knockout mouse model provides an important in vivo model.
McBride EL, Rao A, Zhang G, Hoyne JD, Calco GN, Kuo BC, He Q, Prince AA, Pokrovskaya ID, Storrie B, Sousa AA, Aronova MA, Leapman RD., Comparison of 3D cellular imaging techniques based on scanned electron probes: Serial block face SEM vs. Axial bright-field STEM tomography., J Struct Biol. 2018 Jun;202(3):216-228. doi: 10.1016/j.jsb.2018.01.012. Epub 2018 Feb 1.
Krokowski D, Guan BJ, Wu J, Zheng Y, Pattabiraman PP, Jobava R, Gao XH, Di XJ, Snider MD, Mu TW, Liu S, Storrie B, Pearlman E, Blumental-Perry A, Hatzoglou M., GADD34 Function in Protein Trafficking Promotes Adaptation to Hyperosmotic Stress in Human Corneal Cells., Cell Rep. 2017 Dec 5;21(10):2895-2910. doi: 10.1016/j.celrep.2017.11.027.
Goud B, Liu S, Storrie B., Rab proteins as major determinants of the Golgi complex structure., Small GTPases. 2018 Mar 4;9(1-2):66-75. doi: 10.1080/21541248.2017.1384087. Epub 2018 Jan 29.
Banerjee M, Joshi S, Zhang J, Moncman CL, Yadav S, Bouchard BA, Storrie B, Whiteheart SW., Cellubrevin/vesicle-associated membrane protein-3-mediated endocytosis and trafficking regulate platelet functions., Blood.2017 Dec 28;130(26):2872-2883. doi: 10.1182/blood-2017-02-768176. Epub 2017 Sep 20.
Kamykowski JA, Storrie B., Managing the Introduction of Super-Resolution Microscopy into a Core Facility., Methods Mol Biol. 2017;1663:15-19. doi: 10.1007/978-1-4939-7265-4_2
Storrie B, Whiteheart SW., Editorial: Platelet Secretion., Platelets. 2017 Mar;28(2):107. doi: 10.1080/09537104.2016.1277676.<
Siegel N, Lupashin V, Storrie B, Brooker G., High-magnification super-resolution FINCH microscopy using birefringent crystal lens interferometers., Nat Photonics. 2016 Dec;10:802-808. doi: 10.1038/NPHOTON.2016.207. Epub 2016 Nov 14.
Yadav S, Storrie B., The cellular basis of platelet secretion: Emerging structure/function relationships., Platelets. 2017 Mar;28(2):108-118. doi: 10.1080/09537104.2016.1257786. Epub 2016 Dec 23.
Yadav S, Williamson JK, Aronova MA, Prince AA, Pokrovskaya ID, Leapman RD, Storrie B., Golgi proteins in circulating human platelets are distributed across non-stacked, scattered structures., Platelets. 2017 Jun;28(4):400-408. doi: 10.1080/09537104.2016.1235685. Epub 2016 Oct 18.
Storrie B., A tip of the cap to procoagulant platelets., Blood. 2016 Sep 29;128(13):1668-9. doi: 10.1182/blood-2016-08-730622.
Liu S, Majeed W, Kudlyk T, Lupashin V, Storrie B., Identification of Rab41/6d Effectors Provides an Explanation for the Differential Effects of Rab41/6d and Rab6a/a’ on Golgi Organization., Front Cell Dev Biol. 2016 Mar 1;4:13. doi: 10.3389/fcell.2016.00013. eCollection 2016
Siegel N, Storrie B, Bruce M, Brooker G., CINCH (confocal incoherent correlation holography) super resolution fluorescence microscopy based upon FINCH (Fresnel incoherent correlation holography)., Proc SPIE Int Soc Opt Eng. 2015 Feb 7;9336. pii: 93360S. Epub 2015 Mar 11.
Pokrovskaya ID, Aronova MA, Kamykowski JA, Prince AA, Hoyne JD, Calco GN, Kuo BC, He Q, Leapman RD, Storrie B., STEM tomography reveals that the canalicular system and α-granules remain separate compartments during early secretion stages in blood platelets., J Thromb Haemost. 2016 Mar;14(3):572-84. doi: 10.1111/jth.13225. Epub 2016 Feb 4
Ouseph MM, Huang Y, Banerjee M, Joshi S, MacDonald L, Zhong Y, Liu H, Li X, Xiang B, Zhang G, Komatsu M, Yue Z, Li Z, Storrie B, Whiteheart SW, Wang QJ., Autophagy is induced upon platelet activation and is essential for hemostasis and thrombosis., Blood. 2015 Sep 3;126(10):1224-33. doi: 10.1182/blood-2014-09-598722. Epub 2015 Jul 24.
Storrie B., Defective platelet autocrine signaling in HPS., Blood. 2015 Mar 5;125(10):1515-6. doi: 10.1182/blood-2015-01-616284.
Liu S, Storrie B., How Rab proteins determine Golgi structure., Int Rev Cell Mol Biol. 2015;315:1-22. doi: 10.1016/bs.ircmb.2014.12.002. Epub 2015 Feb 7.
MacDonald L, Baldini G, Storrie B., Does super-resolution fluorescence microscopy obsolete previous microscopic approaches to protein co-localization?, Methods Mol Biol. 2015;1270:255-75. doi: 10.1007/978-1-4939-2309-0_19.
Hickman DA, Syal G, Fausther M, Lavoie EG, Goree JR, Storrie B, Dranoff JA., MCP-1 downregulates MMP-9 export via vesicular redistribution to lysosomes in rat portal fibroblasts., Physiol Rep. 2014 Nov 20;2(11). pii: e12153. doi: 10.14814/phy2.12153. Print 2014 Nov 1.
Majeed W, Liu S, Storrie B., Distinct sets of Rab6 effectors contribute to ZW10–and COG-dependent Golgi homeostasis., Traffic. 2014 Jun;15(6):630-47. doi: 10.1111/tra.12167. Epub 2014 Apr 11.
Liu S, Hunt L, Storrie B., Rab41 is a novel regulator of Golgi apparatus organization that is needed for ER-to-Golgi trafficking and cell growth., PLoS One. 2013 Aug 6;8(8):e71886. doi: 10.1371/journal.pone.0071886. Print 2013.
Micaroni M, Stanley AC, Khromykh T, Venturato J, Wong CX, Lim JP, Marsh BJ, Storrie B, Gleeson PA, Stow JL., Rab6a/a’ are important Golgi regulators of pro-inflammatory TNF secretion in macrophages., PLoS One. 2013;8(2):e57034. doi: 10.1371/journal.pone.0057034. Epub 2013 Feb 21.
Garg TK, Szmania SM, Khan JA, Hoering A, Malbrough PA, Moreno-Bost A, Greenway AD, Lingo JD, Li X, Yaccoby S, Suva LJ, Storrie B, Tricot G, Campana D, Shaughnessy Jr JD Jr, Nair BP, Bellamy WT, Epstein J, Barlogie B, van Rhee F., Highly activated and expanded natural killer cells for multiple myeloma immunotherapy. Haematologica. 2012 Mar 14. [Epub ahead of print]. Free Article. Related citations
Storrie B, Micaroni M, Morgan GP, Jones N, Kamykowski JA, Wilkins N, Pan TH, Marsh BJ Electron Tomography Reveals Rab6 Is Essential to the Trafficking of trans-Golgi Clathrin and COPI-Coated Vesicles and the Maintenance of Golgi Cisternal Number. Traffic. 2012 May;13(5):727-44. doi: 10.1111/j.1600-0854.2012.01343.x. Epub 2012 Mar 14. Related citations
Lisauskas T, Matula P, Claas C, Reusing S, Wiemann S, Erfle H, Lehmann L, Fischer P, Eils R, Rohr K, Storrie B, Starkuviene V., Live-cell assays to identify regulators of ER-to-Golgi trafficking. Traffic. 2012 Mar;13(3):416-32. doi: 10.1111/j.1600-0854.2011.01318.x. Epub 2012 Jan 3. Related citations
Kamykowski J, Carlton P, Sehgal S, Storrie B., Quantitative immunofluorescence mapping reveals little functional coclustering of proteins within platelet α-granules. Blood. 2011 Aug 4;118(5):1370-3. Epub 2011 May 26. Related citations