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Molecular and
Cellular
Neuroscience Laboratory
Some of the techniques used in our lab. (A) Carbon fibre amperometry
recording of exocytosis. The red line indicates
where the cell
has been stimulated. Current spikes indicate the release of catecholamines
from an individual vesicle.
(B) Membrane capacitance measurement
of exocytosis. Exocytosis is elicited by a set of voltage pulses
from -70mV to 0mV.
The membrane capacitance is measured in between
these pulses and increases accordingly. Scale bar in (B) represents
50 pA and 10 sec in (A) and 200 fF and 1 second in (B). (C)
Electron micrograph of a chromaffin cell.
Arrows indicate LDCVs
primed for release at the plasma membrane. (D) Mitochondrial membrane
potential measured using JC-1.
(E) Pancreatic sections stained
with the nuclear stain DAPI (blue) and insulin (pink) to identify
islets.
Our research is focused on the regulation of neuronal communication. Specifically we are interested in identifying specific new molecules which regulate exocytosis of neurotransmitters and the recycling of synaptic vesicles. As well as this we have a significant focus on how neurotransmission is altered in human neurodegenerative diseases. These diseases include Down Syndrome, Alzheimer’s Disease, Huntington’s Disease and Diabetes. We have access to mouse and cell-based models of these diseases, enabling us to investigate the role of specific genes at the whole animal and cellular levels. These studies are carried out using a number of approaches including physiological measurements of exocytosis from single cells with carbon fibre amperometry, or by whole cell capacitance measurements and by the measurement of ion channel currents using patch clamp electrophysiology. We also carry out transfection of cells with GFP-tagged mutant constructs, immunohistochemistry, live cell fluorescence ion imaging, real time RT-PCR, measurement of mitochondrial activity and culturing of primary cells and cell lines.
Investigator
Dr Damien Keating, PhD, ARC Future Fellow
Post-doctoral Researchers
Dr Michael Duffield, Research Associate
Dr Ravinarayan Raghupathi, Research Associate
Dr Heshan Peiris, Research Assistant
Students
Kimberley Mackenzie, PhD Student
Minh-Son To, BMBS/PhD Student
Ravinder Kashmir Singh, MBiotech Student
Leah Zelkas, Honours Student
Contacts
Research Projects
Collaborative Research
Prof Xin-Fu Zhou, Flinders University: Role of
Huntingtin-associated protein 1 in regulating exocytosis.
Dr Melanie Pritchard, Monash University: Defining
the neurological role of Down Syndrome-related and
Alzheimer’s
Disease-related genes using mouse models.
Prof Phil Robinson, Children’s Medical Research Institute,
Sydney: Development of novel small molecule inhibitors
of dynamin, a central controller of receptor-mediated and synaptic
vesicle endocytosis.
Assoc Prof Frederic Meunier,
University of Queensland: Phosphoinositide
regulation of neuroexocytosis.
Dr Nick Spencer, Flinders University: Colonic
5-HT release and gut motility.
Prof Nikolai Petrovsky, Flinders University: Understanding
how mutations in the Alström syndrome-related gene,
Alms1, affect cell communication.
Dr Toby Coates, Queen Elizabeth Hospital: Genetic
regulation of pancreatic beta cell function.
Media Appearances
- Damien Keating - SA Life, Health Funds, October 2010
- Damien Keating - Flinders Journal, Poppies sprout from brain and shark research, August 2010
- Damien Keating - The Adelaide Advertiser, Awards for our smartest brains, August 2010
- Damien Keating - Research Pulse, Flinders rewards its emerging best and brightest, p8, March 2010
- Damien Keating - The Independent Daily, Research achievements rewarded, February 2010
Selected Recent Publications
Spencer NJ, Nicholas SJ, Robinson L, Kyloh M, Flack N, Brookes SJ, Zagorodnyuk VP, Keating DJ (2011) Mechanisms underlying distension-evoked peristalsis in guinea pig distal colon: is there a role for enterochromaffin cells? Am J Physiol Gastrointest Liver Physiol, 301(3):G519-27
Barreto SG, Bazargan M, Zotti M, Hussey DJ, Sukocheva OA, Peiris H, Leong M, Keating DJ, Schloithe AC, Carati CJ, Smith C, Toouli J, Saccone GT (2011) Galanin receptor 3 - a potential target for acute pancreatitis therapy. Neurogastroenterol Motil, 23(3):e141-51
Jentsch TJ, Maritzen T, Keating DJ and Thevenod F (2010) ClC-3 – a granular anion transporter involved in insulin secretion? Cell Metabolism, 12(4):307-8
Keating DJ & Spencer NJ (2010) Controversies Involving the Role of 5-Hydroxytryptamine in Generating Colonic Migrating Motor Complexes: What Is Spontaneous? Gastroenterology, 138:1214-5
Keating DJ and Spencer NJ (2010) Real time recordings of 5-HT release from enterochromaffin (EC) cells reveals their role in the generation and propagation of colonic migrating motor complexes. Gastroenterology, 138:659-70
Pfeffer C, Stein V, Keating DJ, Maier H, Rinke I, Rudhard Y, Hentschke M, Rune G, Jentsch TJ, Hubner CA (2009) NKCC1-Dependent GABAergic Excitation Drives Synaptic Network Maturation During Early Hippocampal Development. J Neurosci, 29:3419-30
Keating DJ, Rychkov GY, Roberts ML (2009) The contribution of voltage-gated Ca2+ currents to K+ channel activation during ovine adrenal chromaffin cell development. Int J Develop Neurosci, 27:357-63
Keating DJ, Dubach D, Zanin MP, Yu Y, Martin K, Zhao YF, Chen C, Porta S, Arbonés ML, Mittaz L, Pritchard MA (2008) DSCR1/RCAN1 regulates vesicle exocytosis and fusion pore kinetics: implications for Down syndrome and Alzheimer's disease. Hum Mol Genet, 17:1020-30
Yu Y, Chu P, Bowser DN, Keating DJ, Harper I, Tkalcevic J, Finkelstein DI, Pritchard MA (2008) Mice deficient for the Down syndrome-related gene Itsn1 exhibit vesicle trafficking abnormalities. Hum Mol Genet, 17: 3281-90
Keating DJ (2008) Mitochondrial dysfunction, oxidative stress, regulation of exocytosis and their relevance to neurodegenerative diseases. J Neurochem, 104: 298-305
Maritzen T, Keating DJ, Neagoe I, Zdebik AA, Jentsch TJ (2008) Role of the vesicular chloride transporter ClC-3 in neuroendocrine tissue. J Neurosci, 28: 10587-98
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