Large-scale networks of coupled oscillators can be inherently difficult to understand due to complex and diverse interactions. Recent developments in connectivity analysis and graphical network theory provide effective ways to probe these interactions. To date, these tools have been widely applied within the field of cognitive neuroscience;however their use has been limited in the study of neural networks with single neuron resolution. The suprachiasmatic nucleus (SCN) is an excellent system where we may now use these methods to probe the functional connectivity of a neural network and reveal organizing principles in populations of coupled oscillators. The SCN is composed of approximately 20,000 neurons which express daily rhythms in firing rate and gene expression. Coupling between oscillating neurons is necessary to maintain synchronous activity and produce a coherent daily output which regulates circadian rhythms including metabolism, hormone release, and sleep-wake cycles. Disruption of these daily rhythms has deleterious effects on many aspects of human physiology including cognition and mood. To understand circadian regulation within the brain, we must understand the intrinsic properties of each SCN oscillator and how these properties are modulated by network interactions. Revealing how these oscillators couple to generate a coherent rhythmic output will have important clinical implications for prevention and treatment of circadian rhythm disorders, including mood disorders.
In Specific Aim #1, we will determine how network interactions alter the circadian behavior of individual SCN neurons. Using state-of-the-art multielectrode array and single-cell imaging technologies in conjunction with genetic and pharmacologic manipulations aimed at blocking specific neurotransmitter pathways, we propose to compare firing rate and gene expression patterns in individual SCN neurons from networks where the native connections are left largely intact (explanted slices) or networks where neurons have been forced to regrow in culture (high density dispersals). We will test the roles of several neurotransmitters in modulating the intrinsic properties of these neurons.
In Specific Aim #2, we will analyze the functional connectivity within these same cultures by measuring correlated activity and mapping communication between SCN neurons over milliseconds to days. We will test the relative roles of different neurotransmitters in mediating this functional connectivity and circadian synchrony. Together, these experiments will elucidate the neural circuits which regulate the amplitude, period, synchrony and precision of circadian rhythms within the mammalian SCN. The proposed experiments have the potential to transform our understanding of SCN connectivity and more generally contribute to our knowledge of complex systems and networks of coupled oscillators. By probing the dynamics of a biologically important network, we will gain enhanced insight into how SCN neurons synchronize;and develop a better understanding of how these networks may be modulated to treat circadian rhythm disorders including mood disorders. 1

Public Health Relevance

Synchronization between oscillating neurons in the mammalian circadian pacemaker is necessary for the maintenance of daily rhythms in physiology and behavior. Disruption of these daily rhythms has serious and deleterious effects on many aspects of human physiology including cognition and mood. Understanding how the neurons of the circadian pacemaker interact to form a coherent output will greatly aid our ability to understand and treat conditions which have significant circadian components such as depression, bipolar disorder, shift work and circadian changes secondary to normal ageing and dementia.

National Institute of Health (NIH)
National Institute of Neurological Disorders and Stroke (NINDS)
Individual Predoctoral NRSA for M.D./Ph.D. Fellowships (ADAMHA) (F30)
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NST-2 Subcommittee (NST)
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Gnadt, James W
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Washington University
Schools of Arts and Sciences
Saint Louis
United States
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Freeman Jr, G Mark; Krock, Rebecca M; Aton, Sara J et al. (2013) GABA networks destabilize genetic oscillations in the circadian pacemaker. Neuron 78:799-806
Freeman Jr, G Mark; Nakajima, Masato; Ueda, Hiroki R et al. (2013) Picrotoxin dramatically speeds the mammalian circadian clock independent of Cys-loop receptors. J Neurophysiol 110:103-8
Freeman Jr, G Mark; Herzog, Erik D (2011) Neuropeptides go the distance for circadian synchrony. Proc Natl Acad Sci U S A 108:13883-4