In the hypothalamus, neurons that synthesize melanin concentrating hormone (MCH) play a key role in the initiation of feeding and in energy homeostasis. MCH antagonists not only reduce obesity, they are also anxiolytic and modulate the sleep-wake state. MCH application in the brain increases feeding and weight gain, whereas loss of the MCH gene causes a decrease. Despite a large amount of fascinating data showing many effects of MCH and MCH neurons, we understand remarkably little about how the neurons act in the brain, and how MCH functions at the cellular level. The primary focus here is to study the MCH neurons, and the actions of MCH at the neuronal level. Whole cell patch clamp recording will be done in transgenic mouse hypothalamic slices containing MCH neurons identifiable by selective expression of GFP. It is surprising that relatively little postsynaptic actions of MCH have been detected. We will test three brain regions that get a high level of MCH axon innervations, the highest level of receptors, and contain a high density of MCH- GFP axons that will be used to target postsynaptic neurons to test MCH responses. These three areas, the hypothalamic dorsomedial nucleus, the medial mammillary nucleus, and locus coeruleus all play key roles in the functions attributed to MCH neurons. A second hypothesis we will test is that heightened activity unmasks MCH actions on postsynaptic targets or presynaptic sites. A third hypothesis tests the view that MCH neurons are a homogeneous group of cells, and suggests that there are two populations of MCH cells, small and large, with different dendritic arbors, axonal projections, and physiological characteristics. We propose to study differences in the dendritic arbors and axonal projections of these cells with a viral vector, dG-vesicular stomatitis virus and pseudorabies virus that express red fluorescent reporters. Ultrastructural differences in the two cell types will be examined with electron microscopy. Differences in active and passive membrane properties and responses to neuromodulators will be studied with whole cell recording in mouse brain slices. In preliminary work, we find that application of the inhibitory amino acid transmitter GABA can result in an unexpected but substantial depolarization of adult MCH neurons. We will test the hypothesis that these GABA actions are not based on Cl- flux, and that they can lead to an increase in spikes in certain physiological states. A current model of energy homeostasis suggests that the orexigenic NPY cells from the actuate nucleus excite MCH neurons. But we find NPY is inhibitory to MCH cells. A more tractable model that we propose to test includes an inhibitory interneuron in the lateral hypothalamus: actuate nucleus NPY inhibits the GABA interneuron, leading to disinhibition of the downstream MCH neuron. We will use a combination of whole cell recording, tract tracing, and immunocytochemistry to test this model. Obesity has become a major health problem leading to increases in heart disease, strokes, mortality and other risks. Studies of the cellular physiology of MCH neurons should provide a better understanding of the role of this system in regulating energy homeostasis, obesity, and anxiety, and will help us understand how we might regulate the MCH system to ameliorate human disease.

Public Health Relevance

Some neurons have a very narrow function. For instance, the magnocellular vasopressin neurons send axons only to the neurohypophysis, and release neuropeptide into the blood to increase water retention in the kidney. In contrast, hypothalamic melanin concentrating hormone (MCH) neurons send axons to over a hundred different brain regions, and appear to exercise a strong modulation of a number of different functions, the foremost of which is modulation of energy homeostasis, body weight, and body fat content. Blocking MCH receptors causes a reduction in food intake, and a decrease in body weight, particularly body fat, and an increase in metabolism. Animals lacking the MCH receptor are resistant to dietary obesity, a growing problem in the USA. In addition, MCH neurons play a role in mental states;modulating MCH actions can reduce anxiety and increase social interaction, and may modulate sleep-wake cycles. We do not yet have a substantive understanding either of how the MCH neuropeptide acts in the brain, nor how the MCH neuron receives and integrates information from other neurons and from the periphery. In today's society, we have both a growing epidemic of obesity that leads to a large number of side effects that reduce quality of life, and increase the cost of health care. Our plan is to unravel some of the complexity surrounding the cellular actions of the MCH peptide, and to test several related hypotheses focusing on how MCH neurons integrate incoming and outgoing information. By better understanding how MCH neurons function at the cellular level, we will be better able to understand how the cell plays such key roles in such a wide variety of functions, and with this knowledge, we will be better able to work through the MCH neuronal system to treat or ameliorate related human health problems.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS048476-08
Application #
8207897
Study Section
Integrative Physiology of Obesity and Diabetes Study Section (IPOD)
Program Officer
Gnadt, James W
Project Start
2004-07-01
Project End
2015-01-31
Budget Start
2012-02-01
Budget End
2013-01-31
Support Year
8
Fiscal Year
2012
Total Cost
$354,791
Indirect Cost
$140,416
Name
Yale University
Department
Neurosurgery
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
Konadhode, Roda Rani; Pelluru, Dheeraj; Blanco-Centurion, Carlos et al. (2013) Optogenetic stimulation of MCH neurons increases sleep. J Neurosci 33:10257-63
Zhang, Xiaobing; van den Pol, Anthony N (2013) Direct inhibition of arcuate proopiomelanocortin neurons: a potential mechanism for the orexigenic actions of dynorphin. J Physiol 591:1731-47
Li, Ying; Xu, Youfen; van den Pol, Anthony N (2013) Reversed synaptic effects of hypocretin and NPY mediated by excitatory GABA-dependent synaptic activity in developing MCH neurons. J Neurophysiol 109:1571-8
Zhang, Xiaobing; van den Pol, Anthony N (2012) Thyrotropin-releasing hormone (TRH) inhibits melanin-concentrating hormone neurons: implications for TRH-mediated anorexic and arousal actions. J Neurosci 32:3032-43
Yao, Yang; Fu, Li-Ying; Zhang, Xiaobing et al. (2012) Vasopressin and oxytocin excite MCH neurons, but not other lateral hypothalamic GABA neurons. Am J Physiol Regul Integr Comp Physiol 302:R815-24
Huang, Hao; Xu, Youfen; van den Pol, Anthony N (2011) Nicotine excites hypothalamic arcuate anorexigenic proopiomelanocortin neurons and orexigenic neuropeptide Y neurons: similarities and differences. J Neurophysiol 106:1191-202
Liu, Meng; Blanco-Centurion, Carlos; Konadhode, RodaRani et al. (2011) Orexin gene transfer into zona incerta neurons suppresses muscle paralysis in narcoleptic mice. J Neurosci 31:6028-40
Fu, Li-Ying; van den Pol, Anthony N (2010) Kisspeptin directly excites anorexigenic proopiomelanocortin neurons but inhibits orexigenic neuropeptide Y cells by an indirect synaptic mechanism. J Neurosci 30:10205-19
van den Pol, Anthony N (2010) Excitatory neuromodulator reduces dopamine release, enhancing prolactin secretion. Neuron 65:147-9
Acuna-Goycolea, Claudio; Obrietan, Karl; van den Pol, Anthony N (2010) Cannabinoids excite circadian clock neurons. J Neurosci 30:10061-6

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