2013 — 2021 |
Ross, Sarah Elizabeth |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Investigating the Neural Circuits of Itch @ University of Pittsburgh At Pittsburgh
DESCRIPTION (provided by applicant): Chronic itch (puritis) is a severe condition that results in severely diminished quality of life. Moreover, this condition is much more widespread than generally appreciated, representing the most common reason to visit a dermatologist, despite the fact that treatments are generally ineffective. However, while itch is initiated in the skin, i is more than just a skin condition. Rather, itch is a complex and poorly understood sensation that is mediated by neural circuits in the periphery, the spinal cord and the brain. Thus, the long-term goal of our research is to gain a better understanding of the neural circuits that mediate itch wit the view of developing more effective therapies for puritis. We previously discovered that the transcription factor Bhlhb5 is required for the survival of a subset of inhibitory interneurons in he spinal cord (which are here termed B5-I neurons) that are required for normal itch sensation; mice lacking these spinal interneurons suffer from persistent pathological itch. Since B5-I neurons are the first component of an itch circuit to be labeled genetically, studying these neurons provides us with a unique opportunity to unravel the neural basis of itch. Importantly, we have recently discovered that B5-I neurons are a specific subset of spinal interneurons that express dynorphin. This finding is important because dynorphin is a kappa opioid receptor (KOR) agonist, and KOR agonists have recently been shown to relieve itch in rodents and man. We therefore hypothesize that B5-I neurons function to inhibit itch, and that they do so in part through the release of dynorphin in the spinal cord. Here we propose to test this hypothesis in through 3 specific aims: Aim 1: Determine the degree to which B5-I neurons are specific to the regulation of itch. Aim 2: Dissect neural circuits in the dorsal horn using B5-I neurons as a molecular handle. Aim 3: Define the role of spinal dynorphin in itch. Results from these experiments are likely to have major clinical implications for people that suffer from chronic itch because our findings will define a key cellular target for the development of future anti-itch therapies and they will provide important insight into the mechanism of KOR-mediated inhibition of itch.
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2013 — 2014 |
Ross, Sarah Elizabeth |
R21Activity Code Description: To encourage the development of new research activities in categorical program areas. (Support generally is restricted in level of support and in time.) |
Using Dual Intersectional Genetics to Understand and Modulate Itch @ University of Pittsburgh At Pittsburgh
DESCRIPTION (provided by applicant): Chronic itch (pruritis) is a widespread condition that severely diminishes quality of life. However, there are few effective treatments for chronic itch, in part because the neural basis for itch remains poorly understood. Thus, the long-term goal of our research is to gain a better understanding of how itch is encoded in the nervous system at the level of specific neural circuits with the view of developing more effective therapies for pruritis. We previously discovered that the transcription factor Bhlhb5 is required for the surviva of a subset of inhibitory interneurons in the spinal cord (which are here termed B5-I neurons) that are required for normal itch sensation; mice lacking these spinal interneurons suffer from persistent pathological itch. These findings imply that B5-I neurons function to inhibit itch; however, the evidence is merely correlative. In this application we propose to use intersectional genetic strategies to manipulate the activity of B5-I neurons in order to establish cause-and-effect relationships between the activity of B5-I neurons and scratching behavior in mice. Here we propose to test this hypothesis through 3 specific aims: 1) Characterize the Bhlhb5-flpO knockin mouse, a key tool for our dual intersection strategy, and use this mouse together with the Ptf1a-cre line to genetically define B5-I neurons. 2) Investigate the functional response properties of B5-I neurons to natural stimulation of the skin and to confirm the ability of pharmacogenetic approaches to specifically manipulate their activity. 3) Use exocytogenetic and pharmacogenetic approaches to delineate function of B5-I neurons in vivo for itch-mediated scratching behavior. Results from these experiments will begin decoding the neural circuits that underlie itch by enabling us to visualize, characterize, and functionally manipulate B5-I neurons in vitro and in vivo. Moreover, the insight gleaned may have major clinical implications for people that suffer from chronic itch.
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2016 — 2020 |
Koerber, H Richard [⬀] Ross, Sarah Elizabeth |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Molecular Genetic Dissection of the Spinal Microcircuits of Wind-Up @ University of Pittsburgh At Pittsburgh
Chronic pain is a debilitating condition that affects one in four Americans, and for which there is a pressing need for safe, effective treatments. Chronic pain patients experience enhanced pain sensations and often experience pain when innocuous stimuli are presented. However, the neural basis for this amplification is poorly understood. Here, we propose to investigate the neural circuit basis for wind-up, a physiological type of central hyperexcitability that may also contribute to persistent pain. The studies we are proposing will begin to identify specific spinal circuitry involved in this amplification, and investigate whether these microcircuits are altered in conditions of injury. This knowledge may elucidate new therapeutic targets for the treatment of pain, which is the long-term goal of research of our program. In the first aim, we will use our novel skin/nerve/DRG/spinal cord preparation combined with optogenetic approaches to examine the involvement of select cell types in wind-up of cutaneous sensory inputs recorded in spinal projection neurons. These studies will examine the roles of specific subsets of cutaneous sensory neurons in wind-up by optogenetic stimulation of their cutaneous projections both in naïve mice and following nerve injury. We will also employ optogenetic strategies to activate or inhibit specific subsets of genetically defined excitatory (neurotensin (Nt)-cre) and inhibitory (nNos-creER) spinal interneurons to determine their roles in this process. In the second aim we will examine potential neural network and/or synaptic mechanisms underlying the wind-up of sensory inputs. In particular, we will test the role of persistent, reverberating currents in wind-up. In addition, investigate which mediators cause the slow depolarizing current that is often observed with wind-up, and determine whether this plays a contributing role. In the third aim, we will use a novel behavioral model of wind-up using temporal summation of cutaneous sensory inputs. Specifically, we have developed a behavioral model of temporal summation in mice using the same optogenetic stimulation that we previously used to induce wind-up in the first aim. This will allow us, for the first time, to make a direct correlation between the physiological phenomenon (wind-up) and a behavioral response to the perception of pain (temporal summation), using place-aversion as a measure of nociception in mice. Completion of the studies proposed in this application will provide new insights into spinal circuitry underlying the processing of sensory information, and how these processes are altered following nerve injury. Importantly could provide potential targets for the development of pharmaceutical therapies. These new therapies could provide for improved treatments for the alleviation of the adverse symptoms of chronic neuropathic pain.
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2021 |
Ross, Sarah Elizabeth Vazquez, Alberto L (co-PI) [⬀] |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Neural Circuit Basis For Neurovascular Coupling @ University of Pittsburgh At Pittsburgh
Abstract Neurovascular coupling (NVC) is the temporal relationship between neural activity and cerebral blood flow (CBF). This neural-evoked hemodynamic response is fundamental to local cerebrovascular homeostasis and is disrupted in cerebrovascular diseases, such as stroke, cerebral amyloid angiopathy, traumatic brain injury, as well as Alzheimer's Disease. The neurons that express neuronal nitric oxide synthase (Nos1) are ideal candidates for the regulation of NVC since nitric oxide (NO) is a very potent vasodilator. Our group has recently developed a Tacr1CreER allele that enables the visualization and manipulation of these neurons. We now have exciting preliminary data supporting the hypothesis that Tacr1 neurons mediate vasodilation. Here, we propose to test this idea through a set of experiments that will: determine the relationship between Tacr1 neurons and blood vessels; examine causality in the regulation of NVC by Tacr1 neurons; and investigate the underlying circuitry. These experiments include correlative studies that will establish whether the structure (place) and function (activity) of Tacr1 neurons positions them to regulate CBF. We will also use optogenetic approaches and laser Doppler flowmetry (LDF) to record CBF in awake behaving mice to test whether Tacr1 neurons necessary and sufficient for vasodilation. Finally, we will use a combination of optogenetic manipulation, GCaMP6f-, and 2P-imaging to elucidate the underlying circuitry of NVC. Overall, our proposal will address a critical gap in knowledge with respect to the specific neural mechanisms that underlie the BOLD signal, which is a widely used, but poorly understood research and clinical tool. Moreover, this insight into NVC is fundamental to our understanding of the pathogenesis of common cerebrovascular diseases and the advancement of pharmacotherapeutics targeting cerebral perfusion.
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