Neurovascular coupling (NVC) is the distinctive process within the cerebral circulation by which local cerebral blood flow (CBF) is precisely directed to active brain regions. NVC is indispensible for all brain functions, reflecting the fact that central neurons have little capacity to store energetic substrates and require prompt delivery of metabolites that are rapidly consumed during synaptic activity. An emerging conceptual paradigm envisions the hundreds of miles of capillaries in the human brain as a sensory web that mediates NVC by detecting elevated neuronal activity and initiating a propagating dilatory signal to produce a local, functional hyperemic response. The overall goals of this proposal are to identify novel sensory mechanisms intrinsic to brain capillary endothelial cells (ECs) that rapidly detect increases in neuronal activity and to elucidate how such signals propagate and act on the cerebral microcirculation to trigger functional hyperemia. We propose the novel mechanistic hypothesis that type 5 metabotropic glutamate receptors (mGluR5s) are present on brain capillary ECs and, when stimulated by glutamate released from astrocytic endfeet, initiate dilation of upstream parenchymal arterioles (PAs). The goal of Aim 1 is to elucidate the intracellular signaling mechanisms initiated by mGluR5s on brain capillary EC that trigger dilation of upstream PAs. We will test the hypothesis that mGluR5 initiates a Gq/11/PLC signaling cascade that leads to increased reactive oxygen species (ROS) generation and Ca2+ influx through TRPA1 channels to trigger dilation of upstream PAs. Proposed studies will use Ca2+ and ROS imaging, patch-clamp electrophysiology of native brain capillary ECs, and a newly developed ex vivo arteriolar-capillary preparation that provide an ideal reduced setting for assessing the role of brain capillaries in regulating the upstream vasculature. The goal of Aim 2 is to elucidate the intercellular signaling mechanisms responsible for conducted dilation of upstream PAs in response to stimulation of mGluR5s on brain capillary ECs. We will test the hypothesis that glutamate binds to mGluR5s, leading to activation of TRPA1 and the generation of intercellular Ca2+ waves that propagate to upstream PAs to signal dilation. Proposed studies will use high-resolution Ca2+ imaging of the cerebral microcirculation and super-resolution microscopy to establish the architecture of Ca2+ signaling complexes in brain capillaries. The goal of Aim 3 is to test the hypothesis that mGluR5s on brain capillary ECs stimulate functional hyperemia in vivo. These studies will use two-photon laser-scanning microscopy to measure mGluR5- and TRPA1- dependent changes in RBC flux and Ca2+ signaling in capillaries in vivo, as well as laser Doppler flowmetry to measure changes in CBF in the somatosensory cortex in response to whisker stimulation. The use of novel EC-specific Ca2+ biosensor mice and EC-specific mGluR5- and TRPA1-knockout mice strengthens our approach in all aims. Anticipated findings will support a new model of NVC in which neuronal activity stimulates glutamate release near capillaries to activate mGluR5s on ECs, which initiate localized increases in blood flow.
The long-term goal of this project is to help to develop new treatments for diseases such as stroke and vascular cognitive impairment that harm the regulation of blood flow in the brain. To accomplish this, we seek to discover a new mechanism that matches blood flow within the brain to regions that are rapidly consuming oxygen and glucose due the activity of neurons. Precise delivery of these substances is essential for all brain function. We will test the hypothesis that receptors located on the walls of capillaries in the brain can detect increases in the activity of neurons and send signals to blood vessel located upstream to cause them to dilate. This process will increase blood flow to the specific areas where oxygen and glucose are needed. We predict that a fuller understanding of this process may eventually lead to the development of new treatment to improve blood flow control in the brain during disease.
| Pires, Paulo Wagner; Earley, Scott (2018) Neuroprotective effects of TRPA1 channels in the cerebral endothelium following ischemic stroke. Elife 7: |
| Zheng, Haifeng; Drumm, Bernard T; Earley, Scott et al. (2018) SOCE mediated by STIM and Orai is essential for pacemaker activity in the interstitial cells of Cajal in the gastrointestinal tract. Sci Signal 11: |
| Pritchard, Harry A T; Pires, Paulo W; Yamasaki, Evan et al. (2018) Nanoscale remodeling of ryanodine receptor cluster size underlies cerebral microvascular dysfunction in Duchenne muscular dystrophy. Proc Natl Acad Sci U S A 115:E9745-E9752 |
| Pritchard, Harry A T; Gonzales, Albert L; Pires, Paulo W et al. (2017) Microtubule structures underlying the sarcoplasmic reticulum support peripheral coupling sites to regulate smooth muscle contractility. Sci Signal 10: |
