? CORE B: TRANSFECTION CORE LABORATORY An important objective of the P01 research proposal is to examine the mechanisms by which the exaggerated exercise pressor reflex (following femoral occlusion) is regulated by pro-inflammatory cytokines (IL-6 and TNF- ?), NaV1.7 and NaV1.8 channels, purinergic (P2X3) channels, prostaglandin (EP4) receptors, and acid sensing ion channels (ASICs) expressed in dorsal root ganglion (DRG) neurons. The responses to contraction of afferents following acute knockdown of myophosphorylase in gastrocnemius muscle will also be investigated. Through Projects 2 and 3, the roles of these channels, receptors and signaling elements will be defined. In order to achieve these aims, the Transfection Core will provide key help through three objectives: 1) the Core will design effective siRNA sequences and provide cDNA clones that code for these `short hair' nucleotide sequences; 2) the Core will perform the in vivo and in vitro transfection of DRG (L4-L5) and gastrocnemius muscle; and 3) the Core will verify protein knockdown employing quantitative RT-PCR (mRNA levels) and Western blotting (protein quantification). These three aims will assist Projects 2 and 3 in testing their specific hypotheses. Overall, the work will lead to a better understanding of the role these proteins play in the exaggerated exercise pressor reflex observed in peripheral artery disease (PAD).
? CORE B: TRANSFECTION CORE LABORATORY In this proposal, the function of the Transfection Core will be to serve as a centralized laboratory that will design and grow the cDNA constructs necessary to silence proteins, deliver genes to neurons (via ultrasound imaging) and skeletal muscle, and measure protein and mRNA of the genes to be silenced. This will enhance the productivity of Projects 2 and 3.
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|Harms, Jonathan; Stone, Audrey J; Kaufman, Marc P (2018) Peripheral µ-opioid receptors attenuate the responses of group III and IV afferents to contraction in rats with simulated peripheral artery disease. J Neurophysiol 119:2052-2058|
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|Farrag, Mohamed; Drobish, Julie K; Puhl, Henry L et al. (2017) Endomorphins potentiate acid-sensing ion channel currents and enhance the lactic acid-mediated increase in arterial blood pressure: effects amplified in hindlimb ischaemia. J Physiol 595:7167-7183|
|Xing, Jihong; Li, Jianhua (2017) Proteinase-Activated Receptor-2 Sensitivity of Amplified TRPA1 Activity in Skeletal Muscle Afferent Nerves and Exercise Pressor Reflex in Rats with Femoral Artery Occlusion. Cell Physiol Biochem 44:163-171|