AAV ACTOne Gi-GPCR Assay Kit-HTR1F

Serotonin Receptor 1F (HTR1F) is a G protein-coupled receptor (GPCR) that belongs to the serotonin receptor family. Like other members of the 5-HT1 receptor subfamily, HTR1F is primarily coupled with the Gi/Go class of G proteins, leading to inhibition of adenylate cyclase and a reduction in cyclic AMP (cAMP) levels within the cell. This reduction in cAMP modulates various intracellular signaling pathways, ultimately influencing neurotransmitter release and neuronal excitability. HTR1F is expressed in the central nervous system (CNS) and some peripheral tissues. It is particularly found in regions associated with migraine and pain modulation, making it a receptor of interest in the treatment of migraines and other types of headaches. The receptor's role in migraine pathophysiology has led to the development of specific agonists that target HTR1F, such as lasmiditan, which is used to treat acute migraine attacks. These drugs work by activating HTR1F, leading to the inhibition of neurotransmitter release that contributes to migraine headaches, without causing the vasoconstriction typically associated with other migraine treatments like triptans. Given its involvement in pain modulation and migraine treatment, HTR1F continues to be a focus of research for developing new therapeutics that target serotonin signaling pathways in the CNS, particularly for conditions that require precise modulation of serotonin activity without affecting vascular function.
This kit uses AAV vectors with a CMV promoter to co-express the HTR1F and cyclic nucleotide-gated (CNG) channel, allowing researchers to conduct high-throughput screening and functional analysis of potential HTR1F-targeting compounds. The kit provides a sensitive and reliable method for evaluating the pharmacological properties of HTR1F drugs, such as agonists and antagonists, in a live-cell environment.

Please contact us at for specific academic pricing.

Background

ACTOne™ is the only high-throughput GPCR screening technology that can directly measure the intracellular changes of the secondary messenger cyclic AMP (cAMP) in living cells, in real-time. It uses a proprietary modified cyclic nucleotide-gated (CNG) channel, which is co-localized with adenylate cyclase at the plasma membrane, as a biosensor of cAMP activity. The CNG channel opens when the cAMP level near the plasma membrane increases, resulting in ion flux and cell membrane depolarization. The influx of cations through the CNG channel can be quantified using fluorescent ion indicators or membrane potential (MP) dyes. It provides information on real time intracellular cAMP changes and is highly sensitive. By combining kinetic and endpoint readouts, we are able to capture and analyze transient responses from endogenous GPCRs and weak responses caused by weak Gs or Gi coupled GPCR activities. Using ACTOne, we are able to detect the subcellular cAMP concentration changes directly caused by GPCR activation. Real-time kinetic readouts minimize artifacts, and provide greater content and more statistically relevant data. The intensity of signal increase caused by GPCR activation is directly related to the receptor number on cell surface. Using ACTOne assay, we were able to detect activities of some endogenous Gs coupled receptors in HEK293 cells that have not been reported in literature. In addition, we have also detected weak Gs coupled activity of a GPCR that was widely considered to be only linked to Gq coupled pathway. The ACTOne assay also provides a useful tool for GPCR de-orphanization.