mProX™ Human MCHR1 Stable Cell Line

Kobayashi Y, Saito Y. "Evaluation of ciliary-GPCR dynamics using a validated organotypic brain slice ." Methods in cell biology, 2023.
The primary cilium, a structural organelle found in most mammalian cells, possesses a unique protein composition that differs from the cytosol and plasma membrane. Its organized environment allows for efficient translation of external cues into internal signals. Recent studies have shown the presence of non-olfactory G protein-coupled receptors (GPCRs), particularly melanin-concentrating hormone receptor 1 (MCHR1), specifically localized to ciliary membranes in various mammalian cell types, including neurons. Using ciliary-specific techniques, evidence has supported the role of ciliary-GPCR signaling in structural changes. However, neuronal dissociation for ciliary-GPCR studies may lead to altered ciliary morphology. Utilizing ex vivo brain slices allows for precise experimental control and examination of individual cells in different brain regions. To investigate ciliary-GPCR dynamics and its interaction with neuropeptides or aminergic activation, a modified method for rat brain slice culture and an immunostaining protocol have been developed and validated.
Pubmed: 36967146   DOI: 10.1016/bs.mcb.2022.09.007

Brewer KM, et al. "Physiological Condition-Dependent Changes in Ciliary GPCR Localization in the ." eNeuro, 2023.
Primary cilia, critical for diverse signaling processes, are found on most cell types, including those in the central nervous system. These cellular appendages play a crucial role in localizing specific G-protein-coupled receptors (GPCRs) and mediating their signaling. While studies in organisms such as Caenorhabditis elegans and Chlamydomonas have implicated the dynamic localization and length and shape changes of cilia in GPCR signaling, it remains unclear if similar mechanisms exist in mammals and under what circumstances. In this study, the authors used two neuronal cilia GPCRs, MCHR1 and NPY2R, as mammalian model ciliary receptors in mice to investigate their ciliary localization under physiological conditions related to their functions in feeding, sleep, and reward. Through unbiased and high-throughput analysis, they observed changes in ciliary length, receptor occupancy, and frequency in response to different conditions and in specific brain regions, suggesting that the dynamic localization of ciliary GPCRs is dependent on the specific properties of the receptor and the cell type in which it is expressed. These findings provide new insights into the molecular mechanisms underlying important behaviors, such as feeding, and could potentially guide future research in this area.
Pubmed: 36849261   DOI: 10.1523/ENEURO.0360-22.2023