[No authors listed]
In Drosophila, two related G-protein-coupled receptors are members of the corticotropin releasing factor (CRF) receptor subfamily. We have previously reported that one of these receptors, encoded by CG8422 is a functional receptor for a diuretic hormone, DH(44). Here, we report that the other CRF receptor subfamily member, encoded by CG12370, is also a receptor for the DH(44) neuropeptide. The lines of evidence to support this identification include increases in cAMP levels due to CG12370 receptor activation and the recruitment of beta-arrestin-GFP to the plasma membrane in response to DH(44) application. We compared these features of the receptors DH44-R2 (encoded by CG12370) and DH44-R1 (encoded by CG8422) and found fundamental differences in signaling, association with the arrestins, and peptide sensitivity. We found that the sensitivity of DH44-R2 to the DH(44) peptide is lower than that of DH44-R1, specifically an estimated EC(50) of 7.98E-07 moll(-1) for DH(44) by DH44-R2 to an EC(50) of 5.12E-09 moll(-1) by DH44-R1 and found that previous reports on the sensitivity of the tubule to DH(44) is in agreement with our measurements of DH44-R2 sensitivity. We employed a specific construct to selectively knock-down DH44-R2 expression and this led to heightened sensitivity to osmotic challenges. The functional characterization of this diuretic hormone receptor in Drosophila demonstrates a high degree of conservation of CRF-like signaling.
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