BIO411H5 Lecture Notes - Lecture 1: Pertussis Toxin, Western Blot, Adenylyl Cyclase
Document Summary
Circadian regulation of intracellular g-protein signalling mediates intercellular synchrony and rhythmicity in the suprachiasmatic nucleus. This paper wanted to identify how cellular clocks in the suprachiasmatic nucleus (scn) communicated in order to have synchronous oscillations. It is hypothesized that g i/o-mediated camp signalling in the scn was under circadian regulation via member(s) of the regulator of g protein signalling (rgs) family. First, dna microanalysis was used to survey all known mouse. Rgs genes for their expression in the scn. Highly expressed genes were selected and further analyzed with quantitative in situ hybridization, leading to rgs16 as the only one to display marked scn-specific expression particularly in the subjective daytime (ct4) of constant darkness (dd). Second, the in vivo function of rgs16 was identified by replacing the whole coding sequence of rgs16 in mice with a lacz promoter. Rgs16 deficiency lengthened the circadian period of behaviour. Third, double in situ hybridization showed both the mrnas.
