Localization of the GoLoco motif carrier regulator of G‐protein signalling 12 and 14 proteins in monkey and rat brain

Abstract

Regulator of G-protein signalling (RGS)12 and -14 proteins possess the RGS domain, Ras-binding domains and the GoLoco motif. Emerging evidence suggests that these proteins are involved in several cellular functions in addition to stimulation of GTPase activity of G-protein alpha subunits. However, our understanding of the role of the two proteins in brain function remains marginal. Here, we have studied the expression pattern of RGS12 and RGS14 proteins in brain at regional, cellular and subcellular levels. Both proteins were expressed throughout the brain regions, including cortex, hippocampus, striatum, thalamus and substantia nigra. The most intense immunostaining for RGS12 was seen in cortex and that of RGS14 was found in striatum. In cortex, RGS12 and RGS14 proteins were associated with pyramidal and nonpyramidal cell types. Apical dendrites of pyramidal cells were also labelled. RGS12 was found in both nuclear and cytoplasmic compartments. In contrast to RGS12 protein, RGS14 was localized in astrocytes in addition to neurons. Pyramidal cells in the CA1 area showed labelling for both RGS proteins. The presence of RGS12 was predominantly nuclear in the striatum of rat brain; however, the labelling of this protein was non-nuclear in adult monkey brain. To our surprise, in 1-month-old monkey brain the immunostaining pattern of the same protein was changed to nuclear. Non-nuclear staining for RGS12 was also evident in thalamus of adult monkey brain; however, in 1-month-old monkey brain, it was seen into two different populations, one with nuclear and the other with cytoplasmic staining. Both RGS12 and RGS14 were exclusively localized at postsynaptic sites of excitatory synapses. Our results demonstrate a highly dynamic expression pattern of RGS12 and RGS14 proteins in the central nervous system, and support the view that these proteins may participate not only in G-protein receptor signalling pathways but also in other cellular activities.

Regulators of G-Protein-Signaling Proteins: Negative Modulators of G-Protein-Coupled Receptor Signaling

“…RGS12 has also been reported to be mostly expressed in trigeminal and dorsal root ganglia (DRG) neurons and muscle in the E14.5 embryonic mouse suggesting a role for mammalian RGS12 in myo-and neurogenesis (Martin-McCaffrey et al, 2005a), and also in the thalamus of adult monkey (Lopez-Aranda et al, 2006). Studies in COS-7 and HEK293 cells, have reported that endogenous or ectopically expressed RGS12 segregates into nuclear locations, thus suggesting its involvement in the regulation of transcription and cell cycle (Chatterjee and Fisher, 2000b;.…”

“…RGS14 is widely expressed in mammal tissues (Buckbinder et al, 1997;Evans et al, 2014;Jia et al, 2012;Lim et al, 2013;Martin-McCaffrey et al, 2005a;Oner et al, 2013;Vellano et al, 2013;Yasui et al, 2013;Zhao et al, 2013), such as brain (Lopez-Aranda et al, 2006;Snow et al, 1997), lymphoid organs (Larminie et al, 2004;Snow et al, 1997) and lung , and cells, such as lymphocytes (Larminie et al, 2004), in pyramidal and nonpyramidal cell types in brain cortex in astrocytes (Lopez-Aranda et al, 2006) and in low rate in platelets (Kim et al, 2006).…”

“…Human RGS12 is expressed in brain (Chatterjee and Fisher, 2000b) and myometrium (O'Brien et al, 2008), while rat RGS12 is expressed at high levels in brain, including cortex, hippocampus, striatum, thalamus, and substantia nigra, as well as in lung (Lopez-Aranda et al, 2006;Snow et al, 1997). RGS12 has also been reported to be mostly expressed in trigeminal and dorsal root ganglia (DRG) neurons and muscle in the E14.5 embryonic mouse suggesting a role for mammalian RGS12 in myo-and neurogenesis (Martin-McCaffrey et al, 2005a), and also in the thalamus of adult monkey (Lopez-Aranda et al, 2006).…”

Prefrontal Inositol Triphosphate Is Molecular Correlate of Working Memory in Nonhuman Primates

“…Homogenates (for active PKA levels) and prepared membrane fractions (for G␣q protein levels) were treated with SDS buffer and processed for Western blot analysis similar to as described previously (Khan et al, 2000(Khan et al, , 2001Ló pez-Aranda et al, 2006). Homogenate proteins (395 g/lane) and membrane proteins (100 g/lane) were separated by 12% SDS-PAGE and transferred onto a polyvinylidene fluoride membrane (GE Healthcare).…”

Regulator of G Protein Signaling 14

“…[15][16][17][18]43 Native RGS14 is found at high levels in brain of various species including mouse, rat, nonhuman primate, and human. 7,15,17,[43][44][45][46] In adult rodents, RGS14 is most highly expressed in brain, specifically in neurons. 7,17,43 Most recently, we have shown that RGS14 mRNA/protein expression is upregulated during postnatal mouse brain development reaching its highest levels in adulthood.…”