The dusp1 immediate early gene is regulated by natural stimuli predominantly in sensory input neurons

Abstract

Many immediate early genes (IEGs) have activity-dependent induction in a subset of brain subdivisions or neuron types. However, none have been reported yet with regulation specific to thalamic-recipient sensory neurons of the telencephalon or in the thalamic sensory input neurons themselves. Here, we report the first such gene, dual specificity phosphatase 1 (dusp1). Dusp1 is an inactivator of mitogen-activated protein kinase (MAPK), and MAPK activates expression of egr1, one of the most commonly studied IEGs, as determined in cultured cells. We found that in the brain of naturally behaving songbirds and other avian species, hearing song, seeing visual stimuli, or performing motor behavior caused high dusp1 upregulation, respectively, in auditory, visual, and somatosensory input cell populations of the thalamus and thalamic-recipient sensory neurons of the telencephalic pallium, whereas high egr1 upregulation occurred only in subsequently connected secondary and tertiary sensory neuronal populations of these same pathways. Motor behavior did not induce high levels of dusp1 expression in the motor-associated areas adjacent to song nuclei, where egr1 is upregulated in response to movement. Our analysis of dusp1 expression in mouse brain suggests similar regulation in the sensory input neurons of the thalamus and thalamic-recipient layer IV and VI neurons of the cortex. These findings suggest that dusp1 has specialized regulation to sensory input neurons of the thalamus and telencephalon; they further suggest that this regulation may serve to attenuate stimulus-induced expression of egr1 and other IEGs, leading to unique molecular properties of forebrain sensory input neurons. INDEXING TERMSmkp1; mkp-1; hvh1; ptpn10; cl100; vision; somatosensory; auditory; motor pathways; brain organization; neural activity; motor behavior; brain evolution; parrot; hummingbird; songbird; ring dove; bird; primary sensory; ZENK In the brain, immediate early genes (IEGs) are genes whose mRNA expression is dependent on neural activity in the absence of new protein synthesis (Greenberg et al., 1986;Flavell and Greenberg, 2008). As such, these genes are used as markers of neural activity to determine relationships between gene regulation and neural firing, and to map functional domains of the (Tischmeyer and Grimm, 1999;Guzowski et al., 2005;Mello and Jarvis, 2008). We have termed this use of IEGs "behavioral molecular brain mapping" (Jarvis, 2004a;Mello and Jarvis, 2008). This approach has been successively used to identify and characterize neural systems involved in perceiving and producing behaviors. For example, in songbirds, hearing-and singing-driven IEG expression helped to discover and/or characterize most nuclei of the vocal learning and auditory pathways, respectively (Fig. 1A,B;Mello et al., 1992;Jarvis and Nottebohm, 1997;Clayton, 2004;Velho et al., 2005;Wada et al., 2006;Pinaud et al., 2008). Likewise, behavioral molecular mapping has recently been used to map visual, somatosensory, and motor pathways in birds ( Fig...

Quantification of developmental birdsong learning from the subsyllabic scale to cultural evolution

“…In addition to NMR technology, various brain imaging technologies at microspatial scales, such as two-photon imaging (77), and mapping of gene expression dynamics across brain regions (78,79) are rapidly evolving. As with fMRI, those techniques are incorporated at different scales into birdsong neuroethology studies to capture different aspects of neural computation, output, and structural plasticity.…”

Radioactive <em>in situ</em> Hybridization for Detecting Diverse Gene Expression Patterns in Tissue

“…X-ray films provide a quick result and analyses of the overall condition of the hybridization. The x-ray film data also reveals broad anatomical resolution and can be used for quantitative analysis 10 . Examples of x-ray film images of late avian embryo heads hybridized with antisense probes for FoxP1 and CoupTF2 gene expression are in Figures 1A and B.…”

“…Radioactive in situ hybridization of mRNA expression is widely used for multiple purposes, including for studying regional tissue organization, cell types, and brain functional activity [2][3][4][5]10,[12][13][14] . The later use is on genes whose mRNA expression in the brain is dependent on increased neural activity, often called activity-dependent genes or immediate early genes.…”

“…The limitations relative to non-radioactive protocols are that the latter are clearer in the cellular resolution and location of the mRNA within the cell, and working with emulsion is very sensitive to manipulation and light. It is possible to combine our method with other methods, such as non-radioactive in situ hybridization to label mRNA expression of more than one gene in the same tissue 10,17,20 . It can be combined with immunocytochemistry to label both RNA and protein expression on the same sample in order to colocalize the mRNA with certain cell types 10 .…”

“…It is possible to combine our method with other methods, such as non-radioactive in situ hybridization to label mRNA expression of more than one gene in the same tissue 10,17,20 . It can be combined with immunocytochemistry to label both RNA and protein expression on the same sample in order to colocalize the mRNA with certain cell types 10 . The protocol modifications necessary for such double labeling experiments are described in the cited references.…”