Identification of regulatory elements modulating PRRXL1/DRG11 transcription during the development of pain-processing neurons
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1
University of Porto, Laboratory of Molecular Cell Biology, Faculty of Medicine and IBMC, Portugal
Pain is a multi-sensory experience associated with potential tissue damage. Pain perception normally depends on signalling the presence of noxious stimuli at the periphery through specialized small diameter neurons of the dorsal root ganglion (DRG) that synapse extensively with marginal zone projecting neurons and substantia gelatinosa interneurons in the spinal cord dorsal horn. The paired-homeodomain transcription factor Prrxl1 (also known as Drg11) has a crucial role in the establishment of synaptic connections between DRG and spinal nociceptive neurons. Nevertheless, our understanding of the transcriptional mechanisms that control the proper spatiotemporal expression of the Prrxl1 gene remains limited. To address this issue, the highly conserved 5'-flanking region of Prrxl1 translation start point was analyzed by cloning entire and partial fragments in a reporter plasmid containing the luciferase gene as a marker of transcriptional induction. The constructs were afterwards transfected in a DRG-derived neuronal cell line (ND7/23) and the enzymatic activity determined. Two distinct regions displaying promoter activity were identified which are suggestive of alternative promoter usage as a mechanism of control of Prrxl1 expression. One of these regions contains a bona fide TATAbox that was validated by Electrophoretic Mobility Shift Assays. Further analysis of this sequence using bioinformatics tools revealed the presence of putative binding sites for bHLH (including neurogenin1) and homeodomain transcription factors (such as Pax2 and Prrxl1 itself), motifs that are highly conserved in others species, namely chick and zebrafish. Altogether, the present results led to the identification of the Prrxl1 promoter and some regulatory motifs that may be implicated in the modulation of Prrxl1 expression.
Conference:
11th Meeting of the Portuguese Society for Neuroscience, Braga, Portugal, 4 Jun - 6 Jun, 2009.
Presentation Type:
Poster Presentation
Topic:
Abstracts
Citation:
Regadas
I,
Monteiro
FA,
Rebelo
S,
Lima
D and
Reguenga
C
(2009). Identification of regulatory elements modulating PRRXL1/DRG11 transcription during the development of pain-processing neurons.
Front. Neurosci.
Conference Abstract:
11th Meeting of the Portuguese Society for Neuroscience.
doi: 10.3389/conf.neuro.01.2009.11.132
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Received:
11 Aug 2009;
Published Online:
11 Aug 2009.
*
Correspondence:
Deolinda Lima, University of Porto, Laboratory of Molecular Cell Biology, Faculty of Medicine and IBMC, Porto, Portugal, limad@med.up.pt