Validating in utero electroporation for the rapid analysis of gene regulatory elements in the murine telencephalon.
With the ultimate goal of understanding how genetic modules have evolved in the telencephalon, we set out to modernize the functional analysis of cross-species cis-regulatory elements in mouse. In utero electroporation is rapidly replacing transgenesis as the method of choice for gain- and loss-of-function studies in the murine telencephalon, but the application of this technique to the analysis of transcriptional regulation has yet to be fully explored and exploited. To empirically define the developmental stages required to target specific populations of neurons in the dorsal telencephalon, or pallium, which gives rise to the neocortex in mouse, we performed a temporal and spatial analysis of the migratory properties of electroporated versus birth-dated cells. Next, we compared the activities of two known Ngn2 enhancers via transgenesis and in utero electroporation, demonstrating that the latter technique more faithfully reports the endogenous telencephalic expression pattern observed in an Ngn2lacZ knock-in line. Finally, we used this approach to test the telencephalic activities of a series of deletion constructs comprised of the zebrafish ER81 upstream regulatory region, allowing us to identify a previously uncharacterized enhancer that displays cross-species activity in the murine piriform cortex and lateral neocortex, yet not in more medial domains of the forebrain. Taken together, our data supports the contention that in utero technology can be exploited to rapidly examine the architecture and evolution of pallial-specific cis-regulatory elements.
Langevin
Lisa Marie
LM
Institute of Maternal and Child Health, HBI, University of Calgary, Calgary, Canada.
Mattar
Pierre
P
Scardigli
Raffaella
R
Roussigné
Myriam
M
Logan
Cairine
C
Blader
Patrick
P
Schuurmans
Carol
C
D000818:Animals; D001483:Base Sequence; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics; D002465:Cell Movement; D017931:DNA Primers / Q000235:genetics; D018274:Electroporation / Q000379:methods*; D004742:Enhancer Elements, Genetic; D005260:Female; D018507:Gene Expression Regulation, Developmental; D005809:Genes, Regulator; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics; D007763:Lac Operon; D051379:Mice; D008822:Mice, Transgenic; D019579:Neocortex / Q000166:cytology / Q000196:embryology / Q000254:growth & development / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics; D011247:Pregnancy; D011994:Recombinant Proteins / Q000235:genetics; D013045:Species Specificity; D013687:Telencephalon / Q000166:cytology / Q000196:embryology* / Q000254:growth & development / Q000378:metabolism; D015027:Zebrafish
D051792:Basic Helix-Loop-Helix Proteins; D017931:DNA Primers; D049452:Green Fluorescent Proteins; D009419:Nerve Tissue Proteins; D011994:Recombinant Proteins; C102938:Neurog2 protein, mouse