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. 2008 Jun;46(6):289-93.
doi: 10.1002/dvg.20393.

Cre recombinase-mediated gene deletion in layer 4 of murine sensory cortical areas

Affiliations

Cre recombinase-mediated gene deletion in layer 4 of murine sensory cortical areas

Guey-Ying Liao et al. Genesis. 2008 Jun.

Abstract

Thalamocortical input to layer 4 carries the major ascending sensory information to the mammalian sensory cortex and is crucial for the function and plasticity of sensory cortical areas. Here we report identification of a Six3-cre transgene that is selectively expressed in layer 4 of sensory cortical areas but not in the thalamus. In the mature somatosensory cortex Cre recombinase expressed from the transgene is able to mediate gene deletion in the overwhelming majority of layer 4 neurons, including GABAergic interneurons. The gene deletion in layer 4 mainly occurs during the first postnatal week. This cre transgene therefore provides a useful tool for examining the role of proteins expressed in layer 4 neurons.

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Figures

Figure 1
Figure 1
Pattern of ROSA26 recombination mediated by Six3-cre#69. Coronal brain sections from adult R26R/+;Six3-cre#69 or R26R/+ mice were stained for β-galactosidase activity (blue staining) and then counterstained with nuclear fast red. Brain images of R26R/+;Six3-cre#69 mice in a–k are arranged in a rostral-caudal order. No blue staining was detected in brain sections from R26R/+ mice (l). Abbreviations: Amy, amygdala; Au, auditory cortex; BS, brainstem; BST, bed nucleus of the stria terminalis; Cbl, cerebellum; DMH, dorsomedial hypothalamus; Hp, hippocampus; M1, primary motor cortex; MPO, medial preoptic nucleus; PAG, periaqueductal gray; Pir, piriform cortex; S1, primary somatosensory cortex; S1BF, S1 cortex barrel field; S1L, S1 cortex limb region; S2, secondary somatosensory cortex; SN, substantia nigra; Stm, striatum; Th, thalamus; V, visual cortex; VMH, ventromedial hypothalamus.
Figure 2
Figure 2
Restriction of the Cre activity from the Six3-cre#69 transgene to layer 4 of the somatosensory cortex. (a) X-gal staining (blue) and NeuN immunohistochemistry (brown) indicate that β-galactosidase activity is restricted to layer 4 of the S1 cortex barrel field. (b) The majority of NeuN-positive cells in layer 4 of the S1 cortex barrel field contain β-galactosidase activity (blue spots). Scale bar, 25 μm. (c–e) A series of confocal microscopy images at a 2-μm interval shows colocalization of NeuN (red) and β-galactosidase (green dots) in layer 4 neurons. Arrows denotes neurons that appear lacking β-galactosidase particles in (d) but do express β-galactosidase, as shown in (c) or (e). Scale bar, 50 μm.
Figure 3
Figure 3
Colocalization of Cre activity with markers for GABAergic interneurons in layer 4 of the S1 cortex barrel field. The activity of β-galactosidase resulting from Cre-mediated recombination of the R26R locus was detectable in layer 4 interneurons expressing somatostatin (a), parvalbumin (b), calretinin (c), and NPY (d). Arrows denote representative neurons expressing β-galactosidase and an interneuron marker. Scale bar, 25 μm.
Figure 4
Figure 4
Time course of Cre-mediated recombination in layer 4 of the sensory cortex. X-gal staining shows the activity of Cre recombinase in layer 4 of the S1 cortex barrel field in R26R/+;Six3-cre#69 mice at postnatal day 2 (P2), 7 (P7), 14 (P14), and 21 (P21). The “4” in panel (a) marks the approximate position of layer 4. Scale bar, 50 μm.

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