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Genetic labeling of neuronal subsets through enhancer trapping in mice.

Kelsch W, Stolfi A, Lois C - PLoS ONE (2012)

Bottom Line: The ability to label, visualize, and manipulate subsets of neurons is critical for elucidating the structure and function of individual cell types in the brain.We have developed an enhancer trap strategy in mammals by generating transgenic mice with lentiviral vectors carrying single-copy enhancer-detector probes encoding either the marker gene lacZ or Cre recombinase.This transgenic strategy allowed us to genetically identify a wide variety of neuronal subpopulations in distinct brain regions.

View Article: PubMed Central - PubMed

Affiliation: Bernstein Center for Computational Neuroscience, CIMH, Medical Faculty Mannheim, University Heidelberg, Heidelberg, Germany.

ABSTRACT
The ability to label, visualize, and manipulate subsets of neurons is critical for elucidating the structure and function of individual cell types in the brain. Enhancer trapping has proved extremely useful for the genetic manipulation of selective cell types in Drosophila. We have developed an enhancer trap strategy in mammals by generating transgenic mice with lentiviral vectors carrying single-copy enhancer-detector probes encoding either the marker gene lacZ or Cre recombinase. This transgenic strategy allowed us to genetically identify a wide variety of neuronal subpopulations in distinct brain regions. Enhancer detection by lentiviral transgenesis could thus provide a complementary method for generating transgenic mouse libraries for the genetic labeling and manipulation of neuronal subsets.

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Labeling of subsets of neurons in thy1mp-cre lines.(A) Sagittal sections of mice carrying a single copy of the thy1mp-cre transgene (lines named FTC.01-FTC.13). A large variety of subsets of neurons were labeled in eight independent mouse lines shown here (bar = 2.5mm). (B) Labeling of olfactory receptor neurons (FTC.07) (bar = 250 um). (C) Labeling of granule cell neurons in the accessory olfactory bulb (FTC.13) (bar = 500um). (D) Sagittal sections of the dorsal hippocampal formation of eight different thy1mp-cre lines reveal recombination restricted to subregions of the hippocampus (CA1, CA3 or dentate gyrus) (bar = 500 um).
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pone-0038593-g004: Labeling of subsets of neurons in thy1mp-cre lines.(A) Sagittal sections of mice carrying a single copy of the thy1mp-cre transgene (lines named FTC.01-FTC.13). A large variety of subsets of neurons were labeled in eight independent mouse lines shown here (bar = 2.5mm). (B) Labeling of olfactory receptor neurons (FTC.07) (bar = 250 um). (C) Labeling of granule cell neurons in the accessory olfactory bulb (FTC.13) (bar = 500um). (D) Sagittal sections of the dorsal hippocampal formation of eight different thy1mp-cre lines reveal recombination restricted to subregions of the hippocampus (CA1, CA3 or dentate gyrus) (bar = 500 um).

Mentions: In order to bias the specificity of Cre expression to neurons, and to reduce the appearance of ubiquitous patterns of recombination, we generated a lentiviral enhancer detection probe carrying cre under the control of the minimal promoter of the thy-1.2 gene, consisting of 310 base pairs upstream from its transcription start site (thy1mp-cre) [16]. The thy-1.2 gene is preferentially expressed in projection neurons in the mouse brain, and its minimal promoter may contain elements that could restrict its activity to neurons [17], [18]. Founder transgenic animals carrying 1–4 insertions of thy1mp-cre were bred to the Cre-dependent GFP reporter lines (Z/EG) to obtain lines with single copy insertions of the enhancer detection probe. We observed a large diversity of restricted recombination patterns in 16 out of 20 lines (Figure 4A). The pattern of recombination was reproducible among animals with the same single insertion of the enhancer probe (Figure S1). In the hippocampus, for example, we observed that recombination patterns were frequently restricted to specific substructures (CA1, CA2 or dentate gyrus, Figure 4D). We also observed transgenic lines that labeled specific neuronal types, such as of olfactory receptor neurons (FTC.07, Figure 4B) and granule cells of the accessory olfactory bulb (FTC.13, Figure 4C). In certain cortical areas, we observed that recombination patterns were restricted to specific layers (Figure 4A).


Genetic labeling of neuronal subsets through enhancer trapping in mice.

Kelsch W, Stolfi A, Lois C - PLoS ONE (2012)

Labeling of subsets of neurons in thy1mp-cre lines.(A) Sagittal sections of mice carrying a single copy of the thy1mp-cre transgene (lines named FTC.01-FTC.13). A large variety of subsets of neurons were labeled in eight independent mouse lines shown here (bar = 2.5mm). (B) Labeling of olfactory receptor neurons (FTC.07) (bar = 250 um). (C) Labeling of granule cell neurons in the accessory olfactory bulb (FTC.13) (bar = 500um). (D) Sagittal sections of the dorsal hippocampal formation of eight different thy1mp-cre lines reveal recombination restricted to subregions of the hippocampus (CA1, CA3 or dentate gyrus) (bar = 500 um).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3369840&req=5

pone-0038593-g004: Labeling of subsets of neurons in thy1mp-cre lines.(A) Sagittal sections of mice carrying a single copy of the thy1mp-cre transgene (lines named FTC.01-FTC.13). A large variety of subsets of neurons were labeled in eight independent mouse lines shown here (bar = 2.5mm). (B) Labeling of olfactory receptor neurons (FTC.07) (bar = 250 um). (C) Labeling of granule cell neurons in the accessory olfactory bulb (FTC.13) (bar = 500um). (D) Sagittal sections of the dorsal hippocampal formation of eight different thy1mp-cre lines reveal recombination restricted to subregions of the hippocampus (CA1, CA3 or dentate gyrus) (bar = 500 um).
Mentions: In order to bias the specificity of Cre expression to neurons, and to reduce the appearance of ubiquitous patterns of recombination, we generated a lentiviral enhancer detection probe carrying cre under the control of the minimal promoter of the thy-1.2 gene, consisting of 310 base pairs upstream from its transcription start site (thy1mp-cre) [16]. The thy-1.2 gene is preferentially expressed in projection neurons in the mouse brain, and its minimal promoter may contain elements that could restrict its activity to neurons [17], [18]. Founder transgenic animals carrying 1–4 insertions of thy1mp-cre were bred to the Cre-dependent GFP reporter lines (Z/EG) to obtain lines with single copy insertions of the enhancer detection probe. We observed a large diversity of restricted recombination patterns in 16 out of 20 lines (Figure 4A). The pattern of recombination was reproducible among animals with the same single insertion of the enhancer probe (Figure S1). In the hippocampus, for example, we observed that recombination patterns were frequently restricted to specific substructures (CA1, CA2 or dentate gyrus, Figure 4D). We also observed transgenic lines that labeled specific neuronal types, such as of olfactory receptor neurons (FTC.07, Figure 4B) and granule cells of the accessory olfactory bulb (FTC.13, Figure 4C). In certain cortical areas, we observed that recombination patterns were restricted to specific layers (Figure 4A).

Bottom Line: The ability to label, visualize, and manipulate subsets of neurons is critical for elucidating the structure and function of individual cell types in the brain.We have developed an enhancer trap strategy in mammals by generating transgenic mice with lentiviral vectors carrying single-copy enhancer-detector probes encoding either the marker gene lacZ or Cre recombinase.This transgenic strategy allowed us to genetically identify a wide variety of neuronal subpopulations in distinct brain regions.

View Article: PubMed Central - PubMed

Affiliation: Bernstein Center for Computational Neuroscience, CIMH, Medical Faculty Mannheim, University Heidelberg, Heidelberg, Germany.

ABSTRACT
The ability to label, visualize, and manipulate subsets of neurons is critical for elucidating the structure and function of individual cell types in the brain. Enhancer trapping has proved extremely useful for the genetic manipulation of selective cell types in Drosophila. We have developed an enhancer trap strategy in mammals by generating transgenic mice with lentiviral vectors carrying single-copy enhancer-detector probes encoding either the marker gene lacZ or Cre recombinase. This transgenic strategy allowed us to genetically identify a wide variety of neuronal subpopulations in distinct brain regions. Enhancer detection by lentiviral transgenesis could thus provide a complementary method for generating transgenic mouse libraries for the genetic labeling and manipulation of neuronal subsets.

Show MeSH
Related in: MedlinePlus