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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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Variability of functional properties in neuronal subsets in thy1mp-cre lines.(A)-(D) Sagittal sections of thy1mp-cre lines (FTC.13 ((A), (B)) and FTC.08 ((C), (D)) reveal changes in the layer-specific expression in the frontal versus occipital cortical areas. In addition, in the occipital cortex the layer-specific expression changed from medial ((A), (C) corresponding to retrosplenial cortex) to lateral ((B), (D) corresponding to visual cortex) (bar = 1 mm). (E) Whole-cell recordings from visual cortex layer 2/3 of thy1mp-cre line FTC.08 were performed from GFP positive and GFP negative neurons in the same animals (n = 7 and 11, respectively). The relationship between the amount of injected current and the frequency of induced action potential was plotted for each recorded cell. GFP positive layer 2/3 neurons from FTC.08 had a smaller variability in their frequency-current relationship than GFP negative layer 2/3 pyramidal neurons in the same animals. (F) shows the adaptation index for GFP positive and GFP negative control layer 2/3 pyramidal neurons from line FTC.08 (n = 7, 11, respectively). The adaptation index indicates the ratio of the second interspike interval (ISI) over the last ISI of a series of pulses (200 ms).
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pone-0038593-g005: Variability of functional properties in neuronal subsets in thy1mp-cre lines.(A)-(D) Sagittal sections of thy1mp-cre lines (FTC.13 ((A), (B)) and FTC.08 ((C), (D)) reveal changes in the layer-specific expression in the frontal versus occipital cortical areas. In addition, in the occipital cortex the layer-specific expression changed from medial ((A), (C) corresponding to retrosplenial cortex) to lateral ((B), (D) corresponding to visual cortex) (bar = 1 mm). (E) Whole-cell recordings from visual cortex layer 2/3 of thy1mp-cre line FTC.08 were performed from GFP positive and GFP negative neurons in the same animals (n = 7 and 11, respectively). The relationship between the amount of injected current and the frequency of induced action potential was plotted for each recorded cell. GFP positive layer 2/3 neurons from FTC.08 had a smaller variability in their frequency-current relationship than GFP negative layer 2/3 pyramidal neurons in the same animals. (F) shows the adaptation index for GFP positive and GFP negative control layer 2/3 pyramidal neurons from line FTC.08 (n = 7, 11, respectively). The adaptation index indicates the ratio of the second interspike interval (ISI) over the last ISI of a series of pulses (200 ms).

Mentions: In several independent transgenic lines, cells with layer-specific recombination in the cortex had the morphology of pyramidal neurons. To examine whether these layer-specific transgenic lines contained subsets of neurons defined by specific electrophysiological properties, we selected the transgenic line FTC.08 for fluorescence-guided whole-cell recordings that had GFP+ neurons in layer 2/3 in the visual cortex (V1 area, Figure 5A-D). To determine the homogeneity of the electrophysiological properties of the GFP+ cells, we compared them to GFP− control neurons in layer 2/3 in the same animals. We observed that in this transgenic line the variability in some of the electrical properties of GFP+ cells, such as frequency–current relationship or adaptation index, was smaller in GFP+ than in GFP− neurons of the same layer (Figure 5E, F, see also Figure S2 for additional data). These observations underscore the usefulness of the enhancer trap approach to identify neuronal populations with defined properties.


Genetic labeling of neuronal subsets through enhancer trapping in mice.

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

Variability of functional properties in neuronal subsets in thy1mp-cre lines.(A)-(D) Sagittal sections of thy1mp-cre lines (FTC.13 ((A), (B)) and FTC.08 ((C), (D)) reveal changes in the layer-specific expression in the frontal versus occipital cortical areas. In addition, in the occipital cortex the layer-specific expression changed from medial ((A), (C) corresponding to retrosplenial cortex) to lateral ((B), (D) corresponding to visual cortex) (bar = 1 mm). (E) Whole-cell recordings from visual cortex layer 2/3 of thy1mp-cre line FTC.08 were performed from GFP positive and GFP negative neurons in the same animals (n = 7 and 11, respectively). The relationship between the amount of injected current and the frequency of induced action potential was plotted for each recorded cell. GFP positive layer 2/3 neurons from FTC.08 had a smaller variability in their frequency-current relationship than GFP negative layer 2/3 pyramidal neurons in the same animals. (F) shows the adaptation index for GFP positive and GFP negative control layer 2/3 pyramidal neurons from line FTC.08 (n = 7, 11, respectively). The adaptation index indicates the ratio of the second interspike interval (ISI) over the last ISI of a series of pulses (200 ms).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3369840&req=5

pone-0038593-g005: Variability of functional properties in neuronal subsets in thy1mp-cre lines.(A)-(D) Sagittal sections of thy1mp-cre lines (FTC.13 ((A), (B)) and FTC.08 ((C), (D)) reveal changes in the layer-specific expression in the frontal versus occipital cortical areas. In addition, in the occipital cortex the layer-specific expression changed from medial ((A), (C) corresponding to retrosplenial cortex) to lateral ((B), (D) corresponding to visual cortex) (bar = 1 mm). (E) Whole-cell recordings from visual cortex layer 2/3 of thy1mp-cre line FTC.08 were performed from GFP positive and GFP negative neurons in the same animals (n = 7 and 11, respectively). The relationship between the amount of injected current and the frequency of induced action potential was plotted for each recorded cell. GFP positive layer 2/3 neurons from FTC.08 had a smaller variability in their frequency-current relationship than GFP negative layer 2/3 pyramidal neurons in the same animals. (F) shows the adaptation index for GFP positive and GFP negative control layer 2/3 pyramidal neurons from line FTC.08 (n = 7, 11, respectively). The adaptation index indicates the ratio of the second interspike interval (ISI) over the last ISI of a series of pulses (200 ms).
Mentions: In several independent transgenic lines, cells with layer-specific recombination in the cortex had the morphology of pyramidal neurons. To examine whether these layer-specific transgenic lines contained subsets of neurons defined by specific electrophysiological properties, we selected the transgenic line FTC.08 for fluorescence-guided whole-cell recordings that had GFP+ neurons in layer 2/3 in the visual cortex (V1 area, Figure 5A-D). To determine the homogeneity of the electrophysiological properties of the GFP+ cells, we compared them to GFP− control neurons in layer 2/3 in the same animals. We observed that in this transgenic line the variability in some of the electrical properties of GFP+ cells, such as frequency–current relationship or adaptation index, was smaller in GFP+ than in GFP− neurons of the same layer (Figure 5E, F, see also Figure S2 for additional data). These observations underscore the usefulness of the enhancer trap approach to identify neuronal populations with defined properties.

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