Mapping of fluorescent protein-expressing neurons and axon pathways in adult and developing Thy1-eYFP-H transgenic mice

Porrero, César; Rubio-Garrido, Pablo; Avendaño, Carlos; Clascá, Francisco

doi:10.1016/j.brainres.2010.05.061

Cited by 167 publications

(159 citation statements)

References 54 publications

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“…This interpretation would be consistent with work in Drosophila showing that DSCAM controls branching and bifurcation of axons in the mushroom body (Wang et al, 2002) and also propose a novel role for DSCAM in pyramidal neuron dendrite development. Whereas Sema3a and Slit/Robo have been shown to promote dendrite outgrowth and branching (Polleux et al, 2000;Fenstermaker et al, 2004;Whitford et al, 2002), our results suggest that DSCAM may have a different function to limit the formation of ectopic apical branches. It is likely that DSCAM works in collaboration with other guidance molecules to orchestrate pyramidal dendrite arborization.…”

Section: Expression Of Dscam In the Cortex Is Developmentally Regulatedmentioning

confidence: 37%

“…In the cortex, Thy1-YFP-H mice specifically express YFP in a subset of layer V pyramidal neurons under control of the Thy1 promoter (Fig. 2I ) (Feng et al, 2000;Porrero et al, 2010). In preparation for FACS, cortical slices from adult transgenic mice were enzymatically dissociated, labeled with PI to mark dead neurons, and sorted to purify YFPϩ and PIϪ neurons, typically yielding 500,000 YFPϩ pyramidal neurons (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Semaphorin 3a (Sema3a) has been shown to orient pyramidal neuron apical dendrites and regulate the length and branching of pyramidal apical and basal dendrite arbors (Polleux et al, 2000, Fenstermaker et al, 2004. Additionally, Slit/Robo signaling has been implicated in dendrite remodeling by mediating an increase in cortical dendrite length and branching (Whitford et al, 2002).…”

Section: Expression Of Dscam In the Cortex Is Developmentally Regulatedmentioning

confidence: 99%

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DSCAM Contributes to Dendrite Arborization and Spine Formation in the Developing Cerebral Cortex

Maynard

Stein

2012

J. Neurosci.

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Down syndrome cell adhesion molecule, or DSCAM, has been implicated in many neurodevelopmental processes including axon guidance, dendrite arborization, and synapse formation. Here we show that DSCAM plays an important role in regulating the morphogenesis of cortical pyramidal neurons in the mouse. We report that DSCAM expression is developmentally regulated and localizes to synaptic plasma membranes during a time of robust cortical dendrite arborization and spine formation. Analysis of mice that carry a spontaneous mutation in DSCAM (DSCAM del17 ) revealed gross morphological changes in brain size and shape in addition to subtle changes in cortical organization, volume, and lamination. Early postnatal mutant mice displayed a transient decrease in cortical thickness, but these reductions could not be attributed to changes in neuron production or cell death. DSCAM del17 mutants showed temporary impairments in the branching of layer V pyramidal neuron dendrites at P10 and P17 that recovered to normal by adulthood. Defects in DSCAM del17 dendrite branching correlated with a temporal increase in apical branch spine density and lasting changes in spine morphology. At P15 and P42, mutant mice displayed a decrease in the percentage of large, stable spines and an increase in the percentage of small, immature spines. Together, our findings suggest that DSCAM contributes to pyramidal neuron morphogenesis by regulating dendrite arborization and spine formation during cortical circuit development.

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Section: Expression Of Dscam In the Cortex Is Developmentally Regulatedmentioning

confidence: 37%

Section: Resultsmentioning

confidence: 99%

Section: Expression Of Dscam In the Cortex Is Developmentally Regulatedmentioning

confidence: 99%

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DSCAM Contributes to Dendrite Arborization and Spine Formation in the Developing Cerebral Cortex

Maynard

Stein

2012

J. Neurosci.

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“…Neuron-specific expression in large parts of the brain is controlled by the regulatory elements of the murine Thy-1.2 gene (thymocyte differentiation antigen 1, CD90) (12,46).…”

Section: Introductionmentioning

confidence: 99%

Redox Indicator Mice Stably Expressing Genetically Encoded Neuronal roGFP: Versatile Tools to Decipher Subcellular Redox Dynamics in Neuropathophysiology

Wagener

Kolbrink

Dietrich

et al. 2016

Antioxidants & Redox Signaling

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Aims: Reactive oxygen species (ROS) and downstream redox alterations not only mediate physiological signaling but also neuropathology. For long, ROS/redox imaging was hampered by a lack of reliable probes. Genetically encoded redox sensors overcame this gap and revolutionized (sub)cellular redox imaging. Yet, the successful delivery of sensor-coding DNA, which demands transfection/transduction of cultured preparations or stereotaxic microinjections of each subject, remains challenging. By generating transgenic mice, we aimed to overcome limiting cultured preparations, circumvent surgical interventions, and to extend effectively redox imaging to complex and adult preparations. Results: Our redox indicator mice widely express Thy1-driven roGFP1 (reduction-oxidation-sensitive green fluorescent protein 1) in neuronal cytosol or mitochondria. Negative phenotypic effects of roGFP1 were excluded and its proper targeting and functionality confirmed. Redox mapping by ratiometric wide-field imaging reveals most oxidizing conditions in CA3 neurons. Furthermore, mitochondria are more oxidized than cytosol. Cytosolic and mitochondrial roGFP1s reliably report cell endogenous redox dynamics upon metabolic challenge or stimulation. Fluorescence lifetime imaging yields stable, but marginal, response ranges. We therefore developed automated excitation ratiometric 2-photon imaging. It offers superior sensitivity, spatial resolution, and response dynamics. Innovation and Conclusion: Redox indicator mice enable quantitative analyses of subcellular redox dynamics in a multitude of preparations and at all postnatal stages. This will uncover cell-and compartment-specific cerebral redox signals and their defined alterations during development, maturation, and aging. Cross-breeding with other disease models will reveal molecular details on compartmental redox homeostasis in neuropathology. Combined with ratiometric 2-photon imaging, this will foster our mechanistic understanding of cellular redox signals in their full complexity. Antioxid. Redox Signal. 25, 41-58.

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“…EGFP was the first fluorescent protein expressed in transgenic mice and remains the most widely used today (43). However, there is a trend toward using fluorescent proteins other than EGFP, e.g., enhanced yellow, blue, and cyan fluorescent protein (EYFP, EBFP, and ECFP) (27,46,59) or the monomeric red fluorescent proteins (mRFPs) (5,45). Excitation and emission frequencies of mRFP-derived fluorescent proteins are significantly higher than those of EGFPderived fluorescent proteins and pH-and Ca 2ϩ -sensitive dyes.…”

Section: Fluorescent Proteinsmentioning

confidence: 99%

Transgenic mice: beyond the knockout

Miller

2011

American Journal of Physiology-Renal Physiology

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mice have had a tremendous impact on biomedical research. Most researchers are familiar with transgenic mice that carry Cre recombinase (Cre) and how they are used to create conditional knockouts. However, some researchers are less familiar with many of the other types of transgenic mice and their applications. For example, transgenic mice can be used to study biochemical and molecular pathways in primary cultures and cell suspensions derived from transgenic mice, cell-cell interactions using multiple fluorescent proteins in the same mouse, and the cell cycle in real time and in the whole animal, and they can be used to perform deep tissue imaging in the whole animal, follow cell lineage during development and disease, and isolate large quantities of a pure cell type directly from organs. These novel transgenic mice and their applications provide the means for studying of molecular and biochemical events in the whole animal that was previously limited to cell cultures. In conclusion, transgenic mice are not just for generating knockouts.Cre; enhanced green fluorescent protein; high-throughput; renal-specific; transgenics TRANSGENIC MICE HAVE REVOLUTIONIZED biomedical research. They permit the manipulation of genetic, molecular, biochemical, and physiological processes in the whole animal with nearly the same specificity seen in cell culture models. Most researchers are familiar with the Cre/loxP system and how it is used to create conditional knockouts. However, most are unfamiliar with the many other transgenic mice and their applications, novel and well-established alike. These novel mice go beyond the knockout and provide the means to study of cellular processes in the whole animal that was traditionally limited to culture models. This review highlights a few of these transgenic mice and their applications.

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Mapping of fluorescent protein-expressing neurons and axon pathways in adult and developing Thy1-eYFP-H transgenic mice

Cited by 167 publications

References 54 publications

DSCAM Contributes to Dendrite Arborization and Spine Formation in the Developing Cerebral Cortex

DSCAM Contributes to Dendrite Arborization and Spine Formation in the Developing Cerebral Cortex

Redox Indicator Mice Stably Expressing Genetically Encoded Neuronal roGFP: Versatile Tools to Decipher Subcellular Redox Dynamics in Neuropathophysiology

Transgenic mice: beyond the knockout

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