scholarly journals Lineage Tracing and Clonal Analysis in Developing Cerebral Cortex Using Mosaic Analysis with Double Markers (MADM)

10.3791/61147 ◽  
2020 ◽  
Author(s):  
Robert Beattie ◽  
Carmen Streicher ◽  
Nicole Amberg ◽  
Giselle Cheung ◽  
Ximena Contreras ◽  
...  
Keyword(s):  
Cerebral Cortex ◽  
Clonal Analysis ◽  
Lineage Tracing ◽  
Mosaic Analysis

scholarly journals Clonal Analysis of Gliogenesis in the Cerebral Cortex Reveals Stochastic Expansion of Glia and Cell Autonomous Responses to Egfr Dosage

Cells ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 2662
Author(s):  
Xuying Zhang ◽  
Christine V. Mennicke ◽  
Guanxi Xiao ◽  
Robert Beattie ◽  
Mansoor A. Haider ◽  
...  
Keyword(s):  
Nervous System ◽  
Epidermal Growth Factor Receptor ◽  
Cerebral Cortex ◽  
Biological Significance ◽  
Growth Factor Receptor ◽  
Clonal Analysis ◽  
Proliferation Capacity ◽  
Mosaic Analysis ◽  
Epidermal Growth ◽  
Genetic Deletion

Development of the nervous system undergoes important transitions, including one from neurogenesis to gliogenesis which occurs late during embryonic gestation. Here we report on clonal analysis of gliogenesis in mice using Mosaic Analysis with Double Markers (MADM) with quantitative and computational methods. Results reveal that developmental gliogenesis in the cerebral cortex occurs in a fraction of earlier neurogenic clones, accelerating around E16.5, and giving rise to both astrocytes and oligodendrocytes. Moreover, MADM-based genetic deletion of the epidermal growth factor receptor (Egfr) in gliogenic clones revealed that Egfr is cell autonomously required for gliogenesis in the mouse dorsolateral cortices. A broad range in the proliferation capacity, symmetry of clones, and competitive advantage of MADM cells was evident in clones that contained one cellular lineage with double dosage of Egfr relative to their environment, while their sibling Egfr-null cells failed to generate glia. Remarkably, the total numbers of glia in MADM clones balance out regardless of significant alterations in clonal symmetries. The variability in glial clones shows stochastic patterns that we define mathematically, which are different from the deterministic patterns in neuronal clones. This study sets a foundation for studying the biological significance of stochastic and deterministic clonal principles underlying tissue development, and identifying mechanisms that differentiate between neurogenesis and gliogenesis.

scholarly journals Persistence of RNAi-Mediated Knockdown inDrosophilaComplicates Mosaic Analysis Yet Enables Highly Sensitive Lineage Tracing

Genetics ◽  
2016 ◽  
Vol 203 (1) ◽  
pp. 109-118 ◽  
Author(s):  
Justin A. Bosch ◽  
Taryn M. Sumabat ◽  
Iswar K. Hariharan
Keyword(s):  
Lineage Tracing ◽  
Highly Sensitive ◽  
Mosaic Analysis

scholarly journals Defining stem cell dynamics and migration during wound healing in mouse skin epidermis

2017 ◽  
Vol 8 (1) ◽  
Author(s):  
Mariaceleste Aragona ◽  
Sophie Dekoninck ◽  
Steffen Rulands ◽  
Sandrine Lenglez ◽  
Guilhem Mascré ◽  
...  
Keyword(s):  
Wound Healing ◽  
Molecular Mechanisms ◽  
Chronic Wound ◽  
Mouse Skin ◽  
Molecular Profiling ◽  
Clonal Analysis ◽  
Lineage Tracing ◽  
Cell Dynamics ◽  
And Migration ◽  
Skin Epidermis

Abstract Wound healing is essential to repair the skin after injury. In the epidermis, distinct stem cells (SCs) populations contribute to wound healing. However, how SCs balance proliferation, differentiation and migration to repair a wound remains poorly understood. Here, we show the cellular and molecular mechanisms that regulate wound healing in mouse tail epidermis. Using a combination of proliferation kinetics experiments and molecular profiling, we identify the gene signatures associated with proliferation, differentiation and migration in different regions surrounding the wound. Functional experiments show that SC proliferation, migration and differentiation can be uncoupled during wound healing. Lineage tracing and quantitative clonal analysis reveal that, following wounding, progenitors divide more rapidly, but conserve their homoeostatic mode of division, leading to their rapid depletion, whereas SCs become active, giving rise to new progenitors that expand and repair the wound. These results have important implications for tissue regeneration, acute and chronic wound disorders.

scholarly journals Variation of human neural stem cells generating organizer states in vitro before committing to cortical excitatory or inhibitory neuronal fates

2019 ◽  
Author(s):  
Nicola Micali ◽  
Suel-Kee Kim ◽  
Marcelo Diaz-Bustamante ◽  
Genevieve Stein-O’Brien ◽  
Seungmae Seo ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cerebral Cortex ◽  
Neural Stem Cells ◽  
Cell Imaging ◽  
Lineage Tracing ◽  
Systematic Analysis ◽  
Human Neural Stem Cells ◽  
Signaling Mechanisms ◽  
Human Telencephalon

SUMMARYBetter understanding the progression of neural stem cells (NSCs) in the developing cerebral cortex is important for modeling neurogenesis and defining the pathogenesis of neuropsychiatric disorders. Here we used RNA-sequencing, cell imaging and lineage tracing of mouse and human in vitro NSCs to model the generation of cortical neuronal fates. We show that conserved signaling mechanisms regulate the acute transition from proliferative NSCs to committed glutamatergic excitatory neurons. As human telencephalic NSCs developed from pluripotency in vitro, they first transitioned through organizer states that spatially pattern the cortex before generating glutamatergic precursor fates. NSCs derived from multiple human pluripotent lines varied in these early patterning states leading differentially to dorsal or ventral telencephalic fates. This work furthers systematic analysis of the earliest patterning events that generate the major neuronal trajectories of the human telencephalon.

scholarly journals Characterization and Stage-Dependent Lineage Analysis of Intermediate Progenitors of Cortical GABAergic Interneurons

2021 ◽  
Vol 15 ◽  
Author(s):  
Shigeyuki Esumi ◽  
Makoto Nasu ◽  
Takeshi Kawauchi ◽  
Koichiro Miike ◽  
Kento Morooka ◽  
...  
Keyword(s):  
Cerebral Cortex ◽  
Lineage Tracing ◽  
Adult Brain ◽  
Inhibitory Neurons ◽  
Gabaergic Interneurons ◽  
Ganglionic Eminence ◽  
Cortical Layers ◽  
Intermediate Progenitors ◽  
Mouse Cerebral Cortex ◽  
Lineage Analysis

Intermediate progenitors of both excitatory and inhibitory neurons, which can replenish neurons in the adult brain, were recently identified. However, the generation of intermediate progenitors of GABAergic inhibitory neurons (IPGNs) has not been studied in detail. Here, we characterized the spatiotemporal distribution of IPGNs in mouse cerebral cortex. IPGNs generated neurons during both embryonic and postnatal stages, but the embryonic IPGNs were more proliferative. Our lineage tracing analyses showed that the embryonically proliferating IPGNs tended to localize to the superficial layers rather than the deep cortical layers at 3 weeks after birth. We also found that embryonic IPGNs derived from the medial and caudal ganglionic eminence (CGE) but more than half of the embryonic IPGNs were derived from the CGE and broadly distributed in the cerebral cortex. Taken together, our data indicate that the broadly located IPGNs during embryonic and postnatal stages exhibit a different proliferative property and layer distribution.

scholarly journals 13-P019 A new tool for lineage tracing and mosaic analysis in zebrafish

2009 ◽  
Vol 126 ◽  
pp. S200
Author(s):  
Claudia Linker ◽  
Russell Collins ◽  
Julian Lewis
Keyword(s):  
Lineage Tracing ◽  
Mosaic Analysis

scholarly journals Tamoxifen-independent recombination of reporter genes limits lineage tracing and mosaic analysis using CreERT2 lines

2019 ◽  
Vol 29 (1) ◽  
pp. 53-68 ◽  
Author(s):  
A. Álvarez-Aznar ◽  
I. Martínez-Corral ◽  
N. Daubel ◽  
C. Betsholtz ◽  
T. Mäkinen ◽  
...  
Keyword(s):  
Cell Population ◽  
Cell Tracking ◽  
Reporter Genes ◽  
Cell Types ◽  
Lineage Tracing ◽  
Cell Labeling ◽  
Activity Levels ◽  
Fluorescent Reporter ◽  
Mosaic Analysis ◽  
Mouse Lines

Abstract The CreERT2/loxP system is widely used to induce conditional gene deletion in mice. One of the main advantages of the system is that Cre-mediated recombination can be controlled in time through Tamoxifen administration. This has allowed researchers to study the function of embryonic lethal genes at later developmental timepoints. In addition, CreERT2 mouse lines are commonly used in combination with reporter genes for lineage tracing and mosaic analysis. In order for these experiments to be reliable, it is crucial that the cell labeling approach only marks the desired cell population and their progeny, as unfaithful expression of reporter genes in other cell types or even unintended labeling of the correct cell population at an undesired time point could lead to wrong conclusions. Here we report that all CreERT2 mouse lines that we have studied exhibit a certain degree of Tamoxifen-independent, basal, Cre activity. Using Ai14 and Ai3, two commonly used fluorescent reporter genes, we show that those basal Cre activity levels are sufficient to label a significant amount of cells in a variety of tissues during embryogenesis, postnatal development and adulthood. This unintended labelling of cells imposes a serious problem for lineage tracing and mosaic analysis experiments. Importantly, however, we find that reporter constructs differ greatly in their susceptibility to basal CreERT2 activity. While Ai14 and Ai3 easily recombine under basal CreERT2 activity levels, mTmG and R26R-EYFP rarely become activated under these conditions and are therefore better suited for cell tracking experiments.

scholarly journals A mosaic analysis system with Cre or Tomato expression in the mouse

2020 ◽  
Vol 117 (45) ◽  
pp. 28212-28220 ◽  
Author(s):  
Qun Wang ◽  
Yen-Yu Lin ◽  
Baojun Zhang ◽  
Jianxuan Wu ◽  
Sumedha Roy ◽  
...  
Keyword(s):  
Somatic Mutations ◽  
Reference Group ◽  
Subsequent Development ◽  
Lineage Tracing ◽  
Clonal Hematopoiesis ◽  
Age Related ◽  
Mosaic Analysis ◽  
Analysis System ◽  
Mutant Cells

Somatic mutations are major genetic contributors to cancers and many other age-related diseases. Many disease-causing somatic mutations can initiate clonal growth prior to the appearance of any disease symptoms, yet experimental models that can be used to examine clonal abnormalities are limited. We describe a mosaic analysis system with Cre or Tomato (MASCOT) for tracking mutant cells and demonstrate its utility for modeling clonal hematopoiesis. MASCOT can be induced to constitutively express either Cre-GFP or Tomato for lineage tracing of a mutant and a reference group of cells simultaneously. We conducted mosaic analysis to assess functions of theId3and/orTet2gene in hematopoietic cell development and clonal hematopoiesis. Using Tomato-positive cells as a reference population, we demonstrated the high sensitivity of this system for detecting cell-intrinsic phenotypes during short-term or long-term tracking of hematopoietic cells. Long-term tracking ofTet2mutant orTet2/Id3double-mutant cells in our MASCOT model revealed a dynamic shift from myeloid expansion to lymphoid expansion and subsequent development of lymphoma. This work demonstrates the utility of the MASCOT method in mosaic analysis of single or combined mutations, making the system suitable for modeling somatic mutations identified in humans.

scholarly journals Massively parallel clonal analysis using CRISPR/Cas9 induced genetic scars

2016 ◽  
Author(s):  
Jan Philipp Junker ◽  
Bastiaan Spanjaard ◽  
Josi Peterson-Maduro ◽  
Anna Alemany ◽  
Bo Hu ◽  
...  
Keyword(s):  
Developmental Biology ◽  
Single Cell ◽  
Clonal Analysis ◽  
Lineage Tracing ◽  
Massively Parallel ◽  
Vertebrate Lineage ◽  
Lineage Trees ◽  
Made In

A key goal of developmental biology is to understand how a single cell transforms into a full-grown organism consisting of many cells. Although impressive progress has been made in lineage tracing using imaging approaches, analysis of vertebrate lineage trees has mostly been limited to relatively small subsets of cells. Here we present scartrace, a strategy for massively parallel clonal analysis based on Cas9 induced genetic scars in the zebrafish.

Sign in / Sign up

Export Citation Format

Share Document