Biochemical and Biophysical Research Communications
Molecular hierarchy in neurons differentiated from mouse ES cells containing a single human chromosome 21
Section snippets
Materials and methods
ES cell culture and construction of ES cell lines containing hChr21. Parental TT2F cells and microcell hybrid clones TT2F/hChr21 were used and maintained on mitomycin C (Sigma) treated G418 resistant mouse embryonic fibroblasts (Gibco) as feeder layers in DMEM (Sigma) supplemented with 15% FBS (Hyclone), 1 mM sodium pyruvate (Gibco), 0.1 mM non-essential amino acids (Gibco), 0.1 mM 2-mercaptoethanol (Sigma), 2 mM l-glutamine (Gibco), and 1000 U/ml LIF (Gibco), with or without 300 μg/ml G418 (Gibco)
Neuron differentiation of TT2F and TT2F/hChr21 clones
ES cells plated under serum free conditions on PA6 mouse stromal cell feeder layer differentiate along the default ectodermal pathway and then along the neuronal pathway as directed by SDIA of PA6. Neuron cells induced to differentiate from ES cells by SDIA were reported to contain naı̈ve neural precursors that respond to patterning signals, directing a full dorsal–ventral range of neuroectodermal derivatives [18]. In our experiment, two independently established TT2F/hChr21 clones 21–10 and
Discussion
Mental retardation, as the most common genetic cause of developmental disability in DS, represents a condition characterized by subnormal intellectual functioning and impaired adaptive behavior that become manifest during developmental years. The relationship between mental retardation and the trisomic condition of DS is complex. Conspicuous morphological abnormalities start to be apparent in brains of newborns and older infants with DS. They have shortened basilar dendrites, a decreased number
Conclusions
Defining how an extra copy of hChr21 results in the phenotype of DS is a special case of the more general problem of explaining how chromosomal imbalance produces abnormalities in morphology and function. No single mechanism can explain the deleterious consequences of aneuploidy. In this study, we found the complex network of molecular changes in neuronal stem cells with extra hChr21 for the defective neuron differentiation mechanism by locus-specific disturbance of chromosome balance and
Acknowledgements
We are grateful to Dr. Shin-ichi Hayashi for PA6 cells. This work was supported in part by the grants from the Ministry of Education, Culture, Sports, Science and Technology of Japan; JSPS Postdoctoral Fellowship for Foreign Researcher P01104; JSPS Grant-in-aid for Scientific Research FY2000; and CUHK Direct Grant for Research 2002.2.024.
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Cited by (25)
Down Syndrome
2013, Neural Circuit Development and Function in the Healthy and Diseased Brain: Comprehensive Developmental NeuroscienceDown Syndrome
2013, Neural Circuit Development and Function in the Heathy and Diseased BrainGene dosage imbalance of human chromosome 21 in mouse embryonic stem cells differentiating to neurons
2011, GeneCitation Excerpt :The differentiating neurons were characterised by dopamine determination, neural specific immunostaining and quantitative RT-PCR analyses. The global gene expression profiling by microarray on the differentiating neurons were previously performed as described (Wang et al., 2004; Kai et al., 2009). Briefly, cultivated ES cells after days 0, 2, 3, 4, and 6 of neuronal differentiation were washed five times in PBS, dissociated using cell dissociation solution (Sigma, St. Louis, MO), and passed through a cell strainer (BD Biosciences, Sparks, MD) to remove any cell clumps.
Mental retardation and associated neurological dysfunctions in Down syndrome: A consequence of dysregulation in critical chromosome 21 genes and associated molecular pathways
2008, European Journal of Paediatric NeurologyCitation Excerpt :On the other hand, quantitative analyses of normal and trisomic tissues allowed identification of transcriptional alteration in DS. Several whole-genome approaches or transcriptome studies have been performed by microarrays or Serial Analysis of Genome Expression (SAGE), to investigate gene expression variations in trisomic tissues compared to the euploid ones using human tissues or cell lines, and also mouse trisomic model tissues and their non-trisomic littermates.91–99 These transcriptome studies have been particularly focused on brain, cerebellum or neuronal cell lines, considering their major interest in the understanding of the mechanisms involved in the MR pathogenesis in DS.91,93–96,99,100
Mental retardation in Down syndrome: From gene dosage imbalance to molecular and cellular mechanisms
2007, Neuroscience Research
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These authors contributed equally to this work.