miRNA
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The study of developmental biology has evolved along with technology, with novel techniques leading to new insights into genetic control of cell growth, differentiation and morphogenesis. What was, at one time, a clearly defined discipline, now defies traditional definitions, merging multiple research perspectives into a single picture. With the increased interest in stem cells for medical research, the subfield of embryology has become the stepping stone to understanding, diagnosing, and treating a host of human diseases. Understanding the molecular mechanisms that regulate cellular proliferation and differentiation is a central theme of modern developmental biology. Microarray technology has paved the way to gaining global perspectives on developmental biology, enabling high throughput analysis of a variety of sample types. The discovery that microRNAs play important roles in regulating gene expression via posttranscriptional repression has revealed a previously unsuspected mechanism controlling development and progenitor-cell function. Traditionally, miRNA expression has been tested using low-throughput techniques such as Northern-blot analysis and real-time PCR, but new developments in microarray technology now enable global profiling of all miRNA genes and their precursors in any sample type.
Jian-Fu Chen and researchers in North Carolina (2005) examined the role of microRNA-1 and microRNA-133 in skeletal muscle proliferation and differentiation through microarray expression analysis using C2C12 myoblasts. Myoblasts were selected based on their ability to mimic skeletal muscle differentiation in vitro, as shown by their induction to terminally differentiated myotubes when serum is withdrawn from the culture medium. The group found that the expression of a few of the miRNAs examined was upregulated in differentiated C2C12 myoblasts or myotubes. This increase in expression of miR-1 and miR-133 in differentiated myoblasts was confirmed by RNA blot analysis. Results demonstrated that miRNA-1 (miR-1) and miRNA-133 (miR-133), which are clustered on the same chromosomal loci, were transcribed together in a tissue-specific manner during development and both have distinct roles in modulating skeletal muscle proliferation and differentiation in vitro and in vivo. These data suggest that two mature miRNAs, derived from the same miRNA polycistron and transcribed together, can carry out distinct biological functions, indicating a molecular mechanism in which miRNAs participate in transcriptional circuits that control skeletal muscle gene expression and embryonic development.
Title: The role of microRNA-1 and microRNA-133 in skeletal muscle proliferation and differentiation.
Authors: Chen JF, Mandel EM, Thomson JM, Wu Q, Callis TE, Hammond SM, Conlon FL, Wang DZ
Journal: Nat Genet, Vol 38, Issue 2: 228-33
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Thomas Andl and colleagues (2006) sought to define the role of the miRNA-processing enzyme, Dicer, in morphorgenesis and maintenance of hair follicles in an attempt to elucidate one of the many aspects of miRNA function in mammalian organogenesis. The group used whole-mount and section in situ hybridization of mouse embryos to identify Dicer in developing hair follicles and microarray analyses of miRNA expression to determine whether miRNAs are expressed in developing mouse skin and hair follicles. To determine whether Dicer is required for development of the hair follicles or epidermis, the group generated an epidermal-specific deletion of the Dicer gene and assessed subsequent phenotypes. Additional experiments carried out by the researchers help substantiate the existence of a previously unsuspected mechanism for posttranscriptional regulation in the skin and pave the way for identification of specific miRNAs required for critical aspects of hair-follicle and epidermal morphogenesis and maintenance.
Title: The miRNA-processing enzyme dicer is essential for the morphogenesis and maintenance of hair follicles.
Authors: Andl T, Murchison EP, Liu F, Zhang Y, Yunta-Gonzalez M, Tobias JW, Andl CD, Seykora JT, Hannon GJ, Millar SE
Journal: Curr Biol, Vol 16, Issue 10: 1041-9
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Manami Amanai and researchers at the RIKEN Center for Developmental Biology in Kobe, Japan use microarray analysis to demonstrate that miRNAs are present in mouse sperm structures that enter the oocyte at fertilization and that there is a restricted role for sperm-borne microRNAs in mammalian fertilization. The researchers demonstrate that the sperm contained a broad profile of miRNAs and a subset of potential mRNA targets, which were expressed in fertilizable metaphase II (mII) oocytes. Interestingly, the levels of sperm-borne miRNA (measured by quantitative PCR) were low relative to those of unfertilized mII oocytes, and fertilization did not alter the mII oocyte miRNA repertoire that included the most abundant sperm-borne miRNAs. Coinjection of mII oocytes with sperm heads plus anti-miRNAs to suppress miRNA function did not perturb pronuclear activation or preimplantation development. In contrast, nuclear transfer by microinjection altered the miRNA profile of enucleated oocytes. These data suggest that sperm-borne prototypical miRNAs play a limited role, if any, in mammalian fertilization or early pre-implantation development.
Title: A restricted role for sperm-borne microRNAs in mammalian fertilization.
Authors: Amanai M, Brahmajosyula M, Perry AC
Journal: Biol Reprod, Vol 75, Issue 6: 877-84
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Title: Zebrafish MiR-430 promotes deadenylation and clearance of maternal mRNAs.
Authors: Giraldez AJ, Mishima Y, Rihel J, Grocock RJ, Van Dongen S, Inoue K, Enright AJ, Schier AF
Journal: Science, Vol 312, Issue 5770: 75-9
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Title: The diverse functions of microRNAs in animal development and disease.
Authors: Kloosterman WP, Plasterk RH
Journal: Dev Cell, Vol 11, Issue 4: 441-50
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Title: New developments in developmental biology.
Authors: Loebel DA, Lewis SL, Rao RS, Nolen LD
Journal: Genome Biol, Vol 6, Issue 13: 364
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Title: Developmental biology. Encountering microRNAs in cell fate signaling.
Authors: Karp X, Ambros V
Journal: Science, Vol 310, Issue 5752: 1288-9
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