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For the TSA treatment experiments, GV-stage oocytes were cultured in MEM-alpha media supplemented with 5% FBS and TSA (100nM) for ~?18 hours S3I-201 cost at 37?��C in an atmosphere of 5% CO2, 5% O2 and 90% N2. The in vitro matured oocytes were then collected for IIF or Western blot analysis. All experiments used for quantification were repeated at least three times. Data were compiled and analyzed using Microsoft Excel 2003. Comparisons were made for the length of spindle, the width of spindle poles, the width of spindle equator, the width of chromosome alignment, the distance of spindle to cortex in Padi6 wild-type versus mutant mature oocytes, and the polyspermic fertilization using unpaired t-test. The threshold of statistical significance was set at P?learn more is transformed from a bud of undifferentiated mesenchyme and overlying ectoderm into a structure with patterned skeletal, muscular and neural tissues. The patterning process happens in a proximal to distal sequence with the cells in the region of the humerus differentiating first followed by the radius/ulna domains and eventually the digits (Martin, 1990). The development of the skeleton has been the primary focus of studies that have established the tetrapod limb as a classical system for studying morphogenesis (Zeller et al., 2009). Mesenchymal cells of the bud condense and differentiate into chondrocytes that secrete the cartilage matrix to form the models of the mature bones. Simultaneously, muscle precursor cells that have migrated into the limb from the dermomyotome of the somites begin to differentiate and coalesce into muscle bundles (Duprez, 2002). The patterning of the somitic derivatives to form particular muscles is determined after their arrival in the limb bud as they are exposed to extrinsic signals in the mesenchyme (Hutcheson et al., 2009?and?Kardon et Quinapyramine al., 2002). Meanwhile axons from motor and sensory neurons innervate the limb in a stereotyped pattern from their point of convergence near the base of the forelimb at the site of the future brachial plexus. Extrinsic signals guide the axons as they enter the limb bud; including the ephrins of the limb mesenchyme that interact with axonal Eph receptors (Luria et al., 2008). The axons that innervate the limb and the progenitors that form the muscles must pass through the proximal limb on their way to more distal domains.