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Objective: In vertebrates, bone morphogenetic proteins (BMPs) and activin signals playmultiple roles in dorso-ventral patterning and development of the spinal cord. Here theinductions of BMP 4 and activin A on embryonic stem cells (ESCs) into dorsal interneuronshave been studied.Materials and Methods: Four different treatments have been used for mESC derivedneural precursors; they include BMP4 (1ng/ml and 10ng/ml), activin A (100 ng/ml), andactivin A+BMP4 (100ng/ml, 10ng/ml). Induction’s effect on expression of specific dorsalinterneuron markers in mature neurons have been evaluated by the use of immunocytochemistryand RT-PCR.Results: Treatment of ESC-derived neural precursors with BMP4, activin A, or bothshowed an increased generation of both dI1 and dI3 interneurons (Lhx2 and Isl-1-positivecells) compared to the control group. However, the synergistic effect in generation of dI3was not observed when both factors were used. Moreover, RT-PCR analysis of differentiatedcells showed the expression of Lhx9, Lhx1, and Isl1, the transcription factors that aremarkers of dI1, dI2, and dI3 interneurons respectively.Conclusion: Our results showed that specific combination of developmental signalingmolecules can direct the differentiation of ESCs into dorsal interneurons. Furthermore,qualitative and quantitative differences in signaling by different members ofthe TGF-β supe