Expression of HAND gene products may be sufficient for the differentiation of avian neural crest-derived cells into catecholaminergic neurons in culture.
Members of the basic helix-loop-helix family of DNA binding proteins have important roles in the development of subpopulations of neural crest-derived neurons. We have cloned the chicken homologues of dHAND (HAND2) and eHAND (HAND1), basic helix-loop-helix DNA binding proteins whose neuronal expression is restricted to sympathetic and enteric neural crest-derived ganglia. Transcripts encoding dHAND and eHAND are expressed in sympathetic ganglia beginning at Hamburger and Hamilton stage 17-18. Antisense blockade of transcripts encoding HAND genes in neural crest-derived cells in vitro results in a significant reduction in neurogenesis. Differentiation of catecholaminergic neurons is also reduced by 52% if the expression of transcripts encoding dHAND and eHAND is reduced using antisense oligonucleotide blockade. The effect on neurogenesis and phenotypic expression of neural crest-derived neurons is specific; blockade of HAND gene expression has no apparent influence on the differentiation in vitro of neural tube-derived neurons. Use of a replication-competent avian retrovirus to constitutively express HAND genes in neural crest-derived cells in vitro, under nonpermissive growth conditions in medium supplemented with 2% chick embryo extract (CEE), induced precocious catecholaminergic differentiation. Constitutive expression of HAND gene products resulted in a significant increase in catecholaminergic differentiation of cells grown in medium supplemented with 10% CEE, a permissive growth condition for catecholaminergic development. These results suggest that the expression by neural crest cells of dHAND and eHAND may be both sufficient and necessary for catecholaminergic phenotypic expression.
Howard
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M
Department of Anatomy, Medical College of Ohio, Toledo, Ohio, 43614, USA. mhoward@mco.edu
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D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins; D002395:Catecholamines / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003370:Coturnix / Q000196:embryology*; D004268:DNA-Binding Proteins / Q000235:genetics*; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000502:physiology; D018507:Gene Expression Regulation, Developmental; D015723:Gene Library; D018257:Helix-Loop-Helix Motifs; D051379:Mice; D009432:Neural Crest / Q000166:cytology* / Q000502:physiology; D009474:Neurons / Q000166:cytology* / Q000502:physiology; D011817:Rabbits; D014157:Transcription Factors / Q000235:genetics*; D014158:Transcription, Genetic; D029961:Zebrafish Proteins
D051792:Basic Helix-Loop-Helix Proteins; D002395:Catecholamines; D004268:DNA-Binding Proteins; D014157:Transcription Factors; D029961:Zebrafish Proteins; C095572:Hand1 protein, mouse; C498766:Hand2 protein, mouse; C410915:hand2 protein, zebrafish; C097341:helix-loop-helix protein, eHAND