A high throughput messenger RNA differential display screen identifies discrete domains of gene expression and novel patterning processes along the developing neural tube.
BACKGROUND
During early development the vertebrate neural tube is broadly organized into the forebrain, midbrain, hindbrain and spinal cord regions. Each of these embryonic zones is patterned by a combination of genetic pathways and the influences of local signaling centres. However, it is clear that much remains to be learned about the complete set of molecular cues that are employed to establish the identity and intrinsic neuronal diversity of these territories. In order to address this, we performed a high-resolution messenger RNA differential display screen to identify molecules whose expression is regionally restricted along the anteroposterior (AP) neuraxis during early chick development, with particular focus on the midbrain and hindbrain vesicles.


RESULTS
This approach identified 44 different genes, with both known and unknown functions, whose transcription is differentially regulated along the AP axis. The identity and ontological classification of these genes is presented. The wide variety of functional classes of transcripts isolated in this screen reflects the diverse spectrum of known influences operating across these embryonic regions. Of these 44 genes, several have been selected for detailed in situ hybridization analysis to validate the screen and accurately define the expression domains. Many of the identified cDNAs showed no identity to the current databases of known or predicted genes or ESTs. Others represent genes whose embryonic expression has not been previously reported. Expression studies confirmed the predictions of the primary differential display data. Moreover, the nature of identified genes, not previously associated with regionalisation of the brain, identifies novel potential mechanisms in that process.


CONCLUSIONS
This study provides an insight into some of the varied and novel molecular networks that operate during the regionalization of embryonic neural tissue and expands our knowledge of molecular repertoire used during development.
Chambers
David
D
MRC Centre for Developmental Neurobiology, King's College London, Guy's Campus, London, SE1 1UL, UK. david.2.chambers@kcl.ac.uk
Mason
Ivor
I
D000818:Animals; D019521:Body Patterning / Q000235:genetics*; D001921:Brain / Q000196:embryology* / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002642:Chick Embryo; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D017403:In Situ Hybridization; D008636:Mesencephalon / Q000196:embryology / Q000378:metabolism; D039842:Neural Cell Adhesion Molecule L1 / Q000235:genetics / Q000378:metabolism; D051762:PAX2 Transcription Factor / Q000235:genetics / Q000378:metabolism; D020648:Peptide Elongation Factor 1 / Q000235:genetics / Q000378:metabolism; D017027:Protein Tyrosine Phosphatases / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis* / Q000378:metabolism; D054633:Receptor-Like Protein Tyrosine Phosphatases, Class 5; D017474:Receptors, Laminin / Q000235:genetics / Q000378:metabolism; D012249:Rhombencephalon / Q000196:embryology / Q000378:metabolism
D002135:Calcium-Binding Proteins; D002352:Carrier Proteins; D016207:Cytokines; D039842:Neural Cell Adhesion Molecule L1; D051762:PAX2 Transcription Factor; D020648:Peptide Elongation Factor 1; D012333:RNA, Messenger; D017474:Receptors, Laminin; C066845:pleiotrophin; D017027:Protein Tyrosine Phosphatases; D054633:Receptor-Like Protein Tyrosine Phosphatases, Class 5