Carboxypeptidase A6 in zebrafish development and implications for VIth cranial nerve pathfinding.
Carboxypeptidase A6 (CPA6) is an extracellular protease that cleaves carboxy-terminal hydrophobic amino acids and has been implicated in the defective innervation of the lateral rectus muscle by the VIth cranial nerve in Duane syndrome. In order to investigate the role of CPA6 in development, in particular its potential role in axon guidance, the zebrafish ortholog was identified and cloned. Zebrafish CPA6 was secreted and interacted with the extracellular matrix where it had a neutral pH optimum and specificity for C-terminal hydrophobic amino acids. Transient mRNA expression was found in newly formed somites, pectoral fin buds, the stomodeum and a conspicuous condensation posterior to the eye. Markers showed this tissue was not myogenic in nature. Rather, the CPA6 localization overlapped with a chondrogenic site which subsequently forms the walls of a myodome surrounding the lateral rectus muscle. No other zebrafish CPA gene exhibited a similar expression profile. Morpholino-mediated knockdown of CPA6 combined with retrograde labeling and horizontal eye movement analyses demonstrated that deficiency of CPA6 alone did not affect either VIth nerve development or function in the zebrafish. We suggest that mutations in other genes and/or enhancer elements, together with defective CPA6 expression, may be required for altered VIth nerve pathfinding. If mutations in CPA6 contribute to Duane syndrome, our results also suggest that Duane syndrome can be a chondrogenic rather than a myogenic or neurogenic developmental disorder.
Lyons
Peter J
PJ
Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, New York, United States of America. peter.lyons@einstein.yu.edu
Ma
Leung-hang
LH
Baker
Robert
R
Fricker
Lloyd D
LD
D000010:Abducens Nerve / Q000201:enzymology* / Q000254:growth & development / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D043422:Carboxypeptidases A / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004195:Disease Models, Animal; D004370:Duane Retraction Syndrome / Q000201:enzymology* / Q000235:genetics / Q000503:physiopathology; D005123:Eye / Q000201:enzymology / Q000294:innervation; D005260:Female; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D015027:Zebrafish / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D029961:Zebrafish Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D029961:Zebrafish Proteins; D043422:Carboxypeptidases A; C555923:carboxypeptidase A6, zebrafish