Appearance of beta 1,4 N-acetylgalactosaminyltransferase (glycosphingolipids GA2/GM2/GD2 synthase) in embryonic chicken vitreous humor during development.
OBJECTIVE
beta 1,4 N-Acetylgalactosaminyltransferase (GalNAcT) is a type II integral membrane protein of the Golgi apparatus that catalyzes the synthesis of the glycosphingolipids GM2, GD2, and GA2. The activity of GalNAcT in chick retinal cells increases 6-fold between embryonic days 7 and 14. Because GalNAcT, like many Golgi glycosyltransferases, is proteolytically cleaved from Golgi membranes to release a soluble form into the culture medium of cells transfected with the cloned human enzyme, we tested whether GalNAcT might be released from embryonic retinal cells into the vitreous humor.


METHODS
Samples of vitreous humor and plasma and extracts of retinal cells were assayed for GalNAcT activity.


RESULTS
The activity of a soluble form of GalNAcT in embryonic chick vitreous humor was nearly undetectable until embryonic day 10, then increased more than six fold until day 16, and remained at that level until birth. The activity was identified as authentic GalNAcT based on a requirement for Mn++, GSL substrate specificity, and product characterization. GalNAcT activity in embryonic plasma was roughly 10% that of the corresponding vitreous humor, suggesting that the plasma was not the source of the activity in the vitreous.


CONCLUSIONS
GalNAcT in embryonic chicken vitreous humor is likely due either to a specific release from neural retinal cells or due to non-specific lysis of these cells during apoptosis associated with the development of the retina. Regardless of the source, GalNAcT in the vitreous humor has the potential to function as a lectin by binding to gangliosides GD3 and GM3 on the surface of retinal cells and, thereby, to influence neuronal development.
Allende
M L
ML
Department of Biological and Biophysical Sciences, School of Dentistry, University of Louisville 40292, USA.
Schwendiman
R M
RM
Young
W W
WW
D000818:Animals; D002457:Cell Extracts / Q000737:chemistry; D002642:Chick Embryo / Q000166:cytology / Q000378:metabolism / Q000502:physiology; D006801:Humans; D017350:N-Acetylgalactosaminyltransferases / Q000032:analysis / Q000097:blood / Q000378:metabolism*; D012160:Retina / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D013379:Substrate Specificity; D013997:Time Factors; D014822:Vitreous Body / Q000196:embryology* / Q000201:enzymology*; D000097763:Polypeptide N-acetylgalactosaminyltransferase
D002457:Cell Extracts; D017350:N-Acetylgalactosaminyltransferases; D000097763:Polypeptide N-acetylgalactosaminyltransferase; C023220:(N-acetylneuraminyl)-galactosylglucosylceramide N-acetylgalactosaminyltransferase