Journal of Neuroscience, Vol 8, 580-592, Copyright © 1988 by Society for Neuroscience
Developmental regulation of ganglioside antigens recognized by the JONES antibody
B Schlosshauer, AS Blum, R Mendez-Otero, CJ Barnstable and M Constantine-Paton
Department of Biology, Yale University, New Haven, Connecticut 06511.
The JONES monoclonal antibody has been immunocytochemically associated with
regions of the developing rat brain where cell and axon migrations are
occurring (Mendez-Otero et al., 1986, 1988). In the present study the
antigens recognized by the JONES antibody were analyzed in a variety of
brain regions and at developmental ages selected to correspond to the
preceding immunocytochemical observations. In accordance with earlier
results from retina, JONES binding could not be detected in SDS gels from
developing brain. Binding of the antibody was, however, prominent in
chloroform/methanol extracts of the same tissues, and it was completely
removed from tissue sections by brief chloroform/methanol form/methanol
treatment. Enzymatic analyses of chloroform/methanol extracts indicated
that the JONES epitope was sensitive to neuraminidase but insensitive to
proteases. Overlay assays on developed high-performance thin-layer
chromatographic plates (HPTLC) indicate that in all regions the JONES
epitope resides on 2 or 3 ganglioside bands, depending on the age examined.
These bands migrate between ganglioside standards GD 1a and GM2 on HPTLC
plates and have been designated GJ1, GJ2, and GJ3, with the higher number
designating the more rapidly migrating species. Occasionally, additional
bands migrating in the range of polysialogangliosides were observed. The
pattern of expression of GJ species was studied in forebrain, retina, and
cerebellar tissue taken from embryonic day 18 (E18), postnatal day 0 (P0),
P7, P14, and adult animals. Both region-specific differences in the
relative prominence of each band and stage-specific differences in the
total amount of the JONES gangliosides were detected. The stage- specific
differences in the amount of JONES antigens are well correlated with the
developmental periods of maximal cell migration in each region. While the
JONES gangliosides are most prominent in forebrain before birth, in they
are most prominent during the first 2 postnatal weeks. In cerebellum, JONES
antigen expression is more pronounced during the 2 periods of cell
migration in this tissue. In retina, the more rapidly migrating GJ3 band
was the most prominent band at all stages examined, and this same band is
retained in the adult. In cerebellum and forebrain GJ3 is also the most
pronounced band during development. However, in contrast to the retina, the
more slowly migrating GJ1 band is retained in adult forebrain and
cerebellum. A variety of non-brain tissues have also been examined for the
presence of the JONES antigens.(ABSTRACT TRUNCATED AT 400 WORDS)