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Journal of Neuroscience, Vol 10, 1014-1024, Copyright © 1990 by Society for Neuroscience
Immunolocalization of G protein alpha-subunits in the Drosophila CNS
WJ Wolfgang, F Quan, P Goldsmith, C Unson, A Spiegel and M Forte
Vollum Institute For Advanced Biomedical Research, Oregon Health Sciences University, Portland 97201.
In order to uncover the role of G proteins in the integrative functioning
and development of the nervous system, we have begun a multidisciplinary
study of the G proteins present in the fruit fly, Drosophila melanogaster.
In this report, we describe the distribution of 3 different G protein
alpha-subunits in the adult Drosophila CNS as determined by
immunocytochemical localization using affinity-purified antibodies
generated to synthetic oligopeptide sequences unique to each alpha-subunit.
Western blot analysis of membranes prepared from Drosophila heads indicates
that antibodies specific for the Drosophila Go alpha and Gs alpha homologs
recognize the appropriate protein species predicted by molecular cloning
(Quan et al., 1989; Thambi et al., 1989). The Gi alpha homolog could not be
detected in head membranes by Western blotting, consistent with the
negligible levels of expression observed for Gi alpha on Northern blots of
head mRNA (Provost et al., 1988). However, a Drosophila Gi alpha fusion
protein could be detected by these antibodies following expression in E.
coli. Immunolocalization studies revealed that the Go alpha and Gs alpha
homologs are expressed at highest levels in neuropils and at intermediate
levels in the cortex of all brain and thoracic ganglion areas. Only the
lamina contained low levels of these alpha-subunits in the CNS.
Additionally, Gs alpha appears to be associated with the cell membranes of
neuronal cell bodies, while Go alpha has a more diffuse distribution,
suggesting its presence in the cytoplasm as well as cell membranes. In
contrast to the wide distribution of Go alpha and Gs alpha, Gi alpha has a
surprisingly restricted distribution in the CNS. It is present at high
levels only in photoreceptor cell terminations, glomerulae of the antennal
lobes, and the ocellar retina. Little or no Gi alpha was detected in other
brain regions or in the thoracic ganglion. Gi alpha, then, appears to be
uniquely associated with some primary sensory afferents and their
terminations, suggesting the presence of specific receptor and/or effector
systems which mediate the transmission of primary sensory information in
Drosophila.
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