The formation and function of chemical synapses is critical for information transfer within the nervous system. This proposal focuses on the functional development of the glutamatergic synapse, utilizing a forward genetic approach at the Drosophila neuromuscular junction (NMJ). With NIH support over the past 5 years. we have systematically mutated the third chromosome of Drosophila (ca. 40% of the genome) and screened to identify 38 mutant complementation groups with severe NMJ synaptic dysfunction phenotypes. All of these 38 complementation groups have been mapped to defined genomic locations using both recombination and deficiency mapping techniques. This proposal is to analyze three of the genes identified in this screen, each of which is required for glutamatergic synaptogenesis during embryonic development. For each gene product, a specific role at the synapse has been hypothesized based on a large body of preliminary experimental work. These hypotheses will be tested in this proposal.This proposal has four specific aims.
Aim I is to test the hypothesis that the mind the gap (mtg) gene encodes a prolyl.4.hydroxylase-a protein required for the formation of a collagen-based, synapse-specific extracellular matrix (ECM) which patterns and maintains postsynaptic glutamate receptor fields.
Aim 2 is to test the hypothesis that the bad reception (brec) gene encodes a novel protein, which is absolutely essential for the post-transcriptional synthesis and membrane insertion of postsynaptic glutamate receptors.
Aim 3 is to test the hypothesis that the slug-a-bed (slab) gene encodes the sole Drosophila ceramidase, which acts to control presynaptic development by determining the ratio of tethered (reserve) to docked (readily releasable) vesicles at presynaptic active zones.
Aim 4 is to continue our forward genetic screens to map, clone and characterize additional genes essential for synaptogenesis.Drosophila has proven to be a powerful system in which to identify genes underpinning fundamental aspects of nervous system development and function. Characterization of genes required for the functional formation of the glutamatergic synapse is particularly relevant to a number of health problems. Abnormal synaptic development and function are associated with inherited neurological diseases, injury, stroke and aging. Through systematic identification of genetic pathways underlying synaptic functional development, the proposed research will provide direct insight into molecular mechanisms related to the treatment of inherited synaptic diseases, age-related decline in synaptic function, and synaptic regeneration following injury.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM054544-08
Application #
6778364
Study Section
Special Emphasis Panel (ZRG1-MDCN-1 (01))
Program Officer
Tompkins, Laurie
Project Start
1997-08-01
Project End
2006-08-31
Budget Start
2004-09-01
Budget End
2005-08-31
Support Year
8
Fiscal Year
2004
Total Cost
$279,339
Indirect Cost
Name
Vanderbilt University Medical Center
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
004413456
City
Nashville
State
TN
Country
United States
Zip Code
37212
Wei, Chunyao; Thatcher, Elizabeth J; Olena, Abigail F et al. (2013) miR-153 regulates SNAP-25, synaptic transmission, and neuronal development. PLoS One 8:e57080
Rohrbough, Jeffrey; Kent, Karla S; Broadie, Kendal et al. (2013) Jelly Belly trans-synaptic signaling to anaplastic lymphoma kinase regulates neurotransmission strength and synapse architecture. Dev Neurobiol 73:189-208
Siller, Saul S; Broadie, Kendal (2012) Matrix metalloproteinases and minocycline: therapeutic avenues for fragile X syndrome. Neural Plast 2012:124548
Dani, Neil; Broadie, Kendal (2012) Glycosylated synaptomatrix regulation of trans-synaptic signaling. Dev Neurobiol 72:2-21
Rushton, Emma; Rohrbough, Jeffrey; Deutsch, Kalie et al. (2012) Structure-function analysis of endogenous lectin mind-the-gap in synaptogenesis. Dev Neurobiol 72:1161-79
Dani, Neil; Nahm, Minyeop; Lee, Seungbok et al. (2012) A targeted glycan-related gene screen reveals heparan sulfate proteoglycan sulfation regulates WNT and BMP trans-synaptic signaling. PLoS Genet 8:e1003031
Broadie, Kendal; Baumgartner, Stefan; Prokop, Andreas (2011) Extracellular matrix and its receptors in Drosophila neural development. Dev Neurobiol 71:1102-30
Siller, Saul S; Broadie, Kendal (2011) Neural circuit architecture defects in a Drosophila model of Fragile X syndrome are alleviated by minocycline treatment and genetic removal of matrix metalloproteinase. Dis Model Mech 4:673-85
Rohrbough, Jeffrey; Broadie, Kendal (2010) Anterograde Jelly belly ligand to Alk receptor signaling at developing synapses is regulated by Mind the gap. Development 137:3523-33
Vijayakrishnan, Niranjana; Phillips, Scott E; Broadie, Kendal (2010) Drosophila rolling blackout displays lipase domain-dependent and -independent endocytic functions downstream of dynamin. Traffic 11:1567-78

Showing the most recent 10 out of 49 publications