» Articles » PMID: 20127822

Immunocytochemical Localization of Synaptic Proteins to Photoreceptor Synapses of Drosophila Melanogaster

Overview
Journal J Comp Neurol
Specialty Neurology
Date 2010 Feb 4
PMID 20127822
Citations 30
Authors
Affiliations
Soon will be listed here.
Abstract

The location of proteins that contribute to synaptic function has been widely studied in vertebrate synapses, far more than at model synapses of the genetically manipulable fruit fly, Drosophila melanogaster. Drosophila photoreceptor terminals have been extensively exploited to characterize the actions of synaptic genes, and their distinct and repetitive synaptic ultrastructure is anatomically well suited for such studies. Synaptic release sites include a bipartite T-bar ribbon, comprising a platform surmounting a pedestal. So far, little is known about the composition and precise location of proteins at either the T-bar ribbon or its associated synaptic organelles, knowledge of which is required to understand many details of synaptic function. We studied the localization of candidate proteins to pre- or postsynaptic organelles, by using immuno-electron microscopy with the pre-embedding method, after first validating immunolabeling by confocal microscopy. We used monoclonal antibodies against Bruchpilot, epidermal growth factor receptor pathway substrate clone 15 (EPS-15), and cysteine string protein (CSP), all raised against a fly head homogenate, as well as sea urchin kinesin (antibody SUK4) and Discs large (DLG). All these antibodies labeled distinct synaptic structures in photoreceptor terminals in the first optic neuropil, the lamina, as did rabbit anti-DPAK (Drosophila p21 activated kinase) and anti-Dynamin. Validating reports from light microscopy, immunoreactivity to Bruchpilot localized to the edge of the platform, and immunoreactivity to SUK4 localized to the pedestal of the T-bar ribbon. Anti-DLG recognized the photoreceptor head of capitate projections, invaginating organelles from surrounding glia. For synaptic vesicles, immunoreactivity to EPS-15 localized to sites of endocytosis, and anti-CSP labeled vesicles lying close to the T-bar ribbon. These results provide markers for synaptic sites, and a basis for further functional studies.

Citing Articles

Proximity labeling reveals interactions necessary to maintain the distinct apical domains of Drosophila photoreceptors.

Sastry L, Rylee J, Mahato S, Zelhof A J Cell Sci. 2024; 137(23).

PMID: 39540276 PMC: 11827603. DOI: 10.1242/jcs.262223.


Tissue distribution of cysteine string protein/DNAJC5 in C. elegans analysed by CRISPR/Cas9-mediated tagging of endogenous DNJ-14.

Barker E, Morgan A, Barclay J Cell Tissue Res. 2024; 396(1):41-55.

PMID: 38403745 PMC: 10997724. DOI: 10.1007/s00441-024-03875-w.


The evolution of synaptic and cognitive capacity: Insights from the nervous system transcriptome of .

Orvis J, Albertin C, Shrestha P, Chen S, Zheng M, Rodriguez C Proc Natl Acad Sci U S A. 2022; 119(28):e2122301119.

PMID: 35867761 PMC: 9282427. DOI: 10.1073/pnas.2122301119.


Genetic regulation of central synapse formation and organization in Drosophila melanogaster.

Duhart J, Mosca T Genetics. 2022; 221(3).

PMID: 35652253 PMC: 9252286. DOI: 10.1093/genetics/iyac078.


Circadian regulation of the Drosophila astrocyte transcriptome.

You S, Yu A, Roberts M, Joseph I, Jackson F PLoS Genet. 2021; 17(9):e1009790.

PMID: 34543266 PMC: 8483315. DOI: 10.1371/journal.pgen.1009790.


References
1.
Conder R, Yu H, Zahedi B, Harden N . The serine/threonine kinase dPak is required for polarized assembly of F-actin bundles and apical-basal polarity in the Drosophila follicular epithelium. Dev Biol. 2007; 305(2):470-82. DOI: 10.1016/j.ydbio.2007.02.034. View

2.
Meinertzhagen I, ONeil S . Synaptic organization of columnar elements in the lamina of the wild type in Drosophila melanogaster. J Comp Neurol. 1991; 305(2):232-63. DOI: 10.1002/cne.903050206. View

3.
Hiesinger P, Scholz M, Meinertzhagen I, Fischbach K, Obermayer K . Visualization of synaptic markers in the optic neuropils of Drosophila using a new constrained deconvolution method. J Comp Neurol. 2000; 429(2):277-88. DOI: 10.1002/1096-9861(20000108)429:2<277::aid-cne8>3.0.co;2-8. View

4.
Stowers R, Megeath L, Gorska-Andrzejak J, Meinertzhagen I, Schwarz T . Axonal transport of mitochondria to synapses depends on milton, a novel Drosophila protein. Neuron. 2002; 36(6):1063-77. DOI: 10.1016/s0896-6273(02)01094-2. View

5.
Tom Dieck S, Altrock W, Kessels M, Qualmann B, Regus H, Brauner D . Molecular dissection of the photoreceptor ribbon synapse: physical interaction of Bassoon and RIBEYE is essential for the assembly of the ribbon complex. J Cell Biol. 2005; 168(5):825-36. PMC: 2171818. DOI: 10.1083/jcb.200408157. View