A versatile genetic tool to study midline glia function in the Drosophila CNS

被引:4
作者
Banerjee, Swati [1 ]
Mino, Rosa E. [1 ,2 ]
Fisher, Elizabeth S. [1 ]
Bhat, Manzoor A. [1 ]
机构
[1] Univ Texas Hlth San Antonio, Ctr Biomed Neurosci, Dept Cellular & Integrat Physiol, 7703 Floyd Curl Dr, San Antonio, TX 78229 USA
[2] UT Southwestern Med Ctr, Dept Cell Biol, 6000 Harry Hines Blvd, Dallas, TX 75235 USA
基金
美国国家卫生研究院;
关键词
Neurexin IV; Wrapper; wrapper-GAL4; Midline glia; Axonal ensheathment; Commissures; SEPTATE JUNCTION; NEUREXIN IV; ORGANIZATION; GUIDANCE; RECEPTOR; DOMAINS; EMBRYOGENESIS; SUPERFAMILY; CONTACTIN; WRAPPER;
D O I
10.1016/j.ydbio.2017.06.010
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
Neuron-glial interactions are crucial for growth, guidance and ensheathment of axons across species. In the Drosophila CNS midline, neuron-glial interactions underlie ensheathment of commissural axons by midline glial (MG) cells in a manner similar to mammalian oligodendrocytes. Although there has been some advance in the study of neuron-glial interactions and ensheathment of axons in the CNS midline, key aspects of axonal ensheathment are still not fully understood. One of the limitations has been the unavailability of MG membrane markers that could highlight the glial processes wrapping the axons. Previous studies have identified two key molecular players from the neuronal and glial cell types in the CNS midline. These are the neuronal transmembrane protein Neurexin IV (Nrx IV) and the membrane-anchored MG protein Wrapper, both of which interact in trans to mediate neuron-glial interactions and ensheathment of commissural axons. In the current study, we attempt to further our understanding of MG biology and try to overcome some of the technical difficulties posed by the lack of a robust MG driver that will specifically allow expression or knockdown of genes in MG. We report the generation of BAC transgenic flies of wrapper-GAL4 and demonstrate how these flies could be used as a genetic tool to understand MG biology. We have utilized the GAL4/UAS system to drive GFP-reporter lines (membrane-bound mCD8-GFP; microtubule-associated tau-GFP) and nuclear lacZ using wrapper-GAL4 to highlight the MG cells and/or their processes that surround and perform axonal ensheathment functions in the embryonic midline. We also describe the utility of the wrapper-GAL4 driver line to down-regulate known MG genes specifically in Wrapper-positive cells. Finally, we validate the functionality of the wrapper-GAL4 driver by rescue of wrapper mutant phenotypes and lethality. Together, these studies provide us with a versatile genetic tool to investigate MG functions and will aid in future investigations where genetic screens using wrapper-GAL4 could be designed to identify novel molecular players at the Drosophila midline and unravel key aspects of MG biology.
引用
收藏
页码:35 / 43
页数:9
相关论文
共 44 条
[1]   New Tools for the Analysis of Glial Cell Biology in Drosophila [J].
Awasaki, Takeshi ;
Lee, Tzumin .
GLIA, 2011, 59 (09) :1377-1386
[2]   Axonal ensheathment and septate junction formation in the peripheral nervous system of Drosophila [J].
Banerjee, S ;
Pillai, AM ;
Paik, R ;
Li, JJ ;
Bhat, MA .
JOURNAL OF NEUROSCIENCE, 2006, 26 (12) :3319-3329
[3]   Organization and function of septate junctions - An evolutionary perspective [J].
Banerjee, Swati ;
Sousa, Aurea D. ;
Bhat, Manzoor A. .
CELL BIOCHEMISTRY AND BIOPHYSICS, 2006, 46 (01) :65-77
[4]   Drosophila Neurexin IV Interacts with Roundabout and Is Required for Repulsive Midline Axon Guidance [J].
Banerjee, Swati ;
Blauth, Kevin ;
Peters, Kimberly ;
Rogers, Stephen L. ;
Fanning, Alan S. ;
Bhat, Manzoor A. .
JOURNAL OF NEUROSCIENCE, 2010, 30 (16) :5653-5667
[5]   A Drosophila neurexin is required for septate junction and blood-nerve barrier formation and function [J].
Baumgartner, S ;
Littleton, JT ;
Broadie, K ;
Bhat, MA ;
Harbecke, R ;
Lengyel, JA ;
ChiquetEhrismann, R ;
Prokop, A ;
Bellen, HJ .
CELL, 1996, 87 (06) :1059-1068
[6]   Molecular organization of axo-glial junctions [J].
Bhat, MA .
CURRENT OPINION IN NEUROBIOLOGY, 2003, 13 (05) :552-559
[7]   Organization and maintenance of molecular domains in myelinated axons [J].
Buttermore, Elizabeth D. ;
Thaxton, Courtney L. ;
Bhat, Manzoor A. .
JOURNAL OF NEUROSCIENCE RESEARCH, 2013, 91 (05) :603-622
[8]   Recombineering Homologous Recombination Constructs in Drosophila [J].
Carreira-Rosario, Arnaldo ;
Scoggin, Shane ;
Shalaby, Nevine A. ;
Williams, Nathan David ;
Hiesinger, P. Robin ;
Buszczak, Michael .
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, 2013, (77)
[9]   Neurofascin is a glial receptor for the paranodin/Caspr-contactin axonal complex at the axoglial junction [J].
Charles, P ;
Tait, S ;
Faivre-Sarrailh, C ;
Barbin, G ;
Gunn-Moore, F ;
Denisenko-Nehrbass, N ;
Guennoc, AM ;
Girault, JA ;
Brophy, PJ ;
Lubetzki, C .
CURRENT BIOLOGY, 2002, 12 (03) :217-220
[10]   Axon-glial interactions at the Drosophila CNS midline [J].
Crews, Stephen T. .
CELL ADHESION & MIGRATION, 2010, 4 (01) :67-71