Expressional Profiling of Carpet Glia in the Developing Drosophila Eye Reveals Its Molecular Signature of Morphology Regulators

• Published in: Frontiers in neuroscience 2019-03, Vol.13, p.244
• Main Authors: Ho, Tsung-Ying, Wu, Wei-Hang, Hung, Sheng-Jou, Liu, Tsunglin, Lee, Yuan-Ming, Liu, Ya-Hsin
• Format: Article
• Language: English
• Subjects: Actin; Adenine; Biotechnology; Blood-brain barrier; carpet glia; Cell adhesion & migration; Cell cycle; Cytology; Cytoskeleton; Deoxyribonucleic acid; DNA; DNA sequencing; Drosophila; Endothelial cells; Endothelium; Extracellular matrix; Gene expression; Gene silencing; Genomes; Genomics; Glial cells; Guanosine triphosphatases; Homeostasis; Insects; Kinases; Morphology; Nervous system; Neuronal-glial interactions; Neuroscience; Next-generation sequencing; Proteins; RNA-mediated interference; subperineurial glia; targeted DamID-seq; Tight junctions; transcriptome
• ISSN: 1662-4548; 1662-453X; 1662-453X
• DOI: 10.3389/fnins.2019.00244

Homeostasis in the nervous system requires intricate regulation and is largely accomplished by the blood-brain barrier (BBB). The major gate keeper of the vertebrate BBB is vascular endothelial cells, which form tight junctions (TJs). To gain insight into the development of the BBB, we studied the carpet glia, a subperineurial glial cell type with vertebrate TJ-equivalent septate junctions, in the developing eye. The large and flat, sheet-like carpet glia, which extends along the developing eye following neuronal differentiation, serves as an easily accessible experimental system to understand the cell types that exhibit barrier function. We profiled transcribed genes in the carpet glia using targeted DNA adenine methyl-transferase identification, followed by next-generation sequencing (targeted DamID-seq) and found that the majority of genes expressed in the carpet glia function in cellular activities were related to its dynamic morphological changes in the developing eye. To unravel the morphology regulators, we silenced genes selected from the carpet glia transcriptome using RNA interference. The gene encoding a GTPase was previously reported as a key regulator of the actin cytoskeleton. The expression of the ( ) gene, encoding a solute carrier transporter in the developing eye, is specific to the carpet glia. The reduced expression of severely disrupted the formation of intact carpet glia, and the silencing impaired the connection between the two carpet glial cells, indicating the pan-cellular and local effects of Rho1 and Path on carpet glial cell morphology, respectively. Our study molecularly characterized a particular subperineurial cell type providing a resource for a further understanding of the cell types comprising the BBB.