skip to main content
US FlagAn official website of the United States government
dot gov icon
Official websites use .gov
A .gov website belongs to an official government organization in the United States.
https lock icon
Secure .gov websites use HTTPS
A lock ( lock ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

Explore Research Products in the PAR
It may take a few hours for recently added research products to appear in PAR search results.


Title: Drosophila as a Model to Study the Blood-Brain Barrier
The Drosophila blood-brain barrier (BBB) has been shown to be largely analogous in structure and function to the vertebrate BBB. Thanks to the genetic tools available for this organism, Drosophila is uniquely suited to study bbb physiology and function, with high relevance for mammalian function. In this chapter we discuss targeting strategies to specifically mark and manipulate BBB cells, how to test BBB integrity, and methods to isolate single-BBB cells.  more » « less
Award ID(s):
1755385
PAR ID:
10323957
Author(s) / Creator(s):
;
Editor(s):
Barichello T.
Date Published:
Journal Name:
Neuromethods
Volume:
142
ISSN:
1940-6045
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
More Like this
  1. ; ; ; ; (, Genes, Brain and Behavior)
    Abstract The blood brain barrier (BBB) has the essential function to protect the brain from potentially hazardous molecules while also enabling controlled selective uptake. How these processes and signaling inside BBB cells control neuronal function is an intense area of interest. Signaling in the adultDrosophilaBBB is required for normal male courtship behavior and relies on male‐specific molecules in the BBB. Here we show that the dopamine receptorD2Ris expressed in the BBB and is required in mature males for normal mating behavior. Conditional adult male knockdown ofD2Rin BBB cells causes courtship defects. The courtship defects observed in geneticD2Rmutants can be rescued by expression of normalD2Rspecifically in the BBB of adult males.DrosophilaBBB cells are glial cells. Our findings thus identify a specific glial function for theDR2receptor and dopamine signaling in the regulation of a complex behavior. 
    more » « less
  2. ; ; ; ; (, Curr Issues Mol Biol)
    Beyond its crucial role as a tight barrier to protect the nervous system, the Blood–Brain Barrier (BBB) is increasingly being recognized for its physiological processes that affect brain function and behavior. In Drosophila melanogaster, the BBB expresses sex-specific transcripts, and a change in the sexual identity of adult BBB cells results in a significant reduction in male courtship behavior. The molecular nature of this BBB/brain interaction and the molecules that mediate it are unknown. Here we feminize BBB cells by targeted expression of the Drosophila female-specific master regulator TraF in otherwise normal males. We examined the effect on RNA expression in dissected brains by RNA sequencing. We find that 283 transcripts change in comparison to normal control males. Transcripts representing cell signaling processes and synaptic communication are enriched, as are hormonal mediators. These transcripts provide a valuable resource for addressing questions about BBB and brain interaction. 
    more » « less
  3. ; ; ; ; ; ; (, PLOS Genetics)
    Wang, Hongyan (Ed.)
    The blood brain barrier (BBB) forms a stringent barrier that protects the brain from components in the circulation that could interfere with neuronal function. At the same time, the BBB enables selective transport of critical nutrients and other chemicals to the brain. Beyond these functions, another recently recognized function is even less characterized, specifically the role of the BBB in modulating behavior by affecting neuronal function in a sex-dependent manner. Notably, signaling in the adult Drosophila BBB is required for normal male courtship behavior. Courtship regulation also relies on male-specific molecules in the BBB. Our previous studies have demonstrated that adult feminization of these cells in males significantly lowered courtship. Here, we conducted microarray analysis of BBB cells isolated from males and females. Findings revealed that these cells contain male- and female-enriched transcripts, respectively. Among these transcripts, nuclear receptor Hr46/Hr3 was identified as a male-enriched BBB transcript. Hr46/Hr3 is best known for its essential roles in the ecdysone response during development and metamorphosis. In this study, we demonstrate that Hr46/Hr3 is specifically required in the BBB cells for courtship behavior in mature males. The protein is localized in the nuclei of sub-perineurial glial cells (SPG), indicating that it might act as a transcriptional regulator. These data provide a catalogue of sexually dimorphic BBB transcripts and demonstrate a physiological adult role for the nuclear receptor Hr46/Hr3 in the regulation of male courtship, a novel function that is independent of its developmental role. 
    more » « less
  4. ; ; (, International Journal of Molecular Sciences)
    The blood–brain barrier (BBB) is a multicellular construct that regulates the diffusion and transport of metabolites, ions, toxins, and inflammatory mediators into and out of the central nervous system (CNS). Its integrity is essential for proper brain physiology, and its breakdown has been shown to contribute to neurological dysfunction. The BBB in vertebrates exists primarily through the coordination between endothelial cells, pericytes, and astrocytes, while invertebrates, which lack a vascularized circulatory system, typically have a barrier composed of glial cells that separate the CNS from humoral fluids. Notably, the invertebrate barrier is molecularly and functionally analogous to the vertebrate BBB, and the fruit fly, Drosophila melanogaster, is increasingly recognized as a useful model system in which to investigate barrier function. The most widely used technique to assess barrier function in the fly is the dye-exclusion assay, which involves monitoring the infiltration of a fluorescent-coupled dextran into the brain. In this study, we explore analytical and technical considerations of this procedure that yield a more reliable assessment of barrier function, and we validate our findings using a traumatic injury model. Together, we have identified parameters that optimize the dye-exclusion assay and provide an alternative framework for future studies examining barrier function in Drosophila. 
    more » « less
  5. ; ; (, Drosophila Research Conference)
    Abstract Flies have an open circulatory system and their Blood Brain Barrier (BBB) surrounds the brain like a tight cap. The fly BBB consists of two layers of glial cells. The outer layer is formed by the Perineurial Glia (PG). The inner BBB layer consists of the Subperineurial Glia (SPG) that form the tight barrier that, like its mammalian counterpart, acts both as a diffusion and a xenobiotic transport barrier. Underneath the SPG lie the neuronal cell bodies. The Drosophila BBB shows the same barrier properties as the mammalian barrier and profiling of Drosophila BBB cells has shown a high degree of molecular conservation. We have previously shown that the Drosophila BBB plays a sex-specific role in regulating behavior. Conditional adult feminization of SPG cells in otherwise normal males leads to significantly reduced courtship. In agreement with this, in a microarray screen of isolated SPG cells, we identified a number of male-enriched transcripts. One of them encodes the dopamine-2 like receptor (D2R). We have found that conditional knockdown of D2R in adult male Drosophila SPG decreases courtship. Likewise, D2R mutant males have courtship defects. They can be rescued by expression of wildtype D2R in the SPG cells of mature adult males, demonstrating a physiological requirement for the receptor in these cells for courtship control4. The D2R receptor is highly conserved. It has been found that it can act via biased signaling (through G protein or b-arrestin) in mammals. We have previously found that signaling through Gao and arrestin in the BBB is required for proper male courtship. Although D2R is best known for signaling through cAMP, we have not found a requirement for cAMP/PKA signaling for courtship. To investigate the signaling pathways downstream of D2R that are responsible for courtship control we have mutagenized D2R proteins and examined their ability to rescue D2R mutants. Based on Peterson et al. we designed proteins capable of G-protein or arrestin biased signaling, respectively, and tested their ability to rescue the courtship defects of D2R mutants. Our data suggest that D2R signaling through b-arrestin is a major mediator of BBB courtship control. 
    more » « less
  • Citation Formats
  • MLA
  • APA
  • Chicago
  • BibTeX
  • Save / Share this Record
  • Send to Email