The Drosophila inebriated-encoded neurotransmitter/osmolyte transporter: dual roles in the control of neuronal excitability and the osmotic stress response.
Water reabsorption by organs such as the mammalian kidney and insect Malpighian tubule/hindgut requires a region of hypertonicity within the organ. To balance the high extracellular osmolarity, cells within these regions accumulate small organic molecules called osmolytes. These osmolytes can accumulate to a high level without toxic effects on cellular processes. Here we provide evidence consistent with the possibility that the two protein isoforms encoded by the inebriated (ine) gene, which are members of the Na+/Cl--dependent neurotransmitter/osmolyte transporter family, perform osmolyte transport within the Malpighian tubule and hindgut. We show that ine mutants lacking both isoforms are hypersensitive to osmotic stress, which we assayed by maintaining flies on media containing NaCl, KCl, or sorbitol, and that this hypersensitivity is completely rescued by high-level ectopic expression of the ine-RB isoform. We provide evidence that this hypersensitivity represents a role for ine that is distinct from the increased neuronal excitability phenotype of ine mutants. Finally, we show that each ine genotype exhibits a "threshold" [NaCl]: long-term maintenance on NaCl-containing media above, but not below, the threshold causes lethality. Furthermore, this threshold value increases with the amount of ine activity. These data suggest that ine mutations confer osmotic stress sensitivity by preventing osmolyte accumulation within the Malpighian tubule and hindgut.
Huang
Xi
X
Department of Biochemistry and Cell Biology, Rice University, Houston, Texas 77251, USA.
Huang
Yanmei
Y
Chinnappan
Raj
R
Bocchini
Claire
C
Gustin
Michael C
MC
Stern
Michael
M
D000818:Animals; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000235:genetics* / Q000502:physiology; D017136:Ion Transport / Q000235:genetics / Q000502:physiology; D026901:Membrane Transport Proteins / Q000235:genetics* / Q000502:physiology; D018377:Neurotransmitter Agents / Q000235:genetics / Q000502:physiology; D050480:Plasma Membrane Neurotransmitter Transport Proteins; D020033:Protein Isoforms / Q000235:genetics / Q000502:physiology; D014882:Water-Electrolyte Balance / Q000235:genetics / Q000502:physiology
D029721:Drosophila Proteins; C102353:INE protein, Drosophila; D026901:Membrane Transport Proteins; D018377:Neurotransmitter Agents; D050480:Plasma Membrane Neurotransmitter Transport Proteins; D020033:Protein Isoforms