Roles of neural communication between the brain and thoracic ganglia in the selection and regulation of the cricket escape behavior.

To survive a predator's attack, prey animals must exhibit escape responses that are appropriately regulated in terms of their moving speed, distance, and direction. Insect locomotion is considered to be controlled by an interaction between the brain, which is involved in behavioral decision-making, and the thoracic ganglia (TG), which are primary motor centers. However, it remains unknown which descending and ascending signals between these neural centers are involved in the regulation of the escape behavior. We addressed the distinct roles of the brain and TG in the wind-elicited escape behavior of crickets by assessing the effects of partial ablation of the intersegmental communications on escape responses. We unilaterally cut the ventral nerve cord (VNC) at different locations, between the brain and TG, or between the TG and terminal abdominal ganglion (TAG), a primary sensory center of the cercal system. The partial ablation of ascending signals to the brain greatly reduced the jumping response rather than running, indicating that sensory information processing in the brain is essential for the choice of escape responses. The ablation of descending signals from the brain to the TG impaired locomotor performance and directional control of the escape responses, suggesting that locomotion in the escape behavior largely depends on the descending signals from the brain. Finally, the extracellular recording from the cervical VNC indicated a difference in the descending activities preceding the escape responses between running and jumping. Our results demonstrated that the brain sends the descending signals encoding the behavioral choice and locomotor regulation to the TG, while the TG seem to have other specific roles, such as in the preparation of escape movement.

A novel role for OATP2A1/SLCO2A1 in a murine model of colon cancer.

Prostaglandin E2 (PGE2) is associated with proliferation and angiogenesis in colorectal tumours. The role of prostaglandin transporter OATP2A1/SLCO2A1 in colon cancer tumorogenesis is unknown. We evaluated mice of various Slco2a1 genotypes in a murine model of colon cancer, the adenomatous polyposis (APC) mutant (Apc ∆716/+) model. Median lifespan was significantly extended from 19 weeks in Slco2a1 +/+/Apc Δ716/+ mice to 25 weeks in Slco2a1 -/-/Apc Δ716/+ mice. Survival was directly related to a reduction in the number of large polyps in the Slco2a1 -/- /Apc ∆716/+ compared to the Slco2a1 +/+/Apc Δ716/+ or Slco2a1 +/-/Apc Δ716/+mice. The large polyps from the Slco2a1 -/- /Apc ∆716/+ mice had significant reductions in microvascular density, consistent with the high expression of Slco2a1 in the tumour-associated vascular endothelial cells. Chemical suppression of OATP2A1 function significantly reduced tube formation and wound-healing activity of PGE2 in human vascular endothelial cells (HUVECs) although the amount of extracellular PGE2 was not affected by an OATP2A1 inhibitor. Further an in vivo model of angiogenesis, showed a significant reduction of haemoglobin content (54.2%) in sponges implanted into Slco2a1 -/-, compared to wildtype mice. These studies indicate that OATP2A1 is likely to promote tumorogenesis by PGE2 uptake into the endothelial cells, suggesting that blockade of OATP2A1 is an additional pharmacologic strategy to improve colon cancer outcomes.

Impact of FDA-Approved Drugs on the Prostaglandin Transporter OATP2A1/SLCO2A1.

To understand interaction of drugs with the prostaglandin transporter OATP2A1/SLCO2A1 that regulates disposition of prostaglandins, we explored the impact of 636 drugs in an FDA-approved drug library on 6-carboxyfluorescein (6-CF) uptake by OATP2A1-expressing HEK293 cells (HEK/2A1). Fifty-one and 10 drugs were found to inhibit and enhance 6-CF uptake by more than 50%, respectively. Effect of the 51 drugs on 6-CF uptake was positively correlated with that on PGE2 uptake (r = 0.64, p < 0.001). Among those, 5 drugs not structurally related to prostaglandins, suramin, pranlukast, zafirlukast, olmesartan medoxomil, and losartan potassium, exhibited more than 90% PGE2 uptake inhibition. Inhibitory affinity of suramin to OATP2A1 was the highest (IC50,2A1 of 0.17 µM), and its IC50 values to MRP4-mediated PGE2 transport (IC50,MRP4) and PGE2 synthesis in human U-937 cells treated with phorbol 12-myristate 13-acetate (IC50,Syn) were 73.6 and 336.7 times higher than IC50,2A1, respectively. Moreover, structure-activity relationship study in 29 nonsteroidal anti-inflammatory drugs contained in the library displayed inhibitory activities of anthranilic acid derivatives, but enhancing effects of propionic acid derivatives. These results demonstrate that suramin is a potent selective inhibitor of OATP2A1, providing a comprehensive information about drugs in clinical use that interact with OATP2A1.

Prostaglandin transporter (OATP2A1/SLCO2A1) contributes to local disposition of eicosapentaenoic acid-derived PGE3.

Eicosapentaenoic acid (EPA)-derived prostaglandin E3 (PGE3) possesses an anti-inflammatory effect; however, information for transporters that regulate its peri-cellular concentration is limited. The present study, therefore, aimed to clarify transporters involved in local disposition of PGE3. PGE3 uptake was assessed in HEK293 cells transfected with OATP2A1/SLCO2A1, OATP1B1/SLCO1B1, OATP2B1/SLCO2B1, OAT1/SLC22A6, OCT1/SLC22A1 or OCT2/SLC22A2 genes, compared with HEK293 cells transfected with plasmid vector alone (Mock). PGE3 uptake by OATP2A1-expressing HEK293 cells (HEK/2A1) was the highest and followed by HEK/1B1, while no significantly higher uptake of PGE3 than Mock cells was detected by other transporters. Saturation kinetics in PGE3 uptake by HEK/2A1 estimated the Km as 7.202 ± 0.595 µM, which was 22 times higher than that of PGE2 (Km=0.331 ± 0.131 µM). Furthermore, tissue disposition of PGE3 was examined in wild-type (WT) and Slco2a1-deficient (Slco2a1(-/-)) mice after oral administration of EPA ethyl ester (EPA-E) when they underwent intraperitoneal injection of endotoxin (e.g., lipopolysaccharide). PGE3 concentration was significantly higher in the lung, and tended to increase in the colon, stomach, and kidney of Slco2a1(-/-), compared to WT mice. Ratio of PGE2 metabolite 15-keto PGE2 over PGE2 concentration was significantly lower in the lung and colon of Slco2a1(-/-) than that of WT mice, suggesting that PGE3 metabolism is downregulated in Slco2a1(-/-) mice. In conclusion, PGE3 was found to be a substrate of OATP2A1, and local disposition of PGE3 could be regulated by OATP2A1 at least in the lung.