title: 17β-Estradiol attenuates the activity of the glutamate transporter type 3 expressed in Xenopus oocytes.
abstract: Estrogen, a neuroactive sex hormone in the brain, enhances neuronal excitability and increases seizures. Glutamate transporters help in limiting the excitatory neurotransmission by uptaking glutamate from the synapses. We investigated the effects of 17β-estradiol on the activity of a glutamate transporter, excitatory amino acid transporter 3 (EAAT3), in Xenopus oocytes. EAAT3 was expressed in Xenopus oocytes by injection of rat EAAT3 mRNA. l-Glutamate (30 μM)-induced membrane currents mediated by EAAT3 were measured using the two-electrode voltage clamp technique. 17β-Estradiol reduced EAAT3 activity in a concentration- and time-dependent manner. 17β-Estradiol (10nM for 72h) significantly decreased V(max) but had no effect on K(m) of EAAT3 for glutamate. When 17β-estradiol treated oocytes were incubated with phorbol-12-myrisate-13-acetate, a protein kinase C (PKC) activator, 17β-estradiol-induced decrease in EAAT3 activity was abolished. Furthermore, in pretreatment of oocytes with chelerythrine or staurosporine, two PKC inhibitors, EAAT3 activity was significantly decreased. However, there was no statistical difference among the 17β-estradiol, PKC inhibitor, or 17β-estradiol plus PKC inhibitor groups. Likewise, wortmannin, a phosphatidylinositol 3-kinase (PI3K) inhibitor, significantly reduced basal EAAT3 activity, but the activity did not differ among the 17β-estradiol, wortmannin, or 17β-estradiol plus wortmannin groups. Estradiol receptor inhibitor, fulvestrant, did not change the reduced EAAT3 activity by 17β-estradiol. Our results suggest that 17β-estradiol decreases EAAT3 activity. PKC and PI3K seem to be involved in this effect, possibly not via estradiol receptors.
authors.affiliation: Department of Anesthesiology and Pain Medicine, Seoul National University Bundang Hospital, Seongnam-si, Gyeonggi-do, South Korea.
authors.forename: Hyo-Seok
authors.initials: HS
authors.lastname: Na
authors.forename: Hee-Pyeong
authors.initials: HP
authors.lastname: Park
authors.forename: Chong-Sung
authors.initials: CS
authors.lastname: Kim
authors.forename: Sang-Hwan
authors.initials: SH
authors.lastname: Do
authors.forename: Zhiyi
authors.initials: Z
authors.lastname: Zuo
authors.forename: Chong-Soo
authors.initials: CS
authors.lastname: Kim
mesh_terms: D000818:Animals; D004305:Dose-Response Relationship, Drug; D004958:Estradiol / Q000494:pharmacology*; D050580:Excitatory Amino Acid Transporter 3 / Q000037:antagonists & inhibitors* / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D009865:Oocytes / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects; D013997:Time Factors; D014981:Xenopus / Q000235:genetics*
chemical_list: D004958:Estradiol; D050580:Excitatory Amino Acid Transporter 3; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011493:Protein Kinase C; D047428:Protein Kinase Inhibitors; C493398:Slc1a1 protein, rat