title: Chloride Accumulators NKCC1 and AE2 in Mouse GnRH Neurons: Implications for GABAA Mediated Excitation.
abstract: A developmental "switch" in chloride transporters occurs in most neurons resulting in GABAA mediated hyperpolarization in the adult. However, several neuronal cell subtypes maintain primarily depolarizing responses to GABAA receptor activation. Among this group are gonadotropin-releasing hormone-1 (GnRH) neurons, which control puberty and reproduction. NKCC1 is the primary chloride accumulator in neurons, expressed at high levels early in development and contributes to depolarization after GABAA receptor activation. In contrast, KCC2 is the primary chloride extruder in neurons, expressed at high levels in the adult and contributes to hyperpolarization after GABAA receptor activation. Anion exchangers (AEs) are also potential modulators of responses to GABAA activation since they accumulate chloride and extrude bicarbonate. To evaluate the mechanism(s) underlying GABAA mediated depolarization, GnRH neurons were analyzed for 1) expression of chloride transporters and AEs in embryonic, pre-pubertal, and adult mice 2) responses to GABAA receptor activation in NKCC1-/- mice and 3) function of AEs in these responses. At all ages, GnRH neurons were immunopositive for NKCC1 and AE2 but not KCC2 or AE3. Using explants, calcium imaging and gramicidin perforated patch clamp techniques we found that GnRH neurons from NKCC1-/- mice retained relatively normal responses to the GABAA agonist muscimol. However, acute pharmacological inhibition of NKCC1 with bumetanide eliminated the depolarization/calcium response to muscimol in 40% of GnRH neurons from WT mice. In the remaining GnRH neurons, HCO3- mediated mechanisms accounted for the remaining calcium responses to muscimol. Collectively these data reveal mechanisms responsible for maintaining depolarizing GABAA mediated transmission in GnRH neurons.
authors.affiliation: Cellular and Developmental Neurobiology Section, NINDS/NIH, Bethesda, Maryland, United States of America.
authors.forename: Carol
authors.initials: C
authors.lastname: Taylor-Burds
authors.affiliation: Cellular and Developmental Neurobiology Section, NINDS/NIH, Bethesda, Maryland, United States of America.
authors.forename: Paul
authors.initials: P
authors.lastname: Cheng
authors.affiliation: Cellular and Developmental Neurobiology Section, NINDS/NIH, Bethesda, Maryland, United States of America.
authors.forename: Susan
authors.initials: S
authors.lastname: Wray
mesh_terms: D000818:Animals; D001639:Bicarbonates / Q000378:metabolism; D002034:Bumetanide / Q000494:pharmacology; D027963:Chloride-Bicarbonate Antiporters / Q000378:metabolism; D002712:Chlorides / Q000378:metabolism*; D005260:Female; D007987:Gonadotropin-Releasing Hormone / Q000378:metabolism*; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D026901:Membrane Transport Proteins / Q000378:metabolism*; D051379:Mice; D009118:Muscimol / Q000494:pharmacology; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D011963:Receptors, GABA-A / Q000378:metabolism*; D012741:Sexual Maturation / Q000187:drug effects / Q000502:physiology; D064506:Solute Carrier Family 12, Member 2 / Q000378:metabolism*
chemical_list: D001639:Bicarbonates; D027963:Chloride-Bicarbonate Antiporters; D002712:Chlorides; D026901:Membrane Transport Proteins; D011963:Receptors, GABA-A; C578878:Slc12a2 protein, mouse; C579495:Slc4a2 protein, mouse; D064506:Solute Carrier Family 12, Member 2; D002034:Bumetanide; D009118:Muscimol; D007987:Gonadotropin-Releasing Hormone