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In this study, we used multiple-label immunocytochemistry to investigate the possibility of changes in the \u03b3-aminobutyric acid (GABA) neurotransmitter system at two key components of the GnRH pulse generator in prenatal T-treated sheep: kisspeptin\/neurokinin B\/dynorphin (KNDy) neurons of the arcuate nucleus, and GnRH neurons in the preoptic area (POA) and mediobasal hypothalamus (MBH). We observed a significant decrease and increase, respectively, in the number of GABAergic synapses onto POA and MBH GnRH neurons in prenatal T-treated ewes; additionally, there was a significant increase in the number of GABAergic inputs onto KNDy neurons. To determine the actions of GABA on GnRH and KNDy neurons, we examined colocalization with the chloride transporters NKCC1 and KCC2, which indicate stimulatory or inhibitory activation of neurons by GABA, respectively. Most GnRH neurons in both POA and MBH colocalized NKCC1 cotransporter whereas none contained the KCC2 cotransporter. Most KNDy neurons colocalized either NKCC1 or KCC2, and 28% of the KNDy population contained NKCC1 alone. Therefore, we suggest that, as in the mouse, GABA in the sheep is stimulatory to GnRH neurons, as well as to a subset of KNDy neurons. Increased numbers of stimulatory GABAergic inputs to both MBH GnRH and KNDy neurons in prenatal T-treated animals may contribute to alterations in steroid feedback control and increased GnRH\/LH pulse frequency seen in this animal model of PCOS.<\/jats:p>","DOI":"10.1210\/en.2019-00137","type":"journal-article","created":{"date-parts":[[2019,8,16]],"date-time":"2019-08-16T12:18:03Z","timestamp":1565957883000},"page":"2529-2542","source":"Crossref","is-referenced-by-count":56,"title":["Prenatal Testosterone Exposure Alters GABAergic Synaptic Inputs to GnRH and KNDy Neurons in a Sheep Model of Polycystic Ovarian Syndrome"],"prefix":"10.1210","volume":"160","author":[{"given":"Danielle T","family":"Porter","sequence":"first","affiliation":[{"name":"Graduate Program in Neuroscience, Department of Neurobiology and Anatomical Sciences, University of Mississippi Medical Center, Jackson, Mississippi"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aleisha 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