The mechanisms that regulate the activity of the neuron specific K+Cl- cotransporter (KCC2) remain poorly understood, despite the critical importance of this transporter in inhibitory synaptic transmission and plasticity. In this thesis I made three novel discoveries which reveal the cellular and molecular mechanisms of KCC2 regulation. First, I assayed the K+Cl- cotransport function of KCC2 under isotonic conditions and determined the molecular domain of the cotransporter required for constitutive Cl- transport in hippocampal neurons (Acton et al 2012). Specifically, I identified the 15 amino acid domain of the C-terminus in neurons that is responsible for the ability of KCC2 to cotransport K+Cl- under basal isotonic conditions, allowing it to remain constitutively active to create the steep Cl- gradient across the neuronal membrane required for synaptic inhibition. Secondly, I investigated a novel KCC2-interacting protein named Neto2 and determined its effect on the postsynaptic action of GABA (Ivakine et al 2013). I have found that Neto2, which is also an auxiliary protein of kainate-type ionotropic receptors, can also regulate the activity of the KCC2. Neto2 is required for neurons to maintain low [Cl-]i and strong synaptic inhibition. Third, I examined the functional relevance of the KCC2:Neto2:KAR multiprotein complex and found that this complex regulates the surface level membrane expression pattern of KCC2 and the stability of the cotransporter in the membrane. Moreover, I have provided the first evidence that the interactions of KCC2:Neto2:GluK2 regulate KCC2 via a PKC-mediated phosphorylation of the cotransporter. Taken together, these results resolve three novel mechanisms of KCC2 regulation: the identity of the key C-terminal domain of KCC2 required for isotonic transport, the functional significance of the KCC2:Neto2 interaction, and the mechanism by which the KCC2:Neto2:KAR complex regulates KCC2 expression and mobility in the neuronal membrane.

Ammonium chloride (NH4Cl) has been the primary preventive modality for struvite urolithiasis in goats. This study investigated the effect of continuous and pulse dose NH4Cl therapeutic regimens on urine pH in ten goats. The initial regimen (feed additive) consisted of 0.007% NH4Cl as a feed additive. Following this week long regime, the two treatment regimens were designed as a standard ten goat cross-over design. The first treatment regimen (continuous) consisted of daily administration of a titrated dosage of NH4Cl for ten days, followed by four days without treatment. The third treatment regimen (pulse) used daily administration of a titrated dose of NH4Cl for three consecutive days followed by four days without treatment for three treatment periods. Ammonium chloride dosages were titrated to result in a urine pH of < 6.5 (target level) prior to commencing treatment regimens. Urine pH was evaluated once daily during feed additive regimen and twice daily during the treatment regimens. A Bayesian methodology was used to determine the daily odds ratios for production of target urine pH during treatment regimens. The odds ratios were also calculated between pulse dosages during the pulse regimen. The feed additive regimen did not result in target urine pH within 7 days. Treatment with the continuous regimen resulted in target pH, however, pH returned to >6.5 within five days, (odds ratio 0.23-1.56 at Treatment Time 10). The odds ratios for each pulse period of the pulse dose regimen were 2.20-7.45, 0.41-1.68 and 1.59-5.62 respectively. The results of this study indicate that variability in response to therapy warrants titrating individual dosages of NH4Cl, continuous therapy results in a loss of effectiveness, and pulse dosage is effective in repeatedly producing a urine pH of <6.5.

Thesis(Master)--İzmir Institute of Technology,İzmir, 2004 / Includes bibliographical references (leaves. 101).

Thesis(M.S.)--Auburn University, 2006. / Abstract. Includes bibliographic references (p.92-99).

Thesis (M.S.)--Rochester Institute of Technology, 1987. / Typescript. Includes bibliographical references (leaves 76-79).

Thesis (Ph. D.)--Clark University, 1922. / Reprinted from an article by Charles A. Kraus and Conral C. Callis published in the Journal of the American Chemical Society, v. 45, no. 11, November 1923.

Thesis (Ph. D.)--University of Chicago, 1939. / Lithoprinted. Includes bibliographical references.

Thesis (Ph. D.)--University of Michigan, 1933. / By J. Gilbert Malone, A.L. Ferguson and L.O. Case. From the Journal of chemical physics, v. 1, December, 1933.

Thesis (M.S. in Materials Science Engineering)--Washington State University. / Includes bibliographical references.

Thesis (Ph. D.)--Clark University, 1922. / Reprinted from an article by Charles A. Kraus and Conral C. Callis published in the Journal of the American Chemical Society, v. 45, no. 11, November 1923.