Journal: Biology Open
Article Title: Ammonia transport mediated by urea transporter A isoforms
doi: 10.1242/bio.061655
Figure Lengend Snippet: Summary of changes in surface pH following ammonia exposure of oocytes expressing wild-type or mutant mUT-A2, mUT-A3 , or mUT-B. (A) Changes in surface pH [ΔpH S(NH3) ] of oocytes expressing wild-type mUT-A2, mUT-A3 , or mUT-B following exposure to 0.5 M NH 3 /NH 4 + . (B) Changes in surface pH [ΔpH S(NH3) ] of water-injected oocytes following exposure to 0.5 M NH 3 /NH 4 + . (C) Surface pH measurements with water-injected ( n =10), water-injected with phloretin ( n =3), UT-A2 WT ( n =6), UT-A2 with phloretin ( n =5), UT-A2 T176V ( n =5), and UT-A2 T338V ( n =6) oocytes following exposure to 0.5 M NH 3 /NH 4 + . (D) Surface pH measurements with water-injected ( n =10), water-injected with phloretin ( n =3), UT-A3 WT ( n =3), UT-A3 with phloretin ( n =3), UT-A3 T246V ( n =4), and UT-A3 T408V ( n =4) oocytes following exposure to 0.5 M NH 3 /NH 4 + . (E) Surface pH measurements with water-injected ( n =10), water-injected with phloretin ( n =3), UT-B WT ( n =4), UT-B WT with phloretin ( n =4), UT-B T172V ( n =4), and UT-B T334V ( n =4) oocytes. Panels F–H on the right display background-subtracted results that yield channel-dependent activity. Oocytes expressing RhCG ( n =5; panels I and K) or AQP2 ( n =7; panels J and L) were used as positive and negative controls for assessing NH 3 transport. Gray circles indicate the values of each oocyte. Colored circles and triangles represent the averages of experiments performed on different days. The horizontal and vertical (error bars) lines on each plot represent the overall mean and the standard error of the mean of each group and condition. A standard two-tailed Student's t -test with Bonferroni correction was used to compare the difference between two means, as indicated above each graph. The significance level was set at P <0.0125.
Article Snippet: Experiments using oocytes injected with hAQP2 or hRhCG employed the same processing and detection protocols used for the UTs, except that a polyclonal anti-AQP2 antibody (Alpha Diagnostics, San Antonio, TX, USA) or a polyclonal antibody raised against the C-terminal region of RhCG ( ) and a goat anti-rabbit secondary antibody conjugated to HRP (AP132P; Millipore, Billerica, MA, USA) were used as reported previously ( , ).
Techniques: Expressing, Mutagenesis, Injection, Activity Assay, Two Tailed Test
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![Summary of changes in surface pH following ammonia exposure of oocytes expressing wild-type or mutant mUT-A2, mUT-A3 , or mUT-B. (A) Changes in surface pH [ΔpH S(NH3) ] of oocytes expressing wild-type mUT-A2, mUT-A3 , or mUT-B following exposure to 0.5 M NH 3 /NH 4 + . (B) Changes in surface pH [ΔpH S(NH3) ] of water-injected oocytes following exposure to 0.5 M NH 3 /NH 4 + . (C) Surface pH measurements with water-injected ( n =10), water-injected with phloretin ( n =3), UT-A2 WT ( n =6), UT-A2 with phloretin ( n =5), UT-A2 T176V ( n =5), and UT-A2 T338V ( n =6) oocytes following exposure to 0.5 M NH 3 /NH 4 + . (D) Surface pH measurements with water-injected ( n =10), water-injected with phloretin ( n =3), UT-A3 WT ( n =3), UT-A3 with phloretin ( n =3), UT-A3 T246V ( n =4), and UT-A3 T408V ( n =4) oocytes following exposure to 0.5 M NH 3 /NH 4 + . (E) Surface pH measurements with water-injected ( n =10), water-injected with phloretin ( n =3), UT-B WT ( n =4), UT-B WT with phloretin ( n =4), UT-B T172V ( n =4), and UT-B T334V ( n =4) oocytes. Panels F–H on the right display background-subtracted results that yield channel-dependent activity. Oocytes expressing RhCG ( n =5; panels I and K) or <t>AQP2</t> ( n =7; panels J and L) were used as positive and negative controls for assessing NH 3 transport. Gray circles indicate the values of each oocyte. Colored circles and triangles represent the averages of experiments performed on different days. The horizontal and vertical (error bars) lines on each plot represent the overall mean and the standard error of the mean of each group and condition. A standard two-tailed Student's t -test with Bonferroni correction was used to compare the difference between two means, as indicated above each graph. The significance level was set at P <0.0125.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_1576/pmc12171576/pmc12171576__biolopen-14-061655-g5.jpg)
