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A , Quantitative RT‐PCR analysis of <t>TACR1</t> , TACR2 , and TACR3 mRNAs in 42 independent APAs. Each dot indicates 1 adenoma. Data are presented as mean±SEM. P =0.012; P <0.0001. B , Comparison of the expression levels of the TACR1 gene between different APA subgroups. classical APAs; nonclassical APAs (left panel) P =0.045; NKD and KCNJ5 (middle panel); CYP11B2‐positive nodules in nonclassical APAs (<2 nodules vs ≥2 nodules, right panel). C through F , Distribution of NK1R immunostaining in APA and adjacent adrenal tissue representative of n=56 independent APAs. C, E, F, NK1R staining in a classical APA and its adjacent adrenal cortex. C , Overall NK1R staining pattern in an APA and the adjacent adrenal cortex. D , Closer view of the NK1R staining in a different classical APA, highlighting areas of staining present in the zona glomerulosa or organized in micronodules within the adrenal tissue adjacent to the adenoma (arrows). E , Close‐up view of NK1R‐positive staining in the ZG and nerve ganglia in the adrenal cortex adjacent to the adenoma). F , High magnification view illustrating the distribution of NK1R staining in a group of adenoma cells. Arrow and arrowhead show membrane and cytoplasmic NK1R staining, respectively. APA indicates aldosterone‐producing adenoma; C, classical; Ca, capsule; G, nerve ganglia; KCNJ5 , KCNJ5 mutation detected; NC, nonclassical; NKD, no KCNJ5 mutation detected; NK1R, neurokinin type 1 receptor; RT‐PCR, reverse transcription polymerase chain reaction; and ZG, zona glomerulosa.
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A , Quantitative RT‐PCR analysis of <t>TACR1</t> , TACR2 , and TACR3 mRNAs in 42 independent APAs. Each dot indicates 1 adenoma. Data are presented as mean±SEM. P =0.012; P <0.0001. B , Comparison of the expression levels of the TACR1 gene between different APA subgroups. classical APAs; nonclassical APAs (left panel) P =0.045; NKD and KCNJ5 (middle panel); CYP11B2‐positive nodules in nonclassical APAs (<2 nodules vs ≥2 nodules, right panel). C through F , Distribution of NK1R immunostaining in APA and adjacent adrenal tissue representative of n=56 independent APAs. C, E, F, NK1R staining in a classical APA and its adjacent adrenal cortex. C , Overall NK1R staining pattern in an APA and the adjacent adrenal cortex. D , Closer view of the NK1R staining in a different classical APA, highlighting areas of staining present in the zona glomerulosa or organized in micronodules within the adrenal tissue adjacent to the adenoma (arrows). E , Close‐up view of NK1R‐positive staining in the ZG and nerve ganglia in the adrenal cortex adjacent to the adenoma). F , High magnification view illustrating the distribution of NK1R staining in a group of adenoma cells. Arrow and arrowhead show membrane and cytoplasmic NK1R staining, respectively. APA indicates aldosterone‐producing adenoma; C, classical; Ca, capsule; G, nerve ganglia; KCNJ5 , KCNJ5 mutation detected; NC, nonclassical; NKD, no KCNJ5 mutation detected; NK1R, neurokinin type 1 receptor; RT‐PCR, reverse transcription polymerase chain reaction; and ZG, zona glomerulosa.
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A , Quantitative RT‐PCR analysis of <t>TACR1</t> , TACR2 , and TACR3 mRNAs in 42 independent APAs. Each dot indicates 1 adenoma. Data are presented as mean±SEM. P =0.012; P <0.0001. B , Comparison of the expression levels of the TACR1 gene between different APA subgroups. classical APAs; nonclassical APAs (left panel) P =0.045; NKD and KCNJ5 (middle panel); CYP11B2‐positive nodules in nonclassical APAs (<2 nodules vs ≥2 nodules, right panel). C through F , Distribution of NK1R immunostaining in APA and adjacent adrenal tissue representative of n=56 independent APAs. C, E, F, NK1R staining in a classical APA and its adjacent adrenal cortex. C , Overall NK1R staining pattern in an APA and the adjacent adrenal cortex. D , Closer view of the NK1R staining in a different classical APA, highlighting areas of staining present in the zona glomerulosa or organized in micronodules within the adrenal tissue adjacent to the adenoma (arrows). E , Close‐up view of NK1R‐positive staining in the ZG and nerve ganglia in the adrenal cortex adjacent to the adenoma). F , High magnification view illustrating the distribution of NK1R staining in a group of adenoma cells. Arrow and arrowhead show membrane and cytoplasmic NK1R staining, respectively. APA indicates aldosterone‐producing adenoma; C, classical; Ca, capsule; G, nerve ganglia; KCNJ5 , KCNJ5 mutation detected; NC, nonclassical; NKD, no KCNJ5 mutation detected; NK1R, neurokinin type 1 receptor; RT‐PCR, reverse transcription polymerase chain reaction; and ZG, zona glomerulosa.
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A , Quantitative RT‐PCR analysis of <t>TACR1</t> , TACR2 , and TACR3 mRNAs in 42 independent APAs. Each dot indicates 1 adenoma. Data are presented as mean±SEM. P =0.012; P <0.0001. B , Comparison of the expression levels of the TACR1 gene between different APA subgroups. classical APAs; nonclassical APAs (left panel) P =0.045; NKD and KCNJ5 (middle panel); CYP11B2‐positive nodules in nonclassical APAs (<2 nodules vs ≥2 nodules, right panel). C through F , Distribution of NK1R immunostaining in APA and adjacent adrenal tissue representative of n=56 independent APAs. C, E, F, NK1R staining in a classical APA and its adjacent adrenal cortex. C , Overall NK1R staining pattern in an APA and the adjacent adrenal cortex. D , Closer view of the NK1R staining in a different classical APA, highlighting areas of staining present in the zona glomerulosa or organized in micronodules within the adrenal tissue adjacent to the adenoma (arrows). E , Close‐up view of NK1R‐positive staining in the ZG and nerve ganglia in the adrenal cortex adjacent to the adenoma). F , High magnification view illustrating the distribution of NK1R staining in a group of adenoma cells. Arrow and arrowhead show membrane and cytoplasmic NK1R staining, respectively. APA indicates aldosterone‐producing adenoma; C, classical; Ca, capsule; G, nerve ganglia; KCNJ5 , KCNJ5 mutation detected; NC, nonclassical; NKD, no KCNJ5 mutation detected; NK1R, neurokinin type 1 receptor; RT‐PCR, reverse transcription polymerase chain reaction; and ZG, zona glomerulosa.
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A , Quantitative RT‐PCR analysis of <t>TACR1</t> , TACR2 , and TACR3 mRNAs in 42 independent APAs. Each dot indicates 1 adenoma. Data are presented as mean±SEM. P =0.012; P <0.0001. B , Comparison of the expression levels of the TACR1 gene between different APA subgroups. classical APAs; nonclassical APAs (left panel) P =0.045; NKD and KCNJ5 (middle panel); CYP11B2‐positive nodules in nonclassical APAs (<2 nodules vs ≥2 nodules, right panel). C through F , Distribution of NK1R immunostaining in APA and adjacent adrenal tissue representative of n=56 independent APAs. C, E, F, NK1R staining in a classical APA and its adjacent adrenal cortex. C , Overall NK1R staining pattern in an APA and the adjacent adrenal cortex. D , Closer view of the NK1R staining in a different classical APA, highlighting areas of staining present in the zona glomerulosa or organized in micronodules within the adrenal tissue adjacent to the adenoma (arrows). E , Close‐up view of NK1R‐positive staining in the ZG and nerve ganglia in the adrenal cortex adjacent to the adenoma). F , High magnification view illustrating the distribution of NK1R staining in a group of adenoma cells. Arrow and arrowhead show membrane and cytoplasmic NK1R staining, respectively. APA indicates aldosterone‐producing adenoma; C, classical; Ca, capsule; G, nerve ganglia; KCNJ5 , KCNJ5 mutation detected; NC, nonclassical; NKD, no KCNJ5 mutation detected; NK1R, neurokinin type 1 receptor; RT‐PCR, reverse transcription polymerase chain reaction; and ZG, zona glomerulosa.
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A , Quantitative RT‐PCR analysis of <t>TACR1</t> , TACR2 , and TACR3 mRNAs in 42 independent APAs. Each dot indicates 1 adenoma. Data are presented as mean±SEM. P =0.012; P <0.0001. B , Comparison of the expression levels of the TACR1 gene between different APA subgroups. classical APAs; nonclassical APAs (left panel) P =0.045; NKD and KCNJ5 (middle panel); CYP11B2‐positive nodules in nonclassical APAs (<2 nodules vs ≥2 nodules, right panel). C through F , Distribution of NK1R immunostaining in APA and adjacent adrenal tissue representative of n=56 independent APAs. C, E, F, NK1R staining in a classical APA and its adjacent adrenal cortex. C , Overall NK1R staining pattern in an APA and the adjacent adrenal cortex. D , Closer view of the NK1R staining in a different classical APA, highlighting areas of staining present in the zona glomerulosa or organized in micronodules within the adrenal tissue adjacent to the adenoma (arrows). E , Close‐up view of NK1R‐positive staining in the ZG and nerve ganglia in the adrenal cortex adjacent to the adenoma). F , High magnification view illustrating the distribution of NK1R staining in a group of adenoma cells. Arrow and arrowhead show membrane and cytoplasmic NK1R staining, respectively. APA indicates aldosterone‐producing adenoma; C, classical; Ca, capsule; G, nerve ganglia; KCNJ5 , KCNJ5 mutation detected; NC, nonclassical; NKD, no KCNJ5 mutation detected; NK1R, neurokinin type 1 receptor; RT‐PCR, reverse transcription polymerase chain reaction; and ZG, zona glomerulosa.
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A , Quantitative RT‐PCR analysis of TACR1 , TACR2 , and TACR3 mRNAs in 42 independent APAs. Each dot indicates 1 adenoma. Data are presented as mean±SEM. P =0.012; P <0.0001. B , Comparison of the expression levels of the TACR1 gene between different APA subgroups. classical APAs; nonclassical APAs (left panel) P =0.045; NKD and KCNJ5 (middle panel); CYP11B2‐positive nodules in nonclassical APAs (<2 nodules vs ≥2 nodules, right panel). C through F , Distribution of NK1R immunostaining in APA and adjacent adrenal tissue representative of n=56 independent APAs. C, E, F, NK1R staining in a classical APA and its adjacent adrenal cortex. C , Overall NK1R staining pattern in an APA and the adjacent adrenal cortex. D , Closer view of the NK1R staining in a different classical APA, highlighting areas of staining present in the zona glomerulosa or organized in micronodules within the adrenal tissue adjacent to the adenoma (arrows). E , Close‐up view of NK1R‐positive staining in the ZG and nerve ganglia in the adrenal cortex adjacent to the adenoma). F , High magnification view illustrating the distribution of NK1R staining in a group of adenoma cells. Arrow and arrowhead show membrane and cytoplasmic NK1R staining, respectively. APA indicates aldosterone‐producing adenoma; C, classical; Ca, capsule; G, nerve ganglia; KCNJ5 , KCNJ5 mutation detected; NC, nonclassical; NKD, no KCNJ5 mutation detected; NK1R, neurokinin type 1 receptor; RT‐PCR, reverse transcription polymerase chain reaction; and ZG, zona glomerulosa.

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Regulation of Aldosterone Secretion by Substance P and the Neurokinin Type 1 Receptor in Aldosterone‐Producing Adenomas

doi: 10.1161/JAHA.125.045539

Figure Lengend Snippet: A , Quantitative RT‐PCR analysis of TACR1 , TACR2 , and TACR3 mRNAs in 42 independent APAs. Each dot indicates 1 adenoma. Data are presented as mean±SEM. P =0.012; P <0.0001. B , Comparison of the expression levels of the TACR1 gene between different APA subgroups. classical APAs; nonclassical APAs (left panel) P =0.045; NKD and KCNJ5 (middle panel); CYP11B2‐positive nodules in nonclassical APAs (<2 nodules vs ≥2 nodules, right panel). C through F , Distribution of NK1R immunostaining in APA and adjacent adrenal tissue representative of n=56 independent APAs. C, E, F, NK1R staining in a classical APA and its adjacent adrenal cortex. C , Overall NK1R staining pattern in an APA and the adjacent adrenal cortex. D , Closer view of the NK1R staining in a different classical APA, highlighting areas of staining present in the zona glomerulosa or organized in micronodules within the adrenal tissue adjacent to the adenoma (arrows). E , Close‐up view of NK1R‐positive staining in the ZG and nerve ganglia in the adrenal cortex adjacent to the adenoma). F , High magnification view illustrating the distribution of NK1R staining in a group of adenoma cells. Arrow and arrowhead show membrane and cytoplasmic NK1R staining, respectively. APA indicates aldosterone‐producing adenoma; C, classical; Ca, capsule; G, nerve ganglia; KCNJ5 , KCNJ5 mutation detected; NC, nonclassical; NKD, no KCNJ5 mutation detected; NK1R, neurokinin type 1 receptor; RT‐PCR, reverse transcription polymerase chain reaction; and ZG, zona glomerulosa.

Article Snippet: Each sample was analyzed in duplicates, and cDNA quantification was normalized to PPIA (cyclophilin) using the ΔCt method and standard curves generated from polyA mRNAs or TACR1 human quantitative PCR template ( HK210362 , Origene, Rockville, USA).

Techniques: Quantitative RT-PCR, Comparison, Expressing, Immunostaining, Staining, Membrane, Mutagenesis, Reverse Transcription Polymerase Chain Reaction, Reverse Transcription, Polymerase Chain Reaction