Immunolocalization of hyperpolarization-activated cationic HCN1 and HCN3 channels in the rat nephron: regulation of HCN3 by potassium diets View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2015-10-29

AUTHORS

Zinaeli López-González, Cosete Ayala-Aguilera, Flavio Martinez-Morales, Othir Galicia-Cruz, Carolina Salvador-Hernández, José Pedraza-Chaverri, Mara Medeiros, Ana Maria Hernández, Laura I. Escobar

ABSTRACT

Hyperpolarization-activated cationic and cyclic nucleotide-gated channels (HCN) comprise four homologous subunits (HCN1–HCN4). HCN channels are found in excitable and non-excitable tissues in mammals. We have previously shown that HCN2 may transport ammonium (NH4+), besides sodium (Na+), in the rat distal nephron. In the present work, we identified HCN1 and HCN3 in the proximal tubule (PT) and HCN3 in the thick ascending limb of Henle (TALH) of the rat kidney. Immunoblot assays detected HCN1 (130 kDa) and HCN3 (90 KDa) and their truncated proteins C-terminal HCN1 (93 KDa) and N-terminal HCN3 (65 KDa) in enriched plasma membranes from cortex (CX) and outer medulla (OM), as well as in brush-border membrane vesicles. Immunofluorescence assays confirmed apical localization of HCN1 and HCN3 in the PT. HCN3 was also found at the basolateral membrane of TALH. We evaluated chronic changes in mineral dietary on HCN3 protein abundance. Animals were fed with three different diets: sodium-deficient (SD) diet, potassium-deficient (KD) diet, and high-potassium (HK) diet. Up-regulation of HCN3 was observed in OM by KD and in CX and OM by HK; the opposite effect occurred with the N-terminal truncated HCN3 in CX (KD) and OM (HK). SD diet did not produce any change. Since HCN channels activate with membrane hyperpolarization, our results suggest that HCN channels may play a role in the Na+–K+-ATPase activity, contributing to Na+, K+, and acid–base homeostasis in the rat kidney. More... »

PAGES

25-40

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s00418-015-1375-6

DOI

http://dx.doi.org/10.1007/s00418-015-1375-6

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1046118829

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/26515056


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31 schema:description Abstract Hyperpolarization-activated cationic and cyclic nucleotide-gated channels (HCN) comprise four homologous subunits (HCN1–HCN4). HCN channels are found in excitable and non-excitable tissues in mammals. We have previously shown that HCN2 may transport ammonium (NH4+), besides sodium (Na+), in the rat distal nephron. In the present work, we identified HCN1 and HCN3 in the proximal tubule (PT) and HCN3 in the thick ascending limb of Henle (TALH) of the rat kidney. Immunoblot assays detected HCN1 (130 kDa) and HCN3 (90 KDa) and their truncated proteins C-terminal HCN1 (93 KDa) and N-terminal HCN3 (65 KDa) in enriched plasma membranes from cortex (CX) and outer medulla (OM), as well as in brush-border membrane vesicles. Immunofluorescence assays confirmed apical localization of HCN1 and HCN3 in the PT. HCN3 was also found at the basolateral membrane of TALH. We evaluated chronic changes in mineral dietary on HCN3 protein abundance. Animals were fed with three different diets: sodium-deficient (SD) diet, potassium-deficient (KD) diet, and high-potassium (HK) diet. Up-regulation of HCN3 was observed in OM by KD and in CX and OM by HK; the opposite effect occurred with the N-terminal truncated HCN3 in CX (KD) and OM (HK). SD diet did not produce any change. Since HCN channels activate with membrane hyperpolarization, our results suggest that HCN channels may play a role in the Na+–K+-ATPase activity, contributing to Na+, K+, and acid–base homeostasis in the rat kidney.
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38 schema:keywords ATPase activity
39 HCN channels
40 HCN1
41 HCN2
42 HCN3
43 HCN3 channels
44 HK
45 Henle
46 Kd
47 N-terminal
48 SD diet
49 TALH
50 abundance
51 acid-base homeostasis
52 activity
53 ammonium
54 animals
55 apical localization
56 ascending limb
57 assays
58 basolateral membrane
59 brush border membrane vesicles
60 changes
61 channels
62 chronic changes
63 cortex
64 diet
65 dietary
66 different diets
67 distal nephron
68 effect
69 high potassium diet
70 homeostasis
71 homologous subunits
72 hyperpolarization
73 immunoblot assay
74 immunofluorescence assays
75 immunolocalization
76 kidney
77 limb
78 localization
79 mammals
80 medulla
81 membrane
82 membrane hyperpolarization
83 membrane vesicles
84 nephron
85 non-excitable tissues
86 opposite effect
87 outer medulla
88 plasma membrane
89 potassium diet
90 potassium-deficient diet
91 present work
92 protein abundance
93 proximal tubules
94 rat distal nephron
95 rat kidney
96 rat nephron
97 regulation
98 results
99 role
100 sodium
101 sodium-deficient diet
102 subunits
103 thick ascending limb
104 tissue
105 tubules
106 vesicles
107 work
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