In situ hybridization and immunolocalization of concentrative and equilibrative nucleoside transporters in the human intestine, liver, kidneys, and placenta

Rajgopal Govindarajan, Aimee H. Bakken, Kelly L. Hudkins, Yurong Lai, F. Javier Casado, Marçal Pastor-Anglada, Chung Ming Tse, Jun Hayashi, Jashvant D. Unadkat

Research output: Contribution to journalArticle

Abstract

To better understand the role of human equilibrative (hENTs) and concentrative (hCNTs) nucleoside transporters in physiology and pharmacology, we investigated the regional, cellular, and spatial distribution of two hCNTs (hCNT1 and hCNT2) and two hENTs (hENT1 and hENT2) in four human tissues. Using in situ hybridization and immunohistochemical techniques, we found that the duodenum expressed hCNT1 and hCNT2 mRNAs in enterocytes and hENT1 and hENT2 mRNAs in crypt cells. In these cells, the hCNT and hENT proteins were predominantly localized in the apical and lateral membrane, respectively. Hepatocytes expressed higher levels of mRNAs of hENT1, hCNT1, and hENT2 than of hCNT2 and expressed all these proteins at hepatocyte cell borders and in the cytoplasm. While the kidney expressed hCNT1 and hCNT2 mRNAs in the proximal tubules, hENT1 and hENT2 mRNAs were present in the distal tubules, glomeruli, endothelial cells, and vascular smooth muscle cells. Proximal tubules adjacent to corticomedullary junctions expressed hENT1, hCNT1, and hCNT2 mRNA. Immunolocalization studies revealed predominant localization of hCNTs in the brush-border membrane of the proximal tubular epithelial cells and hENTs in the basolateral membrane of the distal tubular epithelial cells. Chorionic villi sections of human term placenta expressed mRNAs and proteins for hENT1 and hENT2 but only mRNA for hCNT2. Immunolocalization studies showed presence of hENT1 in the brush-border membrane of the syncytiotrophoblasts. These data are critical for a better understanding of the role of nucleoside transporters in the physiological and pharmacological effects of nucleosides and nucleoside drugs, respectively.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Regulatory Integrative and Comparative Physiology
Volume293
Issue number5
DOIs
StatePublished - Nov 2007

Fingerprint

Nucleoside Transport Proteins
Placenta
Intestines
In Situ Hybridization
Kidney
Messenger RNA
Liver
Membranes
Microvilli
Nucleosides
Hepatocytes
Epithelial Cells
Pharmacology
Chorionic Villi
Proteins
Enterocytes
Trophoblasts
Vascular Smooth Muscle
Duodenum
Smooth Muscle Myocytes

Keywords

  • Expression
  • Human tissue
  • Localization

ASJC Scopus subject areas

  • Physiology

Cite this

In situ hybridization and immunolocalization of concentrative and equilibrative nucleoside transporters in the human intestine, liver, kidneys, and placenta. / Govindarajan, Rajgopal; Bakken, Aimee H.; Hudkins, Kelly L.; Lai, Yurong; Casado, F. Javier; Pastor-Anglada, Marçal; Tse, Chung Ming; Hayashi, Jun; Unadkat, Jashvant D.

In: American Journal of Physiology - Regulatory Integrative and Comparative Physiology, Vol. 293, No. 5, 11.2007.

Research output: Contribution to journalArticle

Govindarajan, Rajgopal ; Bakken, Aimee H. ; Hudkins, Kelly L. ; Lai, Yurong ; Casado, F. Javier ; Pastor-Anglada, Marçal ; Tse, Chung Ming ; Hayashi, Jun ; Unadkat, Jashvant D. / In situ hybridization and immunolocalization of concentrative and equilibrative nucleoside transporters in the human intestine, liver, kidneys, and placenta. In: American Journal of Physiology - Regulatory Integrative and Comparative Physiology. 2007 ; Vol. 293, No. 5.
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AU - Bakken, Aimee H.

AU - Hudkins, Kelly L.

AU - Lai, Yurong

AU - Casado, F. Javier

AU - Pastor-Anglada, Marçal

AU - Tse, Chung Ming

AU - Hayashi, Jun

AU - Unadkat, Jashvant D.

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AB - To better understand the role of human equilibrative (hENTs) and concentrative (hCNTs) nucleoside transporters in physiology and pharmacology, we investigated the regional, cellular, and spatial distribution of two hCNTs (hCNT1 and hCNT2) and two hENTs (hENT1 and hENT2) in four human tissues. Using in situ hybridization and immunohistochemical techniques, we found that the duodenum expressed hCNT1 and hCNT2 mRNAs in enterocytes and hENT1 and hENT2 mRNAs in crypt cells. In these cells, the hCNT and hENT proteins were predominantly localized in the apical and lateral membrane, respectively. Hepatocytes expressed higher levels of mRNAs of hENT1, hCNT1, and hENT2 than of hCNT2 and expressed all these proteins at hepatocyte cell borders and in the cytoplasm. While the kidney expressed hCNT1 and hCNT2 mRNAs in the proximal tubules, hENT1 and hENT2 mRNAs were present in the distal tubules, glomeruli, endothelial cells, and vascular smooth muscle cells. Proximal tubules adjacent to corticomedullary junctions expressed hENT1, hCNT1, and hCNT2 mRNA. Immunolocalization studies revealed predominant localization of hCNTs in the brush-border membrane of the proximal tubular epithelial cells and hENTs in the basolateral membrane of the distal tubular epithelial cells. Chorionic villi sections of human term placenta expressed mRNAs and proteins for hENT1 and hENT2 but only mRNA for hCNT2. Immunolocalization studies showed presence of hENT1 in the brush-border membrane of the syncytiotrophoblasts. These data are critical for a better understanding of the role of nucleoside transporters in the physiological and pharmacological effects of nucleosides and nucleoside drugs, respectively.

KW - Expression

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