Metallothionein isoform 3 and proximal tubule vectorial active transport

Metallothionein isoform 3 and proximal tubule vectorial active transport. Metallothionein isoform 3 (MT-3) is expressed in the proximal tubule cells of the human kidney. The goal of the present study was to further characterize the basal expression of MT-3 in the proximal tubule and to determine if...

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Bibliographic Details
Published in:Kidney international Vol. 61; no. 2; pp. 464 - 472
Main Authors: Kim, Doyeob, Garrett, Scott H., Sens, Mary Ann, Somji, Seema, Sens, Donald A.
Format: Journal Article
Language:English
Published: New York, NY Elsevier Inc 01.02.2002
Nature Publishing
Elsevier Limited
Subjects:
Aged
Biological and medical sciences
Biological Transport, Active - physiology
cell culture
Cell Division - physiology
Cells, Cultured
dome formation
Female
Fundamental and applied biological sciences. Psychology
Gene Expression - physiology
growth inhibitory factor
HK-2 cells
Humans
Kidney Tubules, Collecting - cytology
Kidney Tubules, Collecting - metabolism
Kidney Tubules, Proximal - cytology
Kidney Tubules, Proximal - metabolism
Loop of Henle - cytology
Loop of Henle - metabolism
Male
Metallothionein - genetics
Metallothionein - metabolism
Middle Aged
MT-3
Nerve Tissue Proteins - genetics
Nerve Tissue Proteins - metabolism
proximal tubule
RNA, Messenger - analysis
Transfection
Vertebrates: urinary system
MT-3
proximal tubule
growth inhibitory factor
cell culture
dome formation
HK-2 cells
Human
Renal tubule
Renal function
Messenger RNA
Gene
Established cell line
Isoforming
Reverse transcription polymerase chain reaction
Metallothionein
ISSN:0085-2538, 1523-1755
Online Access:Get full text
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Summary:Metallothionein isoform 3 and proximal tubule vectorial active transport. Metallothionein isoform 3 (MT-3) is expressed in the proximal tubule cells of the human kidney. The goal of the present study was to further characterize the basal expression of MT-3 in the proximal tubule and to determine if MT-3 participates in the maintenance of proximal tubule cell function. Expression of MT-3 mRNA was determined in the intact proximal tubule using microdissection and reverse transcription-polymerase chain reaction (RT-PCR). Basal expression of MT-3 mRNA and protein was determined in cultured human proximal tubule (HPT) cells and an immortalized proximal tubular cell line, HK-2 cells, using RT-PCR and immunoblotting. The MT-3 gene was stably transfected into the HK-2 cell line using the pcDNA3.1/Hygro (+) vector. MT-3 mRNA was detected in the proximal tubule of the in situ kidney with relative expression in excess to that of the β-actin housekeeping gene. The mortal HPT cells were shown to express both MT-3 mRNA and protein and to form domes, while immortal HK-2 cells were shown to have no expression of MT-3 mRNA and protein nor to form domes. The stable transfection of MT-3 in HK-2 restored MT-3 expression and dome formation to the HK-2 cells. MT-3 mRNA is present in the human proximal tubule, and MT-3 expression is involved in the transport function of a human renal cell line that retains properties of the proximal tubule.
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ISSN:0085-2538
1523-1755
DOI:10.1046/j.1523-1755.2002.00153.x