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Novel approach for the detection of tubular cell migration into the interstitium during renal fibrosis in rats.

Nakasatomi M, Maeshima A, Mishima K, Ikeuchi H, Sakairi T, Kaneko Y, Hiromura K, Nojima Y - Fibrogenesis Tissue Repair (2015)

Bottom Line: The number of BrdU-positive cells migrating into the interstitium significantly increased and peaked at 8 days after UUO.Long-term BrdU labeling marked some of the proximal tubular cells and enabled us to detect tubular cell migration into the interstitium after UUO.This simple method might be useful to detect EMT in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine and Clinical Science, Gunma University Graduate School of Medicine, 3-39-15 Showa, Maebashi, 371-8511 Japan.

ABSTRACT

Background: The process of epithelial-mesenchymal transition (EMT), which is generally defined by phenotypic changes of injured tubules such as loss of epithelial markers or acquisition of mesenchymal markers, implies various activating steps, including proliferation, migration, and ability to produce extracellular matrix proteins. We established here a novel approach for the detection of tubular cell migration into the interstitium during renal fibrosis in vivo.

Results: Using an osmotic pump, bromodeoxyuridine (BrdU) was continuously given to 7-week-old Wistar rats for 4 weeks, and BrdU-positive cells were detected by immunostaining. BrdU-positive cells were present in aquaporin-1-positive proximal tubules, but not in the interstitium of BrdU-treated rat kidneys. After unilateral ureteral obstruction (UUO), some BrdU-positive tubular cells protruded from the basement membrane and migrated into the interstitium. Interstitial BrdU-positive cells were co-localized with alpha-smooth muscle actin, fibroblast specific protein-1, vimentin, and type I collagen, but not with CD68 or CD3. No BrdU-positive cells were observed in the interstitium of sham-operated kidneys. The number of BrdU-positive cells migrating into the interstitium significantly increased and peaked at 8 days after UUO.

Conclusions: Long-term BrdU labeling marked some of the proximal tubular cells and enabled us to detect tubular cell migration into the interstitium after UUO. This simple method might be useful to detect EMT in vivo.

No MeSH data available.


Related in: MedlinePlus

Migration of BrdU-positive tubular cells into the interstitium of the UUO kidneys. a Masson’s trichrome staining (a–d) and EVG staining (e–f) of kidney sections. (a, e) Normal kidney; (b, f) contralateral kidney, 10 days; (c, g) UUO kidney, 6 days; and (d, h) UUO kidney, 10 days. Magnification, ×200. b Localization of BrdU-positive cells in the normal (a), contralateral (b), and UUO (c–f) kidneys of rats with long-term BrdU labeling. Magnification, ×1000. Arrowheads indicate BrdU-positive cells protruding from TBM or migrating into interstitium. c Destruction of tubular basement membrane in the UUO kidneys of rats with long-term BrdU labeling. BrdU (red), markers (green), and DAPI (blue). Magnification, ×1000
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Fig3: Migration of BrdU-positive tubular cells into the interstitium of the UUO kidneys. a Masson’s trichrome staining (a–d) and EVG staining (e–f) of kidney sections. (a, e) Normal kidney; (b, f) contralateral kidney, 10 days; (c, g) UUO kidney, 6 days; and (d, h) UUO kidney, 10 days. Magnification, ×200. b Localization of BrdU-positive cells in the normal (a), contralateral (b), and UUO (c–f) kidneys of rats with long-term BrdU labeling. Magnification, ×1000. Arrowheads indicate BrdU-positive cells protruding from TBM or migrating into interstitium. c Destruction of tubular basement membrane in the UUO kidneys of rats with long-term BrdU labeling. BrdU (red), markers (green), and DAPI (blue). Magnification, ×1000

Mentions: Masson’s trichrome staining (Fig. 3a (a–d)) as well as elastin-van Gieson (EVG) staining (Fig. 3a (e–h)) demonstrated the deposition of extracellular matrix in the UUO kidneys at 6 days (Fig. 3a (c, g)) or at 10 days (Fig. 3a (d, h)) after operation, but not in the normal kidneys (Fig. 3a (a, e)) or contralateral kidneys (Fig. 3a (b, f)), demonstrating the establishment of renal fibrosis in this rat UUO model.Fig. 3


Novel approach for the detection of tubular cell migration into the interstitium during renal fibrosis in rats.

Nakasatomi M, Maeshima A, Mishima K, Ikeuchi H, Sakairi T, Kaneko Y, Hiromura K, Nojima Y - Fibrogenesis Tissue Repair (2015)

Migration of BrdU-positive tubular cells into the interstitium of the UUO kidneys. a Masson’s trichrome staining (a–d) and EVG staining (e–f) of kidney sections. (a, e) Normal kidney; (b, f) contralateral kidney, 10 days; (c, g) UUO kidney, 6 days; and (d, h) UUO kidney, 10 days. Magnification, ×200. b Localization of BrdU-positive cells in the normal (a), contralateral (b), and UUO (c–f) kidneys of rats with long-term BrdU labeling. Magnification, ×1000. Arrowheads indicate BrdU-positive cells protruding from TBM or migrating into interstitium. c Destruction of tubular basement membrane in the UUO kidneys of rats with long-term BrdU labeling. BrdU (red), markers (green), and DAPI (blue). Magnification, ×1000
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4496823&req=5

Fig3: Migration of BrdU-positive tubular cells into the interstitium of the UUO kidneys. a Masson’s trichrome staining (a–d) and EVG staining (e–f) of kidney sections. (a, e) Normal kidney; (b, f) contralateral kidney, 10 days; (c, g) UUO kidney, 6 days; and (d, h) UUO kidney, 10 days. Magnification, ×200. b Localization of BrdU-positive cells in the normal (a), contralateral (b), and UUO (c–f) kidneys of rats with long-term BrdU labeling. Magnification, ×1000. Arrowheads indicate BrdU-positive cells protruding from TBM or migrating into interstitium. c Destruction of tubular basement membrane in the UUO kidneys of rats with long-term BrdU labeling. BrdU (red), markers (green), and DAPI (blue). Magnification, ×1000
Mentions: Masson’s trichrome staining (Fig. 3a (a–d)) as well as elastin-van Gieson (EVG) staining (Fig. 3a (e–h)) demonstrated the deposition of extracellular matrix in the UUO kidneys at 6 days (Fig. 3a (c, g)) or at 10 days (Fig. 3a (d, h)) after operation, but not in the normal kidneys (Fig. 3a (a, e)) or contralateral kidneys (Fig. 3a (b, f)), demonstrating the establishment of renal fibrosis in this rat UUO model.Fig. 3

Bottom Line: The number of BrdU-positive cells migrating into the interstitium significantly increased and peaked at 8 days after UUO.Long-term BrdU labeling marked some of the proximal tubular cells and enabled us to detect tubular cell migration into the interstitium after UUO.This simple method might be useful to detect EMT in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine and Clinical Science, Gunma University Graduate School of Medicine, 3-39-15 Showa, Maebashi, 371-8511 Japan.

ABSTRACT

Background: The process of epithelial-mesenchymal transition (EMT), which is generally defined by phenotypic changes of injured tubules such as loss of epithelial markers or acquisition of mesenchymal markers, implies various activating steps, including proliferation, migration, and ability to produce extracellular matrix proteins. We established here a novel approach for the detection of tubular cell migration into the interstitium during renal fibrosis in vivo.

Results: Using an osmotic pump, bromodeoxyuridine (BrdU) was continuously given to 7-week-old Wistar rats for 4 weeks, and BrdU-positive cells were detected by immunostaining. BrdU-positive cells were present in aquaporin-1-positive proximal tubules, but not in the interstitium of BrdU-treated rat kidneys. After unilateral ureteral obstruction (UUO), some BrdU-positive tubular cells protruded from the basement membrane and migrated into the interstitium. Interstitial BrdU-positive cells were co-localized with alpha-smooth muscle actin, fibroblast specific protein-1, vimentin, and type I collagen, but not with CD68 or CD3. No BrdU-positive cells were observed in the interstitium of sham-operated kidneys. The number of BrdU-positive cells migrating into the interstitium significantly increased and peaked at 8 days after UUO.

Conclusions: Long-term BrdU labeling marked some of the proximal tubular cells and enabled us to detect tubular cell migration into the interstitium after UUO. This simple method might be useful to detect EMT in vivo.

No MeSH data available.


Related in: MedlinePlus