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Generation of a tenascin-C-CreER2 knockin mouse line for conditional DNA recombination in renal medullary interstitial cells.

He W, Xie Q, Wang Y, Chen J, Zhao M, Davis LS, Breyer MD, Gu G, Hao CM - PLoS ONE (2013)

Bottom Line: Renal medullary interstitial cells (RMIC) are specialized fibroblast-like cells that exert important functions in maintaining body fluid homeostasis and systemic blood pressure.After recombination (tenascin-C-CreER2(+/-)/ROSA26-lacZ(+/-) mice + tamoxifen), β-gal activity was restricted to the cells in the inner medulla of the kidney, and didn't co-localize with AQP2, consistent with selective Cre recombinase activity in RMICs.Cre activity was not obvious in other major organs or without tamoxifen treatment.

View Article: PubMed Central - PubMed

Affiliation: Gladstone Institute of Virology & Immunology, San Francisco, California, United States of America ; Nephrology Division, Vanderbilt University Medical Center School of Medicine, Nashville, Tennessee, United States of America.

ABSTRACT
Renal medullary interstitial cells (RMIC) are specialized fibroblast-like cells that exert important functions in maintaining body fluid homeostasis and systemic blood pressure. Here, we generated a RMIC specific tenascin-C promoter driven inducible CreER2 knockin mouse line with an EGFP reporter. Similar as endogenous tenascin-C expression, the reporter EGFP expression in the tenascin-C-CreER2(+/-) mice was observed in the inner medulla of the kidney, and co-localized with COX2 but not with AQP2 or AQP1, suggesting selective expression in RMICs. After recombination (tenascin-C-CreER2(+/-)/ROSA26-lacZ(+/-) mice + tamoxifen), β-gal activity was restricted to the cells in the inner medulla of the kidney, and didn't co-localize with AQP2, consistent with selective Cre recombinase activity in RMICs. Cre activity was not obvious in other major organs or without tamoxifen treatment. This inducible RMIC specific Cre mouse line should therefore provide a novel tool to manipulate genes of interest in RMICs.

Show MeSH
High levels of tenascin-C mRNA expression in the mouse renal medullary interstitium.(a, b) Dark field in situ hybridization pictures show high levels of tenascin-C mRNA in the mouse renal medulla with low levels in the renal cortex. Scale bar, 400 µm. (c, d) Bright field in situ hybridization pictures further show tenascin-C mRNA is highly expressed in the renal medullary interstitium. Scale bar, 100 µm.
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pone-0079839-g001: High levels of tenascin-C mRNA expression in the mouse renal medullary interstitium.(a, b) Dark field in situ hybridization pictures show high levels of tenascin-C mRNA in the mouse renal medulla with low levels in the renal cortex. Scale bar, 400 µm. (c, d) Bright field in situ hybridization pictures further show tenascin-C mRNA is highly expressed in the renal medullary interstitium. Scale bar, 100 µm.

Mentions: Our microarray study identified tenascin-C as one of RMIC specific gene products in the kidney (Table 1). Selective expression of tenascin-C mRNA in the renal medullary interstitial cells was further confirmed by in situ hybridization (Figure 1). The data are consistent with published studies showing tenascin-C expression in the renal medullary stroma of the adult mice [10]–[12].


Generation of a tenascin-C-CreER2 knockin mouse line for conditional DNA recombination in renal medullary interstitial cells.

He W, Xie Q, Wang Y, Chen J, Zhao M, Davis LS, Breyer MD, Gu G, Hao CM - PLoS ONE (2013)

High levels of tenascin-C mRNA expression in the mouse renal medullary interstitium.(a, b) Dark field in situ hybridization pictures show high levels of tenascin-C mRNA in the mouse renal medulla with low levels in the renal cortex. Scale bar, 400 µm. (c, d) Bright field in situ hybridization pictures further show tenascin-C mRNA is highly expressed in the renal medullary interstitium. Scale bar, 100 µm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3823583&req=5

pone-0079839-g001: High levels of tenascin-C mRNA expression in the mouse renal medullary interstitium.(a, b) Dark field in situ hybridization pictures show high levels of tenascin-C mRNA in the mouse renal medulla with low levels in the renal cortex. Scale bar, 400 µm. (c, d) Bright field in situ hybridization pictures further show tenascin-C mRNA is highly expressed in the renal medullary interstitium. Scale bar, 100 µm.
Mentions: Our microarray study identified tenascin-C as one of RMIC specific gene products in the kidney (Table 1). Selective expression of tenascin-C mRNA in the renal medullary interstitial cells was further confirmed by in situ hybridization (Figure 1). The data are consistent with published studies showing tenascin-C expression in the renal medullary stroma of the adult mice [10]–[12].

Bottom Line: Renal medullary interstitial cells (RMIC) are specialized fibroblast-like cells that exert important functions in maintaining body fluid homeostasis and systemic blood pressure.After recombination (tenascin-C-CreER2(+/-)/ROSA26-lacZ(+/-) mice + tamoxifen), β-gal activity was restricted to the cells in the inner medulla of the kidney, and didn't co-localize with AQP2, consistent with selective Cre recombinase activity in RMICs.Cre activity was not obvious in other major organs or without tamoxifen treatment.

View Article: PubMed Central - PubMed

Affiliation: Gladstone Institute of Virology & Immunology, San Francisco, California, United States of America ; Nephrology Division, Vanderbilt University Medical Center School of Medicine, Nashville, Tennessee, United States of America.

ABSTRACT
Renal medullary interstitial cells (RMIC) are specialized fibroblast-like cells that exert important functions in maintaining body fluid homeostasis and systemic blood pressure. Here, we generated a RMIC specific tenascin-C promoter driven inducible CreER2 knockin mouse line with an EGFP reporter. Similar as endogenous tenascin-C expression, the reporter EGFP expression in the tenascin-C-CreER2(+/-) mice was observed in the inner medulla of the kidney, and co-localized with COX2 but not with AQP2 or AQP1, suggesting selective expression in RMICs. After recombination (tenascin-C-CreER2(+/-)/ROSA26-lacZ(+/-) mice + tamoxifen), β-gal activity was restricted to the cells in the inner medulla of the kidney, and didn't co-localize with AQP2, consistent with selective Cre recombinase activity in RMICs. Cre activity was not obvious in other major organs or without tamoxifen treatment. This inducible RMIC specific Cre mouse line should therefore provide a novel tool to manipulate genes of interest in RMICs.

Show MeSH