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Variable Expression of Neural Cell Adhesion Molecule Isoforms in Renal Tissue: Possible Role in Incipient Renal Fibrosis.

Marković-Lipkovski J, Životić M, Müller CA, Tampe B, Ćirović S, Vještica J, Tomanović N, Zeisberg M, Müller GA - PLoS ONE (2015)

Bottom Line: Applying qRT-PCR on pure NCAM+ cells population, obtained by laser capture microdissection, significant mRNA over-expression of NCAM140kD isoform was found in NCAM+ cells within incipient IRF (p = 0.004), while NCAM120kD and NCAM180kD isoforms were not changed significantly (p = 0.750; p = 0.704; respectively).Simultaneously, qRT-PCR also showed significant αSMA (p = 0.014) and SLUG (p = 0.004) mRNAs up-regulation within the NCAM+ cells of incipient IRF, as well as highly decreased matrix metalloproteinases (MMP) -2 and -9 mRNAs (p = 0.028; p = 0.036; respectively).Fibroblast growth factor receptor 1 (FGFR1) and α5β1 integrin were extensively expressed on NCAM+ cells within the incipient IRF areas, whereas human epididymis protein-4 (HE4) was found to be present in few NCAM+ cells of both normal and interstitium with incipient fibrosis.

View Article: PubMed Central - PubMed

Affiliation: Institute of Pathology, Medical Faculty, University of Belgrade, Belgrade, Serbia.

ABSTRACT
Rare neural cell adhesion molecule (NCAM) positive cells have been previously described within the normal human adult kidney interstitium, speculating that they could increase in the interstitium with incipient interstitial renal fibrosis (IRF). In the present study, among 93 biopsy samples of various kidney diseases, NCAM+ interstitial cells were detected in 62.4% cases. An increased number of NCAM+ cells was significantly observed only in incipient IRF compared to normal renal tissues and advanced IRF stages (p<0.001), independently of underlying diseases (p = 0.657). All three major NCAM isoforms' RT-PCR bands were visible either in normal or in kidneys with incipient IRF, albeit their mRNA expression levels measured by qRT-PCR were different. Applying qRT-PCR on pure NCAM+ cells population, obtained by laser capture microdissection, significant mRNA over-expression of NCAM140kD isoform was found in NCAM+ cells within incipient IRF (p = 0.004), while NCAM120kD and NCAM180kD isoforms were not changed significantly (p = 0.750; p = 0.704; respectively). Simultaneously, qRT-PCR also showed significant αSMA (p = 0.014) and SLUG (p = 0.004) mRNAs up-regulation within the NCAM+ cells of incipient IRF, as well as highly decreased matrix metalloproteinases (MMP) -2 and -9 mRNAs (p = 0.028; p = 0.036; respectively). However, using double immunofluorescence MMP-9 could still be detectable on the protein level in rare NCAM+ cells within the incipient IRF. Further characterization of NCAM+ cells by double immunofluorescent labeling revealed their association with molecules involved in fibrosis. Fibroblast growth factor receptor 1 (FGFR1) and α5β1 integrin were extensively expressed on NCAM+ cells within the incipient IRF areas, whereas human epididymis protein-4 (HE4) was found to be present in few NCAM+ cells of both normal and interstitium with incipient fibrosis. Heterogeneity of NCAM+ interstitial cells in normal and incipient IRF, concerning molecules related to fibrosis and variable expression of NCAM isoforms, could suggest diverse role of NCAM+ cells in homeostasis and in regulation of renal fibrosis in diseased kidneys.

No MeSH data available.


Related in: MedlinePlus

Expression of FGFR1, HE4 and α5β1 integrin on NCAM positive cells in normal renal interstitium.(A) FGFR1 expression on renal interstitial cells. (B) HE4 expression on renal interstitial cells. (C) Integrin α5β1 expression on renal interstitial cells. (A-C) cryostat sections, immunoperoxidase, x400; (D) FGFR1 expression. (E) HE4 expression. (F) Integrin α5β1 expression. (G-I) NCAM expression. (J) Coexpression FGFR1 and NCAM on the same interstitial cells. (K) Coexpression HE4 and NCAM on the same interstitial cell. (L) coexpression of α5β1 and NCAM on the same interstitial cells. (D-L) cryostat section, immunofluorescent labeling, x600; staining techniques are described in detail under Material and Methods.
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pone.0137028.g005: Expression of FGFR1, HE4 and α5β1 integrin on NCAM positive cells in normal renal interstitium.(A) FGFR1 expression on renal interstitial cells. (B) HE4 expression on renal interstitial cells. (C) Integrin α5β1 expression on renal interstitial cells. (A-C) cryostat sections, immunoperoxidase, x400; (D) FGFR1 expression. (E) HE4 expression. (F) Integrin α5β1 expression. (G-I) NCAM expression. (J) Coexpression FGFR1 and NCAM on the same interstitial cells. (K) Coexpression HE4 and NCAM on the same interstitial cell. (L) coexpression of α5β1 and NCAM on the same interstitial cells. (D-L) cryostat section, immunofluorescent labeling, x600; staining techniques are described in detail under Material and Methods.

Mentions: Using immunoperoxidase staining, presence of few relevant markers for fibrosis has been investigated in renal interstitial cells: FGFR1 (Fig 5A), HE4 (Fig 5B), and α5β1 integrin (Fig 5C). Double immunofluorescence staining was applied to clarify whether NCAM+ renal interstitial cells could coexpress these markers.


Variable Expression of Neural Cell Adhesion Molecule Isoforms in Renal Tissue: Possible Role in Incipient Renal Fibrosis.

Marković-Lipkovski J, Životić M, Müller CA, Tampe B, Ćirović S, Vještica J, Tomanović N, Zeisberg M, Müller GA - PLoS ONE (2015)

Expression of FGFR1, HE4 and α5β1 integrin on NCAM positive cells in normal renal interstitium.(A) FGFR1 expression on renal interstitial cells. (B) HE4 expression on renal interstitial cells. (C) Integrin α5β1 expression on renal interstitial cells. (A-C) cryostat sections, immunoperoxidase, x400; (D) FGFR1 expression. (E) HE4 expression. (F) Integrin α5β1 expression. (G-I) NCAM expression. (J) Coexpression FGFR1 and NCAM on the same interstitial cells. (K) Coexpression HE4 and NCAM on the same interstitial cell. (L) coexpression of α5β1 and NCAM on the same interstitial cells. (D-L) cryostat section, immunofluorescent labeling, x600; staining techniques are described in detail under Material and Methods.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4556687&req=5

pone.0137028.g005: Expression of FGFR1, HE4 and α5β1 integrin on NCAM positive cells in normal renal interstitium.(A) FGFR1 expression on renal interstitial cells. (B) HE4 expression on renal interstitial cells. (C) Integrin α5β1 expression on renal interstitial cells. (A-C) cryostat sections, immunoperoxidase, x400; (D) FGFR1 expression. (E) HE4 expression. (F) Integrin α5β1 expression. (G-I) NCAM expression. (J) Coexpression FGFR1 and NCAM on the same interstitial cells. (K) Coexpression HE4 and NCAM on the same interstitial cell. (L) coexpression of α5β1 and NCAM on the same interstitial cells. (D-L) cryostat section, immunofluorescent labeling, x600; staining techniques are described in detail under Material and Methods.
Mentions: Using immunoperoxidase staining, presence of few relevant markers for fibrosis has been investigated in renal interstitial cells: FGFR1 (Fig 5A), HE4 (Fig 5B), and α5β1 integrin (Fig 5C). Double immunofluorescence staining was applied to clarify whether NCAM+ renal interstitial cells could coexpress these markers.

Bottom Line: Applying qRT-PCR on pure NCAM+ cells population, obtained by laser capture microdissection, significant mRNA over-expression of NCAM140kD isoform was found in NCAM+ cells within incipient IRF (p = 0.004), while NCAM120kD and NCAM180kD isoforms were not changed significantly (p = 0.750; p = 0.704; respectively).Simultaneously, qRT-PCR also showed significant αSMA (p = 0.014) and SLUG (p = 0.004) mRNAs up-regulation within the NCAM+ cells of incipient IRF, as well as highly decreased matrix metalloproteinases (MMP) -2 and -9 mRNAs (p = 0.028; p = 0.036; respectively).Fibroblast growth factor receptor 1 (FGFR1) and α5β1 integrin were extensively expressed on NCAM+ cells within the incipient IRF areas, whereas human epididymis protein-4 (HE4) was found to be present in few NCAM+ cells of both normal and interstitium with incipient fibrosis.

View Article: PubMed Central - PubMed

Affiliation: Institute of Pathology, Medical Faculty, University of Belgrade, Belgrade, Serbia.

ABSTRACT
Rare neural cell adhesion molecule (NCAM) positive cells have been previously described within the normal human adult kidney interstitium, speculating that they could increase in the interstitium with incipient interstitial renal fibrosis (IRF). In the present study, among 93 biopsy samples of various kidney diseases, NCAM+ interstitial cells were detected in 62.4% cases. An increased number of NCAM+ cells was significantly observed only in incipient IRF compared to normal renal tissues and advanced IRF stages (p<0.001), independently of underlying diseases (p = 0.657). All three major NCAM isoforms' RT-PCR bands were visible either in normal or in kidneys with incipient IRF, albeit their mRNA expression levels measured by qRT-PCR were different. Applying qRT-PCR on pure NCAM+ cells population, obtained by laser capture microdissection, significant mRNA over-expression of NCAM140kD isoform was found in NCAM+ cells within incipient IRF (p = 0.004), while NCAM120kD and NCAM180kD isoforms were not changed significantly (p = 0.750; p = 0.704; respectively). Simultaneously, qRT-PCR also showed significant αSMA (p = 0.014) and SLUG (p = 0.004) mRNAs up-regulation within the NCAM+ cells of incipient IRF, as well as highly decreased matrix metalloproteinases (MMP) -2 and -9 mRNAs (p = 0.028; p = 0.036; respectively). However, using double immunofluorescence MMP-9 could still be detectable on the protein level in rare NCAM+ cells within the incipient IRF. Further characterization of NCAM+ cells by double immunofluorescent labeling revealed their association with molecules involved in fibrosis. Fibroblast growth factor receptor 1 (FGFR1) and α5β1 integrin were extensively expressed on NCAM+ cells within the incipient IRF areas, whereas human epididymis protein-4 (HE4) was found to be present in few NCAM+ cells of both normal and interstitium with incipient fibrosis. Heterogeneity of NCAM+ interstitial cells in normal and incipient IRF, concerning molecules related to fibrosis and variable expression of NCAM isoforms, could suggest diverse role of NCAM+ cells in homeostasis and in regulation of renal fibrosis in diseased kidneys.

No MeSH data available.


Related in: MedlinePlus