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Peptidylarginine deiminase type 2 is over expressed in the glaucomatous optic nerve.

Cafaro TA, Santo S, Robles LA, Crim N, Urrets-Zavalia JA, Serra HM - Mol. Vis. (2010)

Bottom Line: To determine levels of Peptidyl arginine deiminase 2 (PAD2) and its product protein-bound citrulline in cadaver eyes that suffered from normal tension glaucoma (NTG) compared to primary open angle glaucoma (POAG), and controls.We report over expression of PAD2 and higher levels of its product protein-bound citrulline in the optic nerve of normal tension glaucoma patients (NTG).This is the first report demonstrating that like in POAG, NTG also possesses elevated levels of both PAD2 and protein-bound citrulline.

View Article: PubMed Central - PubMed

Affiliation: CIBICI, Faculty of Chemistry, National University of Córdoba, Argentina.

ABSTRACT

Purpose: To determine levels of Peptidyl arginine deiminase 2 (PAD2) and its product protein-bound citrulline in cadaver eyes that suffered from normal tension glaucoma (NTG) compared to primary open angle glaucoma (POAG), and controls.

Methods: Western analysis, ELISA, and immunohistochemical analysis were performed with human tissues.

Results: We report over expression of PAD2 and higher levels of its product protein-bound citrulline in the optic nerve of normal tension glaucoma patients (NTG).

Conclusions: This is the first report demonstrating that like in POAG, NTG also possesses elevated levels of both PAD2 and protein-bound citrulline.

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Related in: MedlinePlus

Representative immunohistochemical analyses of NTG (70 F) and control (72F) donor optic nerve sections. A: Immunoreactivity for protein-bound citrulline detected by a rabbit polyclonal antibody to citrulline after monoxime modification as described in methods. B: Merged image of anti-citrulline with DAPI. C: Control donor section probed for immunoreacitivty with anti-citrulline. D: Merged image of anti-citrulline and DAPI for control donor section. E: Densitometric analysis of immunohistochemical detection of protein-bound citrulline in NTG eyes. The data was analyzed using Image J software.
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f4: Representative immunohistochemical analyses of NTG (70 F) and control (72F) donor optic nerve sections. A: Immunoreactivity for protein-bound citrulline detected by a rabbit polyclonal antibody to citrulline after monoxime modification as described in methods. B: Merged image of anti-citrulline with DAPI. C: Control donor section probed for immunoreacitivty with anti-citrulline. D: Merged image of anti-citrulline and DAPI for control donor section. E: Densitometric analysis of immunohistochemical detection of protein-bound citrulline in NTG eyes. The data was analyzed using Image J software.

Mentions: Elevated PAD2 was detected in normal tension glaucoma donors as well as in POAG donors (Figure 1A and Figure 2A,B). Commensurate with elevated PAD2, elevated protein-bound citrulline (Figure 1B and Figure 2C,D) was also detected in NTG optic nerve compared to control. Activity measurement showed elevated PAD2 in protein extract derived from NTG and POAG donors compared to control (Figure 3). Elevated PAD2 activity is commensurate with elevated PAD2 immunoreactivity as well as that for protein-bound citrulline (Figure 1). We also investigated protein-bound citrulline immunoreactivity (product of PAD2 activity) using immunohistochemistry in optic nerve sections of NTG eyes (Figure 4A,B). The immunohistochemical comparison of protein-bound citrulline immunoreactivity, commensurate with western (Figure 1B) and ELISA (Figure 2) analyses showed elevated immunoreactivity in the NTG compared to control eyes. All sections subjected to immunohistochemical analyses were also subjected to relative intensity measurement with after normalization of measurement area which showed relatively elevated levels of protein-bound citrulline in NTG eyes compared to controls (Figure 4C).


Peptidylarginine deiminase type 2 is over expressed in the glaucomatous optic nerve.

Cafaro TA, Santo S, Robles LA, Crim N, Urrets-Zavalia JA, Serra HM - Mol. Vis. (2010)

Representative immunohistochemical analyses of NTG (70 F) and control (72F) donor optic nerve sections. A: Immunoreactivity for protein-bound citrulline detected by a rabbit polyclonal antibody to citrulline after monoxime modification as described in methods. B: Merged image of anti-citrulline with DAPI. C: Control donor section probed for immunoreacitivty with anti-citrulline. D: Merged image of anti-citrulline and DAPI for control donor section. E: Densitometric analysis of immunohistochemical detection of protein-bound citrulline in NTG eyes. The data was analyzed using Image J software.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2927370&req=5

f4: Representative immunohistochemical analyses of NTG (70 F) and control (72F) donor optic nerve sections. A: Immunoreactivity for protein-bound citrulline detected by a rabbit polyclonal antibody to citrulline after monoxime modification as described in methods. B: Merged image of anti-citrulline with DAPI. C: Control donor section probed for immunoreacitivty with anti-citrulline. D: Merged image of anti-citrulline and DAPI for control donor section. E: Densitometric analysis of immunohistochemical detection of protein-bound citrulline in NTG eyes. The data was analyzed using Image J software.
Mentions: Elevated PAD2 was detected in normal tension glaucoma donors as well as in POAG donors (Figure 1A and Figure 2A,B). Commensurate with elevated PAD2, elevated protein-bound citrulline (Figure 1B and Figure 2C,D) was also detected in NTG optic nerve compared to control. Activity measurement showed elevated PAD2 in protein extract derived from NTG and POAG donors compared to control (Figure 3). Elevated PAD2 activity is commensurate with elevated PAD2 immunoreactivity as well as that for protein-bound citrulline (Figure 1). We also investigated protein-bound citrulline immunoreactivity (product of PAD2 activity) using immunohistochemistry in optic nerve sections of NTG eyes (Figure 4A,B). The immunohistochemical comparison of protein-bound citrulline immunoreactivity, commensurate with western (Figure 1B) and ELISA (Figure 2) analyses showed elevated immunoreactivity in the NTG compared to control eyes. All sections subjected to immunohistochemical analyses were also subjected to relative intensity measurement with after normalization of measurement area which showed relatively elevated levels of protein-bound citrulline in NTG eyes compared to controls (Figure 4C).

Bottom Line: To determine levels of Peptidyl arginine deiminase 2 (PAD2) and its product protein-bound citrulline in cadaver eyes that suffered from normal tension glaucoma (NTG) compared to primary open angle glaucoma (POAG), and controls.We report over expression of PAD2 and higher levels of its product protein-bound citrulline in the optic nerve of normal tension glaucoma patients (NTG).This is the first report demonstrating that like in POAG, NTG also possesses elevated levels of both PAD2 and protein-bound citrulline.

View Article: PubMed Central - PubMed

Affiliation: CIBICI, Faculty of Chemistry, National University of Córdoba, Argentina.

ABSTRACT

Purpose: To determine levels of Peptidyl arginine deiminase 2 (PAD2) and its product protein-bound citrulline in cadaver eyes that suffered from normal tension glaucoma (NTG) compared to primary open angle glaucoma (POAG), and controls.

Methods: Western analysis, ELISA, and immunohistochemical analysis were performed with human tissues.

Results: We report over expression of PAD2 and higher levels of its product protein-bound citrulline in the optic nerve of normal tension glaucoma patients (NTG).

Conclusions: This is the first report demonstrating that like in POAG, NTG also possesses elevated levels of both PAD2 and protein-bound citrulline.

Show MeSH
Related in: MedlinePlus