Peptidylarginine Deiminase Isoforms Are Differentially Expressed in the Anagen Hair Follicles and Other Human Skin Appendages Rachida Nachat, Marie-Claire.

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Peptidylarginine Deiminase Isoforms Are Differentially Expressed in the Anagen Hair Follicles and Other Human Skin Appendages Rachida Nachat, Marie-Claire Méchin, Marie Charveron, Guy Serre, Jacques Constans, Michel Simon Journal of Investigative Dermatology Volume 125, Issue 1, Pages 34-41 (July 2005) DOI: 10.1111/j.0022-202X.2005.23763.x Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Expression of peptidylarginine deiminase type 1 (PAD1) in hair follicles. Cryosections of human scalp were analyzed by indirect immunofluorescence (double staining) and confocal observations with DG3-10, a monoclonal antibody directed against desmoglein 1 and 2 (DSG1-2), G36-19, a monoclonal antibody directed against corneodesmosin (CDSN), and with affinity-purified anti-PAD1 antibodies. (A) A longitudinal section of a hair follicle double stained with the anti-PAD1 and DG3.10 antibodies. (B, C) Higher magnification of parts of the section shown in (A). (D–F) A transverse section of a hair follicle stained with G36-19 and anti-PAD1 antibodies. (F) Image superimposition of CDSN/PAD1 labeling. The red arrow in (A) and the double-headed red arrows in (A) and (B) denote the site of abrupt cornification of Henle and Huxley layers, respectively. The white open arrowhead in (A) shows a stained Flügelzellen cell. He, Henle layer; He*, cornified Henle layer; Hu, Huxley layer; iCu, IRS cuticle; cl, companion layer; dp, dermal papilla. Scale bar=150 μm (A), =75 μm (B), and =50 μm (C–F). Journal of Investigative Dermatology 2005 125, 34-41DOI: (10.1111/j.0022-202X.2005.23763.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Expression of peptidylarginine deiminase type 3 (PAD3) and desmoglein 1 and 2 (DSG1-2) in hair follicles. Cryosections of the human scalp were analyzed by indirect immunofluorescence (double (A–D) or simple (E–G) staining) and confocal observations with DG3-10 and/or affinity-purified anti-PAD3(B3) antibodies. Longitudinal (A) and cross (B–D) sections of hair follicles analyzed by double staining are shown. The position of each transverse section on the longitudinal section is indicated as follows: 1—B, 2—C, 3—D. The red arrow and the double-headed red arrows in (A) denote the site of abrupt cornification of Henle and Huxley layers, respectively. Note the concomitant abrupt disappearance of PAD3 detection. The arrows in E show the labeling at the periphery of granules. The white open arrowhead in (A), (C), and (G) shows a PAD3-stained Flügelzellen cell. M, medulla; the other abbreviations are the same as in Figure 1. Scale bar=150 μm (A and F), =75 μm (B–D), =50 μm (E), and =10 μm (G). Journal of Investigative Dermatology 2005 125, 34-41DOI: (10.1111/j.0022-202X.2005.23763.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Expression of peptidylarginine deiminase type 3 (PAD3) and corneodesmosin (CDSN) in hair follicles. Cryosections of the human scalp were analyzed by indirect immunofluorescence (double staining) and confocal observations with the anti-PAD3(B3) and G36-19 antibodies. Longitudinal (A) and transverse (B–D) sections are shown. The position of each transverse section on the longitudinal section is indicated as follows: 1—B, 2—C, 3—D. The red arrow and the double-headed red arrows in (A) denote the site of abrupt cornification of Henle and Huxley layers, respectively, accompanied by the abrupt disappearance of PAD3 detection. The white open arrowheads in (A) show stained Flügelzellen cells. The abbreviations are the same as in Figure 1. Scale bar=150 μm (A) and =75 μm (B–D). Journal of Investigative Dermatology 2005 125, 34-41DOI: (10.1111/j.0022-202X.2005.23763.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Expression of peptidylarginine deiminase type 3 (PAD3) and trichohyalin (THH) in hair follicles. Cryosections of the human scalp were analyzed by indirect immunofluorescence with the anti-PAD3(B3) and anti-THH AE-16 antibodies. Staining with anti-PAD3(B3) (A) and AE-16 (B) of tangential sections of hair follicles. (C) Image superimposition of THH/PAD3 labelings. The double-headed red arrows in (A) denote the site of abrupt cornification of Huxley (Hu) layer. The white arrows show the co-location of PAD3 and THH. Scale bar=150 μm. Journal of Investigative Dermatology 2005 125, 34-41DOI: (10.1111/j.0022-202X.2005.23763.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Expression of peptidylarginine deiminase type 1 (PAD1) and type 2 (PAD2) in arrector pili muscles. Cryosections of human scalp were analyzed by indirect immunofluorescence and confocal microscopy with anti-PAD1 (A), anti-PAD2(C2) (D) and anti-α-actin (B and E) antibodies. (C and F) Image superimposition of α-actin/PAD1 and α-actin/PAD2 labeling, respectively. Scale bar=100 μm (A–C) and =75 μm (D–E). Journal of Investigative Dermatology 2005 125, 34-41DOI: (10.1111/j.0022-202X.2005.23763.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 6 Expression of peptidylarginine deiminase type 1 (PAD1) and type 2 (PAD2) in sweat glands. Cryosections of the human scalp were analyzed by indirect immunofluorescence and confocal microscopy with anti-PAD1 (A), anti-PAD2(C2) (D), and anti-α-actin (B and E) antibodies. (C and F) Image superimposition of α-actin/PAD1 and α-actin/PAD2 labeling, respectively. Scale bar=75 μm. Journal of Investigative Dermatology 2005 125, 34-41DOI: (10.1111/j.0022-202X.2005.23763.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 7 Location of deiminated proteins in hair follicles (HF). Longitudinal cryosections of hair follicles were analyzed by immunoperoxidase with AMC antibodies specific to chemically modified citrullines. Note the appearance of deiminated proteins (red) first in the Henle layer (blue arrows in A and B), then in the cuticle (double-headed blue arrows in B and C) and finally in the Huxley layer (red arrows in B and C) of the inner root sheath. Deiminated proteins were also localized in the medulla (M) of the HF. Note the black pigment deposited in the hair follicles (A and B). Scale bar=100 μm. Journal of Investigative Dermatology 2005 125, 34-41DOI: (10.1111/j.0022-202X.2005.23763.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions