Thuyen Nguyen, Maria L. Wei Journal of Investigative Dermatology

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Presentation transcript:

Hermansky–Pudlak HPS1/pale ear Gene Regulates Epidermal and Dermal Melanocyte Development Thuyen Nguyen, Maria L. Wei Journal of Investigative Dermatology Volume 127, Issue 2, Pages 421-428 (February 2007) DOI: 10.1038/sj.jid.5700566 Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 ep and le mice have hypopigmented ears, tails and paws. Control le heterozygote (le/+), le homozygous mutant (le/le), control ep heterozygote (ep/+), ep homozygous mutant (ep/ep). Eight-week-old littermates are shown. Journal of Investigative Dermatology 2007 127, 421-428DOI: (10.1038/sj.jid.5700566) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Decreased tyrosinase activity in tail epidermis and dermis of 4-week-old ep mutants. Tail tissue from 4-week-old mutant and heterozygote littermates were stained with DOPA to detect tyrosinase activity and whole-mount tissue sections were examined by light microscopy. ep/+, wild-type heterozygote littermate; ep/ep, ep mutant littermate. (a) Control tail epidermis, (C57BL/6). Note organization of melanocytes into “clusters’' surrounded by areas relatively sparsely populated by melanocytes. Bar=100μm. Brown pigment surrounding black melanocytes is due to melanocytes out of the plane of focus. (b) Tail epidermis examined by light microscopy, comparing ep/+ with ep/ep. Bar=100μm. Arrows, hair follicles out of plane of focus. Representative interfollicular melanocytes are boxed. (c) Number of melanocytes detected in ep/ep versus ep/+ tail epidermis at original magnification × 20. Five fields, each field focusing on a melanocyte “cluster”, were counted and averaged. (d) Number of melanocytes detected in ep/ep versus ep/+ tail dermis. Journal of Investigative Dermatology 2007 127, 421-428DOI: (10.1038/sj.jid.5700566) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Developmental delay in onset of pigmentation in ep mice. Postgestational day 1–9 (P1–P9) pups shown. For each pair, wild-type ep/+ heterozygote is on the left; mutant ep/ep littermate is on the right. Note that although dorsal back fur pigment gradually darkens to almost wild-type hue, tail, ears, and paws remain hypopigmented. Journal of Investigative Dermatology 2007 127, 421-428DOI: (10.1038/sj.jid.5700566) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Delayed onset of tyrosinase activity in neonatal ep tail interfollicular epidermis. Tail tissue was harvested from post-gestational day 1 (P1) through post-gestational day 9 (P9) mice, epidermis and dermis separated and stained with DOPA and whole-mount tissue sections examined by light microscopy. (a) ep/+, wild-type heterozygote; ep/ep, mutant homozygous ep. Arrows, hair follicles out of plane of focus. Representative interfollicular melanocytes are boxed. Original magnification × 20. (b) Delayed onset of tyrosinase activity and rapid decrease in the number of melanocytes in ep/ep tail neonatal epidermis. P1, P<0.0001; P4, P=0.8; P6, P=0.03; P9, P<0.001. Journal of Investigative Dermatology 2007 127, 421-428DOI: (10.1038/sj.jid.5700566) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Decreased numbers of melanocytes in neonatal ep interfollicular tail dermis. (a) Tail tissue was harvested and treated as in Figure 4. (a) ep/+, wild-type heterozygote; note reticular pattern of interfollicular melanocytes; follicular melanocytes are out of the plane of focus. ep/ep, mutant ep; P1, note absence of reticularly arrayed melanocytes, only follicular melanocytes are visible. Arrows, hair bulbs. Representative interfollicular melanocytes are boxed. Original magnification × 20. (b) Decreased numbers of melanocytes in the dermis of ep/ep neonatal tail. P1, P<0.0001; P4, P=0.01; P6, P<0.01; P9, P<0.01. Journal of Investigative Dermatology 2007 127, 421-428DOI: (10.1038/sj.jid.5700566) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 6 Biogenesis of melanosomes in ep tail interfollicular epidermis is markedly impaired. (a) Transmission electron microscopy of tail epidermis. Stage I melanosomes have intraluminal vesicles (middle inset). Stage II melanosomes have intralumenal striations, (left inset). Stage III have evidence of pigment deposition and stage IV have pigment filling the lumen, obscuring intralumenal contents. Aberrant appearing undefined melanosomal types are vesicles similar in size to melanosomes with granular internal content, without discernable intralumenal vesicles, striations, or pigment (right inset). Nuc, melanocyte nucleus; Ker, neighboring keratinocyte. Bar=0.5μm. (b) Percentages of stage I–IV melanosomes in ep homozygotes and controls. (c) Number of melanosomes/unit area. (d) Average size of fully pigmented stage IV melanosomes. Journal of Investigative Dermatology 2007 127, 421-428DOI: (10.1038/sj.jid.5700566) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 7 Marked decrease in tyrosinase activity in back neonatal interfollicular epidermis. Back skin tissue was harvested and treated as described in Figure 4. (a) Whole-tissue mount of epidermis examined by light microscopy. Representative interfollicular melanocytes boxed. Follicles can be seen as objects out of the plane of focus, arrows. ep/+, wild-type heterozygote; ep/ep, mutant ep. Original magnification × 20. (b) Numbers of interfollicular melanocytes in back neonatal epidermis. P1-P9, P<0.001. Journal of Investigative Dermatology 2007 127, 421-428DOI: (10.1038/sj.jid.5700566) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 8 Significant decrease in tyrosinase activity in ep back neonatal dermis at P1. Back skin tissue was harvested and treated as described in Figure 4. (a) Whole-tissue mount of dermis examined by light microscopy. ep/+, wild-type heterozygote; ep/ep, mutant ep. Arrows, hair follicles out of plane of focus. Representative interfollicular melanocyte is boxed. Original magnification × 20. (b) Numbers of interfollicular melanocytes in back neonatal dermis. Journal of Investigative Dermatology 2007 127, 421-428DOI: (10.1038/sj.jid.5700566) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions