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Research Topics in Medical Cell Biology

Regulation of Pigmentation

AIMS
Where and how melanogenesis occurs Regulation of constitutive pigmentation / MC-1R Regulation of facultative pigmentation / UVR

 What determines phenoype?

Oxy/deoxy haemoglobin Carotenoids Melanin

What controls Pigmentation?

> 125 distinct genes
- development of melanoblasts - differentiation & survival melanocytes

> 25 genes
- biogenesis & function of melanosomes / proteins Critical enzymes include: - Tyrosinase
- TRP-1 (tyrosinase related protein -1) - DCT (DOPAchrometautomerase)

 Critical structural proteins:

- Pmel17 - MART 1 Structural maturation melanosomes

Mutations: enzymes & structural proteins:

- inherited pigmentary disorders - eg Albinism: Tyrosinase dysfunction

Melanocytes
Specialise: in the synthesis of melanin Derived: melanoblasts Visible phenotype: Accurate migration, distribution & functioning Mblasts / Mcyte Location: - basal layer epidermis - connected to keratinocytes, fibroblasts - hair follicle: bulb / ORS sebaceous gland
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 Location of Melanocytes in the Epidermis

keratinocyte melanocyte melanin

fibroblast

Location of Melanocytes in the Hair Follicle

E-Mc

IFD-Mc SG-Mc

ORS-Mc

BMc DP 8

 Melanogenesis
where? melanosomes 3 distinct melanins: - Eumelanin: Brown/black - Pheomelanin: Red/yellow melanosomes mature: - Transferred cite synthesis = perikaryon dendrites - neighbouring keratinocytes
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Melanocyte structure

pheomelanosome

eumelanosome

dendrite

premelanosome Nucleus

Melanocyte
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 Melanogenesis
Tyrosinase L-Tyrosine
6BH4 /O2

Tyrosinase L-DOPA DOPAquinone + Cysteine CysteinylDOPA

L-Tyrosine DHI DOPAchrome Tyrosinase Indole-5,6quinone DHICA LeucoDOPAchrome DCT Indole-5,6-quinonecarboxylic acid TRP-1 1,4 Benzothiazinyl-alanine

DHI melanin Dark brown/black

EUMELANIN Brown/Black

DHICA melanin Light Brown

PHEOMELANIN Yellow/Red
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Why do differences in human skin colour exist?

-Tyrosinase activity -No. & size of Msomes -Transfer of mature Msomes Kcytes

Caucasian skin / melanosomes:

- smaller / less melanin - several Msomes are transferred Kcytes - Msomes degraded in lower epidermal layers
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 Black skin / melanosomes:

- Larger / more melanin - Msomes transferred individually Kcytes - Msomes degraded in upper epidermal layers

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UVR
Light
K K K M K

UVR
Dark
K

K M

BM
F F F F

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 Constitutive and Facultative Pigmentation Constitutive: - Genetically determined

- Absence of external influences

Facultative: - pigmentation in response stimuln

- UVR major regulator

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Constitutive Pigmentation
Migration of melanoblasts Survival & differentiation melanocytes Expression enzymatic/structural constituents Msomes Synthesis of eu- pheomelanin Transport of Msomes dendrites Transfer Msomes keratinocytes

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 Melanocortin -1 Receptor
Major ctrl point in regulating human pigmentation Regulates quantity & quality of melanins

Agonists MC-1R
-MSH & ACTH (precursor peptide POMC) Synthesised: Skin cells / Mcytes & Kcytes Activation MC-1R - stimulates expression of melanogenic cascade - synthesis of eumelanin
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-MSH & ACTH

Mcyte dendricity & proliferation expression of MC-1R gene

Melanogenesis Dendricity

Proliferation

Antagonist MC-1R
Agouti signalling protein (ASP) - stimulates synthesis of pheomelanin
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 Mechanism of Action: MC-1R

cAMP 2nd messenger system
-MSH/ACTH
MC-1R Ac PKA
CREB CRE

cAMP

MITF

MITF
MBOX

Tyrosinase
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Melanogenesis

What is special about this phenoype?

A. Caused by MC-1R polymorphisms

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 MC-1R polymorphisms
Allelic variants / associated with red hair/fair skin - Arg151Cys, Arg160Trp, Asp294His Loss of function MC-1R: - affects -MSH/ACTH binding - subsequent signalling Highly associated with: - poor tanning - risk of melanoma
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What Causes Tanning?

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 Facultative Pigmentation
Regulated by: UVA radiation / tanning reaction Immediate tanning - Occurs within mins of exposure & persists for hrs persistent skin darkening - Lasts several days - oxidation and polymerisation of existing melanin - redistribution of existing melanosomes

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Delayed tanning - Occurs several days after UVR exposure - Activation of melanocyte function - Mcyte proliferation & dendricity - MITF expression & - downstream melanogenic proteins: Pmel17 MART-1 Tyrosinase Melanogenesis TRP-1 DCT
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 EM and EK respond to UV exposure: - synthesis and secretion of -MSH & ACTH - expression and function of MC-1R - Enhances Mcyte responses to -MSH & ACTH Other melanogenic factors secreted by EK - SCF (stem cell factor) - NGF (nerve growth factor)

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Summary
Melanogenesis occurs in melanosomes Melanin consists of distinct forms: - Eumelanin: brown/black - Pheomelanin: yellow/red Constitutive pigmentation: - Genetically determined / abs of external influences Facultative pigmentation: - pigmentation due to a physiological factor
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ratio imp

 References
Yamaguchi Y et al., (2007). The regulation of human skin pigmentation. The Journal of Biological Chemistry 282 (4): 27557-61 Miyamura Y et al., (2006). Regulation of human skin pigmentation and responses to ultra violet radiation. Pig Cell Res 20: 2-13 Abdel-Malek Z and Kadekaro AL (2007). Human pigmentation: Its regulation by ultraviolet light and by endocrine and paracrine factors. Nordlund JJ etal., (Eds.). Oxford University Press, Chapter 20.

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