Cent. Eur. J. Biol.

• 8(4) • 2013 • 315-330

DOI: 10.2478/s11535-013-0141-1

Central European Journal of Biology

Heterogeneity of neural crest-derived

melanocytes
Review Article

Miroslawa Cichorek*, Malgorzata Wachulska, Aneta Stasiewicz

Department of Embryology,
Medical University of Gdansk,
80-210 Gdansk, Poland
Received 05 July 2012; Accepted 27 December 2012

Abstract: The majority of melanocytes originate from the neural crest cells (NCC) that migrate, spread on the whole embryo’s body
to form elements of the nervous system and skeleton, endocrinal glands, muscles and melanocytes. Human melanocytes
differentiate mainly from the cranial and trunk NCC. Although melanocyte development has traditionally been associated with the
dorsally migrating trunk NCC, there is evidence that a part of melanocytes arise from cells migrating ventrally. The ventral NCC
differentiate into neurons and glia of the ganglia or Schwann cells. It has been suggested that the precursors for Schwann cells
differentiate into melanocytes. As melanoblasts travel through the dermis, they multiply, follow the process of differentiation and
invade the forming human fetal epidermis up to third month. After birth, melanocytes lose the ability to proliferate, except the
hair melanocytes that renew during the hair cycle. The localization of neural crest-derived melanocytes in non-cutaneous places
e.g. eye (the choroid and stroma of the iris and the ciliary body), ear (cells of the vestibular organ, cochlear stria vascularis),
meninges of the brain, heart seems to indicate that repertoire of melanocyte functions is much wider than we expected e.g. the
protection of tissues from potentially harmful factors (e.g. free radicals, binding toxins), storage ions, and anti-inflammatory action.

Keywords: Neural crest cells • Melanocytes origin • Skin melanocytes • Ocular melanocytes • Ear melanocytes • Epidermal melanin unit
© Versita Sp. z o.o.

tube during neurulation (formation of the neural tube,

1. Introduction
The melanocyte biology has fascinated scientists of
different disciplines for years, especially according to
the basic function of these cells – production of the
skin, hair and eye pigment [1-3]. Nowadays these cells
seemed to play many other functions in the organism.
The cells broadly classified as melanocytes (cells
able to synthesize melanins) form a heterogeneous
group of cells referring to their embryonic origin and
anatomical location. There are attempts to group
pigment-producing cells into classical (cutaneous)
and non-classical (non-cutaneous) melanocytes [4,5].
The embryonic origin of these two groups is different,
cutaneous melanocytes develop from the neural crest
cells (NCC) while non-cutaneous from NCC and neural
tube cells (e.g. retinal pigment epithelium, RPE).
This review will focus on a group of melanocytes
differentiating from the NCC – an embryonic population
that develops from the dorsal part of the forming neural
primordium for central nervous system) [6,7]. Neural
crest cells (NCC) leave the epithelium of the neural
tube, migrate to many specific places in the body, where
they differentiate into a remarkable variety of cells as
neurons, glial cells, endocrine cells, chondroblasts,
osteoblasts, smooth muscles and melanocytes
[7-10]. Migration occurs along the well-defined
pathways which determines in part what type of cells
NCC will form [8,10].
Melanocytes differentiate from all NCC populations
but the molecular pathways of development are well
characterized only for the trunk NCC [8,11,12]. Except
the skin and eye melanocytes are present in ear,
nervous system and probably other places e.g. heart
[5]. The remarkable feature of the melanocytes is
the melanin production. Melanin protects cells in the
epidermis against sun-induced damage by ultraviolet
radiation (UVR) but what is/are its function/s in other
place e.g. ear, meninges is not clear [11,12].

* E-mail: cichorek@gumed.edu.pl
315
 Heterogeneity of neural crest-derived melanocytes

Apart from being the melanin factories, melanocytes
are immunocompetent, immunomodulatory and
secretory cells [13-15]. They produce and release
neurotransmitters (acetylocholine, catecholamines),
hormones (adrenocorticotropin hormone, ACTH;
melanocyte stimulating hormone, α-MSH) and many
cytokines [16-21]. Melanocytes as cells presenting
antigens and secreting cytokines for lymphocytes, are
active parts of the immune response in the skin [13].
They cooperate with neighboring cells as epidermal
keratinocytes, dermal fibroblasts and work in an
organized regulatory network for the maintenance of
epidermal homeostasis [22]. How this process looks
like in other than epidermis places e.g. stria vascularis
cells of the cochlear duct secreting endolymph or
cardiomyocytes, is completely not understood.
Melanocytes in the inner ear are necessary for the
hearing process although the mechanism is not well
defined [23]. Melanocyte presence, not only in the
epidermis and hair but also in the nervous system and
in some organs, emphasizes the alternative functions of
these cells that are still poorly understood [24,25]. The
developing knowledge of the melanocytes biology give
ability to understanding the melanoma development,
human pigmentation disorders as vitiligo, greying of
hair or even problems with hearing. In this review, the
current knowledge about human melanocytes of the
neural crest-origin and their heterogeneity is described.
2. The origin and development of
melanocytes
2.1 Neural crest – embryonic structure of the
melanocyte origin precursors
Precursors for melanocytes develop from the neural
crest [6]. The basic biology of the neural crest
development is very similar among model organisms
but small differences do exist. Specifically, we know very
little about the molecular development of neural crest
during the early human embryogenesis [7,9]. Neural
crest originates from neuroectoderm, developing from
ectoderm under inductive influence of the notochord,
during the neurulation process (Figure 1). Differentiation
of the neuroectoderm gives neural tube (future central
nervous system elements) and neural crest cells, so
named because of their site of origin, the crest of the
closing neural tube formed from two lateral strips of the
neural plate (Figure 1) [7,8]. NCC change the morphology
from epithelial to mesenchyme and as the result they get
ability to movement at a 4-week old human embryo [7,9].
They migrate out from neuroepithelium and locate above
the neural tube underneath surface ectoderm. NCC
are initially multipotent but gradually become lineage–
restricted in developmental potential (Figure 1) [10].
This potential is determined by anatomical localization
along the anterior-posterior axis. Traditionally there
are distinguishable four NCC populations: cranial,

Figure 1. Differentiation of the ectoderm into surface ectoderm (future epidermis) and neuroectoderm (neural tube and neural crest cells) at
4-weeks human embryo. The neuroectodermal cells proliferate, form neural plate that folds and changes into neural groove (A). The
neural groove fuses from the middle part into both ends and the neural tube forms (B). During this neurulation process, cells from edges
of the neural groove separate from neural tube as independent population of embryonic cells - neural crest cells (NCC), located above
the neural tube (future brain and spinal cord) and beneath surface ectoderm (future epidermis) (B).

316
M. Cichorek et al.
trunk, vagal and sacral, cardiac. Multipotency of NCC
is well known e.g. cranial NCC form most of cartilages
and bones of the face and head, whereas trunk NCC
form elements of the peripheral nervous system,
medulla of the adrenal glands. These cells proliferate,
and start to express distinct molecular markers. Each
NCC population activates different molecular pathways
that produce glial cells, neurons, mesenchyme cells,
melanocytes [9,11,12]. There are conflicting results of
experiments concerning the problem – at what stage
NCC get features of melanoblasts – precursors to
melanocytes [8,9,12] is not clear.
2.2 NCC groups for melanoblast development
NCC of all populations can give rise to melanocytes
although the trunk NCC give most of melanocytes
in the skin. In the region of the future embryo’s head
melanocytes come from the cranial NCC population
that biology is different from others. Head NCC
together with head mesoderm form the mesenchyme
(ectomesenchyme), which gives the skull, muscles,
dermis of the head [7]. The cranial NCC begin the
migration before the closure of the cranial neuropore,
while in the spinal cord of the neural tube (future trunk),
NCC detach after the neural groove fusion. There are
two basic ways of the trunk NCC migration dorsally
or ventrally away from the neural tube (Figure 2). The
different ways of NCC migration and their fate depend
Figure 2. The basic pathways of the trunk neural crest cell (NCC) migration during early embryonic time. Dorsal cells (1) migrate between the
surface ectoderm and somites, finally develop mainly into melanocytes of the epidermis and hair. Ventral cells (2) migrate between
neural tube and somites, give elements of the peripheral nervous system, medulla of the adrenal glands and melanocytes of the skin.
on the extracellular matrix molecules and cells they
meet on the migratory pathways [7-9].
The dorsolaterally migrating cells follow a path between
ectoderm and somites and become melanocytes
(Figure 2). The ventrally migrating cells follow a way
between the neural tube and somites giving rise to the
peripheral nervous system and some endocrine cells
e.g. adrenal medulla (Figure 2) [7,9]. Which of these
two NCC populations migrate first in human is not
clear, in mouse first migrate dorsal NCC but in chick
ventral NCC [8]. Although melanocyte development
has traditionally been associated with this dorsal
way of NCC migration, there is evidence that a part
of melanocytes arise from cells migrating ventrally
and then along the nerves that terminate in the skin
[26-28].
Recent studies pointed that Schwann cells, forming
myelin sheath around peripheral nerves, when cultured
with melanocyte medium develop into precursors able
to give glial cells and melanocytes [29]. NCC migrating
ventrally either differentiate into neurons and glial cells
of the spinal and autonomic ganglia or Schwann cells.
It has been suggested that a population of NCC cells,
nerve sheath stem cells (precursors for Schwann cells),
could differentiate into melanocytes of the epidermis
[28,30]. The recent finding that in human fetuses
the prenatal nevi develop from dermal not epidermis
melanocytes supports this thesis [30].These findings
317

suggested that the precursors for Schwann cells arrive
near epidermis along cutaneous nerves, may respond
to factors secreted by epidermal cells and differentiate
into melanocytes [28,30]. The control of melanocyte
functions by the cutaneous nervous system has been
poorly understood although communication between the
nervous system and epidermal melanocytes has been
shown [31]. The increasing evidence that epidermal
melanocytes molecularly differ from dermal melanocytes
might reflects dual origin of the skin melanocytes [4].
2.3 Melanoblast differentiation into
melanocytes
As melanoblasts travel through the dermis, they multiply
and follow the process of differentiation. These cells
finally except the skin (epidermis, hair follicles) reside
the eye (choroid, iris) and some internal sites, such
as meninges, the inner ear (stria vascularis), mucous
membranes (oral cavity) [29]. What do we know about
human melanoblast migration into all these places?
The only knowledge about this process comes from the
epidermal melanoblasts. During the human development
melanoblast migration into embryo’s surface ectoderm
takes place at a 6-8-weeks old embryo and after the
additional month most of these cells are located in the
epidermis and hair follicles [30,32,33]. Thus after this
time majority of melanoblasts has been suggested to
enter the skin. However, some authors do not exclude
the possibility that a part of melanoblasts might stay in
the dermis [33,34].
Epidermal melanoblasts proliferate, differentiate
together with the fetus growth when the surface of the
epidermis increases but the proliferation of epidermal
melanocytes has not yet been fully characterized. The
basic molecular elements of melanocyte differentiation
are:
- special receptors in the plasma membrane as ephrin
receptors (EphR), endothelin receptors (especially
EdnRB2), tyrosine kinase receptor KIT (c-Kit), receptor
for the beta catenins (Frizzled receptor),
- transcription factors as SOX10, PAX3 and MITF
(Microphthalmia-associated transcription factor)
- melanogenic enzymes e.g. tyrosinase-related
protein 2 (TRP2), tyrosinase (TYR) [9,12,35].
The most important growth factors regulating
melanoblasts development from the NCC are
Wnt proteins, stem cell factor (SCF, c-Kit ligand),
endothelins, ephrins, bone morphogenetic proteins
(BMP) [35-38]. Ephrins and SCF bind to receptors
that have a cytoplasmic tyrosine kinase domain and
activate the MAPK- signaling pathway (Ras/Raf/
MEK/ERK). Early melanoblast development requires
the presence of the SCF and its receptor c-Kit. SCF
318
Heterogeneity of neural crest-derived melanocytes
is secreted by dermal and epidermal cells during
melanoblast migration from the region of the neural
tube through the dermis into epidermis. SCF is a factor
required for survival of melanoblasts [35,38]. Except
for the MAPK-signaling pathway, the Notch-signaling
and Wnt-signaling are essential for melanocyte
development [38,39]. Like ephrins, Notch receptor
requires the contact with another cell because the
ligand for this receptor is a plasma membrane protein
(Delta protein, Jagged, Serrate). After ligand binding
intracellular domain of Notch receptor is cleaved by
protease and migrates to nucleus where regulates
transcription factors activity [39]. Studies in several
model organisms suggest that components of the
Wnt/Frizzled protein/β-catenin-signaling pathway are
required for induction of melanocyte fate [40]. The
ephrin and endothelin receptors allow melanoblasts to
migrate along extracellular matrix containing ephrins
and endothelins secreted by embryo’s cells [8].
Spaces between tissue layers and cells are filled
with a rich extracellular matrix (ECM) formed by many
fibrillar proteins as collagens, fibronectin. Cell adhere to
one another or ECM using adhesion molecules such as
cadherins and integrins. These elements also participate
in the melanocyte development [41,42].
Most of the above mentioned main melanocytic
differentiation elements induce expression of the
MITF, a protein functioning as a master of melanocyte
proliferation, differentiation and survival. MITF induces
expression of many genes that products are necessary
for melanocytes e.g. antiapoptotic BCL-2 protein
necessary for melanocytes survival, tyrosinase main
enzyme for melanin production [35,43].
The main melanocyte differentiation process is the
creation of melanogenesis pathways. As the result
of this process, in the melanocyte cytoplasm new
organelles called melanosomes (dark vesicles) appear
and a cell forms many dendritic processes, gets a
star-like shape. Melanogenesis is a cytotoxic process
and as a way of defense against the death possibility
melanocyte increases the level of antiapoptotic protein
BCL-2 [44,45].
Melanocyte is a highly differentiated cell and as
such loose the ability to proliferate during the maturation
process. In the adult skin, the only renewing melanocyte
groups are the hair melanocytes which cyclically, as hair
are lost, rebuilt the number [46]. Epidermal melanocytes
are a very stable population which proliferate extremely
rare at the physiological conditions [46-48]. One
known reason for this longevity is the resistance of
melanocytes to apoptosis, particularly resulting from a
high constitutive level of antiapoptotic BCL-2 [44]. It is
consider that in terminally differentiated cells the cell
M. Cichorek et al.
cycle is inhibited by several mechanisms: increased level
of cyclin dependent kinase inhibitors (p16, p27, p21),
retinoblastoma protein (pRB) hypophosphorylation,
decreased level of cyclin D1 [47,48].
Most pigment cells, in all parts of the body, seem to
be constant in number and function until approximately
the middle ages (the fourth or fifth decade of life) [49].
Thereafter, the number of melanocytes in the skin and
hair decreases, approximately 10 percent per decade
[49,50]. Melanocytes from older people are larger, more
dendritic and have decreased tyrosinase activity [45].
In vitro, fetal melanocytes proliferate better than adults
and melanocytes from patients with the prematurely
aging disorder [48,51]. Furthermore, the reason of
decreased number of melanocytes in older people could
be apoptosis of terminally differentiated melanocytes
[48,52]. It is stressed that senescence maintenance
melanocytes with damaged DNA and simultaneously
can produce viable melanocytes predisposed to
malignant transformation into melanoma cells [48,53].
The process of hair greying is the result of melanocyte
stem cells death [46].
3. Melanogenesis
3.1 Melanin synthesis
Melanogenesis is a process of pigment melanin
synthesis within membrane-bound organelles termed
melanosomes [54,55]. A distribution of melanin is
responsible not only for skin, hair and eye pigmentation,
Figure 3. The simplified scheme of the melanin synthesis in melanocytes during melanogenesis. The basic substrate tyrosine under influence of
the basic enzymes as tyrosinase (TYR), tyrosine-related protein 1 (TRP1) and 2 (TRP2) changes into a polymer of melanin, a mixture of
pigments named eumelanin and pheomelanin.
but also for photoprotection (UV light absorption),
reactive oxygen species (ROS) neutralization and ions
storage [50,56-59].
Melanosomes produce three kinds of melanin –
pheomelanin and two types of eumelanins. Pheomelanin
is red to yellow and it is responsible for red hair and
freckles. The most abundant form of melanin is a highly
heterogenous dark brown to black, insoluble – eumelanin
[57,60]. Eumelanin has antioxidant properties, while
pheomelanin exhibits phototoxic prooxidant action
[55]. Melanogenesis requires a number of specific
enzymes for melanin synthesis and structural proteins
for building the fibrillar stroma for this process in the
melanosomes. Critical enzymes are tyrosinase (TYR),
tyrosinase-related protein 1 (TRP1) and tyrosinase-
related protein 2 (TRP2, DOPAchrome tautomerase
DCT) [55,61,62]. The most important structural proteins
inside melanosomes are Pmel17 (gp100) and MART1
(melanoma-associated antigen recognized by T cells)
[45,63].
The first step, common for all melanin kinds, involve
hydroxilation of tyrosine to
3-4-dihydroxyphenylalanine (L-DOPA) and
oxidation of L-DOPA to DOPAquinone. Both of which
are catalyzed by tyrosinase, a crucial enzyme for the
melanin biosynthetic pathway [50,55,64]. The availability
of substrates and the function of melanogenetic
enzymes decide which kind of melanin is produced.
DOPAquinone in the presence of cysteine creates 3- or
5-cysteinylDOPA which then oxidize and polymerize,
giving rise to yellow/red pheomelanin (Figure 3) [45,65].
319
 Heterogeneity of neural crest-derived melanocytes

In the absence of cysteine, DOPAquinone undergoes
spontaneously non-enzymatic intramolecular cyclization
and futher rearrangment, yielding DOPAchrome. This
process happens very fast [66-68]. DOPAchrome gives
eumelanins by two pathways – nonenzymatic and
enzymatic, with use tyrosine related proteins (TRPs).
In the first case DOPAchrome undergoes spontaneous
decarboxylation, becoming 5,6-dihydroxyindole (DHI),
which rapidly oxidizes and polymerizes to form dark
brown/black, DHI-melanin (Figure 3) [66,68,69].
In the second pathway TRP2 (DCT, DOPAchrome
tautomerase) tautomerizes DOPAchrome to form DHI-
2-carboxylic acid (DHICA) [70]. Next, TRP1 catalyzes
DHICA oxidation and polymerization and form light
brown, DHICA-melanin. When DOPAchrome is
present, TRP-2 or metal ion (as copper or zinc) favor
the formation of DHICA instead of DHI (Figure 3)
[65,69].
The pigmentation is mostly genetically determined, but
the skin melanocytes activity is under control of signals
from neighboring keratinocytes as well as autocrine
signals and environmental factors such as UV radiation
[58,71]. The secretion of keratinocyte-derived factors is
increased by UV radiation, but it is also evident that UV
can directly stimulate melanin production and melanocyte
dendricity [58,71]. Melanin production is stimulated mainly
by alpha melanocyte stimulating hormone (α-MSH) that
influences on melanocyte through the melanocortin 1
receptor (MC1R) [58,71,72]. In regulation of the melanin
synthesis signal molecules and transcription factors,
such as MITF and cAMP, play an important roles [64,73].
In the group of melanogenic inhibitors are sphingolipids,
BMP-4 (Bone Morphogenetic Protein) and
tetrahydrobiopterin [64,74,75]. During melanin synthesis
many cytotoxic agents are formed e.g. hydrogen
peroxide, quinones. To avoid the interaction between
them and other cytosolic components, the process of
melanization takes place in the encircled by membrane
structures – melanosomes [54].
3.2 Melanosome biogenesis
Melanosome biogenesis include four stages (I–IV),
which are determined by their quality, quantity, structure
and arrangement of the melanin produced (Figure 4).
They are often separated into “early” (Stage I and II) and
“late” melanosomes (Stage II and IV) according to lack
or presence of pigment [50,58,76,77].
The formation of stage I is still an open question [64].
The early melanosomes show the content of proteins
derived from the endoplasmatic reticulum, coated vesicles,
lysosomes and endosomes [76,78]. These melanosomes
are spherical vacuoles with no internal structural
components. In stage II, the visible fibrillar matrix formed
by glycoproteins (Pmel17, MART-1), melanosomes
elongates, tyrosinase is present [71,77,79]. In stage III,
melanosomes begin melanin production that polymerizes

Figure 4. The melanosome formation and maturation during the melanin production by a melanocyte. The melanogenesis takes place in a special
organelles named melanosomes. As first develops a vesicle (Stage I) which builds inside a fibrillar matrix formed by glycoproteins
(Pmel17, MART-1) and gets tyrosinase and other enzymes of melanogenesis (Stage II). The melanosome produces melanin, that
polymerizes and settles on the internal fibrils (Stage III). In the last Stage IV melanosome fulfills melanin. Each type of the melanin is
synthesized in a separated melanosomes.

320
M. Cichorek et al.
and settle on the internal fibrills net. Melanosomes have
elliptical or ellipsoidal shapes. In the last stage IV melanin
fulfills melanosome. Each type of melanin is synthesized in
a separated melanosomes [58,72]. The eumelanosomes
are large, elliptical, and contain a fibrillar matrix required
for eumelanin synthesis. The pheomelanosomes are
smaller, spherical in shape, and their matrix has loose
structure. Both types of melanosomes might be present in
a single melanocyte [50]. Most of melanosomal proteins
are glycosylated. They undergo early glycosylation in
the endoplasmic reticulum (ER) and finally in the Golgi
apparatus. In the next step, proteins are trafficked to early
or late endosomes and then to the melanosomes. The
tyrosinase uses this mechanism to reach melanosomes
[76,77,79]. Aforementioned melanogenesis process
concerns the melanin production in the epidermal
melanocytes but whether it goes the same way in
melanocytes at other places is not known.
The skin melanocytes (epidermal, hair) transfer
mature melanosomes to the dendrical processes ends
and to keratinocytes. Transport is arranged on the long
axis of the dentrites under control of cytoskeleton [64,77].
The exact mechanism of the melanosomes transfer to
keratinocytes is not well understood, it could follow by
many ways: fusion of plasma membranes, exocytosis,
transfer by membrane vesicles or cytophagocytosis
[64,77]. Human skin contains mixture of all types of
melanins [45,80].
Figure 5. Melanocytes localization in the epidermis and hair. The epidermal melanocytes are located among the basal layer (stratum basalis) of a
stratified squamos keratinized epithelium (A). The hair melanocytes are between cells covering the hair papilla in the hair bulb (B). Stem
cells for melanocytes are located in the region named the hair bulge.
4. Melanocytes in the human skin
4.1 Melanocytes in the epidermis
The mature epidermal melanocytes, as small dendritic
cells with melanosomes, are located in a basal layer of
the epidermis (Figure 5) [81]. Melanocyte cooperates
with 30–40 associated keratinocytes and form the
epidermal melanin unit together [58,82,83]. About
1200 melanocytes exist per mm2 of the skin independent
of the human race [84]. Cutaneous pigmentation
does not depend on melanocyte number, but the
intensity of melanogenesis and melanin distribution in
keratinocytes. Homeostasis between these two type
of cells is maintained by cell-cell adhesion structures
and by auto- and paracrine factors secreted by skin
cells [15,58,64,77]. The result of different disturbances
of these interactions can be uncontrolled proliferation
of the melanocytes and the nevi or melanoma
development [85]. Under the normal conditions all of
the melanin produced is carried away from melanocytes
into keratinocytes. Joshi et al. described the physical
contact between melanocyte and keratinocytes as the
ligand-receptor type interaction causing intracellular
calcium signal in keratinocytes necessary for melanin
transfer [86]. Melanin granules are accumulated in
keratinocytes as a cup above the nucleus, it protects
keratinocytes’ DNA against UV radiation and gives
color of the skin [50,58,87].
321
 Heterogeneity of neural crest-derived melanocytes

Epidermal melanocytes cooperate not only with
keratinocytes but also with fibroblasts located in
the dermis [76,88]. Melanocytes, keratinocytes and
fibroblasts communicate with each other by secreted
factors e.g. growth factors, hormones, inflammatory
mediators and cell-cell contacts (E-cadherin, integrins)
[45,83,89,90]. Hormones secreted by keratinocytes
regulate melanocyte proliferation, melanogenesis
and dendrites growth [50,58]. The signaling pathways
cross-talk between keratinocytes and melanocytes is
a part of epidermal complex network involved in the
maintenance of skin homeostatis and its basic elements
are presented in Figure 6 [35,50,58,91]. Proliferation
and pigmentation of melanocytes are under influence
of the factors secreted also by fibroblasts [50,88].
In vitro studies pointed that secreted by the dermis
fibroblasts Dickkopf-related protein 1 (DKK1) decreases
melanocytes growth and melanin production while
neuroregulin (NRG1) increases melanin production
[45,88]. Thus, paracrine and autocrine molecules
secreted by keratinocytes, fibroblast and melanocytes
are necessary for the regulation of pigment production
in response to environmental factors and also to control
topological differences in the skin color [88,92]. In
addition, these factors influence not only the growth
and pigmentation of melanocytes, but also their shape,
dendricity, mobility and adhesive properties [50,93].
It is stressed that in the melanocyte-keratinocytes
unit the pigment cell is very active. Melanocytes are
able to secrete a number of signal molecules (cytokines,
growth factors, hormones) targeting keratinocytes and
other epidermal cells [15,50,58]. Melanocytes are
important components of the skin immune system [13].
Inflammatory mediators can directly affect melanocytes
functions and may cause hypo- or hyperpigmentation of
the skin [94].
The neuroendocrine capabilities of melanocytes are
of special importance in the human skin homeostasis

Figure 6. The graphical presentation of the basic cross-talk between melanocytes and keratinocytes in the epidermis. The melanocytes
proliferation, differentiation, melanogenesis are under control of surrounding keratinocytes. Melanocyte and up to 40 keratinocytes
form the epidermal melanin unit. CF stem cell factor; bFGF basic fibroblast growth factor; GM-CSF granulocyte-macrophage colony-
stimulating factor; ET-1 endothelin 1; a-MSH melanocyte-stimulating hormones; PGE2 prostaglandin E2; PGF2a prostaglandin
F2a; NGF nerve growth factor; c-Kit tyrosine kinase receptor; FGFR1/2 fibroblast growth factor receptor; GM-CSFR granulocyte-
macrophage colony-stimulating factor receptor; ETBR endothelin B receptor; MC1R melanocortin 1 receptor; EP1/EP3/FP prostanoid
receptors; NGFR nerve growth factor receptor; MAPK mitogen-activated protein (MAP) kinases ; PKC Protein kinase C; PKA Protein
kinase A; PLC phospholipase C; TYR tyrosinase; TRP1 tyrosinase-related protein 1; TRP2 tyrosinase-related protein 2; MITF-M
Melanocyte-specific MITF (Microphthalmia-associated transcription factor) isoform; CRE cAMP response elements; CREB cAMP
response element-binding.

322
M. Cichorek et al.
[14,15]. Cumulative evidence provided that these cells
are elements of cholinergic system and hypothalamic-
pitiutary-thyroid axis [16,17,22,95]. Thus, skin
melanocytes, as the mediators of the humoral and
neural signals, coordinate not only cutaneous but
also a systemic (peripherial, global) homeostasis. The
melanocytes with melanosomes – unique organelle/
messengers – participate in the regulation of skin
functions including its protective, bioregulatory and
sensory properties [17,21,22,95].
4.2 Hair follicle melanocytes
Melanocytes are responsible for either skin or hair
pigmentation. They are located in the proximal bulb of
each hair follicle among the epithelial cells of the hair
papilla at the apex. Melanocytes dendritic processes
enter between hair keratinocytes and form the hair
melanin units (Figure 5) [96,97]. Hair pigmentation is the
result of the structural and functional interactions between
follicular melanocytes, matrix keratinocytes and dermal
papilla fibroblasts. Process of hair pigmentation includes
the melanogenic activity of follicular melanocytes, the
transfer of melanin granules into keratinocytes that form
the pigmented hair shaft [96-98]. Hair melanocytes are
larger and more dendritic than the epidermal ones,
including mainly mature Stage IV melanosomes but it
is considered that melanin transport to the keratinocytes
in the growing hair shaft is similar to the epidermal [97].
The mechanism of the melanin synthesis and its
control in the hair is similar to epidermal melanocytes
but it is stated that hair follicles melanocytes are
more sensitive to aging influences than the epidermal
melanocytes, leading to hair graying [97]. It seems that
substances derived from fibroblast of dermal papilla (e.g.
insulin-like growth factor IGF-1, keratinocyte growth
factor KGF, hepatocyte growth factor HGF, noggin
and stem cell factor SCF) have special significance
for control division and differentiation of the hair matrix
keratinocytes and differentiation, proliferation and
activity of melanocytes during the anagen hair growth
phase [96,99,100].
Epidermal melanocytes proliferate rarely, while hair
melanocytes divide repeatedly, migrate and undergo
maturation in every hair cycle [46]. Melanogenesis
process takes place only during the anagen stage
(growing phase) of the hair growth cycle; pigment
formation is turned-off in catagen (regressing phase)
and absent in telogen (resting phase). Additionally it is
marked that during catagen completely differentiated
bulbar melanocytes die through apoptosis but with the
development of the new hair new melanocytes appear.
So, except for high melanogenic melanocytes in the hair
bulb a minor population of poorly differentiated pigment
cells with inactive way of melanin biosynthesis also
exist, the melanocyte stem cells (MelSCs) [46,96,101].
MelSC are located in the hair follicle bulge together
with hair follicle stem cells (HFSCs; precursors for
keratinocytes). The bulge area is a part of outer root
sheath that provides the insertion point for arector pili
muscle and points the bottom of the permanent portion
of hair follicles (Figure 5). Each time a hair is lost the
hair follicle regenerates and MelSC are activated
under control of Wnt signaling [102]. After stimulation
these cells migrate from bulge into a new hair bulb and
differentiate into mature melanocytes [33,103]. MelSCs
are a melanocyte reservoir not only for hair but also for
epidermis [46]. The understanding of MelSCs biology
is the key for explanation the mechanisms of hair
pigmentation, greying and skin disorders e.g. vitiligo
[46,97].
5. Melanocytes in places other than
skin
As the result of prenatal migration melanocytes are
located in the ear, eye, nervous system, and heart. In
these organs melanocytes are not related to keratinocytes
– the main cells affected pigment cells of the skin.
Pigment cells functioning in the microenvironment of
ear (stria vascularis of cochlear duct), eye (choroid, iris,
ciliary body), brain (leptomeninges) and heart are linked
probably with their impact with local cells/structures. It
must be emphasized that biology of these melanocytes
is less known than skin melanocytes, especially in the
context of the cell cooperation [50].
5.1 Eye
Melanocytes of NCC origin exist in the choroid and
stroma of the iris and the ciliary body. Pigment is also
produced in the retinal pigment epithelium (RPE) and
epithelia of the iris and ciliary body, but the origin of
these cells is the optic cup not the neural crest. Unlike
epidermal melanocytes, ocular melanocytes are in
contact only with each other, and they do not transfer their
melanosomes. Mature melanocytes remain quiescent
and do not produce melanin in adults [5]. Ocular
melanocytes are embeded in the connective tissue
and are probably under influence of different cells e.g.
fibroblasts, endothelial cells, smooth muscles, neurons
[104-106].These localizations probably determines the
role of ocular melanocytes. In the anterior part of the
iris, which is exposed to sunlight and UV radiation, the
photoprotecting effect of melanin dominates. Deeply
localized ocular melanocytes are exposed to high oxygen
tension and melanin seems to protect cells against
323

oxidative damage as free radical scavenger [105,107].
Studies of patients with the iris melanogenesis defects
pointed to abnormal adrenergic innervation of the iris
[108]. Relationship of melanocytes with blood vessels,
nerve fibers and the presence of melanocytes groups
around and along an extraocular muscle, may indicate
a functional role as an ion reservoir [104]. Sharif et al.,
mentioning about intercellular communication within iris
melanogenic process, indicate the histamine receptor
expression on the human isolated iridial fibroblasts and
the presence of endothelin-1 (ET-1), prostaglandin E2
(PGE2) on the ocular melanoblast [106]. Higa et al.
described the presence of melanocytes in an additional
section of the eye, corneal limbus, transitional area
between cornea and sclera [109]. These pigment cells
were MART-1 (melanoma antigen recognized by T-cells)
and tyrosinase positive, have multiple processes and
exist as sporadic cells among basal epithelial cells.
Thus, melanocytes and epithelial cells in the limbus may
form a functional network similar to the melanin unit of
the skin [109].
5.2 Ear
The relationship between pigmentation and hearing
has been noticed by Alfonso Corti in 19th century
[110]. The presence of melanocytes are required
for normal ear development, a deficiency of these
cells in Waardenburg’s Syndrome (WS) resulted in
hypopigmentation and deafness. In another situation,
albinism, the melanin absence does not influence the
hearing process. Thus, melanocytes not melanin is
necessary for the ear development [23]. It is known that
melanocytes appear in almost all parts of the inner ear,
such as the cochlear duct, stria vascularis, vestibular
organ in the region surrounding the cristae and maculae,
semicircular canals [5].
The stria vascularis of the cochlear duct is composed
of a stratified epithelium, between the cells of which
runs a dense capillary network. The melanocytes
form intermediate layer in the stria vascularis. They
are the starry-shaped cells, have globular melanin
granules [23]. The key function of stria vascularis is the
production of endolymph and the creation of the positive
endocochlear potential through the control of the ionic
balance. Melanocytes seem to participate in this last
function. Melanocytes as a biological reservoir for
calcium and potassium ions could regulate the content
of these ions in the endolymph, that is important for the
hearing physiology [5,59,111]. The strial melanocytes
activity is controlled by the same factors as in the
epidermis e.g. MITF, c-Kit, SOX 10, PAX3 [23,112,113].
It has been observed that in humans the cochlear
melanin level increases with age [114]. The animal studies
324
Heterogeneity of neural crest-derived melanocytes
indicated that noise induces cochlea hyperpigmentation
and melanosomes transfer to marginal cells [5,23,115].
It seems that melanocytes prevent noise-induced
damage of the inner ear but the molecular mechanism
of the stress-induced melanization is unknown. The
melanin in ear, as a biological reservoir for ions, can
bind to ototoxic drugs [116]. Noise and ototoxic drugs
induce oxidative stress; high level of the reactive
oxygen substances (ROS) can destroy sensory cells in
the cochlea. Thus melanin as the scavenger of ROS
protects sensory cells from damage [23,117].
5.3 Heart
Recently, in animals studies, the presence of
melanocytes in the heart has been described [24,118].
During mammalian embryogenesis neural crest cells
are involved in the outflow part of the heart formation.
Thus, the presence of the melanocyte cells of the NCC
origin is not a surprise [118]. Melanocytes are situated
in valves, atrial and intraventricular septa [24]. These
melanocytes in heart have some structural differences
in comparison to other melanocytes e.g. have a lot of
caveloae along the plasma membranes [5]. Cardiac
melanocytes are present only in four-chambered hearts
[118]. The functions of the heart melanocytes remain
unclair.
In human heart tissue in the atrium and pulmonary
vein walls are cells with dopachrometautomerase activity
(DCT, TRP2; melanogenesis enzyme, melanoblast’s
marker) [119]. Detailed analysis showed that these
DCT-positive cells also express tyrosinase and MITF,
but further examinations are necessary to explain if
these cells are melanocytes or cardiomyocytes able
to melanin synthesis [119]. Levin et al. observed that
the presence of DCT-positive cardiac melanocyte-like
cells within the pulmonary veins and atrium contribute
to atrialarrhythmogenesis in mouse [119]. This incidate
a new research direction for human atrial arrhythmias
and melanocytes.
5.4 Nervous system
In the central nervous system melanocytes of the NCC
origin are distributed in the leptomeninges over the
ventrolateral surface of the medulla oblongata of the
human brain [25,120]. These dendritic cells contain
melanosomes [5]. It is indicated that these melanocytes
could play neuroendocrine functions e.g. secretion
of opioids. The authors suggest the possibility that
melanocytes are involved in the control of the respiratory
rhytm and sleeping regulation [120]. The human
brain has some places with neurons able to produce
neuromelanin but these neurons developed from neural
tube not NCC [120,121].
M. Cichorek et al.
Summarizing, main functions of the melanocytes
localized outside skin are: the protection of tissues from
potentially harmful factors (e.g. free radicals, binding
toxins), storage ions and anti-inflammatory action.
6. Stem cells for human melanocytes
in adults
Melanocytes proliferate rarely but some situations point
that in the skin are present precursors for melanocytes
e.g. repigmentation of the skin of vitiligo patients after
phototherapy [122].
What do we know about the presence of melanoblasts
in the adult skin?
The first documented reservoir for melanocytes stem
cells was the bulge area of the hair follicles either in
mouse or human (Figure 5) [103,123]. The bulge region
contains pluripotential, morphologically undifferentiated
cells which develop into keratinocyte progenitors and
melanoblasts not only for hair but also for epidermis
[46].
However, there are growing evidence that the dermis
is also a reservoir of melanocytes [33,122,124]. Where
are the melanoblasts located in the dermis? Answer
to this question is not satisfactory enough. Recent
developmental studies using model organisms and
lineage tracing have been able to trace melanocytes
References
[1] Fitzpatrick T., Becker S.W., Lerner A.B., Montgomery
H., Tyrosinase in human skin: Demonstration of its
presence and its role in human melanin formation,
Science, 1950, 112, 223–225
[2] Nordlund J.J., Boissy R.E., Hearing V.J., King R.A,
Ortonne J.P., The Pigmentary System. Physiology
and Pathophysiology, Oxford University Press,
New York, 1998
[3] Halaban R., Hebert D.N., Fisher D.E., Biology of
Melanocytes, In: Freedberg I.M., Wolff K., Austen
K.F., Goldsmith L.A., Katz S.I. (Eds.), Fitzpatrick’s
Dermatology in General Medicine, McGraw-Hill,
New York, 2003
[4] Aoki H., Yamada Y., Hara A., Kunisada T., Two
distinct types of mouse melanocyte: differential
signaling requirement for the maintenance of
non-cutaneous and dermal versus epidermal
melanocytes, Development., 2009, 136, 2511-
2521
arising from migration of a multipotent precursor cell
along nerve projections. It is probable that there are
cells with stem cell properties located in the cutaneous
nerves [26,28]. These cells are retained in a stem
cell-like state until the signal send by the end of the
cutaneous nerve promotes these cells to differentiate
into melanocytes [125]. But there are also observations
that some melanoblasts from the dorsal way of NCC
migration stay in the dermis after the end of epidermis
inoculation, up to the second trimester during fetal time
[33,126,127]. Whether they survive in the human adult
dermis is still an open problem.
There are difficulties in the identification of
melanoblasts in the adults skin because of the lack
of a defined set of melanoblast markers. There are
some markers determining the melanocyte fate of
NCC e.g. dopachrometautomerase (TRP2/DCT),
MITF, c-Kit.
Epidermis and dermis are places for the melanocytes
reservoir and give opportunity for a new look at the
melanoma heterogeneity, a tumor which develops
from melanocytes. There is no agreement if the mature
melanocytes or cells from earlier developmental stages
may follow the tumoricidal transformation [34,128-130].
Whiteman et al. divides melanoma(s) into melanoma
arising from epithelial melanocytes (origin from dorsal
migration way) and not associated with epithelia (origin
from Schwann precursors cells) [129].
[5] Colombo S., Berlin I., Delmas V., Larue L., Classical
and nonclassical melanocytes in vertebrates,
In: Borovansky J., Riley P.A., (Eds.), Melanins
and melanosomes. Biosynthesis, biogenesis,
physiological and pathological functions, Willey-
Blackwell, Weinheim, 2011
[6] Rawles M.E., Origin of pigment cells from neural
crest in the mouse embryo, Physiol. Zool., 1947,
20, 248-270
[7] O’Rahilly R., Müller F., The development of the neural
crest in the human, J. Anat., 2007, 211, 335-351
[8] Harris M.L., Erickson C.A., Lineage specification in
neural crest cell path finding, Dev. Dyn., 2007, 236,
1-19
[9] Betters E., Liu Y., Kjaeldgaard A., Sundström E.,
García-Castro MI., Analysis of early human neural
crest development, Dev. Biol., 2010, 344, 578-592
[10] Theveneau E., Mayor R., Neural crest delamination
and migration: From epithelium-to-mesenchyme
325

transition to collective cell migration, Dev. Biol.,
2012, 366, 34-54
[11] Le Douarin N.M., Creuzet S., Couly G., Dupin
E., Neural Crest Cell plasticity and its limits,
Development, 2004, 131, 4637-4650
[12] Ernfors P., Cellular origin and developmental
mechanisms during the formation of skin
melanocytes, Exp. Cell Res., 2010, 316, 1397-
1407
[13] Lu Y., Zhu W.Y., Tan C., Yu G.H., Gu J.X.,
Melanocytes are potential immunocompetent
cells: evidence from recognition of immunological
characteristics of cultured human melanocytes,
Pigment Cell Res., 2002, 15, 454-460
[14] Slominski A., Paus R., Are L-tyrosine and L-dopa
hormone-like bioregulators?, J. Theor. Biol., 1990,
143, 123-138
[15] Slominski A., Paus R., Schadendorf D.,
Melanocytes as “sensory” and regulatory cells in
the epidermis, J. Theor. Biol., 1993, 164, 103-120
[16] Schallreuter K.U., Lemke K.R., Pittelkow M.R.,
Wood J.M., Körner C., Malik R., Catecholamines
in human keratinocyte differentiation, J. Invest.
Dermatol., 1995, 104, 953-957
[17] Grando S.A., Pittelkow M.R., Schallreuter K.U.,
Adrenergic and cholinergic control in the biology of
epidermis: physiological and clinical significance.,
J. Invest. Dermatol., 2006, 126, 1948-1965
[18] Slominski A., Wortsman J., Luger T., Paus R.,
Solomon S., Corticotropin releasing hormone and
proopiomelanocortin involvement in the cutaneous
response to stress, Physiol. Rev., 2000, 80, 979-1020
[19] Slominski A., Wortsman J., Neuroendocrinology of
the skin, Endocr. Rev., 2000, 21, 457-487
[20] Slominski A., Wortsman J., Tobin D.J., The
cutaneous serotoninergic/melatoninergic system:
securing a place under the sun, FASEB J., 2005,
19, 176-194
[21] Slominski A., Neuroendocrine activity of the
melanocyte, Exp. Dermatol., 2009, 18, 760-763
[22] Slominski A.T., Zmijewski M.A., Skobowiat C.,
Zbytek B., Slominski R.M., Steketee J.D., Sensing
the environment: regulation of local and global
homeostasis by the skin’s neuroendocrine system,
Adv. Anat. Embryol. Cell Biol., 2012, 212, 1-115
[23] Tachibana M., Sound needs sound melanocytes to
be heard, Pigment Cell Res., 1999, 12, 344-354
[24] Mjaatvedt C.H., Kern C.B., Norris R.A., Fairey S.,
Cave C.L., Normal distribution of melanocytes in
the mouse heart, Anat. Rec. Discov. Mol. Cell.
Evol. Biol., 2005, 285, 748–757
[25] Goldgeier M,H,, Klein L.E., Klein-Angerer S.,
Moellmann G., Nordlund J.J., The distribution of
326
Heterogeneity of neural crest-derived melanocytes
melanocytes in the leptomeninges of the human
brain., J. Invest. Dermatol., 1984, 82, 235-238
[26] Cramer S.F., The histogenesis of acquired
melanocytic nevi-based on a new concept of
melanocytic differentiation, Am. J. Dermatopathol.,
1984, 6, 289-298
[27] Cramer S.F., Stem cells for epidermal melanocytes-
a challenge for students of dermatology, Am. J.
Dermatopathol., 2009, 31, 331-341
[28] Adameyko I., Lallemend F., Aquino J.B., Pereira J.A.,
Topilko P., Muller T., et al., Schwann cell precursors
from nerve innervation are a cellular origin of
melanocytes in skin, Cell, 2009, 139, 366–379
[29] Dupin E., Real C., Glavieux-Pardanaud C., Vaigot
P., LeDourain N.M., Reversal of developmental
restrictions in neural crest cells lineages; transition
from Schwann cells to glial-melanocytic precursors
in vitro, Proc. Natl. Acad. Sci. USA, 2003, 100,
5229-5233
[30] Cramer S.F., Fesyuk A., On the development
of neurocutaneous units-implications for the
histogenesis of congenital, acquired, and dysplastic
nevi, Am. J. Dermatopathol., 2012, 34, 60-81
[31] Hara M., Toyoda M., Yaar M., Bhawan J., Avila
E.M., Penner I.R., et al., Innervation of melanocytes
in human skin, J. Exp. Med., 1996, 184, 1385-1395
[32] Holbrook K.A., Underwood R.A., Vogel A.M., Gown
A.M., Kimball H., The appearance, density and
distribution of melanocytes in human embryonic
and fetal skin revealed by anti melanoma
monoclonal antibody HMB-45, Anat. Embryol.,
1989, 180, 443-455
[33] Gleason B.C., Crum C.P., Murphy G.F.,
Expression patterns of MITF during human
cutaneous embryogenesis: evidence for bulge
epithelial expression and persistence of dermal
melanoblasts, J. Cutan. Pathol., 2008, 35, 615-622
[34] Zabierowski S.E., Fukunaga-Kalabis M., Li L.,
Herlyn M., Dermis-derived stem cells: a source of
epidermal melanocytes and melanoma? Pigment
Cell Melanoma Res., 2011, 24, 422-429
[35] Hirobe T., How are proliferation and differentiation of
melanocytes regulated?, Pigment Cell Melanoma
Res., 2011, 24, 462-478
[36] Cooper C.D., Raible D.W., Mechanisms for
reaching the differentiated state: Insights from
neural crest-derived melanocytes, Semin. Cell
Dev. Biol., 2009, 20, 105-110
[37] Sommer L., Generation of melanocytes from neural
crest cells, Pigment Cell Melanoma Res., 2011, 24,
411-421
[38] Hari L., Miescher I., Shakhova O., Suter U., Chin L.,
Taketo M., et al., Temporal control of neural crest
M. Cichorek et al.
lineage generation by Wnt/β-catenin signaling,
Development, 2012, 139, 2107-2117
[39] Cornell R.A., Eisen J.S., Notch in the pathway:
the roles of Notch signaling in neural crest
development, Semin. Cell Dev. Biol., 2005, 16,
663-672
[40] Raible D.W., Ragland J.W., Reiterated Wnt and
BMP signals in neural crest development, Semin.
Cell Dev. Biol., 2005, 16, 673-682
[41] Lin J.Y., Fisher D.E., Melanocyte biology and skin
pigmentation, Nature, 2007, 445, 843-850
[42] Clay M.R., Halloran M.C., Regulation of cell
adhesions and motility during initiation of neural
crest migration, Curr. Opin. Neurobiol., 2011, 21,
17-22
[43] Vance K.W., Goding C.R., The transcription
network regulating melanocyte development and
melanoma, Pigment Cell Res., 2004, 17, 318-
325
[44] McGill G.G., Horstmann M., Widlund H.R., Du J.,
Motyckova G., Nishimura E.K. et al., Bcl2 regulation
by the melanocyte master regulator Mitf modulates
lineage survival and melanoma cell viability, Cell,
2002, 109, 707-718
[45] Yamaguchi Y., Brenner M., Hearing V.J., The
regulations of skin pigmentation, J. Biol. Chem.,
2007, 13, 1-11
[46] Nishimura E.K., Melanocyte stem cells: a
melanocyte reservoir in hair follicles for hair and
skin pigmentation, Pigment Cell Melanoma Res.,
2011, 24, 401-404
[47] Bennet D.C., Medrano E.E., Molecular regulation
of melanocyte senescence, Pigment Cell Res.,
2002, 15, 242-250
[48] Rizos H., Becker T.M., Holland E.A., Cell cycle
regulation in the melanocyte, In: Thomson J.F.,
Morton D.L., Kroon B.B.R., Textbook of Melanoma,
Martin Dunitz Taylor & Francis Group, London,
2004
[49] Nordlund J.J., The lives of pigment cells, Dermatol.
Clin., 1986, 4, 407-418
[50] Costin G.E., Hearing V.J., Human skin pigmentation:
melanocytes modulate skin color in response to
stress, FASEB J., 2007, 21, 976-994
[51] Haddad M.M., Xu W., Medrano E.E., Aging in
epidermal melanocytes: cell cycle genes and
melanins, J. Investig. Dermatol. Symp. Proc.,
1998, 3, 36-40
[52] Selzer E., Schlagbauer-Wadl H., Okamoto I.,
Pehamberger H., Pötter R., Jansen B., Expression
of Bcl-2 family members in human melanocytes, in
melanoma metastases and in melanoma cell lines,
Melanoma Res., 1998, 8, 197-203
[53] Campisi J., The role of cellular senescence in skin
aging, J. Investig. Dermatol. Symp. Proc., 1998, 3,
1-5
[54] Seiji H., Fitzpatrick T.B., The reciprocal relationship
between melanization and tyrosinase activity in
melanosomes (melanin granules), J. Biochem.,
1961, 49, 700-706
[55] Simon D.J., Peles D., Wakamatsu K., Ito S., Current
challenges in understanding melanogenesis:
bridging chemistry, biological control, morphology
and function, Pigment Cell Melanoma Res., 2009,
22, 563-579
[56] Pathak M.A., Rilley F.C., Fitzpatrick T.B.,
Melanogenesis in human skin following exposure
to long-wave ultraviolet and visible light., J. Invest.
Dermatol., 1962, 39, 435-443
[57] Riley P.A., Melanin., Int. J. Biochem. Cell Biol.,
1997, 29, 1235-1239
[58] Slominski A., Tobin D.J., Shibahara S., Wortsman
J., Melanin pigmentation in mammalian skin and its
hormonal regulation, Physiol. Rev., 2004, 84, 1155-
1228
[59] Bush W.D., Simon J.D., Quantification of Ca(2+)
binding to melanin supports the hypothesis that
melanosomes serve a functional role in regulating
calcium homeostasis, Pigment Cell Res., 2007,
20, 134–139
[60] Ito S., High-performance liquid chromatography
(HPLC) analysis of eu- and pheomelanin in
melanogenesis control, J. Invest. Dermatol., 1993,
100, 166–171
[61] Fitzpatrick T.B., Miyamoto M., Ishikawa K., The
evolution of concepts of melanin biology, Arch.
Dermatol., 1967, 96, 305-323
[62] Hearing V.J., Tsukamoto K., Enzymatic control
of pigmentation in mammals, FASEB J., 1991, 5,
2902-2909
[63] Hoashi T., Watabe H., Muller J., Yamaguchi Y.,
Vieira W.D., Hearing V.J., MART-1 is required
for the function of the melanosomal matrix
protein PMEL17/GP100 and the maturation of
melanosomes, J. Biol. Chem., 2005, 280, 14006-
14016
[64] Park H.Y., Kasmadaki M., Gilchrest Y.B.A., Cellular
mechanisms regulating melanogenesis, Cell Mol.
Life Sci., 2009, 66, 1493-1506
[65] Ito S., The IFPCS presidential lecture: a chemist’s
view of melanogenesis, Pigment Cell Res., 2003,
16, 230–236
[66] Pawelek J.M., After dopachrome?, Pigment Cell
Res., 1991, 4, 53–62
[67] Olivares C., Jiménez-Cervantes C., Lozano
J.A. Solano F., García-Borrón J.C., The
327

5,6-dihydroxyindole-2-carboxylic acid (DHICA)
oxidase activity of human tyrosinase, Biochem. J.,
2001, 354, 131-139
[68] Ito S., Wakamatsu K., Chemistry of mixed
melanogenesis-pivotal roles of dopaquinone,
Photochem. Photobiol., 2008, 84, 582-592
[69] Del Marmol V., Beerman F., Tyrosinase and related
proteins in mammalian pigmentation, FEBS Lett.,
1996, 381, 165-168
[70] Korner A.M., Pawelek J., DOPAchrome
conversion: a possible control point in melanin
biosynthesis, J. Invest. Dermatol., 1980, 75, 192–
195
[71] Hearing V.J., Determination of melanin synthetic
pathway, J. Invest. Dermatol., 2011, 131, 8-11
[72] Schiaffino M.V., Signalling pathways in melanosome
biogenesis and pathology, Int. J. Biochem. Biol.,
2010, 42, 1094-1104
[73] Goding C.R., Mitf from neural crest to melanoma:
signal transduction and transcription in the
melanocyte lineage, Genes. Dev., 2000, 14, 1712-
1728
[74] Park H.Y., Wu C., Yaar M., Stachur C.M., Kosmadaki
M., Gilchrest B.A., Role of BMP-4 and its signaling
pathways in cultured human melanocytes, Int. J.
Cell Biol., 2009, 2009, doi:10.1155/2009/750482
[75] Schallreuter K.U., Wood J.M., Pittelkow M.R.,
Gütlich M., Lemke K.R., Rödl W., et al., Regulation
of melanin biosynthesis in the human epidermis
by tetrahydrobiopterin, Science, 1994, 263, 1444-
1446
[76] Hearing V.J., Biogenesis of pigment granules: a
sensitive way to regulate melanocyte function, J.
Dermatol. Sci., 2005, 37, 3-14
[77] Watabe H., Valencia J.C., Le Pape E., Yamaguchi
Y., Nakamura M., Rouzaud F., Involvement of
dynein and spectrin with early melanosome
transport and melanosomal protein trafficking, J.
Invest. Dermatol., 2008, 128, 162-173
[78] Raposo G., Tenza D., Murphy D.M., Berson
J.F., Marks M.S., Distinct protein sorting and
localization to premelanosomes, melanosomes,
and lysosomes in pigmented melanocytic cells, J.
Cell Biol., 2001, 152, 809-824
[79] Kushimoto T., Basrur V., Valencia J., Matsunaga
J., Vieira W. D., Ferrans V.J., et al., A model for
melanosome biogenesis based on the purification
and analysis of early melanosomes, Proc. Natl.
Acad. Sci. USA., 2001, 98, 10698–10703
[80] Kondo T., Hearing V.J., Update on the regulation
of mammalian melanocyte function and skin
pigmentation, Expert. Rev. Dermatol., 2011, 6, 97-
108
328
Heterogeneity of neural crest-derived melanocytes
[81] Busam K.J., Charles C., Lee G., Halpern A.C.,
Morphologic features of melanocytes, pigment
keratinocytes, and melanophages by in vivo
confocal scanning laser microscopy, Mod. Pathol.,
2001, 14, 862-868
[82] Fitzpatrick T.B., Breathnach A.S., The epidermal
melanin unit system, Dermatol. Wochenschr.,
1963, 147, 481-489
[83] Haass N.K., Smalley K.S., Li L., Herlyn M.,
Adhesion, migration and communications in
melanocytes and melanoma, Pigment Cell Res.,
2005, 18, 150-159
[84] Miot L.D., Miot H.A., Silva M.G., Marques M.E.,
Physiopathology of melasma, An. Bras. Dermatol.,
2009, 84, 623-635
[85] Rickelt S., Franke W.W., Doerflinger Y., Goerdt
S., Brandner J.M., Peitsch W.K., Subtypes of
melanocytes and melanoma cells distinguished by
their intercellular contacts: heterotypic adherent
junctions, adhesive associations, and dispersed
desmoglein 2 glycoproteins, Cell Tissue Res.,
2008, 334, 401-422
[86] Joshi P.G., Nair N., Begum G., Joshi N.B., Sinkar
V.P., Vora S., Melanocyte-keratinocyte interaction
induces calcium signaling and melanin transfer to
keratinocytes, Pigment Cell Res., 2007, 20, 380-
384
[87] Plonka P.M., Passeron T., Brenner D.J., Tobin S.,
Shibahara S., Thomas A., What are melanocytes
really doing all day long…?, Exp. Dermatol., 2009,
18, 799-819
[88] Choi W., Kolbe L., Hearing V.J., Characterization
of the bioactive motif of neuregulin-1, a fibroblast-
derived paracrine factor that regulates the
constitutive color and the function of melanocytes
in human skin, Pigment Cell Melanoma Res., 2012,
25, 1-5
[89] Lee A.Y., Role of keratinocytes in the development
of vitiligo, Ann. Dermatol., 2012, 24, 115-125
[90] Tabone-Eglinger S., Wehrle-Haller M., Aebischer
N., Jacquier M.C., Wehrle-Haller B., Membrane
bound Kit ligand regulates melanocyte adhesion
and survival, providing physical interaction with an
intraepithelial niche, FASEB J., 2012, 26, 3738-
3753
[91] Hirobe T., Role of keratinocyte-derived
factors involved in regulating the proliferation
and differentiation of mammalian epidermal
melanocytes, Pigment Cell Res., 2004, 18, 2-12
[92] Imokawa G., Autocrine and paracrine
regulation of melanocytes in human skin and
in pigmentary disorders, Pigment Cell Res.,
2004, 17, 96-110
M. Cichorek et al.
[93] Wang Z., Coleman D.J., Bajaj G., Liang X.,
Ganguli-Indra G., Indra AK., RXRα ablation in
epidermal keratinocytes enhances UVR-induced
DNA damage, apoptosis, and proliferation
of keratinocytes and melanocytes, J. Invest.
Dermatol., 2011, 131, 177-187
[94] Taylor S., Grimes P., Lim J., Im S., Lui H.,
Postinflammatory hyperpigmentation, J. Cutan.
Med. Surg., 2009, 13, 183-191
[95] Slominski A., Zmijewski M.A., Pawelek J.,
L-tyrosine and L-dihydroxyphenylalanine as
hormone-like regulators of melanocyte functions,
Pigment Cell Melanoma Res., 2012, 25, 14-27
[96] Commo S., Bernard B. A., Melanocyte
subpopulation turnover during the human hair
cycle: an immunohistochemical study, Pigment
Cell Res., 2000, 13, 253-259
[97] Slominski A., Wortsman J., Plonka P.M.,
Schallreuter K.U., Paus R., Tobin D.J., Hair
Follicle Pigmentation, J. Invest. Dermatol., 2005,
124, 13-21
[98] Commo S., Gaillard O., Thibaut S., Bernard B.A.,
Absence of TRP-2 in melanogenic melanocytes of
human hair, Pigment Cell Res., 2004, 17, 488-497
[99] Randall V.A., Androgens and hair growth,
Dermatol. Ther., 2008, 21, 314-328
[100] Slominski A., Paus R., Melanogenesis is coupled
to murine anagen: toward new concepts for
the role of melanocytes and the regulation
of melanogenesis in hair growth, J. Invest.
Dermatol., 1993, 101, 90-97
[101] Slominski A., Paus R., Plonka P., Chakraborty A.,
Maurer M., Pruski D., et al., Melanogenesis during
the anagen-catagen-telogen transformation of
the murine hair cycle, J. Invest. Dermatol., 1994,
102, 862-869
[102] Goding C.R., Melanocytes; the new black, Int. J.
Biochem. Cell Biol., 2007, 39, 275-279
[103] Nishimura E., Jordan S.A., Oshima H., Yoshida
H., Osawa M., Moriyama M., et al., Dominant
role of the niche in melanocyte stem-cell fate
determination, Nature, 2002, 416, 854-860
[104] Wan der Werf F., Baljet B., Otto A.J., Pigment-
containing cells in extraocular tissues of the
primate, Doc. Ophthalmol., 1992, 81, 357-368
[105] Hu D.N., Simon J., Sarna T., Role of ocular
melanin in ophtalmic physiology and pathology,
Phatochem. Phatobiol., 2008, 84, 639-644
[106] Sharif N.A., Crider J.Y., Intracellular signaling in
human iridial fibroblasts and iridial melanocytes
in response to prostaglandins, endothelin,
isoproterenol, and other pharmacological agents,
Curr. Eye Res., 2011, 36, 310-320
[107] Sarna T.J., Properties and function of the ocular
melanin - a photobiophysical view, Photochem.
Photobiol. B., 1992, 12, 215-258
[108] Melamed S., Lahav M., Sandbank U., Yassur Y.,
Ben-Sira I., Fuch’s heterochromic iridocyclitis:
an electron microscopic study of the iris, Invest.
Ophthalmol. Vis. Sci., 1978, 17, 1193-1199
[109] Higa K., Shimmura S., Miyashita H., Shimazaki
J., Tsubota K., Melanocytes in the corneal limbus
interact with K19-positive basal epithelial cells,
Exp. Eye Res., 2005, 81, 218-223
[110] Corti A., Recherche sur l`organe des l`ouie des
mammiferes. Premiere partie 2 Z. wiss, Zoologie,
1851, 3, 109-169
[111] Meyer zum Gottesberge A.M., Calcium dependent
intercellular interaction of the neural crest
derivate-melanocytes and the epithelial cells of
the vestibular organ, Acta. Otolaryngol., 1995,
520, 360-361
[112] Tachibana M., Cochlear melanocytes and MITF
signaling, J. Invest. Dermatol. Symp. Proc., 2001,
6, 95-98
[113] Price E.R., Fisher D.E., Sensorineural deafness
and pigmentation genes: melanocytes and the
Mitf transcriptional network, Neuron, 2001, 30,
15-18
[114] Suzuki T., Nomoto Y., Nakagawa T., Kuwahat
N., Ogawa H., Suzuki Y., et al., Age-dependent
degeneration of the stria vascularis in human
cochleae, Laryngoscope, 2006, 116, 1846-1850
[115] Wassif G.A., El Begermy M., Age-related changes
of the cochlear lateral wall (stria vascularis) in the
guinea pigs: an ultrastructural study, Egypt J.
Histol., 2008, 31, 332-340
[116] Meyer zum Gottesberge A.M., Physiology and
pathophysiology of inner ear melanin, Pigment
Cell Res., 1988, 1, 238-249
[117] Henderson D., McFadden S.L., Liu C.C., Hight N.,
Zheng X.Y., The role of antioxidants in protection
from impulse noise, Ann. N. Y. Acad. Sci., 1999,
884, 368-380
[118] Brito F.C., Kos L., Timeline and distribution of
melanocyte precursors in the mouse heart,
Pigment Cell Melanoma Res., 2008, 21, 464-470
[119] Levin M.D., Lu M.M., Petrenko N.B., Hawkins
B.J., Gupta T.H., Lang D., et al., Melanocyte-like
cells in the heart and pulmonary veins contribute
to atrial arrhythmia triggers, J. Clin. Invest., 2009,
119, 3420-3436
[120] Takeda K., Takahashi N.H., Shibahara S.,
Neuroendocrine function of melanocytes: beyond
the skin-deep melanin maker, Tohoku J. Exp.
Med., 2007, 211, 201-221
329

[121] Zecca L., Bellei Ch., Costi P., Albertini A., Monzani
E., Casella L., New melanic pigments in the
human brain that accumulate in aging and block
environmental toxic metals, Proc. Natl. Acad. Sci.
USA, 2008, 105, 17567-17572
[122] Davids L.M., du Toit E., Kidson S.H., Todd G., A
rare repigmentation pattern in a vitiligo patient:
a clue to an epidermal stem-cell reservoir of
melanocytes?, Clin. Exp. Dermatol., 2009, 34,
246-248
[123] Nishimura E.K., Granter S.R., Fisher D.E.,
Mechanisms of hair graying; incomplete
melanocyte stem cell maintance in the niche,
Science, 2005, 307, 720-724
[124] Toma J.G., McKenzie I.A., Bagli D., Miller F.D.,
Isolation and characterization of multipotent skin-
derived precursors from human skin, Stem Cells,
2005, 23, 727-737
[125] Adameyko I., Lallemend F., Furlan A., Zinin
N., Aranda S., Kitambi S.S., et al., Sox2 and
Mitf cross-regulatory interactions consolidate
330
Heterogeneity of neural crest-derived melanocytes
progenitor and melanocyte lineages in the
cranial neural crest, Development, 2012, 139,
397-410
[126] Suder E., Bruzewicz S., Melanocytes of fetal
dermis - studies with anti-HMB-45 antibody, Med.
Sci. Monit., 2004, 10, 229-232
[127] Dupin E., Sommer L., Neural crest progenitors
and stem cells: From early development to
adulthood, Dev. Biol., 2012, 366, 83-95
[128] Schatton T., Frank M.H., Cancer stem cells and
human malignant melanoma, Pigment Cell
Melanoma Res., 2007, 21, 39-55
[129] Whiteman D.C., Pavan W.J., Bastian B.C., The
melanomas: a synthesis of epidemiological,
clinical, histopathological, genetic, and biological
aspects, supporting distinct subtypes, casual
pathways, and cells of origin, Pigment Cell
Melanoma Res., 2011, 24, 879-897
[130] Hoek K.S., Goding C.R., Cancer stem cells versus
phenotype-switching in melanoma, Pigment Cell
Melanoma Res., 2010, 23, 746-759