Nigel A. Hibberts

The Hair Follicle: A Paradoxical Androgen Target Organ

Androgens are the main regulator of normal human hair growth. After puberty, they promote transformation of vellus follicles, producing tiny, unpigmented hairs, to terminal ones, forming larger pigmented hairs, in many areas, e.g. the axilla. However, they have no apparent effect on the eyelashes, but can cause the opposite transformation on the scalp leading to the replacement of terminal hairs by vellus ones and the gradual onset of androgenetic alopecia. This paradox appears to be an unique hormonal effect. Hair follicles are mainly epithelial tissues, continuous with the epidermis, which project into the dermis. A mesenchyme-derived dermal papilla enclosed within the hair bulb at the base controls many aspects of follicle function. In the current hypothesis for androgen regulation, the dermal papilla is also considered the main site of androgen action with androgens from the blood binding to receptors in dermal papilla cells of androgen-sensitive follicles and causing an alteration of their production of paracrine factors for target cells e.g. keratinocytes. Studies of cultured dermal papilla cells from sites with different responses to androgens in vivo have confirmed the paradoxical responses. All dermal papilla cells from androgen-sensitive sites contain low capacity, high affinity androgen receptors. However, only some cells formed 5α-dihydrotestosterone, e.g. beard but not axillary cells, in line with hair growth in 5α-reductase deficiency. Incubation with androgens also stimulated the mitogenic capacity of beard cell media, but inhibited that produced by scalp cells. This suggests that the paradoxical differences are due to differential gene expression within hair follicles, presumably caused during embryogenesis.

Tyrosine hydroxylase isoenzyme I is present in human melanosomes: a possible novel function in pigmentation

Abstract: Both human epidermal melanocytes and keratinocytes have the full capacity for de novo synthesis of 6(R) L‐erythro 5,6,7,8, tetrahydrobiopterin, the essential cofactor for the rate limiting step in catecholamine synthesis, via tyrosine hydroxylase. Catecholamine synthesis has been demonstrated in proliferating keratinocytes of the epidermis in human skin. This study presented herein identified for the first time the expression of tyrosine hydroxylase isozyme I mRNA within the melanocyte. The location of the enzyme was demonstrated in both the cytosol and melanosomes of human epidermal melanocytes, using immunohistochemistry and immunofluorescence double staining as well as immunogold electron microscopy. High‐performance liquid chromatography (HPLC) analysis of pure melanosomal extracts from the human melanoma cell line, FM94, confirmed the production of L‐dopa within these organelles. In addition, enzyme activities for both tyrosine hydroxylase and tyrosinase were measured in the same preparations, by following the catalytic release of tritiated water from L‐[3,5‐3H]tyrosine. The melanosomal membrane location of tyrosine hydroxylase together with tyrosinase implies a coupled interaction, where L‐dopa production facilitates the activation of tyrosinase. Our results support a direct function for tyrosine hydroxylase in the melanosome via a concerted action with tyrosinase to promote pigmentation.

Hormones and hair growth: Variations in androgen receptor content of dermal papilla cells cultured from human and red deer (Cervus Elaphus) hair follicles

Many hair follicles produce different types of hair in response to environmental changes or the mammals age, that are translated to the follicle by hormones. Androgens cause many changes, such as transforming vellus follicles producing insignificant hairs on the face to terminal beard ones at puberty or the reverse on the scalp. In male red deer the breeding season rise in androgens causes the annual production of a mane on the neck that is lost during the spring. Because the dermal papilla situated at the base of the hair follicle is important in determining the type of hair produced, androgens may act via the dermal papilla. Therefore, primary cell lines of dermal papilla cells from human and red deer follicles with different responses to androgens have been established. Specific saturable androgen receptors were present in all human papilla cells examined, with higher levels in cells from androgen-dependent follicles, e.g., beard than in control, non-balding scalp cells. In preliminary investigations of red deer, androgen receptors were only present in cells derived from mane follicles and were undetectable in flank or spring neck follicles. These similar results from both species support the hypothesis that androgens are acting on hair follicles via the dermal papilla. They also suggest that dermal papilla cells are potentially useful models for investigating the mechanism of androgen action because cultured cells appear to retain differences that relate to the androgen responsiveness of their parent follicle. The red deer seems particularly interesting in view of the much shorter hair-growth cycle than human scalp or beard follicles.

Stem cell factor/c-Kit signalling in normal and androgenetic alopecia hair follicles

Androgens stimulate many hair follicles to alter hair colour and size via the hair growth cycle; in androgenetic alopecia tiny, pale hairs gradually replace large, pigmented ones. Since stem cell factor (SCF) is important in embryonic melanocyte migration and maintaining adult rodent pigmentation, we investigated SCF/c-Kit signalling in human hair follicles to determine whether this was altered in androgenetic alopecia. Quantitative immunohistochemistry detected three melanocyte-lineage markers and c-Kit in four focus areas: the epidermis, infundibulum, hair bulb (where pigment is formed) and mid-follicle outer root sheath (ORS). Colocalisation confirmed melanocyte c-Kit expression; cultured follicular melanocytes also exhibited c-Kit. Few ORS cells expressed differentiated melanocyte markers or c-Kit, but NKI/beteb antibody, which also recognises early melanocyte-lineage antigens, identified fourfold more cells, confirmed by colocalisation. Occasional similar bulbar cells were seen. Melanocyte distribution, concentration and c-Kit expression were unaltered in balding follicles. Androgenetic alopecia cultured dermal papilla cells secreted less SCF, measured by ELISA, than normal cells. This identifies three types of melanocyte-lineage cells in human follicles. The c-Kit expression by dendritic, pigmenting, bulbar melanocytes and rounded, differentiated, non-pigmenting ORS melanocytes implicate SCF in maintaining pigmentation and migration into regenerating hair bulbs. Less differentiated, c-Kit-independent cells in the mid-follicle ORS stem cell niche and occasionally in the bulb, presumably a local reserve for long scalp hair growth, implicate other factors in activating stem cells. Androgens appear to reduce alopecia hair colour by inhibiting dermal papilla SCF production, impeding bulbar melanocyte pigmentation. These results may facilitate new treatments for hair colour changes in hirsutism, alopecia or greying.

A comparison of the culture and growth of dermal papilla cells from hair follicles from non-balding and balding (androgenetic alopecia) scalp

Male pattern baldness is a common, androgen‐dependent skin problem in adult men which is not well understood, although androgens are believed to act on the hair follicle via the mesenchyme‐derived dermal papilla situated in the middle of the hair follicle bulb. Since dermal papilla cells retain specific characteristics in culture, such as hair‐growth promoting ability and appropriate features of the mechanism of androgen action, dermal papilla cells from follicles undergoing androgen‐stimulated miniaturization may provide a useful in vitro model system. Therefore, dermal papilla cells have been derived from intermediate follicles from balding and nearly clinically normal sites of men with androgenetic alopecia. Balding dermal papillae were much smaller than non‐balding ones and grew much less well under normal growth conditions. Supplementing the medium with human serum, rather than fetal calf serum, increased both the yield of established cultures and the number and health of the dermal papilla cells produced. Non‐balding cells also grew faster in human serum. Balding cells retained the normal fibroblastic shape and aggregative behaviour of dermal papilla cells, but always grew less well than non‐balding cells. Nearly clinically normal dermal papillae were similar, or slightly smaller, in size to non‐balding ones, but their growth resembled balding cells. Since balding dermal papilla cells can be cultured, though with much greater difficulty than non‐balding ones, and exhibit differing growth characteristics to non‐balding cells, they merit further investigation which may Increase our understanding of. and ability to control, androgenetic alopecia.

In situ and in vitro evidence for DCoH/HNF-1α transcription of tyrosinase in human skin melanocytes

Human epidermal melanocytes hold the full capacity for autocrine de novo synthesis/regulation/recycling of the essential cofactor 6-tetrahydrobiopterin (6BH(4)) for conversion of L-phenylalanine via phenylalanine hydroxylase to L-tyrosine and for production of L-Dopa via tyrosine hydroxylase to initiate both pigmentation and catecholamine synthesis in these neural crest-derived cells. Earlier we have demonstrated pterin-4a-carbinolamine dehydratase (PCD) mRNA and enzyme activities in epidermal melanocytes and keratinocytes. This protein dimerises also the transcription factor hepatocyte nuclear factor 1 (HNF-1), leading to activation of multiple genes. This study demonstrates for the first time DCoH/HNF-1 alpha expression and transcriptional activity in human epidermal melanocytes in vitro and in situ and identified tyrosinase, the key enzyme for pigmentation, as a new transcriptional target. Specific binding of DCoH/HNF-1 complex to the human tyrosinase promoter was confirmed by gel shift analysis. These results provide a novel mechanism in the regulation of skin pigmentation.

Ability to culture dermal papilla cells from red deer (Cervus elaphus) hair follicles with differing hormonal responses in vivo offers a new model for studying the control of hair follicle biology

Red deer stags annually grow two distinct seasonal coats, a winter coat and a summer coat; in addition, they produce a mane during the breeding season when plasma testosterone levels are high, which is replaced by the short neck hairs of the summer coat when testosterone levels are low. As two very different hair types are produced from the same follicle under hormonal regulation, they offer an interesting model for studying the effects of hormones, particularly androgens, on mammalian hair growth. Since the dermal papilla of the hair follicle has a regulatory function and is probably the site of androgen action, we have investigated whether cells from the dermal papilla can be readily cultured from various types of red deer follicles; as the follicular connective tissue sheath may regenerate a new papilla in vivo, this was also examined. Individual dermal papillae and lower portions of the connective tissue sheath were microdissected from mane and flank follicles of red deer stags during the winter breeding season and from the summer coat during the nonbreeding season. Primary cultures were established from isolated dermal papillae, connective tissue sheath and dermal explants, subcultured and reestablished after freezing. Deer dermal papilla cells resembled sheep cells; they displayed a polygonal shape and irregular organisation, but did not form aggregates in contrast to human and rat vibrissa cells. Connective tissue sheath cell morphology was intermediate between that of dermal papilla cells and dermal fibroblasts. However, all three cell types derived during the breeding season grew at a much faster rate than the same cells derived during the nonbreeding season. Therefore, primary cell lines can be fairly readily derived from deer hair follicles. Since the red deer stag offers both androgen-dependent neck (mane) and control flank follicles in the breeding season, plus control nonbreeding season neck follicles, this means that stag follicular cells, particularly the dermal papilla cells, appear to offer a unique novel model system for the study of the hormonal regulation of hair growth.