Destruction of the arrector pili muscle and fat infiltration in androgenic alopecia.

BACKGROUND: Androgenic alopecia (AGA) is the most common hair loss condition in men and women. Hair loss is caused by follicle miniaturization, which is largely irreversible beyond a certain degree of follicular regression. In contrast, hair loss in telogen effluvium (TE) is readily reversible. The arrector pili muscle (APM) connects the follicle to the surrounding skin. OBJECTIVES: To compare histopathological features of the APM in AGA and TE. METHODS: Archival blocks of 4-mm scalp punch biopsies from eight patients with AGA and five with TE were obtained. New 4-mm biopsies from five normal cases were used as controls. Serial 7-µm sections were stained with a modified Masson's trichrome stain. 'Reconstruct' software was used to construct and evaluate three-dimensional images of the follicle and APM. RESULTS: The APM degenerated and was replaced by adipose tissue in all AGA specimens. Remnants of the APM remained attached to the hair follicle. There was no fat in the normal skin specimens. Fat was seen in two of five TE specimens but could be attributed to these patients also showing evidence of AGA. Quantitative analysis showed that muscle volume decreased and fat volume increased significantly (P < 0·05) in AGA compared with controls. CONCLUSIONS: APM degeneration and replacement with fat in AGA has not previously been described. The underlying mechanism remains to be determined. However, we speculate that this phenomenon might be related to depletion of stem or progenitor cells from the follicle mesenchyme, explaining why AGA is treatment resistant.

Hair loss in women: medical and cosmetic approaches to increase scalp hair fullness.

Androgenetic alopecia affects both men and women. In men it produces male pattern hair loss with bitemporal recession and vertex baldness. In women it produces female pattern hair loss (FPHL) with diffuse alopecia over the mid-frontal scalp. FPHL occurs as a result of nonuniform hair follicle miniaturization within follicular units. Diffuse alopecia is produced by a reduction in the number of terminal fibres per follicular unit. Baldness occurs only when all hairs within the follicular units are miniaturized and is a relatively late event in women. The concepts of follicular units and primary and secondary hair follicles within follicular units are well established in comparative mammalian studies, particularly in sheep. However, discovery of these structures in the human scalp hair and investigation of the changes in follicular unit anatomy during the development of androgenetic alopecia have provided a clearer understanding of the early stages of androgenetic alopecia and how the male and female patterns of hair loss are related. FPHL is the most common cause of alopecia in women and approximately one-third of adult caucasian women experience hair loss. The impact of FPHL is predominantly psychological. While men anticipate age-related hair loss, hair loss in women is usually unexpected and unwelcome at any age. Treatment options to arrest hair loss progression and stimulate partial hair regrowth for FPHL include the androgen receptor antagonists spironolactone and cyproterone acetate, the 5α-reductase inhibitor finasteride and the androgen-independent hair growth stimulator minoxidil. These treatments appear to work best when initiated early. Hair transplantation should be considered in advanced FPHL that is resistant to medical treatments. Hair transplantation requires well-preserved hair growth over the occipital donor area. The psychological impact of FPHL may also be reduced by cosmetic products that improve the appearance of the hair. These agents work to minimize hair fibre breakage, improve hair volume or conceal visible bald scalp.

Epidermal Cells Expressing Putative Cell Markers in Nonglabrous Skin Existing in Direct Proximity with the Distal End of the Arrector Pili Muscle.

Inconsistent with the view that epidermal stem cells reside randomly spread along the basal layer of the epidermal rete ridges, we found that epidermal cells expressing stem cell markers in nonglabrous skin exist in direct connection with the distal end of the arrector pili muscle. The epidermal cells that express stem cell markers consist of a subpopulation of basal keratinocytes located in a niche at the lowermost portion of the rete ridges at the distal arrector pili muscle attachment site. Keratinocytes in the epidermal stem cell niche express K15, MCSP, and α6 integrin. α5 integrin marks the distal end of the APM colocalized with basal keratinocytes expressing stem cell markers located in a well-protected and nourished environment at the lowermost point of the epidermis; these cells are hypothesized to participate directly in epidermal renewal and homeostasis and also indirectly in wound healing through communication with the hair follicle bulge epithelial stem cell population through the APM. Our findings, plus a reevaluation of the literature, support the hierarchical model of interfollicular epidermal stem cell units of Fitzpatrick. This new view provides insights into epidermal control and the possible involvement of epidermal stem cells in nonmelanoma skin carcinogenesis.

Identification of differentially expressed genes during a wool follicle growth cycle induced by prolactin.

The wool follicles of New Zealand Wiltshire sheep can be induced to undergo growth cycles by manipulating circulating prolactin levels. Altered patterns of gene expression through this cycle were examined using differential display, and nine sequence tags for differentially expressed genes were isolated. Four of these tags were identified as fragments of known genes, encoding a wool keratin, KRTAP3.2, a desmosome component, desmoglein 1, an epithelial cell marker, stratifin, and a protein kinase, Clk3. All four genes were shown to be downregulated in telogen skin compared with anagen. In situ hybridization showed that all had localization patterns which included cells that are absent in telogen. The stratifin tag was used to clone a cDNA that incorporated a complete open-reading frame for ovine stratifin. Ovine stratifin is similar to the human form, showing only six single residue differences in the predicted amino acid sequence. Stratifin probably acts as a regulator of other proteins involved in trichocyte cell cycling and differentiation. Clk3 is involved in regulating RNA splicing. KRTAP3.2 and Dsg1 both play structural roles in hair follicles. The other five tags, including two representing genes that were upregulated during catagen, could not be identified by homology. Differential display is an effective means of identifying genes involved in follicle function and, potentially, of genes controlling the growth cycle.

Myogenic differentiation of dermal papilla cells from bovine skin.

Cells from the dermal papilla and dermal sheath of hair follicles exhibit pronounced plasticity in vitro, being capable of adopting fat, bone, hematopoietic, and nerve cell phenotypes. In this study, we show that bovine dermal papilla cells (DPC) are also capable of undergoing skeletal muscle differentiation. DiI labeled DPC incorporated into myotubes when co-cultured with differentiating C(2)C(12) myoblasts. Bovine-specific PCR assays showed that the muscle markers MyoD and myogenin were up-regulated, confirming that the DPC had adopted a myogenic gene expression program. Nine clonal lines of DPC underwent both adipogenic and myogenic differentiation, demonstrating the multipotency of individual cells. Primary populations of both DPC and extra-follicular dermal fibroblasts were also capable of both adipogenic and myogenic differentiation. However, on myogenic differentiation, cells derived from dermal papillae expressed higher levels of myogenin than primary fibroblasts derived from extra-follicular dermis, suggesting that papilla cells undergo myogenesis more efficiently. This result shows that populations of fibroblastic cells derived from different anatomical sites within the skin are not equivalent with respect to their plasticity. Cultured DPC and dermal fibroblasts both expressed Pax3, a marker for the dermomyotome which represents a common embryological origin of muscle and dermis. Quantitative PCR showed that Pax3 expression levels before myogenic induction correlated with myogenin expression levels after myogenesis. These results suggest that a degree of dedifferentiation may underlie the plasticity of dermal cells in vitro, and that this plasticity may be predicted, at least in part, by levels of Pax3 expression.

Purification and characterization of the candidate prohormone-processing enzyme SPC3 produced in a mouse L cell line.

SPC3 is a member of a growing family of mammalian subtilisin-like serine proteases which play a probable role in proprotein maturation. In this study we have prepared a mouse L cell line stably expressing rat SPC3 cDNA and characterized the recombinant SPC3 protein secreted into the medium. Three molecular forms of recombinant SPC3 were identified with molecular masses of 86, 75, and 64 kDa. NH2-terminal sequence analysis indicated that all three forms were cleaved following the sequence -Arg107-Arg-Lys-Arg110, indicating removal of an SPC3 prosequence. All three molecular forms showed a 3-4-kDa decrease in molecular mass following incubation with endoglycosidase F. Two SPC3 carboxyl-terminal-directed antisera recognized only the 86-kDa molecular form of recombinant SPC3, demonstrating that COOH-terminal truncation of SPC3 protein is responsible for the different molecular mass forms. Recombinant SPC3 had a pH optimum of 6.0 and was stimulated by calcium, with maximum activity at 10 mM. Recombinant SPC3 was inhibited most effectively by the thiol-reactive reagent p-hydroxymecuribenzoate and the heavy metal chelators EDTA and EGTA. Recombinant SPC3 was also inhibited by alpha 1-antitrypsin Pittsburgh as well as wild type alpha 1-antitrypsin and antithrombin III. The inhibitor specificities revealed using these high molecular mass serpins differ from those reported for other members of the subtilisin-like serine protease family and may be able to be exploited to distinguish between closely related members of this new enzyme superfamily. Studies of cleavage specificity using tri- and tetrapeptidyl coumarins that contained pairs of basic residues indicated that tetrapeptide substrates that contained an S4 Arg residue as part of an -Arg-X-Lys/Arg-Arg motif were the most effective synthetic peptide substrates. Recombinant SPC3 also cleaved human proalbumin following the Arg-Gly-Val-Phe-Arg-Arg prosequence. Circulating human proalbumin variants that contained a mutation at either of the basic amino acids adjacent to the cleavage site were not cleaved by recombinant SPC3. Recombinant SPC3 was also able to cleave after a single arginine residue in chicken proalbumin following the Arg-Asn-Leu-Gln-Arg-Phe-Ala-Arg prosequence. These results define the primary structure requirements for cleavage by recombinant SPC3 and remain consistent with a role for SPC3 in proprotein/prohormone maturation.