Oxidative enzymes in the sebaceous glands of the domestic cat

Synopsis The distribution and activities of several oxidative enzymes in various regions of the sebaceous glands of the domestic cat have been studied. The results obtained emphasize the outstanding importance of NADP-linked dehydrogenases for lipogenesis during sebum production. In particular, the reactions for glucose-6-phosphate dehydrogenase were very strong. Among the NAD-linked dehydrogenases investigated, lactate dehydrogenase showed strong activity in the peripheral cells of the sebaceous gland. The reactions for cytochrome oxidase and succinate dehydrogenase were weaker.     

Acne

The cause of acne is still obscure, but genetic predisposition, sebaceous overactivity, overgrowth of bacterial flora and exposure to comedogenic substances are all significant factors.Acne lesions occur mainly in sebaceous follicles, which are characterized by deep follicular canals and large sebaceous glands. The associated seborrhea is not due to a circulatory excess of androgens but may be caused by a local amplification of androgenic activity. This, in turn, may be due to large numbers of androgen receptors and a high concentration of enzymes such as 17β-hydroxysteroid dehydrogenase, within the sebaceous gland itself. Hyperkeratosis of the retention type in the pilary infrainfundibulum obstructs the outflow of sebum and keratin flakes. This favors the proliferation of Propionibacterium acnes which may initiate inflammation in microcomedos and lead to formation of pustules, papules or nodules.Topical therapy with tretinoin, benzoyl peroxide and antibiotics such as clindamycin is widely used today. Oral tetracyclines and other chemotherapeutic agents remain necessary in severe cases.     

Comparative responsiveness of supracaudal and other sebaceous glands in male and female guinea pigs to hormones

Different types of sebaceous glands in guinea pigs were chosen to study their comparative responsiveness to steroid hormones. Glands selected were (1) Sebaceous glands associated with rudimentary hair in the supracaudal gland; (2) Free sebaceous glands of the nipple; (3) Sebaceous glands associated with the hair. The results showed distinct differences in sensitivity among these different sebaceous glands and according to the sex of the experimental animal. The most responsive of the glands in males was the supracaudal gland; in the female the most responsive glands were the sebaceous glands of the nipple. Sebaceous glands associated with normal hair were relatively insensitive to changes in hormonal level. In all cases testosterone propionate was more potent in stimulating any of the sebaceous glands than progesterone. After gonadectomy, various sebaceous glands showed unequal states of depletion due to the different rates of differentiation and disintegration of their cells into sebum.     

Fine structure of the gular gland of the free-tailed bat Tadarida brasiliensis

The gular gland of the bat Tadarida brasiliensis is a specialized sebaceous gland located in the skin of the suprasternal region of adult males. It consists of an aggregation of simple branched tubulo-acinar gland units, the number of which varies seasonally. Each acinus is composed of densely packed sebaceous cells at various stages of differentiation. Acinar basal cells and cells of the epithelium of the ducts can differentiate into sebaceous cells. Two main changes appear in the cytoplasm concurrent with the sebaceous transformation: the differentiation of cytoplasmic organelles and the deposition of lipid material. The appearance of a different type of mitochondrion and the development of large numbers of ribosomes and polyribosomes can be recognized in the cytoplasm at an early stage of differentiation. Concomitant with the deposition of significant numbers of lipid droplets, the cells develop abundant agranular endoplasmic reticulum occurring mainly as scattered tubular cisternae. These at times form whorls surrounding lipid droplets. At later stages, the cisternae of the agranular endoplasmic reticulum often occur in crystalline arrays between secretory oil droplets. The roles of the different cytoplasmic organelles, especially in relation to the production of sebum, are discussed.     

Comparative studies of the ultrastructure of the sebaceous gland

Three-dimensional reconstructions and ultrastructural evidence on the sebaceous glands of man and domestic animals indicate that sebum is produced from a column of developing and degenerating cell populations derived from peripheral progenitor cells at the base of the lobe in a manner analogous to hair growth. The remainder of the peripheral cells apparently have no direct involvement with sebocyte production; those towards the neck of the gland contribute keratin to the secretory product.     

Cidea Control of Lipid Storage and Secretion in Mouse and Human Sebaceous Glands

Sebaceous glands are skin appendages that secrete sebum onto hair follicles to lubricate the hair and maintain skin homeostasis. In this study, we demonstrated that Cidea is expressed at high levels in lipid-laden mature sebocytes and that Cidea deficiency led to dry hair and hair loss in aged mice. In addition, Cidea-deficient mice had markedly reduced levels of skin surface lipids, including triacylglycerides (TAGs) and wax diesters (WDEs), and these mice were defective in water repulsion and thermoregulation. Furthermore, we observed that Cidea-deficient sebocytes accumulated a large number of smaller-sized lipid droplets (LDs), whereas overexpression of Cidea in human SZ95 sebocytes resulted in increased lipid storage and the accumulation of large LDs. Importantly, Cidea was highly expressed in human sebaceous glands, and its expression levels were positively correlated with human sebum secretion. Our data revealed that Cidea is a crucial regulator of sebaceous gland lipid storage and sebum lipid secretion in mammals and humans.     

Correlation of Propionibacterium acnes Populations with the Presence of Triglycerides on Nonhuman Skin

The skins of mice, rats, rabbits, sheep, guinea pigs, and dogs were cultured for Propionibacterium acnes. Only the sebaceous regions (perianal gland) of guinea pigs harbored a significant P. acnes population. Analysis of the lipid from this region revealed a significant percentage of triglycerides, compounds lacking in the sebum of the other animals.     

Identification and characterization of ABCB1-mediated and non-apoptotic sebum secretion in differentiated hamster sebocytes

Sebaceous glands secrete sebum onto the skin surface in a holocrine manner and as such a thin lipid layer is formed as a physiological barrier. In the present study, extracellular level of triacylglycerols (TG), a major sebum component, as well as intracellular TG accumulation was augmented in insulin-differentiated hamster sebocytes (DHS). The DHS exhibited phosphatidylserine exposure in an apoptosis-independent manner. In addition, intracellular ATP level and membrane-transporter activity using a substrate, Rhodamine 123, were highly detectable in the DHS rather than in the undifferentiated hamster sebocytes. A membrane-transporter activating reagent, 2'(3')-O-(4-benzoylbenzoyl) adenosine 5'-triphosphate (BzATP), enhanced transporter activity, extracellular TG level, and phosphatidylserine exposure in the DHS. Both transporter activity and TG secretion were suppressed by R-verapamil, a potent membrane-transporter inhibitor, in the BzATP-treated and untreated DHS. Furthermore, the gene expression and production of ATP-binding cassette subfamily B member 1 (ABCB1) were augmented in the DHS. ABCB1 was also detectable in sebaceous glands in the skin of hamsters. Moreover, the cell-differentiation- and BzATP-augmented transporter activity and TG secretion were dose-dependently inhibited by adding not only an ABCB1 antibody but also a selective inhibitor of ABCB1, PSC833. Thus, these results provide novel evidence that ABCB1 is involved in sebum secretion in the DHS, which is associated with non-apoptotic phosphatidylserine exposure and the increased level of intracellular ATP. These findings should accelerate the understanding of sebum secretion occurring in a holocrine-independent manner in sebaceous glands, and may contribute to the development of therapies for sebaceous gland disorders such as acne, seborrhea, and xerosis.     

Acid hydrolases in the perinuclear secretion granules of the rat preputial gland. A light and electron microscope study

Synopsis Four acid hydrolases in the secretory cells and the sebum of the preputial sebaceous gland of the rat were incestigated cytochemically. A strong -glucuronidase activity was found to occur in the matrix of the perinuclear secretion granules, whereas the granule crystalloids were unreactive. The distribution of acid phosphatase at the light microscope level was similar, though the intensity of the reaction was lower and the number of positive granules smaller. By electron microscopy, the final reaction product of acid phosphatase occurred in patches at the periphery of the granule matrix, as well as in the vesicles adjoining the Golgi stacks, from which the perinuclear granules seemed to arise. In the sebum, the two hydrolases occurred in the background material between the unstained crystalloid masses. There was noN-acetyl--glucosaminidase or aryl sulphatase activity in the gland. The perinuclear granules appear to be secretory lysosomes which, after discharge from the disaggregating cell, release their acid hydrolases into the sebum.     

Normal fur development and sebum production depends on fatty acid 2-hydroxylase expression in sebaceous glands

2-Hydroxylated fatty acid (HFA)-containing sphingolipids are abundant in mammalian skin and are believed to play a role in the formation of the epidermal barrier. Fatty acid 2-hydroxylase (FA2H), required for the synthesis of 2-hydroxylated sphingolipids in various organs, is highly expressed in skin, and previous in vitro studies demonstrated its role in the synthesis of HFA sphingolipids in human keratinocytes. Unexpectedly, however, mice deficient in FA2H did not show significant changes in their epidermal HFA sphingolipids. Expression of FA2H in murine skin was restricted to the sebaceous glands, where it was required for synthesis of 2-hydroxylated glucosylceramide and a fraction of type II wax diesters. Absence of FA2H resulted in hyperproliferation of sebocytes and enlarged sebaceous glands during hair follicle morphogenesis and anagen (active growth phase) in adult mice. This was accompanied by a significant up-regulation of the epidermal growth factor receptor ligand epigen in sebocytes. Loss of FA2H significantly altered the composition and physicochemical properties of sebum, which often blocked the hair canal, apparently causing a delay in the hair fiber exit. Furthermore, mice lacking FA2H displayed a cycling alopecia with hair loss in telogen. These results underline the importance of the sebaceous glands and suggest a role of specific sebaceous gland or sebum lipids, synthesized by FA2H, in the hair follicle homeostasis.     

1H NMR-based lipidomics of rodent fur: species-specific lipid profiles and SCD1 inhibitor-related dermal toxicity

A method is described that allows noninvasive identification and quantitative assessment of lipid classes present in sebaceous excretions in rodents. The method relies on direct high-field proton NMR analysis of common group lipid protons in deuterated organic solvent extracts of fur. Extracts from as little as 15 mg of fur from rat, mouse, and hamster provided acceptable results on a 600 MHz NMR equipped with a cryogenically cooled proton-observe probe. In rats, sex- and age-related differences in lipid composition are larger than differences in fur collected from various body regions within an individual and much larger than interanimal differences in age- and sex-matched specimens. The utility of this method to noninvasively monitor drug-induced sebaceous gland atrophy in rodents is demonstrated in rats dosed with a stearoyl-CoA desaturase 1 (SCD1) inhibitor. In this model, a 35% reduction in sebum lipids, extracted from fur, was observed. Finally, structural elucidation of cholesta-7,24-dien-3β-ol ester as the most prominent, previously unidentified sebum sterol ester in male Syrian hamsters is described. The utility of this method for drug and cosmetic safety and efficacy assessment is discussed.     

Development and homeostasis of the sebaceous gland

The important role of epidermal appendages especially the sebaceous gland has only recently been recognized. In particular, it has been convincingly shown that normal development and maintenance of the sebaceous gland are required for skin homeostasis since atrophic sebaceous glands and disturbances in sebaceous lipid composition result in major defects of the physiological barrier and maintenance of the skin. Consequently, it is important to unravel the signaling network controlling proper sebaceous lineage differentiation in mammalian skin and to understand the underlying mechanisms leading to severe skin diseases, including abnormal proliferation and differentiation of the gland, defects of the lipid metabolism and barrier, as well as sebaceous tumor formation. Over the last years, results from transgenic and knock out mouse models manipulating distinct signaling pathways in the skin as well as the detailed analysis of human sebaceous gland-derived cell lines provided new insights into crucial mediators balancing proliferation and differentiation of the sebaceous gland. Here, we discuss our current knowledge of in vivo mechanisms of sebaceous gland development, maintenance and disorders and highlight recent contributions to the field of sebaceous gland biology.     

Identification of two novel human acyl-CoA wax alcohol acyltransferases: members of the diacylglycerol acyltransferase 2 (DGAT2) gene superfamily

The esterification of alcohols such as sterols, diacylglycerols, and monoacylglycerols with fatty acids represents the formation of both storage and cytoprotective molecules. Conversely, the overproduction of these molecules is associated with several disease pathologies, including atherosclerosis and obesity. The human acyl-CoA:diacylglycerol acyltransferase (DGAT) 2 gene superfamily comprises seven members, four of which have been previously implicated in the synthesis of di- or triacylglycerol. The remaining 3 members comprise an X-linked locus and have not been characterized. We describe here the expression of DGAT2 and the three X-linked genes in Saccharomyces cerevisiae strains virtually devoid of neutral lipids. All four gene products mediate the synthesis of triacylglycerol; however, two of the X-linked genes act as acyl-CoA wax alcohol acyltransferases (AWAT 1 and 2) that predominantly esterify long chain (wax) alcohols with acyl-CoA-derived fatty acids to produce wax esters. AWAT1 and AWAT2 have very distinct substrate preferences in terms of alcohol chain length and fatty acyl saturation. The enzymes are expressed in many human tissues but predominate in skin. In situ hybridizations demonstrate a differentiation-specific expression pattern within the human sebaceous gland for the two AWAT genes, consistent with a significant role in the composition of sebum.     

Further Observations on the Pathogenesis of Acne

The composition of the lipid of the forehead skin surface and the sebum excretion rate were determined in 217 subjects and controls. Acne was associated with an increase in serum excretion rate and in the squalene and wax and sterol esters in surface lipid. The changes in sebum excretion rate and squalene were statistically significant only in women, but acne in men was associated with a significant decrease in the free fatty acid content or surface lipid.The increased amounts of squalene and wax esters may lead to pilosebaceous obstruction in acne subjects. The increased sebum excretion rate in acne may ensure increased production of sebaceous triglyceride substrate available for lipolysis to irritant free fatty acids, which may then mediate the inflammatory changes of acne.     

Mammalian wax biosynthesis. II. Expression cloning of wax synthase cDNAs encoding a member of the acyltransferase enzyme family

Wax monoesters are synthesized by the esterification of fatty alcohols and fatty acids. A mammalian enzyme that catalyzes this reaction has not been isolated. We used expression cloning to identify cDNAs encoding a wax synthase in the mouse preputial gland. The wax synthase gene is located on the X chromosome and encodes a member of the acyltransferase family of enzymes that synthesize neutral lipids. Expression of wax synthase in cultured cells led to the formation of wax monoesters from straight chain saturated, unsaturated, and polyunsaturated fatty alcohols and acids. Polyisoprenols also were incorporated into wax monoesters by the enzyme. The wax synthase had little or no ability to synthesize cholesteryl esters, diacylglycerols, or triacylglycerols, whereas other acyltransferases, including the acyl-CoA:monoacylglycerol acyltransferase 1 and 2 enzymes and the acyl-CoA:diacylglycerol acyltransferase 1 and 2 enzymes, exhibited modest wax monoester synthesis activities. Confocal light microscopy indicated that the wax synthase was localized in membranes of the endoplasmic reticulum. Wax synthase mRNA was abundant in tissues rich in sebaceous glands such as the preputial gland and eyelid and was present at lower levels in other tissues. Coexpression of cDNAs specifying fatty acyl-CoA reductase 1 and wax synthase led to the synthesis of wax monoesters. The data suggest that wax monoester synthesis in mammals involves a two step biosynthetic pathway catalyzed by fatty acyl-CoA reductase and wax synthase enzymes.     

Histology and cytochemistry of human skin. IX. The distribution of non-specific esterases

1. In the epidermis non-specific esterase activity outlines a strongly reactive band between the stratum granulosum and the stratum corneum. In the epidermis of the palm, there is no such esterase-rich band. 2. The outer sheath of active hair follicles has strong enzyme activity. The degenerating hair bulb in catagen follicles is very strongly reactive, and clusters of cells around the hair club in quiescent follicles are rich in enzyme activity. 3. Strong enzyme activity is found in young sebaceous cells, while decaying sebaceous cells and newly formed sebum are unreactive. Old sebum, however, is very intensely reactive. 4. Only the "dark" cells of eccrine sweat glands show a reaction; the "clear" cells are negative. 5. The cells of axillary apocrine glands abound in enzyme.     

Structure and function of pilosebaceous units on the heads of small mammals (Insectivora, Chiroptera, Rodentia)

Haffner, M. 2000. Structure and function of pilosebaceous units on the heads of small mammals (Insectivora, Chiroptera, Rodentia). —Acta Zoologica (Stockholm) 81 : 195–203 The cranial integument of 32 species was examined for increased sebum production and structures serving sebum storage and expulsion by means of light and scanning electron microscopy. The head is rich in sebaceous glands in all species. Some have no further specializations other than regions with enlarged sebum production. These regions either need extra protection per se or they are situated where they can be used for fur grooming directly or be easily reached by the forepaws. The water shrew stores sebum that it can possibly apply as a water‐repellent lipid coating for the fur if required. All vespertilionid bats not only have huge glandular complexes with storage chambers, but also structures for the retaining and application of sebum. Stabilizing elements reflect the extent of external pressure on the specialized pilary canal during sebum discharge. Originally simple hair follicles evolved highly adapted pilosebaceous complexes that possibly produce scent themselves or at least a carrier substance for scent secreted by the many apocrine glands on the snout.     

HISTOLOGY AND CYTOCHEMISTRY OF HUMAN SKIN : IX. THE DISTRIBUTION OF NON-SPECIFIC ESTERASES

1. In the epidermis non-specific esterase activity outlines a strongly reactive band between the stratum granulosum and the stratum corneum. In the epidermis of the palm, there is no such esterase-rich band. 2. The outer sheath of active hair follicles has strong enzyme activity. The degenerating hair bulb in catagen follicles is very strongly reactive, and clusters of cells around the hair club in quiescent follicles are rich in enzyme activity. 3. Strong enzyme activity is found in young sebaceous cells, while decaying sebaceous cells and newly formed sebum are unreactive. Old sebum, however, is very intensely reactive. 4. Only the "dark" cells of eccrine sweat glands show a reaction; the "clear" cells are negative. 5. The cells of axillary apocrine glands abound in enzyme.     

Sexual hormones in human skin.

The skin locally synthesizes significant amounts of sexual hormones with intracrine or paracrine actions. The local level of each sexual steroid depends upon the expression of each of the androgen- and estrogen-synthesizing enzymes in each cell type, with sebaceous glands and sweat glands being the major contributors. Sebocytes express very little of the key enzyme, cytochrome P450c17, necessary for synthesis of the androgenic prohormones dehydroepiandrosterone and androstenedione, however, these prohormones can be converted by sebocytes and sweat glands, and probably also by dermal papilla cells, into more potent androgens like testosterone and dihydrotestosterone. Five major enzymes are involved in the activation and deactivation of androgens in skin. Androgens affect several functions of human skin, such as sebaceous gland growth and differentiation, hair growth, epidermal barrier homeostasis and wound healing. Their effects are mediated by binding to the nuclear androgen receptor. Changes of isoenzyme and/or androgen receptor levels may have important implications in the development of hyperandrogenism and the associated skin diseases such as acne, seborrhoea, hirsutism and androgenetic alopecia. On the other hand, estrogens have been implicated in skin aging, pigmentation, hair growth, sebum production and skin cancer. Estrogens exert their actions through intracellular receptors or via cell surface receptors, which activate specific second messenger signaling pathways. Recent studies suggest specific site-related distribution of ERalpha and ERbeta in human skin. In contrast, progestins play no role in the pathogenesis of skin disorders. However, they play a major role in the treatment of hirsutism and acne vulgaris, where they are prescribed as components of estrogen-progestin combination pills and as anti-androgens. These combinations enhance gonadotropin suppression of ovarian androgen production. Estrogen-progestin treatment can reduce the need for shaving by half and arrest progression of hirsutism of various etiologies, but do not necessarily reverse it. However, they reliably reduce acne. Cyproterone acetate and spironolactone are similarly effective as anti-androgens in reducing hirsutism, although there is wide variability in individual responses.