An apocrine mechanism delivers a fully immunocompetent exocrine secretion.

Apocrine secretion is a recently discovered widespread non-canonical and non-vesicular secretory mechanism whose regulation and purpose is only partly defined. Here, we demonstrate that apocrine secretion in the prepupal salivary glands (SGs) of Drosophila provides the sole source of immune-competent and defense-response proteins to the exuvial fluid that lies between the metamorphosing pupae and its pupal case. Genetic ablation of its delivery from the prepupal SGs to the exuvial fluid decreases the survival of pupae to microbial challenges, and the isolated apocrine secretion has strong antimicrobial effects in "agar-plate" tests. Thus, apocrine secretion provides an essential first line of defense against exogenously born infection and represents a highly specialized cellular mechanism for delivering components of innate immunity at the interface between an organism and its external environment.

Xanthine oxidoreductase mediates membrane docking of milk-fat droplets but is not essential for apocrine lipid secretion.

KEY POINTS: Xanthine oxidoreductase (XOR) modulates milk lipid secretion and lactation initiation. XOR is required for butyrophilin1a1 clustering in the membrane during milk lipid secretion. XOR mediates apical membrane reorganization during milk lipid secretion. Loss of XOR delays milk fat globule secretion. XOR loss alters the proteome of milk fat globules. ABSTRACT: Apocrine secretion is utilized by epithelial cells of exocrine glands. These cells bud off membrane-bound particles into the lumen of the gland, losing a portion of the cytoplasm in the secretion product. The lactating mammary gland secretes milk lipid by this mechanism, and xanthine oxidoreductase (XOR) has long been thought to be functionally important. We generated mammary-specific XOR knockout (MGKO) mice, expecting lactation to fail. Histology of the knockout glands showed very large lipid droplets enclosed in the mammary alveolar cells, but milk analysis showed that these large globules were secreted. Butyrophilin, a membrane protein known to bind to XOR, was clustered at the point of contact of the cytoplasmic lipid droplet with the apical plasma membrane, in the wild-type gland but not in the knockout, suggesting that XOR mediates 'docking' to this membrane. Secreted milk fat globules were isolated from mouse milk of wild-type and XOR MGKO dams, and subjected to LC-MS/MS for analysis of protein component. Proteomic results showed that loss of XOR leads to an increase in cytoplasmic, cytoskeletal, Golgi apparatus and lipid metabolism proteins associated with the secreted milk fat globule. Association of XOR with the lipid droplet results in membrane docking and more efficient retention of cytoplasmic components by the secretory cell. Loss of XOR then results in a reversion to a more rudimentary, less efficient, apocrine secretion mechanism, but does not prevent milk fat globule secretion.

Sweat gland progenitors in development, homeostasis, and wound repair.

The human body is covered with several million sweat glands. These tiny coiled tubular skin appendages produce the sweat that is our primary source of cooling and hydration of the skin. Numerous studies have been published on their morphology and physiology. Until recently, however, little was known about how glandular skin maintains homeostasis and repairs itself after tissue injury. Here, we provide a brief overview of sweat gland biology, including newly identified reservoirs of stem cells in glandular skin and their activation in response to different types of injuries. Finally, we discuss how the genetics and biology of glandular skin has advanced our knowledge of human disorders associated with altered sweat gland activity.

Multipotent nestin-positive stem cells reside in the stroma of human eccrine and apocrine sweat glands and can be propagated robustly in vitro.

Human skin harbours multiple different stem cell populations. In contrast to the relatively well-characterized niches of epidermal and hair follicle stem cells, the localization and niches of stem cells in other human skin compartments are as yet insufficiently investigated. Previously, we had shown in a pilot study that human sweat gland stroma contains Nestin-positive stem cells. Isolated sweat gland stroma-derived stem cells (SGSCs) proliferated in vitro and expressed Nestin in 80% of the cells. In this study, we were able to determine the precise localization of Nestin-positive cells in both eccrine and apocrine sweat glands of human axillary skin. We established a reproducible isolation procedure and characterized the spontaneous, long-lasting multipotent differentiation capacity of SGSCs. Thereby, a pronounced ectodermal differentiation was observed. Moreover, the secretion of prominent cytokines demonstrated the immunological potential of SGSCs. The comparison to human adult epidermal stem cells (EpiSCs) and bone marrow stem cells (BMSCs) revealed differences in protein expression and differentiation capacity. Furthermore, we found a coexpression of the stem cell markers Nestin and Iα6 within SGSCs and human sweat gland stroma. In conclusion the initial results of the pilot study were confirmed, indicating that human sweat glands are a new source of unique stem cells with multilineage differentiation potential, high proliferation capacity and remarkable self renewal. With regard to the easy accessibility of skin tissue biopsies, an autologous application of SGSCs in clinical therapies appears promising.

Sympathetic skin responses in adult humans during sequential swallowing.

STUDY AIMS: Autonomic changes, especially those of sympathetic skin responses (SSR), during sequential water swallowing (SWS) have not been systematically investigated. This study aims to electrophysiologically examine these autonomic changes (SSR and heart rate) that occur during 50 ml sequential water swallowing from a cup. MATERIALS AND METHODS: Fifty-eight normal healthy adults were included in the study. Their submental muscle activity, respiratory activity, heart rate changes, and sympathetic skin responses were recorded during 50 ml water swallowing. In addition, we requested subjects to imagine drinking water as they did just before. The same recordings were performed during this imagination period. RESULTS: SSR appeared at the beginning and at the end of SWS in 52% of subjects. A first sympathetic skin response was evoked at the onset of SWS, and a second one appeared 8.6±1.7 seconds after the first one and at the end of swallowing. Similar double SSRs were also obtained during imagination in most investigated subjects (33 out of 35 of selected subjects in a total group of 58 subjects). Swallowing tachycardia was observed during the SWS-associated apnea period, but not during the imagination period. Heart rate significantly increased during the SWS-associated apnea period. CONCLUSION: The first SSR that appeared at the onset of swallowing is likely related to arousal. The appearance of a second response is a novel finding, which is probably related to the activity of subtil corticosubcortical networks. While discrete/single swallows can be used to evoke SSRs, SWS is unlikely to be clinically useful in its current form. In contrast, swallowing tachycardia could be a useful tool to examine dysphagic patients.

Sweaty skin, background and assessments.

Overproduction of sweat, sweaty skin, and body malodors are felt unpleasant by many people in diverse social groups. This review summarizes the functional aspects of the eccrine, apocrine, and apoeccrine sweat glands as encountered in health and disease. A series of measuring methods are conveniently used to explore any trouble in sweat production. These include the gravimetric method, water evaporation quantification, skin staining procedures, dye injections, casting replicas, average electrometric assessments, and skin capacitance mapping / imaging (SCM). Qualitative assessment is provided by collection of sweat and volatile compounds followed by chromatographic analysis. At present, SCM appears to be the most valuable method for assessing the number of active glands and their individual sweat production by the eccrine and apoeccrine glands.

Both type-I hemidesmosomes and adherens-type junctions contribute to the cell-substratum adhesion system in myoepithelial cells.

Myoepithelial cells present in exocrine glands cause secretion from the glands by contraction. They have mixed characteristics with regard to cytoskeletal elements, containing both epithelial-type intermediate filaments and smooth muscle-type myofilaments. For further characterization, myoepithelial cells from bovine apocrine sweat glands and tracheal glands were here examined with special attention to the cell-substratum adhesion system. Immunofluorescence microscopy using a panel of antibodies against adherens-type junctional and hemidesmosomal proteins demonstrated two types of cell-substratum junctions in myoepithelial cells from both glands. Type-I hemidesmosomes (HDs) consisting of plectin, BP230, integrin alpha6beta4, and BP180 were thus observed as punctate arrays longitudinally arranged along myoepithelial cell surfaces, while adherens-type junctions were similarly evident as linear rib-like structures. Double-label immunofluoresence revealed the two junctions to be distributed in a mutually exclusive or independent manner. Electron microscopy further demonstrated that apocrine myoepithelial cells surround secretory epithelial cells completely, without any gaps, HDs being abundant along the basement membrane, but with no distinct structures in the inter-hemidesmosomal regions. Immunoelectron microscopy, however, revealed an interhemidesmosomal localization of vinculin, pointing to the existence of adherens-type junctions. Secretory epithelial cells in tracheal glands were found not to be completely covered with myoepithelial cells, so that more than half of them are directly attached to the basement membrane, where they form type II-HDs lacking BP230 and BP180, but no detectable adherens junctions, like epidermal basal cells and sebaceous gland cells. These observations demonstrate that, in addition to their cytoskeleton, myoepithelial cells have both epithelial- and smooth muscle-type cell-substratum adhesion structures, i.e. HDs and dense plaque-like adherens junctions.

Apocrine secretory mechanism: recent findings and unresolved problems.

Cell secretion is an important physiological process that ensures smooth metabolic activities, tissue repair and growth and immunological functions in the body. It occurs when the intracellular secretory materials are released to the exterior; these may be in the form of lipids, protein or mucous and may travel through a duct system or via blood to reach the target organ. To date three types of secretory mechanisms have been characterized, they include apocrine, holocrine and exocytosis. Apocrine secretion occurs when the release of secretory materials is accompanied with loss of part of cytoplasm. The secretory materials may be contained in the secretory vesicles or dissolved in the cytoplasm that is lost during secretion. In holocrine secretion, the entire cell is secreted into the glandular lumen, and it is presumed that the intended secretory materials are contained in the cell cytoplasm. Exocytosis is the most commonly occurring type of secretion; here the secretory materials are contained in the secretory vesicles and released without loss of cytoplasm. Apocrine secretory mechanisms have not been properly discussed; for example the biochemical and physiological pathways that regulate apocrine secretory process are not clearly known. Similarly, the plasma membrane dynamics during apocrine secretion has not been researched. In other glands morphological features during apocrine secretion have not been documented. The current paper reviews what is known about apocrine secretion, recent findings and highlights on the unresolved areas for future research.

The biological actions of estrogens on skin.

There is still extensive disparity in our understanding of how estrogens exert their actions, particularly in non-reproductive tissues such as the skin. Although it has been recognized for some time that estrogens have significant effects on many aspects of skin physiology and pathophysiology, studies on estrogen action in skin have been limited. However, estrogens clearly have an important function in many components of human skin including the epidermis, dermis, vasculature, hair follicle and the sebaceous, eccrine and apocrine glands, having significant roles in skin aging, pigmentation, hair growth, sebum production and skin cancer. The recent discovery of a second intracellular estrogen receptor (ERbeta) with different cell-specific roles to the classic estrogen receptor (ERalpha), and the identification of cell surface estrogen receptors, has provided further challenges to understanding the mechanism of estrogen action. It is now time to readdress many of the outstanding questions regarding the role of estrogens in skin and improve our understanding of the physiology and interaction of steroid hormones and their receptors in human skin. Not only will this lead to a better understanding of estrogen action, but may also provide a basis for further interventions in pathological processes that involve dysregulation of estrogen action.

Structure and function of human sweat glands studied with histochemistry and cytochemistry.

The basic structure and the physiological function of human sweat glands were reviewed. Histochemical and cytochemical techniques greatly contributed the elucidation of the ionic mechanism of sweat secretion. X-ray microanalysis using freeze-dried cryosections clarified the level of Na, K, and Cl in each secretory cell of the human sweat gland. Enzyme cytochemistry, immunohistochemistry and autoradiography elucidated the localization of Na,K-ATPase. These data supported the idea that human eccrine sweat is produced by the model of N-K-2Cl cotransport. Cationic colloidal gold localizes anionic sites on histological sections. Human eccrine and apocrine sweat glands showed completely different localization and enzyme sensitivity of anionic sites studied with cationic gold. Human sweat glands have many immunohistochemical markers. Some of them are specific to apocrine sweat glands, although many of them stain both eccrine and apocrine sweat glands. Histochemical techniques, especially immunohistochemistry using a confocal laser scanning microscope and in situ hybridization, will further clarify the relationship of the structure and function in human sweat glands.

[Morphology of sweat glands]. / Morphologie des glandes sudorales.

There are two types of sweat glands: eccrine glands, which do not show cytological changes during secretion and apocrine glands, characterised by decapitation secretion, in which part of the cell is pinched off and released into the lumen. Eccrine glands play a major role in thermoregulation and electrolyte balance. They are present everywhere in the human skin and are composed of a secretory portion, an intradermal duct and an upper intraepidermal part, called acrosyringium. The acrosyringium has a unique symmetrical and helicoidal course, which length is correlated to the thickness of the epidermis. Apocrine glands are located only on genital, axillary and mammary areas, where they are always connected to a hair follicle. Their exact role in humans is unknown. A third type of intermediate sweat glands, the apoeccrine glands, was recently described in axillary areas. Sweat glands can be involved in various inflammatory processes and can lead to a large range of both benign and malignant tumors.

Quantification of the response of equine apocrine sweat glands to beta2-adrenergic stimulation.

The aim of the present study was to characterise the quantitative sweating response of the horse to beta2-adrenergic stimulation. The sweating responses of 6 horses to the randomised infusion of 8 different adrenaline concentrations (0.025, 0.05, 0.075, 0.1, 0.2, 0.4, 1.0 or 2.0 microg/kg bwt/min), was investigated. Sweating rate (SR) and skin temperature (TSK) on the neck (N) and gluteal region (G), and plasma adrenaline and noradrenaline concentrations were measured. Peak SR was approximately 15 (N) and approximately 9 g/m2/min (G) during infusion of both 1.0 and 2.0 microg/kg bwt/min adrenaline. Sweat produced per nmol/l plasma adrenaline peaked during the infusion of 0.075 microg/kg bwt/min adrenaline. Higher adrenaline infusion concentrations resulted in a progressive decrease in the amount of sweat produced per nmol/l plasma adrenaline and a plateau of 6 g/m2/(nmol/l) plasma adrenaline was reached for infusions between 1.0 and 2.0 microg/kg bwt/min. Peak SR were far lower than we have previously reported during exercise. There was no evidence of sweat gland fatigue or vasoconstriction during infusion, suggesting saturation of sweat gland beta2 receptors. We conclude that sweating in the horse is under dual control from a combination of hormonal and neural mechanisms.

Peroxisomes in the apocrine sweat glands of the human axilla and their putative role in pheromone production.

The products of the human apocrine axillary glands contain volatile steroids which act as pheromones. The steroidal structure of these pheromones implies that the axillary glands should be able to synthesize cholesterol which is the essential precursor of these molecules. Since important steps in cholesterol synthesis are localized within peroxisomes, we investigated the occurrence and the putative role of peroxisomes in the axillary glands at protein and mRNA levels by immunocytochemistry, Western blotting, and RT-PCR. Numerous peroxisomes were localized in the cells of the apocrine glands by immunocytochemistry, and the presence of catalase was confirmed by Western blotting and RT-PCR. Additionally, RT-PCR revealed the presence of mRNAs of two peroxisome-associated enzymes of the cholesterol biosynthetic pathway, mevalonate kinase and farnesyl diphosphate synthase. The results suggest that the peroxisomes in the human apocrine axillary glands may play a pivotal role in the biosynthesis of pheromones.

Why do we have apocrine and sebaceous glands?

The secretions of sebaceous and apocrine glands fulfil an important thermoregulatory role in cold-stressed and heat-stressed hunter-gatherers. In hot conditions the secretions emulsify eccrine sweat and thus encourage the formation of a sweat sheet and discourage the formation and loss of sweat drops from the skin. In colder conditions sebum changes its nature and repels rain from skin and hair.

Cultivation of epithelia from the secretory coil of the ovine apocrine gland: evidence of secretory cell function and ductal morphogenesis in vitro.

The secretory coil of the ovine apocrine gland is composed predominantly of two cell types, secretory cells lining the lumen and myoepithelial cells adjacent to the basement membrane. The glands synthesize a number of hormones and growth factors, but analysis of the functions of these molecules may be hampered by the mixing of apocrine and sebaceous secretions in the pilary canal. The purpose of this study was to isolate the glands and devise simple culture procedures to facilitate investigations of secretory cell function. The most successful approach involved microdissection of the secretory coils individually from skin biopsies and culture in Dulbecco's modified Eagle's medium. After 1-2 wk in medium, cell outgrowths were seen from explants. These consisted predominantly of populations of epithelial cells, many containing granules. Smaller granules were usually concentrated around the cell nuclei and accumulated lipophilic dyes. Large granules were unreactive. Western analysis showed that cells in culture synthesized nerve growth factor-like peptides, a feature consistent with one of the functions of the gland in vivo. When isolated secretor, coils were explanted to culture dishes coated with matrigel, highly compact, multilayered masses of cells grew out. Subsequently, tubular structures formed. The observations suggest that some differentiated functions of gland cells were retained in vitro and that the procedures described provide a system for the study, of apocrine secretions in isolation from those of other skin glands.

Apocrine secretion--fact or artifact?

In this review, the history of apocrine secretion and the essential categories are briefly mentioned and fused into a more generally applicable terminology. Using the coagulating gland of the male rat as a model, the mechanisms of apocrine secretion, the participation of the cytoskeleton in the formation of the apocrine blebs ("aposomes") and the structure of the secretory proteins, as well as the hormonal regulation of their biosynthesis are described. Apocrine secreted proteins share the following peculiarities: (i) Their biosynthesis and post-translational modification (including an unusual form of glycosylation) take place in the cytoplasm. (ii) Intracellular transport proceeds without participation of the endomembrane system, the Golgi apparatus and secretion granules. (iii) Blood serum derived transsudated albumin entering the secretory cells functions as a carrier of the apocrine-released proteins. Some common molecular features are specific for the apocrine-synthesized proteins studied so far by our group: (a) Their primary sequence is synthesized without a signal peptide. (b) Their N-terminus is blocked by acetylation. (c) The substituting glycanes are neither O- nor N-linked. (d) At least one of the apocrine-synthesized proteins (secretory transglutaminase) contains a glycerol-phosphoinositol (GPI-) anchor. There are a number of still open questions in apocrine secretion, pertaining to (I) the intracellular transport and targeting of the proteins, (II) the coordination of simultaneously occurring apocrine and merocrine secretion in several of the apocrine glands, (III) the biosynthesis of the apical membrane proteins surrounding the aposomes and (IV) the repair mechanisms of the apical cell pole following the release of the aposomes. In conclusion, apocrine release is not an artifact but rather an alternative extrusion mechanism of soluble and membrane-associated proteins, usually linked with sex- or reproductive-related glands, such as the prostate, the mammary glands, apocrine sweat glands or epididymis.

Serum albumin as a potential carrier for the apocrine secretion of proteins in the rat coagulating gland.

A protein of 66k was purified to homogeneity from the total secretion of rat coagulating gland. Its close structural relationship to serum albumin was demonstrated by N-terminal amino acid sequence analysis, proteolytic fingerprinting and Western blotting studies using polyclonal antibodies raised against the 66k protein and rat serum albumin. Immunofluorescence staining showed that the 66k protein was localised in the cytoplasm of coagulating gland epithelial cells from which it is released via apocrine blebs. Performing immunoelectron microscopy, the 66k protein was by no means detectable in the endoplasmic reticulum and the Golgi apparatus. Reverse transcription-PCR, Northern blotting studies and in situ hybridisation experiments demonstrated that mRNA of albumin is not expressed by coagulating gland epithelial cells. Therefore, intravascular albumin should be transferred into the epithelial cells of the rat coagulating gland followed by secretion via aposomes. Furthermore, overlay blots proved that the 66k protein binds to the apocrine proteins carbonic anhydrase II and secretory transglutaminase and vice versa. In contrast, no binding was evident to the merocrine 115k protein and to cytoplasmic resident proteins e.g. lactate dehydrogenase. These findings point to the assumption that serum albumin taken up from extracellular sources could function as a selective carrier for cytoplasmic proteins destined for apocrine secretion.