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Adrenomedullin: A novel melanocyte dendrite branching factor
G Model DESC 2848 No. of Pages 4
Journal of Dermatological Science xxx (2015) xxx–xxx
Contents lists available at ScienceDirect
Journal of Dermatological Science journal homepage: www.jdsjournal.com
Correspondence Adrenomedullin: A novel melanocyte dendrite branching factor
Human melanocytes produce melanin in the epidermis, and skin color and pigmented spots are greatly influenced by the level of melanin accumulation. Many melanocyte-stimulating factors including endothelin, prostaglandin E2, and nitric oxide have been identified to be involved in melanin accrual in human skin. As these factors are all secreted by keratinocytes and stimulate blood vessel formation [1–3], we hypothesized that adrenomedullin (ADM), which displays similar features, is a potent stimulator of melanogenesis. ADM is secreted from human endothelial cells and keratinocytes and is known to have many effects including angiogenesis [4]. In the present study, we examined the association between ADM and melanin accumulation. We found that ADM is a new melanocyte dendrite branching factor that functions in a paracrine manner, and detected high levels of ADM in pigmented lesions of human epidermis, including lentigo senilis and UVinduced pigmentation. This article describes the results of these in vitro and in vivo experiments. Fig. 1a and b shows representative effects of ADM on morphological changes in melanocytes. ADM increased the number of melanocyte dendrites and its effect was much stronger than other melanocyte activation factors (Fig. 1c). Moreover, ADM increased melanin synthesis in melanocytes (Fig. 1d), but did not have a proliferative effect (data not shown). The number of dendrites, as well as melanin synthesis, is thought to be critical for the pigment to be accumulated in human skin, because the presence of multiple dendrites on melanocyte cells allows melanin to be distributed to many keratinocytes. It has been reported that one melanocyte can distribute melanin to as many as 36 keratinocytes [5]. Hence, ADM seems to be a crucial factor for melanin accumulation in human skin. The importance of UV-irradiated keratinocytes as ADMsecreting cells and the significance of ADM as a dendriteproducing factor were demonstrated by three in vitro experiments (the scheme of these experiments is described in Fig. 1e): (1) addition of medium from UV-irradiated keratinocytes to melanocytes caused dendrite production that was inhibited by siRNA knockdown of ADM in keratinocytes (Fig. 1f), whereas medium from UV-irradiated melanocytes did not induce morphological changes (data not shown); (2) induction of ADM protein in UV-irradiated keratinocytes was observed (Fig. 1g); (3) the knockdown of RAMP 2 (receptor activity modifying protein 2), which forms part of ADM receptor [6], inhibited dendrite production caused by the hormone (Fig. 1h). These results demonstrated that ADM is a paracrine factor for melanocytes that induces dendrite production. Thus, ADM is a unique dendrite branching factor and may be important for melanin accumulation
in pigmented lesions. In addition, both ADM mRNA and protein were induced in UV-irradiated keratinocytes (Fig. 1f and g), suggesting it may also have a pivotal role in UV-induced pigmentation. Our in vitro studies suggested the importance of ADM in human skin; therefore, we performed in vivo immunostaining of pigmented lesions and UV-induced pigmentation of human skin. ADM was present in the epidermis and was increased in pigmented lesions compared with perilesional skin areas (Fig. 2a and b). Similar results were observed between tissues obtained from two patients. ADM protein was highly expressed in UV-irradiated human skin in erythema (2 days after UV-irradiation) and pigmentation (5 days after UV-irradiation; Fig. 2c–e). Similar results were observed from immunostaining of two tissue samples from two healthy volunteers. Both the in vivo and in vitro experiments strongly suggest the importance and significance of ADM on pigmentation in human skin. This study identified ADM as a novel factor that increased the number of melanocyte dendrites. ADM is unique because it induces dendrite branching to a higher degree and up-regulates melanin synthesis. Moreover, high levels of ADM were detected in pigmented human skin, including lentigo senilis and UV-induced pigmentation, compared with normal tissue. ADM, which consists of 52 amino acids, was first identified as a hypotensive peptide in the adrenal medulla. It is a pluripotent peptide with numerous physiological roles including vasodilation, neurotransmission and growth modulation[4]. Homozygous ADM knockout mice die in utero and embryos are characterized by poorly developed vitelline vessels and extreme hydrops fetalis, while heterozygous ADM mice are fertile and survive until adulthood but develop arterial hypertension [7], and ADM over-expressing mice develop low blood pressure [8]. ADM has been detected in keratinocytes and melanocytes in skin [9] and is able to stimulate keratinocyte and fibroblast proliferation [10]. We have not studied the mechanism(s) of ADM-induced dendrite neogenesis at this stage, but the intercellular differences in the reaction might be the key to understanding the mechanism. As seen in figure 1b, dendrite neogenesis does not occur at the same time in all melanocytes. In the future, we will aim to study the cause of these intercellular differences using time-lapse imaging. In reaction to a changing environment and internal milieu, human skin generates many biologically active compounds that maintain homeostasis. Exposure of skin to UV irradiation induces a complicated melanin accumulation system. A potential study limitation was the low number of human samples. However, using in vitro and in vivo methods, this study demonstrated that ADM was induced by UV and promoted the accrual of melanin. Thus, ADM shows similarities to other factors involved in UV-induced
http://dx.doi.org/10.1016/j.jdermsci.2015.06.001 0923-1811/ ã 2015 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.
Please cite this article in press as: T. Motokawa, et al., Adrenomedullin: A novel melanocyte dendrite branching factor, J Dermatol Sci (2015), http://dx.doi.org/10.1016/j.jdermsci.2015.06.001
G Model DESC 2848 No. of Pages 4
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Correspondence / Journal of Dermatological Science xxx (2015) xxx–xxx
Fig. 1. ADM increases melanocyte dendrite numbers and melanin synthesis in vitro. We used normal human melanocytes (Kurabo, Osaka, Japan) in Medium 254 (Kurabo) with MHGS additive factors (Kurabo) excluding bovine pituitary extract and insulin. (a) Melanocytes without ADM. (b) Morphological changes and increased dendrite numbers in melanocytes 12 h after addition of 1.0 mM ADM. (c) Effects on dendrite numbers after the addition of ADM or other melanocyte stimulating factors (1.0 mM for ADM, MSH, EDN, and 0.1 mM for PGE due to cytotoxicity). MSH: alpha-melanocyte stimulating hormone, EDN: endothelin, PGE: prostaglandin E2. (d) Effects on melanin
Please cite this article in press as: T. Motokawa, et al., Adrenomedullin: A novel melanocyte dendrite branching factor, J Dermatol Sci (2015), http://dx.doi.org/10.1016/j.jdermsci.2015.06.001
G Model DESC 2848 No. of Pages 4
Correspondence / Journal of Dermatological Science xxx (2015) xxx–xxx
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Fig. 2. Levels of ADM are increased in pigmented lesions. (a) Lentigo senilis lesions (45-year-old, male, Japanese). Excised tissues included pigmented lesions and perilesional skin. Brightfield image of skin tissue. Melanin accumulation was observed on the left side (dotted area). (b) Fluorescence immunostaining for ADM (green). Concentrations of ADM were higher in pigmented lesions than in non-pigmented lesions. (c) UV-irradiated human skin. Each volunteer was irradiated with UVB (equivalent to 2 minimal erythema doses; 101.2–193.2 mJ/cm2) (FL 20 SE30 lamp, Toshiba, Tokyo, Japan). Tissues, including UV-irradiated lesions and non-irradiated lesions (dotted circles), were excised during erythema (2 days after irradiation) and pigmentation (5 days after irradiation). (d, e) Fluorescence immunostaining of ADM (green). Concentrations of ADM were higher in UV-irradiated lesions during erythema and pigmentation. Anti-human ADM antibody (Abcam, Cambridge, UK) in 10% Block Ace (DS Pharma Biomedical, Osaka, Japan) and a commercial biotin-free catalyzed amplification system (Dako Japan, Tokyo, Japan) were used to detect ADM according to the manufacturer's instructions.
melanin accumulation, and should therefore be regarded as a kind of hormone reactive to UV rays. It is very interesting to note that many melanocyte stimulating factors, including ADM, are also vascular endothelial factors that lead to angiogenesis. We do not know the reason for this, but many studies that focused on the association between vascular endothelial factors and pigmentation might provide clues for understanding this link. Here, we demonstrate for the first time an association between ADM and melanin accumulation. We also show that ADM is involved in dendrite branching and melanin synthesis. These findings may contribute to both the development of basic and etiological research, because melanocyte activators are often
related to pigment disorders. Finally, this novel role of ADM might provide a useful tool to study the mechanisms behind the creation of melanocyte dendrite morphology. Conflict of interest The authors have no conflict of interest to declare. Appendix A. Supplementary data Supplementary data associated with this article can be found, in the online version, at http://dx.doi.org/10.1016/j. jdermsci.2015.06.001.
synthesis in melanocytes. Significant increase of melanin synthesis was observed after 72 h of incubation with 2–4 mM ADM. (e) Scheme of three in vitro experiments to demonstrate the importance of ADM as a paracrine factor. (f) ADM secreted by keratinocytes induces dendrite production in melanocytes. UVB (65 mJ/cm2)-induced dendrite production was inhibited by ADM knockdown (siADM5 and siADM6) in keratinocytes. (g) Induction of ADM protein in keratinocytes by UV irradiation was measured using the ADM (1-52; Human) Enzyme Immunoassay Kit (Phoenix Pharmaceuticals, Burlingame, CA, USA). Significant increase of ADM protein in the medium from UV-irradiated keratinocytes. (h) The knockdown of RAMP2, which forms part of ADM receptor, limited the number of dendrites increased by ADM.
Please cite this article in press as: T. Motokawa, et al., Adrenomedullin: A novel melanocyte dendrite branching factor, J Dermatol Sci (2015), http://dx.doi.org/10.1016/j.jdermsci.2015.06.001
G Model DESC 2848 No. of Pages 4
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Correspondence / Journal of Dermatological Science xxx (2015) xxx–xxx
References [1] D. Salani, V. Di Castro, M.R. Nicotra, L. Rosano, R. Tecce, A. Venuti, et al., Role of endothelin-1 in neovascularization of ovarian carcinoma, Am. J. Pathol. 157 (2000) 1537–1547. [2] M. Ziche, A. Parenti, F. Ledda, P. Dell'Era, H.J. Granger, C.A. Maggi, et al., Nitric oxide promotes proliferation and plasminogen activator production by coronary venular endothelium through endogenous bFGF, Circ Res 80 (1997) 845–852. [3] F. Finetti, R. Solito, L. Morbidelli, A. Giachetti, M. Ziche, S. Donnini, Prostaglandin E2 regulates angiogenesis via activation of fibroblast growth factor receptor-1, J Biol Chem 283 (2008) 2139–2146. [4] J.P. Hinson, S. Kapas, D.M. Smith, Adrenomedullin, a multifunctional regulatory peptide, Endocr Rev 21 (2000) 138–167. [5] Fitzpatrick TB, Breathnach AS. [the Epidermal Melanin Unit System]. Dermatol Wochenschr 147: 481-489, 1963. [6] S. Kamitani, M. Asakawa, Y. Shimekake, K. Kuwasako, K. Nakahara, T. Sakata, The RAMP2/CRLR complex is a functional adrenomedullin receptor in human endothelial and vascular smooth muscle cells, FEBS Lett 448 (1999) 111–114. [7] K.M. Caron, O. Smithies, Extreme hydrops fetalis and cardiovascular abnormalities in mice lacking a functional Adrenomedullin gene, Proc Natl Acad Sci U S A 98 (2001) 615–619. [8] T. Shindo, H. Kurihara, K. Maemura, Y. Kurihara, T. Kuwaki, T. Izumida, et al., Hypotension and resistance to lipopolysaccharide-induced shock in transgenic mice overexpressing adrenomedullin in their vasculature, Circulation 101 (2000) 2309–2316.
Martinez, T.H. Elsasser, C. Muro-Cacho, T.W. Moody, M.J. Miller, C.J. Macri, et al., Expression of adrenomedullin and its receptor in normal and malignant human skin: a potential pluripotent role in the integument, Endocrinology 138 (1997) 5597–5604. [10] G. Albertin, G. Carraro, P.P. Parnigotto, M.T. Conconi, A. Ziolkowska, L.K. Malendowicz, et al., Human skin keratinocytes and fibroblasts express adrenomedullin and its receptors, and adrenomedullin enhances their growth in vitro by stimulating proliferation and inhibiting apoptosis, Int J Mol Med 11 (2003) 635–639.
Tomonori Motokawa PhD* , Takahiro Miwa Mayu Mochizuki Minako Toritsuka Aya Sakata Masaaki Ito Skin Research Department, POLA Chemical Industries, Inc., Yokohama, Japan * Corresponding author. Skin Research Department, POLA Chemical Industries, Inc., 560 Kashio-cho, Totsuka-ku, Yokohama 2440812, Japan. Fax: +81 45 826 7249.
[9] A.
Please cite this article in press as: T. Motokawa, et al., Adrenomedullin: A novel melanocyte dendrite branching factor, J Dermatol Sci (2015), http://dx.doi.org/10.1016/j.jdermsci.2015.06.001
Journal of Dermatological Science xxx (2015) xxx–xxx
Contents lists available at ScienceDirect
Journal of Dermatological Science journal homepage: www.jdsjournal.com
Correspondence Adrenomedullin: A novel melanocyte dendrite branching factor
Human melanocytes produce melanin in the epidermis, and skin color and pigmented spots are greatly influenced by the level of melanin accumulation. Many melanocyte-stimulating factors including endothelin, prostaglandin E2, and nitric oxide have been identified to be involved in melanin accrual in human skin. As these factors are all secreted by keratinocytes and stimulate blood vessel formation [1–3], we hypothesized that adrenomedullin (ADM), which displays similar features, is a potent stimulator of melanogenesis. ADM is secreted from human endothelial cells and keratinocytes and is known to have many effects including angiogenesis [4]. In the present study, we examined the association between ADM and melanin accumulation. We found that ADM is a new melanocyte dendrite branching factor that functions in a paracrine manner, and detected high levels of ADM in pigmented lesions of human epidermis, including lentigo senilis and UVinduced pigmentation. This article describes the results of these in vitro and in vivo experiments. Fig. 1a and b shows representative effects of ADM on morphological changes in melanocytes. ADM increased the number of melanocyte dendrites and its effect was much stronger than other melanocyte activation factors (Fig. 1c). Moreover, ADM increased melanin synthesis in melanocytes (Fig. 1d), but did not have a proliferative effect (data not shown). The number of dendrites, as well as melanin synthesis, is thought to be critical for the pigment to be accumulated in human skin, because the presence of multiple dendrites on melanocyte cells allows melanin to be distributed to many keratinocytes. It has been reported that one melanocyte can distribute melanin to as many as 36 keratinocytes [5]. Hence, ADM seems to be a crucial factor for melanin accumulation in human skin. The importance of UV-irradiated keratinocytes as ADMsecreting cells and the significance of ADM as a dendriteproducing factor were demonstrated by three in vitro experiments (the scheme of these experiments is described in Fig. 1e): (1) addition of medium from UV-irradiated keratinocytes to melanocytes caused dendrite production that was inhibited by siRNA knockdown of ADM in keratinocytes (Fig. 1f), whereas medium from UV-irradiated melanocytes did not induce morphological changes (data not shown); (2) induction of ADM protein in UV-irradiated keratinocytes was observed (Fig. 1g); (3) the knockdown of RAMP 2 (receptor activity modifying protein 2), which forms part of ADM receptor [6], inhibited dendrite production caused by the hormone (Fig. 1h). These results demonstrated that ADM is a paracrine factor for melanocytes that induces dendrite production. Thus, ADM is a unique dendrite branching factor and may be important for melanin accumulation
in pigmented lesions. In addition, both ADM mRNA and protein were induced in UV-irradiated keratinocytes (Fig. 1f and g), suggesting it may also have a pivotal role in UV-induced pigmentation. Our in vitro studies suggested the importance of ADM in human skin; therefore, we performed in vivo immunostaining of pigmented lesions and UV-induced pigmentation of human skin. ADM was present in the epidermis and was increased in pigmented lesions compared with perilesional skin areas (Fig. 2a and b). Similar results were observed between tissues obtained from two patients. ADM protein was highly expressed in UV-irradiated human skin in erythema (2 days after UV-irradiation) and pigmentation (5 days after UV-irradiation; Fig. 2c–e). Similar results were observed from immunostaining of two tissue samples from two healthy volunteers. Both the in vivo and in vitro experiments strongly suggest the importance and significance of ADM on pigmentation in human skin. This study identified ADM as a novel factor that increased the number of melanocyte dendrites. ADM is unique because it induces dendrite branching to a higher degree and up-regulates melanin synthesis. Moreover, high levels of ADM were detected in pigmented human skin, including lentigo senilis and UV-induced pigmentation, compared with normal tissue. ADM, which consists of 52 amino acids, was first identified as a hypotensive peptide in the adrenal medulla. It is a pluripotent peptide with numerous physiological roles including vasodilation, neurotransmission and growth modulation[4]. Homozygous ADM knockout mice die in utero and embryos are characterized by poorly developed vitelline vessels and extreme hydrops fetalis, while heterozygous ADM mice are fertile and survive until adulthood but develop arterial hypertension [7], and ADM over-expressing mice develop low blood pressure [8]. ADM has been detected in keratinocytes and melanocytes in skin [9] and is able to stimulate keratinocyte and fibroblast proliferation [10]. We have not studied the mechanism(s) of ADM-induced dendrite neogenesis at this stage, but the intercellular differences in the reaction might be the key to understanding the mechanism. As seen in figure 1b, dendrite neogenesis does not occur at the same time in all melanocytes. In the future, we will aim to study the cause of these intercellular differences using time-lapse imaging. In reaction to a changing environment and internal milieu, human skin generates many biologically active compounds that maintain homeostasis. Exposure of skin to UV irradiation induces a complicated melanin accumulation system. A potential study limitation was the low number of human samples. However, using in vitro and in vivo methods, this study demonstrated that ADM was induced by UV and promoted the accrual of melanin. Thus, ADM shows similarities to other factors involved in UV-induced
http://dx.doi.org/10.1016/j.jdermsci.2015.06.001 0923-1811/ ã 2015 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.
Please cite this article in press as: T. Motokawa, et al., Adrenomedullin: A novel melanocyte dendrite branching factor, J Dermatol Sci (2015), http://dx.doi.org/10.1016/j.jdermsci.2015.06.001
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Correspondence / Journal of Dermatological Science xxx (2015) xxx–xxx
Fig. 1. ADM increases melanocyte dendrite numbers and melanin synthesis in vitro. We used normal human melanocytes (Kurabo, Osaka, Japan) in Medium 254 (Kurabo) with MHGS additive factors (Kurabo) excluding bovine pituitary extract and insulin. (a) Melanocytes without ADM. (b) Morphological changes and increased dendrite numbers in melanocytes 12 h after addition of 1.0 mM ADM. (c) Effects on dendrite numbers after the addition of ADM or other melanocyte stimulating factors (1.0 mM for ADM, MSH, EDN, and 0.1 mM for PGE due to cytotoxicity). MSH: alpha-melanocyte stimulating hormone, EDN: endothelin, PGE: prostaglandin E2. (d) Effects on melanin
Please cite this article in press as: T. Motokawa, et al., Adrenomedullin: A novel melanocyte dendrite branching factor, J Dermatol Sci (2015), http://dx.doi.org/10.1016/j.jdermsci.2015.06.001
G Model DESC 2848 No. of Pages 4
Correspondence / Journal of Dermatological Science xxx (2015) xxx–xxx
3
Fig. 2. Levels of ADM are increased in pigmented lesions. (a) Lentigo senilis lesions (45-year-old, male, Japanese). Excised tissues included pigmented lesions and perilesional skin. Brightfield image of skin tissue. Melanin accumulation was observed on the left side (dotted area). (b) Fluorescence immunostaining for ADM (green). Concentrations of ADM were higher in pigmented lesions than in non-pigmented lesions. (c) UV-irradiated human skin. Each volunteer was irradiated with UVB (equivalent to 2 minimal erythema doses; 101.2–193.2 mJ/cm2) (FL 20 SE30 lamp, Toshiba, Tokyo, Japan). Tissues, including UV-irradiated lesions and non-irradiated lesions (dotted circles), were excised during erythema (2 days after irradiation) and pigmentation (5 days after irradiation). (d, e) Fluorescence immunostaining of ADM (green). Concentrations of ADM were higher in UV-irradiated lesions during erythema and pigmentation. Anti-human ADM antibody (Abcam, Cambridge, UK) in 10% Block Ace (DS Pharma Biomedical, Osaka, Japan) and a commercial biotin-free catalyzed amplification system (Dako Japan, Tokyo, Japan) were used to detect ADM according to the manufacturer's instructions.
melanin accumulation, and should therefore be regarded as a kind of hormone reactive to UV rays. It is very interesting to note that many melanocyte stimulating factors, including ADM, are also vascular endothelial factors that lead to angiogenesis. We do not know the reason for this, but many studies that focused on the association between vascular endothelial factors and pigmentation might provide clues for understanding this link. Here, we demonstrate for the first time an association between ADM and melanin accumulation. We also show that ADM is involved in dendrite branching and melanin synthesis. These findings may contribute to both the development of basic and etiological research, because melanocyte activators are often
related to pigment disorders. Finally, this novel role of ADM might provide a useful tool to study the mechanisms behind the creation of melanocyte dendrite morphology. Conflict of interest The authors have no conflict of interest to declare. Appendix A. Supplementary data Supplementary data associated with this article can be found, in the online version, at http://dx.doi.org/10.1016/j. jdermsci.2015.06.001.
synthesis in melanocytes. Significant increase of melanin synthesis was observed after 72 h of incubation with 2–4 mM ADM. (e) Scheme of three in vitro experiments to demonstrate the importance of ADM as a paracrine factor. (f) ADM secreted by keratinocytes induces dendrite production in melanocytes. UVB (65 mJ/cm2)-induced dendrite production was inhibited by ADM knockdown (siADM5 and siADM6) in keratinocytes. (g) Induction of ADM protein in keratinocytes by UV irradiation was measured using the ADM (1-52; Human) Enzyme Immunoassay Kit (Phoenix Pharmaceuticals, Burlingame, CA, USA). Significant increase of ADM protein in the medium from UV-irradiated keratinocytes. (h) The knockdown of RAMP2, which forms part of ADM receptor, limited the number of dendrites increased by ADM.
Please cite this article in press as: T. Motokawa, et al., Adrenomedullin: A novel melanocyte dendrite branching factor, J Dermatol Sci (2015), http://dx.doi.org/10.1016/j.jdermsci.2015.06.001
G Model DESC 2848 No. of Pages 4
4
Correspondence / Journal of Dermatological Science xxx (2015) xxx–xxx
References [1] D. Salani, V. Di Castro, M.R. Nicotra, L. Rosano, R. Tecce, A. Venuti, et al., Role of endothelin-1 in neovascularization of ovarian carcinoma, Am. J. Pathol. 157 (2000) 1537–1547. [2] M. Ziche, A. Parenti, F. Ledda, P. Dell'Era, H.J. Granger, C.A. Maggi, et al., Nitric oxide promotes proliferation and plasminogen activator production by coronary venular endothelium through endogenous bFGF, Circ Res 80 (1997) 845–852. [3] F. Finetti, R. Solito, L. Morbidelli, A. Giachetti, M. Ziche, S. Donnini, Prostaglandin E2 regulates angiogenesis via activation of fibroblast growth factor receptor-1, J Biol Chem 283 (2008) 2139–2146. [4] J.P. Hinson, S. Kapas, D.M. Smith, Adrenomedullin, a multifunctional regulatory peptide, Endocr Rev 21 (2000) 138–167. [5] Fitzpatrick TB, Breathnach AS. [the Epidermal Melanin Unit System]. Dermatol Wochenschr 147: 481-489, 1963. [6] S. Kamitani, M. Asakawa, Y. Shimekake, K. Kuwasako, K. Nakahara, T. Sakata, The RAMP2/CRLR complex is a functional adrenomedullin receptor in human endothelial and vascular smooth muscle cells, FEBS Lett 448 (1999) 111–114. [7] K.M. Caron, O. Smithies, Extreme hydrops fetalis and cardiovascular abnormalities in mice lacking a functional Adrenomedullin gene, Proc Natl Acad Sci U S A 98 (2001) 615–619. [8] T. Shindo, H. Kurihara, K. Maemura, Y. Kurihara, T. Kuwaki, T. Izumida, et al., Hypotension and resistance to lipopolysaccharide-induced shock in transgenic mice overexpressing adrenomedullin in their vasculature, Circulation 101 (2000) 2309–2316.
Martinez, T.H. Elsasser, C. Muro-Cacho, T.W. Moody, M.J. Miller, C.J. Macri, et al., Expression of adrenomedullin and its receptor in normal and malignant human skin: a potential pluripotent role in the integument, Endocrinology 138 (1997) 5597–5604. [10] G. Albertin, G. Carraro, P.P. Parnigotto, M.T. Conconi, A. Ziolkowska, L.K. Malendowicz, et al., Human skin keratinocytes and fibroblasts express adrenomedullin and its receptors, and adrenomedullin enhances their growth in vitro by stimulating proliferation and inhibiting apoptosis, Int J Mol Med 11 (2003) 635–639.
Tomonori Motokawa PhD* , Takahiro Miwa Mayu Mochizuki Minako Toritsuka Aya Sakata Masaaki Ito Skin Research Department, POLA Chemical Industries, Inc., Yokohama, Japan * Corresponding author. Skin Research Department, POLA Chemical Industries, Inc., 560 Kashio-cho, Totsuka-ku, Yokohama 2440812, Japan. Fax: +81 45 826 7249.
[9] A.
Please cite this article in press as: T. Motokawa, et al., Adrenomedullin: A novel melanocyte dendrite branching factor, J Dermatol Sci (2015), http://dx.doi.org/10.1016/j.jdermsci.2015.06.001