Proteomic analysis of immature murine melanocytes at different stages of maturation: A crucial role for calreticulin

Kawase, Ayumi; Kushimoto, Tsuneto; Kawa, Yoko; Ohsumi, Kayoko; Nishikawa, Hiroyuki; Kawakami, Tamihiro; Mizoguchi, Masahiro; Soma, Yoshinao

doi:10.1016/j.jdermsci.2007.07.012

Cited by 6 publications

(3 citation statements)

References 27 publications

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“…The method for preparation of melanosomes from B16 was adapted from previously described method [9,10]. B16 cells were harvested with trypsin/EDTA and collected by centrifugation.…”

Section: Resultsmentioning

confidence: 99%

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Three-Dimensional Analysis of Melanosomes Isolated from B16 Melanoma Cells by Using Ultra High Voltage Electron Microscopy

Akazaki¹,

Takahashi²,

Nakano³

et al. 2014

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Melanosomes, isolated by centrifugal separation from culture broth of B16 melanoma cells derived from mouse, were observed by scanning electron microscopy (SEM), and by transmission electron microscopy (TEM). Some interesting structural features were found inside and outside of the melanosomes. By SEM observation, the melanosomes were ellipsoid shape, their surface was not smooth and was covered with rough substructure, 10 to 20 nm particles. By TEM, uneven structure and micro particles were observed in the melanosomes. Furthermore, three-dimensional analysis was tried by using the ultra-high voltage electron microscopy (UHVEM). Micrographs of the melanosomes were taken at various tilted angles by UHVEM, after preparing 500 nm thickness specimens stained with lead citrate. From the micrographs collected, the three-dimensional structures were reconstructed by using i-mode software. Melanin stained by lead and non stained parts was clearly observed in the reconstructed structure. Non stained parts were round, regular size, and distributed widely in the melanosomes.

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“…The method for preparation of melanosomes from B16 was adapted from previously described method [9,10]. B16 cells were harvested with trypsin/EDTA and collected by centrifugation.…”

Section: Resultsmentioning

confidence: 99%

“…After centrifugation, pellets including melanosomes were precipitated [9,10]. Pellets were suspended twice by distilled water and preserved at −80˚C, temporary.…”

Section: Preparation 1 Of Melanosomementioning

confidence: 99%

Three-Dimensional Analysis of Melanosomes Isolated from B16 Melanoma Cells by Using Ultra High Voltage Electron Microscopy

Akazaki¹,

Takahashi²,

Nakano³

et al. 2014

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“…Microscopy and flow cytometry provide an immediate, easy method to study some functional parameters and cell morphology. Fluorochrome‐conjugated antibodies are widely applied to visualise and quantify the expression of some surface or internal melanocyte markers . Novel approaches, based on fluorescence and confocal microscopy on slide‐cultured cells, provide interesting information, such as the phenotypic characterisation [c‐Kit, ETBR (Invitrogen, Milano, Italy), tyrosinase, MITF expression] of cultured vitiligo melanocytes (Fig.…”

Section: New Tools and Trends: The Fantastic World Of The Moving Pictmentioning

confidence: 99%

In vitro research on vitiligo: strategies, principles, methodological options and common pitfalls

Dell’Anna

Cario‐André

Bellei

et al. 2012

Experimental Dermatology

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Understanding the cellular and molecular mechanisms leading to melanocyte loss in vitiligo is a mandatory step in improving the overall management of vitiligo patients. Until now, the study of vitiligo was characterised by a fragmented approach, and it is very hard to share and compare the data obtained by the different teams. The scenario mirrors the pathogenic puzzle, but it delays a true productive focus on the disease. The in vitro research is based on different models, ranging from monolayer cell culture of epidermal and dermal cells or 3D reconstructed skin to histological data, gene expression, computer simulation profile. For each model, several different (biochemical, phenotypic, immunological) aspects have been considered, increasing the mass of data difficult to be merged. Our purpose was to provide a practical synopsis of consolidated and advanced possibilities in the study of vitiligo, showing how data have been poorly shared until now. Following a short overview of the background of the disease, the approaches, ranging from basic cell biology to molecular and 'omics' studies, are summarised. New fluorescent probes and techniques open new possibilities for functional studies. Next, intracellular and superficial markers of the melanocytes, the main involved cells, are listed. Moving the focus from the epidermal level to the systemic and subcellular ones, this review aims to propose innovative multidisciplinary options for the vitiligo understanding. This paper focuses on the major practical and theoretical questions to be solved. It may be the basis for a more coordinated and productive approach to the biological question.

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In Vitro Study of Vitiligo

Dell’Anna

Cario‐André²

2019

Vitiligo

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Proteomic analysis of immature murine melanocytes at different stages of maturation: A crucial role for calreticulin

Cited by 6 publications

References 27 publications

Three-Dimensional Analysis of Melanosomes Isolated from B16 Melanoma Cells by Using Ultra High Voltage Electron Microscopy

Three-Dimensional Analysis of Melanosomes Isolated from B16 Melanoma Cells by Using Ultra High Voltage Electron Microscopy

In vitro research on vitiligo: strategies, principles, methodological options and common pitfalls

In Vitro Study of Vitiligo

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