Merkel Cell-Nerve Cell Interaction Undergoes Formation of a Synapse-like Structure in a Primary Culture

Shimohira-Yamasaki, Mineko; Toda, Shuji; Narisawa, Yutaka; Sugihara, Hajime

doi:10.1247/csf.31.39

Cited by 19 publications

(30 citation statements)

References 24 publications

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“…during culture. To allow detachment of the epidermal layer of skin samples, each skin sample was incubated overnight in 0.5 ml of dispase I solution at 4 1C (Shimohira-Yamasaki et al, 2006), whereby the epidermis and dermis became easily separable. The control was similarly incubated overnight.…”

Section: Preparation Of Skin Sections For Organ Culturementioning

confidence: 99%

Neuronal Nitric Oxide Synthase in Epidermis Is Involved in Cutaneous Circulatory Response to Mechanical Stimulation

Ikeyama

Denda

Tsutsumi

et al. 2010

Journal of Investigative Dermatology

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The source of nitric oxide (NO) in the cutaneous circulation remains controversial. We hypothesized that epidermis might generate NO in response to mechanical stimulation. In hairless mouse (HR-1) skin organ culture, mechanical stimulation resulted in NO release, which declined within 30 minutes after cessation. A similar NO release occurred in a reconstructed skin model containing only keratinocytes and fibroblasts and was suppressed after detachment of the epidermal layer. Moreover, the stimulation-induced NO release was significantly lower in skin organ culture from neuronal NO synthase knockout (nNOS-KO) mice, compared with wild-type (WT) mice. Mechanical stimulation of skin organ cultures from HR-1, nNOS-KO, endothelial NOS-KO (eNOS-KO), and WT mice caused an enlargement of cutaneous lymphatic vessels. The enlargement was significantly lower after detachment of the epidermal layer than in normal skin samples and was significantly lower for nNOS-KO than for WT mice. Skin blood flow in nNOS-KO mice after stimulation was significantly lower than in WT mice. eNOS-KO mice also showed lower responses than WT mice, and the difference was similar to that in the case of nNOS-KO mice. These results are consistent with the idea that NO generated by epidermal nNOS has a significant role in the cutaneous circulatory response to mechanical stimulation.

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Section: Preparation Of Skin Sections For Organ Culturementioning

confidence: 99%

Neuronal Nitric Oxide Synthase in Epidermis Is Involved in Cutaneous Circulatory Response to Mechanical Stimulation

Ikeyama

Denda

Tsutsumi

et al. 2010

Journal of Investigative Dermatology

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“…Since then, there has been no clear evidence for pre-and post-synaptic densities associated with the Merkel cell-neurite complex in micrographs. However, it is important to note that very recently Shimohira-Yamasaki et al (2006) obtained a synapse-like structure in a primary culture of Merkel cells and nerve cells. Furthermore, when quinacrine-loaded Merkel cells are selectively eliminated by light irradiation, the mechanoreceptive function remains (Mills and Diamond 1995).…”

Section: Development and The Merkel Cell-neurite Complex As A Mechanomentioning

confidence: 99%

Localization of Merkel cells in the monkey skin: An anatomical model

Güçlü

Mahoney

Pawson

et al. 2008

Somatosensory & Motor Research

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The Merkel cell-neurite complex is considered to be one class of mechanoreceptors in the skin. Merkel cells are innervated by slowly adapting type I (SAI) tactile nerve fibers. In this paper, the detailed distribution of Merkel cells is studied by immunohistochemical labeling of the monkey (Macaca fascicularis) digital glabrous skin. Specific morphometric variables (density of intermediate epidermal ridges and Merkel cells, distance between skin surface and ridge tips and bases, maximum and average cell counts per ridge, distance between cells and ridges) were measured by a combination of light/fluorescence microscopy and computer-image analysis. The morphometric results were similar for each digit of the monkey's hand. Next, the anatomical data were used to form a three-dimensional reconstruction of the Merkel-cell distribution in the fingertip skin. A patch of the distal-pad surface was then computationally flattened to obtain the two-dimensional distribution of Merkel cells. Based on previous anatomical and physiological data, SAI fibers were simulated to innervate clusters of Merkel cells in the distal-pad surface. On average, 28 cells were innervated by a single fiber. The resulting anatomical model may be used to estimate the population response of SAI fibers by incorporating spike generation.

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“…These differentiated PC12 cells have been used as a model for sensory or autonomic neurons (14)(15)(16). We developed an in vitro model of human skin explant, which can be considered as an equivalent of injured skin, co-cultured with PC12 cells differentiated into neurons.…”

mentioning

confidence: 99%

Effect of human skin explants on the neurite growth of the PC12 cell line

Lebonvallet

Pennec

Gall

et al. 2013

Experimental Dermatology

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The skin is a densely innervated organ. After a traumatic injury, such as an amputation, burn or skin graft, nerve growth and the recovery of sensitivity take a long time and are often incomplete. The roles played by growth factors and the process of neuronal growth are crucial. We developed an in vitro model of human skin explants co-cultured with a rat pheochromocytoma cell line differentiated in neuron in presence of nerve growth factor (NGF). This model allowed the study of the influence of skin explants on nerve cells and nerve fibre growth, probably through mediators produced by the explant, in a simplified manner. The neurite length of differentiated PC12 cells cocultured with skin explants increased after 6 days. These observations demonstrated the influence of trophic factors produced by skin explants on PC12 cells.Key words: neuron -neuronal growth -skin Accepted for publication 18 January 2013Nerve growth following a traumatic injury, such as an amputation, burns or skin graft, takes a long time and is often incomplete. Neurotrophins and semaphorins have been implicated in the guidance of neuronal growth (1,2) through interactions with each other to regulate the motility of the sensory neuronal growth cone. However, the mechanisms of the interactions between the peripheral neurons and the keratinocytes/epidermis/skin remain unclear. To study these interactions under normal physiological and injured conditions, the development of in vitro co-culture models is important. At present, models enabling the study of neuron-skin interactions are rare (3), with the majority involving either primary sensory neurons or a neuronal cell line co-cultured with keratinocytes (4-6), reconstructed skin (7), a skin explant (8) or lesional skin (9). Furthermore, the neurons used in these models are limited due to ethical problems involved in harvesting primary neurons and by the necessity to select an appropriate cell line. Today, adult stem cells could be small sources of neurons or other cellular types for regenerative medicine and tissue engineering, but they are not co-cultured (10,11).PC12 is a versatile rat pheochromocytoma cell line that can be differentiated into neurons with sensory or autonomic characteristics using NGF (12,13). These differentiated PC12 cells have been used as a model for sensory or autonomic neurons (14-16). We developed an in vitro model of human skin explant, which can be considered as an equivalent of injured skin, co-cultured with PC12 cells differentiated into neurons.PC12 cells were co-cultured with human skin explants (E) until 10 days. Skin explants (three donors, one for each experiment) were cut using a 6-mm diameter biopsy punch and placed in 12-well culture plate with one skin explant per well. The skin explants were co-cultured with PC12 cells for more than 10 days at an airliquid interface, and the medium was changed every 2 days. The PC12 cells, which were obtained from the ATCC (ATCC, CRL-1721), were grown in DMEM-F12 media (Lonza, BE12-719F) supplemented with 10% ca...

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Merkel Cell-Nerve Cell Interaction Undergoes Formation of a Synapse-like Structure in a Primary Culture

Cited by 19 publications

References 24 publications

Neuronal Nitric Oxide Synthase in Epidermis Is Involved in Cutaneous Circulatory Response to Mechanical Stimulation

Neuronal Nitric Oxide Synthase in Epidermis Is Involved in Cutaneous Circulatory Response to Mechanical Stimulation

Localization of Merkel cells in the monkey skin: An anatomical model

Effect of human skin explants on the neurite growth of the PC12 cell line

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