Patterns of neural differentiation in melanomas.

BACKGROUND: Melanomas, highly malignant tumors arise from the melanocytes which originate as multipotent neural crest cells during neural tube genesis. The purpose of this study is to assess the pattern of neural differentiation in relation to angiogenesis in VGP melanomas using the tumor as a three dimensional system. METHODS: Tumor-vascular complexes [TVC] are formed at the tumor-stroma interphase, by tumor cells ensheathing angiogenic vessels to proliferate into a mantle of 5 to 6 layers [L1 to L5] forming a perivascular mantle zone [PMZ]. The pattern of neural differentiation is assessed by immunopositivity for HMB45, GFAP, NFP and synaptophysin has been compared in: [a] the general tumor [b] tumor-vascular complexes and [c] perimantle zone [PC] on serial frozen and paraffin sections. STATISTICAL ANALYSIS: ANOVA: Kruskal-Wallis One Way Analysis of Variance; All Pairwise Multiple Comparison Procedures [Tukey Test]. RESULTS: The cells abutting on the basement membrane acquire GFAP positivity and extend processes. New layers of tumor cells show a transition between L2 to L3 followed by NFP and Syn positivity in L4&L5. The level of GFAP+vity in L1&L2 directly proportionate to the percentage of NFP/Syn+vity in L4&L5, on comparing pigmented PMZ with poorly pigmented PMZ. Tumor cells in the perimantle zone show high NFP [65%] and Syn [35.4%] positivity with very low GFAP [6.9%] correlating with the positivity in the outer layers. DISCUSSION: From this study it is seen that melanoma cells revert to the embryonic pattern of differentiation, with radial glial like cells [GFAP+ve] which further differentiate into neuronal positive cells [NFP&Syn+ve] during angiogenic tumor-vascular interaction, as seen during neurogenesis, to populate the tumor substance.

The melanocyte photosensory system in the human skin.

The pigment cells form the largest population of neural crest cells to migrate into the epidermis and hair follicle along each dermatomic area from the neural folds. The melanopsin system responsible for photoentrainment, was isolated from the photosensitive dermal melanophores of frogs Xenopus laevis responding to light. Melanocytes form a photoresponsive network which reads the environmental seasonal variations in the light cycles in the same manner. The present work was undertaken to study the organization of this system by: I. Experimental assessment of photoresponse and II. Evidence of an organized system of photoreception in the skin. Melanocytes, in whole skin organ cultures and epidermal strips, from margin of vitiligo in G2 phase show prominent dendricity, and express pigment, biogenic amines and hormones on UV exposure. The photoresponse depends on the photosensitive enzymes NAT/HIOMT and dopaoxidase. Melanocytes interact with adjacent keratinocytes, dermal capillaries, and nerve endings. The melanocyte network reads the diurnal and seasonal photophase by the melatonin/serotonin switch like the pineal. Sleep disorders and winter depression are corrected by phototherapy utilising this mechanism. Melanocytes showing photoactivity, aplasia, hypoplasia and hyperplasia, and interactive keratinocytes occupy the trigeminal, brachial and lumbosacral dermatomes, zones of high embryonic induction, forming an ectodermal placodal system. Melanin units and hair follicles serve as photoreceptors. Migration of active melanocytes to defined areas is evident in pigment patterns in guinea pigs. This study identifies defined photoreceptor melanocyte/epidermal domains which read the seasonal photophase and control the sleep waking cycle in response to the environmental light. I. Whole skin organ cultures, and epidermal strips from margin of vitiligo in G2 phase are exposed to UV and IR to study sequential and dose response of marginal melanocytes, using histochemistry, immunohistochemistry to assess pigment, biogenic amines and hormones on UV exposure. II. Dermatomic Distributions: Detailed maps of melanocyte photoresponse in 356 biopsies, lesions in 297 vitiligo, 100 melanosis, 165 melanomas 142 leprosy and 442 basal cell/keratinocytes lesions were assessed for patterns of dermatomic distribution. Embryonal melanocyte migration along dermatomes was assessed in 285 guinea pigs from an inbred colony having black, brown and white patches.

Embryonic vasculogenesis in nodular melanomas and tumour differentiation.

The relationship of vasculogenic mimicry to pigment in nodular vertical growth phase [VGP] cutaneous melanomas is assessed in this study. 10 nodules each from 27 tumors, 15 pigmented and 12 amelanotic were sampled in proportion to the pigment level. Serial frozen and paraffin sections subjected to HE, Reticulin, PAS to assess the vascular pattern; Dopa Oxidase and Immunopositivity for HMB45, LN5 [laminin 5] & integrin[α(5)ß(1)], and EM [electron microscopy] to identify Weibel-Palade bodies within endothelial cells. The vascular pattern, pigment and the immunopositivity was mapped to assess the percentage VM [vasculogenic sinusoids] vs INC [incorporated microvasculature]. In pigmented melanomas, INC from pre-existing stromal vessels is predominant. Amelanotic melanomas show embryonic vasculogenic mimicry, a self-propagating system of spaces within the sheets of tumors cells. Both INC and VM co-exist in tumors with both amelanotic and melanotic nodules. In areas with VM, loci of LN5 and α(5)ß(1) integrin positive cells appear within the proliferating columns, positivity in these cells suggesting a switch to a more aggressive form. Irregular spaces appear lined by tumor cells, with initial hemopoeitic activity, coalesce and interlink into tubular networks. Spaces lined by tumor cells extend into an intricate network which then connects with the angiogenetic system. The tumor cells lining the vasculogenic spaces are positive for LN5, α(5)ß(1) integrin. Statistically, INC is significantly higher in pigmented melanomas, whereas amelanotic melanomas show significantly higher VM. Pigmentation is correlated positively with INC and negatively with VM. INC and VM are negatively correlated with each other.