Effects of Ultraviolet Irradiation on the Cell Cycle

Cultured mouse Cloudman melanoma cells, EMT6 breast carcinoma cells, and 3T3 fibroblasts all accumulated in the G2/M phase of the cell cycle when exposed to UVB radiation. The effects of UVB were maximal at 20–30 mJ/cm² for all three cell lines, and could be observed by flow cytometry as early as 12 hr post irradiation. It has been known since the mid-1970s that MSH receptor binding activity is highest on Cloudman melanoma cells when they are in the G2/M phase of their cycle. Here we show that either UVB irradiation or synchronization of Cloudman cells with colchicine results in a stimulation of MSH binding within 24 hr following treatment, a time when both treatments have resulted in accumulation of cells in the G2/M phase of the cycle. Furthermore, the two treatments performed together on the melanoma cells stimulated MSH receptor activity to the same extent as either treatment performed separately, suggesting that each may be influencing MSH receptor activity solely through a G2/M accumulation of cells. Together, these results raise the possibility that an increase in the number of cells in the G2 phase of the cell cycle is a generalized cellular response to injury, such as UV irradiation. However, in the case of pigment cells this response includes a mechanism for increasing melanin formation, i.e., increased MSH receptor activity. Should this be the case, similar G2/M “injury responses” of other cell types might be expected, consistent with their differentiated phenotypes.     

The Amphibian Kupffer Cells Build and Demolish Melanosomes: An Ultrastructural Point of View

This ultrastructural research was carried out to investigate the nature of the liver pigment cells of anuran and caudate amphibians, the pattern of melanosome ontogenesis, and the demolition processes of old melanosomes. We demonstrate that these liver pigment cells are able to internalize zymosan particles and latex beads; therefore, being professional phagocytes, they, as liver resident macrophages, can be classified as Kupffer cells (KCs). They show “melanosomogenesis centers” in which several maturation stages of premelanosomes are visible; the premelanosomes are formed by two principal components: a filamentous structure that will constitute the “inner” area of the melanosome and a vesicular component, budding from the Trans Golgi Network and that carries enzymes, which will constitute the “cortical area” of the melanosome. Thus the KCs, thanks to the presence of the “melanosomogenesis centers,” are also melanosome producing cells. They are also able to demolish melanosomes by heterophagocytosis and, probably, also by autophagocytosis. In conclusion, we propose a classification of vertebrate pigment cells.     

Melanins in Physiological Conditions Protect Against Lipoperoxidation. A Study on Albino and Pigmented Xenopus

The susceptibility to lipoperoxidation in liver of albino and pigmented Xenopus laevis Daudin, has been studied. Albino Xenopus liver was richer in polyunsaturated fatty acids (PUFAs) than the pigmented one; moreover, it was also richer in mitochondrial superoxide dismutase (MnSOD) and in reduced glutathione (GSH). The thiobarbituric acid-reactive substances (TBARS) values were more abundant in the albino tissue compared to the pigmented tissue both during spontaneous and Fe⁺⁺ induced lipoperoxidation. Therefore, when isolated and purified melanin, in physiological quantities, was added to albino tissue, the TBARS values drastically decreased. Thus, melanin, in our experimental conditions, protects the albino tissue even more than SOD and GSH do. Melanin, in our opinion, acts as an antioxidant, because it is able to scavenge O₂^?.     

Isolation and characterization of eight polymorphic microsatellite markers in Aphanius fasciatus (Teleostei: Cyprinodontidae)

The first set of eight polymorphic microsatellites markers was successfully optimized from a partial genomic library enriched for an AC motif of the killifish Aphanius fasciatus Nardo, 1827 (Teleostei, Cyprinodontidae), a brackish‐water fish widely distributed along the coasts of the Mediterranean Sea. The developed loci proved to be polymorphic, displaying from two to six alleles per locus with expected heterozygosity ranging from 0.277 to 0.752. All loci were under Hardy–Weinberg equilibrium, except Af8 that showed significant heterozygotes deficiency, and there was no evidence of linkage associations between any pair of loci. Markers reported here will be potentially useful in monitoring of genetic population structure in this species.     

Exploring the soluble proteome of Tobacco Bright Yellow‐2 cells at the switch towards different cell fates in response to heat shocks

Tobacco (Nicotiana tabacum) Bright Yellow‐2 (TBY‐2) cells undergo different fates when exposed for 10 minutes to heat stresses of different severity. A 35 °C treatment causes a homeostatic response (HRE) allowing cells to cope with the stress; 55 °C triggers processes leading to programmed cell death (PCD), which is complete after 72 h. We have used a proteomic approach to gain insight into the molecular mechanisms defining the fate of TBY‐2 cells induced by these two heat stresses. Tandem mass spectrometry (MS/MS) and two‐dimensional electrophoresis (2‐DE) analysis revealed little overlap of differentially‐accumulated proteins: the different severities of heat treatment induced the modulation of specific proteins, some of which are responsible for different cell fates. When the imposed heat shock is beyond a certain threshold, the overall reduced metabolism may be the result of a series of events involving gene expression and oxidative damage that would lead to PCD. Our data suggest that the down‐accumulation of several proteins involved in cellular redox homeostasis could provide, until now, an unappreciated contribution to understanding how many partners are involved in promoting the redox impairment leading to PCD. Moreover post‐translational modifications seem to play important regulatory roles in the adaptation of TBY‐2 cells to different intensities of heat stress.     

Geographical and morphological variation within and between colour phases in Coris julis (L. 1758), a protogynous marine fish

The possible differences between sexes in patterns of morphological variation in geographical space have been explored only in gonochorist freshwater species. We explored patterns of body shape variation in geographical space in a marine sequential hermaphrodite species, Coris julis (L. 1758), analyzing variation both within and between colour phases, through the use of geometric morphometrics and spatially‐explicit statistical analyses. We also tested for the association of body shape with two environmental variables: temperature and chlorophyll a concentration, as obtained from time‐series of satellite‐derived data. Both colour phases showed a significant morphological variation in geographical space and patterns of variation divergent between phases. Although the morphological variation was qualitatively similar, individuals in the initial colour phase showed a more marked variation than individuals in the terminal phase. Body shape showed a weak but significant correlation with environmental variables, which was more pronounced in primary specimens. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 104 , 148–162.