Fish and amphibians have pigment cells that generate colorful skins important for signaling, camouflage, thermoregulation and protection against ultraviolet radiation (UVR). However, many animals also have pigment cells inside their bodies, on their internal organs and membranes. In contrast to external pigmentation, internal pigmentation is remarkably little studied and its function is not well known. Here, we tested genotoxic effects of UVR and its effects on internal pigmentation in a neotropical frog, Physalaemus nattereri. We found increases in body darkness and internal melanin pigmentation in testes and heart surfaces and in the mesenterium and lumbar region after just a few hours of UVR exposure. The melanin dispersion in melanomacrophages in the liver and melanocytes in testes increased after UV exposure. In addition, the amount of melanin inside melanomacrophages cells also increased. Although mast cells were quickly activated by UVR, only longer UVR exposure resulted in genotoxic effects inside frogs, by increasing the frequency of micronuclei in red blood cells. This is the first study to describe systemic responses of external UVR on internal melanin pigmentation, melanomacrophages and melanocytes in frogs and thus provides a functional explanation to the presence of internal pigmentation.
Glyphosate (GLY) is the most used herbicide worldwide and its effects on anurans are well known. Pollutants can cause physiological and morphological effects. Therefore, this study evaluated the effects of GLY on hepatic melanomacrophages as a response to environmental stressors. Three treatments were exposed to different concentrations of pure GLY (100, 1000, and 10,000 μg g(-1), respectively), and there was also a control group. After the experimental time, liver and blood were analyzed. Melanomacrophages (MMCs) were located between the hepatocyte cordons, close to sinusoids. GLY increased the melanin area in MMCs of Leptodactylus latinasus exposed since lowest concentration until highest concentration. GLY also changed the occurrence of hepatic catabolism pigments into melanomacrophages and erythrocyte nuclear abnormalities; therefore, it can interfere with the hepatic metabolism. In conclusion, GLY promotes alterations in the hepatic tissue and erythrocyte nuclear abnormalities. Furthermore, MMCs may be useful as morphological responses of GLY effects.
In amphibians, pigmented cells are present in several organs, composing an extracutaneous pigmentary system. Seventeen species from two families were studied to develop a protocol for pigmentary classification. The amount and distribution of these cells are variable, allowing the establishment of anatomical patterns for visceral pigmentation in anuran testes.
Benzo[a]pyrene (BaP) is a bio-accumulative toxic compound found in the atmosphere, water, and soil that may affect the life cycle of amphibians. In this study, a few contamination biomarkers, such as hepatic melanomacrophages (MMs), mast cells, erythrocyte micronuclei (MN) and white blood cells were used to determine how BaP acts in these cells in the anurans Physalaemus cuvieri and Leptodactylus fuscus. Animals of both species were divided into three treatment groups: 1 day, 7 days and 13 days, subcutaneously injected 2 mg/kg BaP diluted in mineral oil and control group with only mineral oil. After 7 days, BaP caused the frequency of MN to increase in both species while reducing melanin area. The micronucleus frequency increased due to the genotoxicity of BaP, while the decreasing melanin area may be related to the inhibition of tyrosinase activity, an enzyme responsible for regulating melanogenesis, decreasing the synthesis of melanin. The mast cell density increased in all groups and in both species as a response to the inflammatory action of BaP. These cells respond to nonspecific inflammatory effects leading, therefore, to this response in all treatments. The percentage of leukocytes remained unchanged probably due to great intraspecific variability. Additionally, the leukocyte profiles of both species were characterized and the differences were attributed to extrinsic factors. In short, BaP can affect the integrity of several organs and tissues, and cell functions leading to the conclusion that this compound is hepatotoxic, genotoxic and immunotoxic for anurans.
Melanomacrophages are the pigmented cells present in the hematopoietic organs. Besides melanin, hemosiderin and lipofuscin are also observed in the melanomacrophages. For the liver, however, numerous studies relate these cells to immunological and metabolic functions. Therefore, the aim of this study is to evaluate the hepatic metabolism by quantifying melanin, hemosiderin and lipofuscin in the anuran Eupemphix nattereri submitted to varying thermal conditions. E. nattereri adult males were separated into three groups, as follows: (i) five animals in the control group were kept at room temperature (27 C); (ii) 30 animals were submitted to hyperthermic (35.1 C); and (iii) 30 to hypothermic (18.9 ) conditions. In each experiment, the animals were analyzed and separated into two different treatments:(1) immediately after undergoing the stress; and, (2) after recovering from the stress caused by the stimulus, at three distinct times (12 hr, 24 hr, and 48 hr). Both hyperthermia and hypothermia decreased hepatic pigmentation after thermal stress. The recovered animals of both experimental treatments showed as much pigmentation as the control animals. Thermal stress alters hemosiderin and lipofuscin as well, which may be related to liver function catabolism. In conclusion, liver pigmentation decreased due to temperature variation and duration of thermal stimulation to which the animals were exposed. The increase in temperature rather than hypothermia led to more drastic physiological disorders. In this study, we observed that thermal stress for a short period compromises the morphology and liver function, as observed by the changing pigmentation of melanomacrophages. These analyses can be used as biomarkers of environmental effects. Anat Rec,
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