Melanophore Density Research Articles

Effect of stocking density on gonadal development, pigmentation and survival of Carassius carassius (Crucian carp) in a biofloc system

The present study was carried out to investigate the optimum stocking density for gonadal development, pigmentation, and survival of Carassius carassius stocked at varying stocking densities under a biofloc-based system. C. carassius (25 ± 0.15 g) were randomly distributed in sixteen tanks, each with 250 L water at the stocking densities viz. 1 kg/m3, 2 kg/m3, 3 kg/m3, 4 kg/m3 (10, 20, 30 and 40 fish/250 L). The carbon‑nitrogen ratio of 15:1 was maintained using wheat flour as a carbon source. The best gonad performance in terms of absolute fecundity (4449.98 ± 115.636), relative fecundity (114.67 ± 2.770), GSI (5.62 ± 0.018 and 2.93 ± 0.014 for females & males respectively), and advanced maturation stages (vitellogenic oocytes and seminiferous tubules with spermatozoa) was observed in the density treatment having 20 fish/250 L. The skin carotenoid content showed an increasing trend with maximum content (8.11 ± 0.018 μg/g)) recorded for the fish stocked at 20/250 L. The scales of the fish from the same treatment presented a higher concentration of yellow orange pigments giving an overall golden hue to the scales while as the density and dispersion of melanophores increased with an increase in stocking density giving an overall darker coloration to the fish, indicating a stressful condition at higher stocking density. The survival rate of fish showed no significant difference (p≥0.05) between the various density groups. The results of this experiment elucidate that Carassius carassius could be cultured with better gonad performance, increased carotenoid content, and high survival at a stocking density of 2 kg/m3 in a biofloc system.

The genetic basis of divergent melanic pigmentation in benthic and limnetic threespine stickleback

Pigmentation is an excellent trait to examine patterns of evolutionary change because it is often under natural selection. Benthic and limnetic threespine stickleback (Gasterosteus aculeatus) exhibit distinct pigmentation phenotypes, likely an adaptation to occupation of divergent niches. The genetic architecture of pigmentation in vertebrates appears to be complex. Prior QTL mapping of threespine stickleback pigmentation phenotypes has identified several candidate loci. However—relative to other morphological phenotypes (e.g., spines or lateral plates)—the genetic architecture of threespine stickleback pigmentation remains understudied. Here, we performed QTL mapping for two melanic pigmentation traits (melanophore density and lateral barring) using benthic-limnetic F2 crosses. The two traits mapped to different chromosomes, suggesting a distinct genetic basis. The resulting QTLs were additive, but explained a relatively small fraction of the total variance (~6%). QTLs maps differed by F1 family, suggesting variation in genetic architecture or ability to detect loci of small effect. Functional analysis identified enriched pathways for candidate loci. Several of the resulting candidate loci for pigmentation, including three loci in enriched pathways (bco1, sulf1, and tyms) have been previously indicated to affect pigmentation in other vertebrates. These findings add to a growing body of evidence suggesting pigmentation is often polygenic.

Assessing Colouration in Ichthyology: Insights from the Melanophore Index Perspective

Fish colouration significantly influences various aspects of their biology, including communication, camouflage, and mate selection. Among the pigment cells responsible for generating colours, melanophores are particularly significant due to their ability to produce dark pigments called melanin. The Melanophore Index (MI) quantitatively evaluates melanophore density and distribution across different fish species. This comprehensive review delves into the significance of fish colouration, the biology of melanophores, methods for assessing MI, factors influencing MI variations, and the ecological and evolutionary implications of MI in fish populations. By synthesizing current research and highlighting gaps in knowledge, this review aims to provide a foundational understanding of assessing fish colouration through the lens of the Melanophore Index.

Developmental and Cellular Basis of Vertical Bar Color Patterns in the East African Cichlid Fish Haplochromis latifasciatus.

The East African adaptive radiations of cichlid fishes are renowned for their diversity in coloration. Yet, the developmental basis of pigment pattern formation remains largely unknown. One of the most common melanic patterns in cichlid fishes are vertical bar patterns. Here we describe the ontogeny of this conspicuous pattern in the Lake Kyoga species Haplochromis latifasciatus. Beginning with the larval stages we tracked the formation of this stereotypic color pattern and discovered that its macroscopic appearance is largely explained by an increase in melanophore density and accumulation of melanin during the first 3 weeks post-fertilization. The embryonal analysis is complemented with cytological quantifications of pigment cells in adult scales and the dermis beneath the scales. In adults, melanic bars are characterized by a two to threefold higher density of melanophores than in the intervening yellow interbars. We found no strong support for differences in other pigment cell types such as xanthophores. Quantitative PCRs for twelve known pigmentation genes showed that expression of melanin synthesis genes tyr and tyrp1a is increased five to sixfold in melanic bars, while xanthophore and iridophore marker genes are not differentially expressed. In summary, we provide novel insights on how vertical bars, one of the most widespread vertebrate color patterns, are formed through dynamic control of melanophore density, melanin synthesis and melanosome dispersal.

Bar, stripe and spot development in sand-dwelling cichlids from Lake Malawi

BackgroundMelanic patterns such as horizontal stripes, vertical bars and spots are common among teleost fishes and often serve roles in camouflage or mimicry. Extensive research in the zebrafish model has shown that the development of horizontal stripes depends on complex cellular interactions between melanophores, xanthophores and iridophores. Little is known about the development of horizontal stripes in other teleosts, and even less is known about bar or spot development. Here, we compare chromatophore composition and development of stripes, bars and spots in two cichlid species of sand-dwellers from Lake Malawi—Copadichromis azureus and Dimidiochromis compressiceps.Results(1) In D. compressiceps, stripes are made of dense melanophores underlaid by xanthophores and overlaid by iridophores. Melanophores and xanthophores are either loose or absent in interstripes, and iridophores are dense. In C. azureus, spots and bars are composed of a chromatophore arrangement similar to that of stripes but are separated by interbars where density of melanophores and xanthophores is only slightly lower than in stripes and iridophore density appears slightly greater. (2) Stripe, bar and spot chromatophores appear in the skin at metamorphosis. Stripe melanophores directly differentiate along horizontal myosepta into the adult pattern. In contrast, bar number and position are dynamic throughout development. As body length increases, new bars appear between old ones or by splitting of old ones through new melanophore appearance, not migration. Xanthophore and iridophore distributions follow melanophore patterns. (3) Metamorphic pigmentation arises in cichlids in a fashion similar to that described in zebrafish: melanophore progenitors derived from the medial route of neural crest migration migrate from the vicinity of the neural tube to the skin during metamorphosis.ConclusionThe three pigment cell types forming stripes, bars and spots arise in the skin at metamorphosis. Stripes develop by differentiation of melanophores along horizontal myosepta, while bars do not develop along patent anatomical boundaries and increase in number in relation with body size. We propose that metamorphic melanophore differentiation and migratory arrest upon arrival to the skin lead to stripe formation, while bar formation must be supported by extensive migration of undifferentiated melanophores in the skin.

Searching integrated strategies for the evaluation of the physiological status in fish fed functional diets: The example of SDPP in gilthead sea bream (Sparus aurata)

In this study we evaluated the effects of high dietary arachidonic acid (ARA) levels on prostaglandin E2 (PGE2) and E3 (PGE3) production and their effect on two morphogenetic processes occurring during metamorphosis: the establishment of the juvenile pigmentation pattern and eye migration and remodeling of cranial bones. In this sense, Senegalese sole larvae were fed from 2 to 50 days post-hatch (dph) with live prey enriched with an experimental emulsion containing high levels of ARA (ARA-H; 10.2 and 7.1% TFA in enriched rotifer and Artemia, respectively) versus a reference commercial enriching product (Algamac 3050®, AGM; 1.0 and 1.4% TFA in enriched rotifer and Artemia, respectively). High dietary ARA levels did not affect larval growth performance at 50 dph, but significantly induced malpigmentation (81.4 ± 7.5%, versus 0.9 ± 0.3% in larvae fed the AGM diet). This malpigmentation was linked to the higher prostaglandin E2 (PGE2) levels observed in pseudo-albino fish as compared to normally pigmented individuals. The PGE2 levels were higher in normally pigmented specimens fed the ARA-H diet than in those fed the AGM diet. The effects of ARA on normally pigmented fish fed the AGM diet and pseudo-albino specimens fed the ARA-H diet were evaluated by means of the density of melanophores and the texture and image segmentation analyses in the dorsal skin of post-metamorphic fish. The skin of pseudo-albino specimens had a more uniform and homogeneous melanophore pattern than normally pigmented fish. Melanophores in pseudo-albino specimens were less abundant and not so aggregated in patches as they were in normally pigmented ones, whereas their shape differed (round vs. dendritic) suggesting their inability to disperse melanin. In addition, fish fed the ARA-H diet presented a higher percentage of cranial deformities (95.1 ± 1.5%) than those fed the control diet (1.9 ± 1.9%) that was significantly and negatively correlated with the incidence of normally-pigmented animals (R2 = − 0.88, P < 0.001). Cranial deformities in pseudo-albino fish were associated with an impaired migration of the eye from the ocular side (the right eye), whereas the left eye migrated from the blind side into the ocular side almost normally. The effects of high dietary ARA levels in the eye migration and cranial bone remodeling processes in post-metamorphic larvae were evaluated by means of the staining of cranial skeletal elements. Pseudo-albino fish showed higher interocular distance and head height than normally pigmented individuals, a different disposition of the eyes with regard to the vertebral column and mouth axes, and a distinct osteological development of some skeletal structures from the neuro- and splanchnocranium, in relation to high dietary ARA contents and high PGE2 production. These results brought new information about possible nutritional forcing factors and physiological mechanisms of pigmentary disorders and impaired eye migration, which are current major bottlenecks in flatfish aquaculture.

Regulation, constraints and benefits of colour plasticity in a mimicry system

Rapid colour change is used in aggressive interactions, ontogenetic transitions, nuptial displays and to prevent detection and/or recognition from predators or prey. The underlying mechanisms, constraints and benefits of colour change are often unclear, but examining such factors offers insights into phenotypic plasticity. Here, we investigated the mechanisms behind how an aggressive reef fish mimic (bluestriped fangblenny Plagiotremus rhinorhynchos) changes colour rapidly (1-5 min) between mimetic and other colour forms. Black with one neon blue dorsal stripe (mimic), black with two neon blue stripes, brown, olive and orange forms differed in melanophore density. Fish skin biopsies were modulated in vitro by hormones, and smaller fangblennies changed coloration more rapidly than larger fish suggesting that the ability to change colour is diminished as fish get larger. Individuals may be limited by differences in pigment cell densities to change colour between extreme colour forms (black to orange); therefore, longer morphological changes may also occur or fangblennies may exhibit dimorphic populations. Behavioural observations suggest that small black and orange individuals were equally successful in attacking passing fish to feed on dermal tissue/scales, indicating that deceptive strategies used by each colour form may deliver equal fitness benefits. The present study demonstrates for the first time how fangblennies change colour and highlights that colour plasticity offers important adaptive advantage; however, physiological constraints should also be considered.

Evaluation of the Effect of Sidoarjo Mud on Aquatic Life Using Chromatophores and the Microstructure of Fish Scales

The Sidoarjo mud is the first visible phenomenon of a mud volcano that occurs in a human settlement and which is subsequently channelled into a river. Clay, aluminium and iron were reported to be the dominant contaminants that could possibly come into contact with and accumulate on the surface of local fish and initiate alteration in scale microstructure. The aim of this study was to evaluate the extent of water body contamination in the Sidoarjo mud by evaluating the chromatophore density and microstructure deformation of fish scales that act as biomarkers. Scale samples were obtained from caged Mozambique tilapia (Oreochromis mossambicus) fish that were placed downstream and upstream of the Sidoarjo mud spillway pipes. With respect to melanophore density, it was found that the scales of fish exposed in the downstream section were significantly lower in chromatophores ( 100 chr/mm²). This study suggested that the density of chromatophores was closely related to the concentration of total suspended solids (r = 0.69), which was possibly enhanced by iron (r = 0.56). Using scanning electron microscopy analysis, some deformation, i.e. irregularity of spherule shape and increasing pits in the space between ridges, were observed.

A gene expression study of dorso-ventrally restricted pigment pattern in adult fins of Neolamprologus meeli, an African cichlid species.

Fish color patterns are among the most diverse phenotypic traits found in the animal kingdom. Understanding the molecular and cellular mechanisms that control in chromatophore distribution and pigmentation underlying this diversity is a major goal in developmental and evolutionary biology, which has predominantly been pursued in the zebrafish model system. Here, we apply results from zebrafish work to study a naturally occurring color pattern phenotype in the fins of an African cichlid species from Lake Tanganyika. The cichlid fish Neolamprologus meeli displays a distinct dorsal color pattern, with black and white stripes along the edges of the dorsal fin and of the dorsal half of the caudal fin, corresponding with differences in melanophore density. To elucidate the molecular mechanisms controlling the differences in dorsal and ventral color patterning in the fins, we quantitatively assessed the expression of 15 candidate target genes involved in adult zebrafish pigmentation and stripe formation. For reference gene validation, we screened the expression stability of seven widely expressed genes across the investigated tissue samples and identified tbp as appropriate reference. Relative expression levels of the candidate target genes were compared between the dorsal, striped fin regions and the corresponding uniform, grey-colored regions in the anal and ventral caudal fin. Dorso-ventral expression differences, with elevated levels in both white and black stripes, were observed in two genes, the melanosome protein coding gene pmel and in igsf11, which affects melanophore adhesion, migration and survival. Next, we predicted potential shared upstream regulators of pmel and igsf11. Testing the expression patterns of six predicted transcriptions factors revealed dorso-ventral expression difference of irf1 and significant, negative expression correlation of irf1 with both pmel and igsf11. Based on these results, we propose pmel, igsf11 and irf1 as likely components of the genetic mechanism controlling distinct dorso-ventral color patterns in N. meeli fins.

Embryonic developmental process governing the conspicuousness of body stripes and blue tail coloration in the lizard Plestiodon latiscutatus.

The combination of body stripes and vivid blue tail color has independently evolved in different lizard families. To understand how and when lizards developed this coloration, we microscopically compared the embryonic development of pigment cells in two island populations of Plestiodon latiscutatus that exhibit either striped and blue tailed or inconspicuously striped and blue tailed juveniles, based on the newly determined 12 normal developmental stages of embryos from shortly after egg laying to just before hatching. We focus on the role of the melanophores in the body stripe and the role of iridophore morphotypes in the extent of blue tail coloration. The melanophore density in the black background region was higher in lizards with vivid stripes than in lizards with drab stripes. Iridophores started to fill the dermal space that was not yet occupied by melanophores, which resulted in a higher iridophore density in stripes than in the inter-stripe regions. We also discovered that iridophores with thin platelets reflecting blue structural coloration appeared at a specific region in the tail at stage 11. The position of the tail where iridophores emerged coincided with the boundary area separating anterior brown and green colored tail from posterior blue colored tail after hatching.

Pigment cell mechanism of postembryonic stripe pattern formation in the Japanese four-lined snake.

Postembryonic changes in the dermal and epidermal pigment cell architecture of the striped and nonstriped morph of the Japanese four-lined snake Elaphe quadrivirgata were examined to reveal stripe pattern formation after hatching. The striped and nonstriped morphs were distinguishable at the hatching, suggesting that the basis of stripe pattern was formed during embryonic development. In the striped morph, the color of stripes changed from red-brown in juveniles to vivid dark-brown in adults, and density of dermal melanophore increased much more in the stripe than background dorsal scales with growth. This increase in density of dermal melanophore was accompanied not only by the increased epidermal melanophore density but also by the change in vertical structures of dermal melanophore. By contrast, the density of epidermal and dermal melanophore evenly increased over the dorsal scales in the nonstriped morph. Thus, the increased vividness of the stripe pattern after hatching is achieved through localized increase of melanophore density particularly in the stripe region but not over the whole dorsal scales.

Comparison of pigment cell ultrastructure and organisation in the dermis of marble trout and brown trout, and first description of erythrophore ultrastructure in salmonids.

Skin pigmentation in animals is an important trait with many functions. The present study focused on two closely related salmonid species, marble trout (Salmo marmoratus) and brown trout (S. trutta), which display an uncommon labyrinthine (marble-like) and spot skin pattern, respectively. To determine the role of chromatophore type in the different formation of skin pigment patterns in the two species, the distribution and ultrastructure of chromatophores was examined with light microscopy and transmission electron microscopy. The presence of three types of chromatophores in trout skin was confirmed: melanophores; xanthophores; and iridophores. In addition, using correlative microscopy, erythrophore ultrastructure in salmonids was described for the first time. Two types of erythrophores are distinguished, both located exclusively in the skin of brown trout: type 1 in black spot skin sections similar to xanthophores; and type 2 with a unique ultrastructure, located only in red spot skin sections. Morphologically, the difference between the light and dark pigmentation of trout skin depends primarily on the position and density of melanophores, in the dark region covering other chromatophores, and in the light region with the iridophores and xanthophores usually exposed. With larger amounts of melanophores, absence of xanthophores and presence of erythrophores type 1 and type L iridophores in the black spot compared with the light regions and the presence of erythrophores type 2 in the red spot, a higher level of pigment cell organisation in the skin of brown trout compared with that of marble trout was demonstrated. Even though the skin regions with chromatophores were well defined, not all the chromatophores were in direct contact, either homophilically or heterophilically, with each other. In addition to short-range interactions, an important role of the cellular environment and long-range interactions between chromatophores in promoting adult pigment pattern formation of trout are proposed.

Senegalese sole (Solea senegalensis) metamorphic larvae are more sensitive to pseudo-albinism induced by high dietary arachidonic acid levels than post-metamorphic larvae

Abstract High dietary levels of arachidonic acid (ARA) and its relative proportions with eicosapentaenoic acid (EPA), fed during early larval stages, have been associated with malpigmentation in various flatfish species. This study investigated whether the nutritional induction of pigmentary disorders at larval stages was related to a specific larval period of increased sensitivity to ARA in Senegalese sole (Solea senegalensis Kaup, 1858). Senegalese sole larvae were fed high dietary ARA levels during pre- and pro-metamorphosis (2–15 dph) and/or post-metamorphosis (15–50 dph). Larval tissues reflected the dietary fatty acid composition. Malpigmentations were significantly related to elevated dietary and larval ARA contents and ARA/EPA ratio. This study reports evidence for a “pigmentation window”, with a higher larval sensitivity to dietary ARA during pre- and pro-metamorphosis than post-metamorphosis. High dietary ARA fed to larvae during pre-metamorphosis enhanced survival, but did not affect growth nor eye migration. The aspect and density of melanophores in the skin of the ocular side of ARA-induced pseudo-albinos were significantly reduced in comparison to normally pigmented individuals, even more in the pseudo-albino fish fed high dietary ARA levels during the pre-metamorphic stage. Pseudo-albino fish fed high dietary ARA levels during post-metamorphosis showed higher concentrations of 2- and 3-series prostaglandins (PGE2 and PGE3) than normally pigmented specimens fed the same diets. An increased sensitivity to ARA-induced malpigmentations has been identified at pre-metamorphosis and early metamorphosis in Senegalese sole. Supplying high dietary ARA amounts imbalanced the dietary ARA/EPA ratio and disrupted the relative concentrations of derived PGE2 and PGE3, resulting in 20 to 81.7% pseudo-albino individuals, depending on the dietary treatment. The administration of high levels of dietary ARA at different developmental stages did not only affect the incidence of animals with pigmentary disorders, but it also affect the melanophore density and skin aspect in normally pigmented and pseudoalbino fish as image segmentation and texture analyses indicated.

The effects of dietary arachidonic acid on Senegalese sole morphogenesis: A synthesis of recent findings

In this study we evaluated the effects of high dietary arachidonic acid (ARA) levels on prostaglandin E 2 (PGE 2 ) and E 3 (PGE 3 ) production and their effect on two morphogenetic processes occurring during metamorphosis: the establishment of the juvenile pigmentation pattern and eye migration and remodeling of cranial bones. In this sense, Senegalese sole larvae were fed from 2 to 50 days post-hatch (dph) with live prey enriched with an experimental emulsion containing high levels of ARA (ARA-H; 10.2 and 7.1% TFA in enriched rotifer and Artemia , respectively) versus a reference commercial enriching product (Algamac 3050®, AGM; 1.0 and 1.4% TFA in enriched rotifer and Artemia , respectively). High dietary ARA levels did not affect larval growth performance at 50 dph, but significantly induced malpigmentation (81.4 ± 7.5%, versus 0.9 ± 0.3% in larvae fed the AGM diet). This malpigmentation was linked to the higher prostaglandin E2 (PGE 2 ) levels observed in pseudo-albino fish as compared to normally pigmented individuals. The PGE 2 levels were higher in normally pigmented specimens fed the ARA-H diet than in those fed the AGM diet. The effects of ARA on normally pigmented fish fed the AGM diet and pseudo-albino specimens fed the ARA-H diet were evaluated by means of the density of melanophores and the texture and image segmentation analyses in the dorsal skin of post-metamorphic fish. The skin of pseudo-albino specimens had a more uniform and homogeneous melanophore pattern than normally pigmented fish. Melanophores in pseudo-albino specimens were less abundant and not so aggregated in patches as they were in normally pigmented ones, whereas their shape differed (round vs. dendritic) suggesting their inability to disperse melanin. In addition, fish fed the ARA-H diet presented a higher percentage of cranial deformities (95.1 ± 1.5%) than those fed the control diet (1.9 ± 1.9%) that was significantly and negatively correlated with the incidence of normally-pigmented animals (R 2 = − 0.88, P < 0.001). Cranial deformities in pseudo-albino fish were associated with an impaired migration of the eye from the ocular side (the right eye), whereas the left eye migrated from the blind side into the ocular side almost normally. The effects of high dietary ARA levels in the eye migration and cranial bone remodeling processes in post-metamorphic larvae were evaluated by means of the staining of cranial skeletal elements. Pseudo-albino fish showed higher interocular distance and head height than normally pigmented individuals, a different disposition of the eyes with regard to the vertebral column and mouth axes, and a distinct osteological development of some skeletal structures from the neuro- and splanchnocranium, in relation to high dietary ARA contents and high PGE 2 production. These results brought new information about possible nutritional forcing factors and physiological mechanisms of pigmentary disorders and impaired eye migration, which are current major bottlenecks in flatfish aquaculture. • High dietary arachidonic acid levels induced pseudo-albinism in Senegalese sole. • Pseudo-albinism was significantly correlated with impaired eye migration. • The process of cranial bone remodeling was affected in pseudo-albino fish. • High ARA levels increased PGE 2 production in pseudo-albino deformed fish.

Developmental changes in melanophores and their asymmetrical responsiveness to melanin-concentrating hormone during metamorphosis in barfin flounder (Verasper moseri)

Barfin flounder larvae exhibit unique black coloration, as well as left–right asymmetry in juvenile stage as in other flatfish. In this study, we first assessed the changes in melanophores with development and then investigated their responsiveness to melanin-concentrating hormone (MCH) during metamorphosis. Larval-type melanophores appeared on both sides of the body before metamorphosis, whereas adult-type melanophores appeared only on the ocular side after metamorphosis. Even in the individuals of this species displaying black coloration, the density of larval-type melanophores was similar to that in transparent larvae of other species. However, unlike in transparent larvae, larval-type melanophores completely dispersed in the black larvae of this species. Therefore, the black coloration during larval stages was mainly due to dispersion, and not the density, of larval-type melanophores. In vitro MCH treatment revealed, for the first time, the responsiveness of melanophores in larval stages. On the ocular side, larval-type melanophores aggregated against MCH during larval stages, while, in the larvae at later metamorphic stages and in juveniles, larval-type melanophores did not aggregate, although aggregation of adult-type melanophores was noted. In contrast, on the blind side, the responsiveness of larval-type melanophores to MCH was consistently present from larval to juvenile stages. The metamorphic transition of MCH responsiveness from larval- to adult-type melanophores only on the ocular side suggests the larval (therefore, immature) nature of the blind side skin. We propose that the inhibited development, and thus the retention of the larval-type skin leads to the formation of the blind side characteristics and is the central mechanism for the flatfish asymmetry.

Comparison of UV-B tolerance between wild and hatchery-reared juveniles of red sea bream (Pagrus major) and black sea bream (Acanthopagrus schlegeli)

The amount of ultraviolet (UV)-B radiation reaching the sea surface has increased due to ozone depletion. Several laboratory studies have highlighted the negative impacts of UV radiation on fish using hatchery-reared specimens. However, potential differences in UV tolerance between wild and hatchery-reared fish have been given little consideration. Wild and reared juveniles of red sea bream and black sea bream were exposed to one of four different UV-B radiation levels (1.8; 1.1; 0.4; 0 W/m2) for 4 h. Survival rate was measured every 2 h for a period of 24 h (red sea bream) or 48 h (black sea bream) following exposure. Wild and reared juvenile red sea bream were characterized by similar survival rate, with survival declining to almost 0 % 24 h after exposure at the 1.1 and 1.8 W/m2 levels. In black sea bream, wild individuals showed significantly higher survival than reared fish in levels 1.1 and 1.8 W/m2. Melanophore density was also measured since melanin absorbs UV radiation. Wild black sea bream showed higher melanophore density compared to reared individuals, while no such difference was observed in red sea bream. We conclude that wild black sea bream juveniles acquire higher UV tolerance partly by increasing melanophore density through exposure to UV radiation. Our results indicate that the predicted impacts of UV radiation on fish populations solely based on experimentation with hatchery-reared specimens may be overestimated for some species.

Effects of Androgens on the Development of Nuptial Coloration and Chromatophores in the BitterlingRhodeus ocellatus ocellatus

Sexually mature male bitterlings, Rhodeus ocellatus ocellatus, exhibit distinct nuptial color, whereas females maintain a body color similar to that of juveniles. In the present study, body color and chromatophores were compared between male and female bitterlings, and the effects of androgens on body color and chromatophore densities were examined in females to clarify the role of androgen in the development of nuptial coloration and chromatophores. Males showed green, blue, and red color in specific regions of their skin and red color on the dorsal and caudal fins; females showed a subdued silver body color. For chromatophores, small greenish-type iridophores were observed in the green color region in the skin of males, whereas females had large spindle-shaped silvery-type iridophores in corresponding regions. Many erythrophores were observed in males in blue and red color regions in the skin and red color regions in the fins, but females possessed xanthophores in corresponding regions. The melanophore density of the skin was not different between males and females, but the distribution of melanophores in the fins was different between them. Treatment with 11-ketotestosterone or methyltestosterone induced male-type nuptial coloration in the female skin and fins. The distribution of chromatophores in androgen-treated females was similar to that in sexually mature males: an increase in the number of greenish-type iridophores and erythrophores was also observed in the skin. These results indicate that androgen induces male-type nuptial coloration in the bitterling and that the responses of chromatophores to androgen differ with the type and distribution site of the chromatophores.

Mechanisms of pigmentation loss in subterranean fishes

Troglobitic (exclusively subterranean) organisms usually present, among their apomorphies related to the subterranean life (troglomorphisms), the regression of eyes and melanic pigmentation. The degree of regression varies among species, from a slight reduction to the complete loss of eyes and dark pigmentation, without a taxonomic correlation. While mechanisms of eye reduction have been intensively investigated in some troglobites such as the Mexican blind tetra characins, genus Astyanax, and the European salamander, Proteus anguinus, few studies have focused on pigmentation. The Brazilian subterranean ichthyofauna distinguishes not only by the species richness (23 troglobitic fishes so far known) but also by the variation in the degree of reduction of eyes and pigmentation. This study focused on Brazilian fishes completely devoid of melanic pigmentation: the characiform Stygichthys typhlops (Characidae) and the siluriforms Ancistrus formoso (Loricariidae), Rhamdiopsis sp.1 (Heptapteridae; from caves in the Chapada Diamantina, Bahia) and Rhamdiopsis sp. 2 (cave in Campo Formoso, Bahia). In order to investigate if such depigmentation is the result of blockage in some step in the melanogenesis, in vitro tests of administration of L-DOPA were done, using caudal-fin fragments extracted from living fish. Except for Rhamdiopsis sp. 2, all the studied species were DOPA(+), i.e., melanin was synthesized after L-DOPA administration. This indicates these fish do have melanophores but they are unable to convert L-tyrosine to L-DOPA. On the other hand, Rhamdiopsis sp. 2, like the albino specimens of Trichomycterus itacarambiensis previously studied (which correspond to one third of the population), are DOPA(-), either because the block of melanin synthesis occurs downstream in melanogenesis, which is probably the case with T. itacarambiensis (monogenic system in view of the phenotypic discontinuity), or because the so-called albinos do no possess melanophores. The physiological loss in the ability to synthesize melanin, apparently caused by different genetic processes in DOPA(+) and in DOPA(-) fishes, may co-exist in subterranean populations with a decrease in the density of melanophores, as observed in the pigmented two thirds of T. itacarambiensis population, a morphological reduction apparently controlled by polygenic systems producing a continuous phenotypic variation.

Adult‐type pigment cells, which color the ocular sides of flounders at metamorphosis, localize as precursor cells at the proximal parts of the dorsal and anal fins in early larvae

Flounders form left-right asymmetry in body coloration during metamorphosis through differentiation of adult-type melanophores and xanthophores on the ocular side. As the first step in investigating the formation of flounder body coloration asymmetry, in this study, we aimed to determine where the precursors of adult-type chromatophores distribute in larvae before metamorphosis. In Paralichthys olivaceus and Verasper variegatus, GTP cyclohydrolase 2 (gch2), a common marker of melanoblasts and xanthoblasts, was found to be transiently expressed in cells located along the bilateral skeletal muscles at the basal parts of the dorsal and anal fins of premetamorphic larvae. When V. variegatus larvae were fed with a strain of Artemia collected in Brazil, this gch2 expression was abolished and the differentiation of adult-type melanophores was completely inhibited, while the density of larval melanophores was not affected. In a cell trace test in which the cells at the basal part of the dorsal fin were labeled with DiI at the premetamorphic stage, adult-type melanophores labeled with DiI were found in the skin on the ocular side after metamorphosis. These data suggest that, in flounder larvae, adult-type melanophores are distributed at the basal parts of the dorsal and anal fins as unpigmented precursor cells.

Regeneration of neural crest derivatives in the Xenopus tadpole tail.

BackgroundAfter amputation of the Xenopus tadpole tail, a functionally competent new tail is regenerated. It contains spinal cord, notochord and muscle, each of which has previously been shown to derive from the corresponding tissue in the stump. The regeneration of the neural crest derivatives has not previously been examined and is described in this paper.ResultsLabelling of the spinal cord by electroporation, or by orthotopic grafting of transgenic tissue expressing GFP, shows that no cells emigrate from the spinal cord in the course of regeneration.There is very limited regeneration of the spinal ganglia, but new neurons as well as fibre tracts do appear in the regenerated spinal cord and the regenerated tail also contains abundant peripheral innervation.The regenerated tail contains a normal density of melanophores. Cell labelling experiments show that melanophores do not arise from the spinal cord during regeneration, nor from the mesenchymal tissues of the skin, but they do arise by activation and proliferation of pre-existing melanophore precursors. If tails are prepared lacking melanophores, then the regenerates also lack them.ConclusionOn regeneration there is no induction of a new neural crest similar to that seen in embryonic development. However there is some regeneration of neural crest derivatives. Abundant melanophores are regenerated from unpigmented precursors, and, although spinal ganglia are not regenerated, sufficient sensory systems are produced to enable essential functions to continue.