A Fluorescent Chromatophore Changes the Level of Fluorescence in a Reef Fish

Body coloration plays a major role in fish ecology and is predominantly generated using two principles: a) absorbance combined with reflection of the incoming light in pigment colors and b) scatter, refraction, diffraction and interference in structural colors. Poikilotherms, and especially fishes possess several cell types, so-called chromatophores, which employ either of these principles. Together, they generate the dynamic, multi-color patterns used in communication and camouflage. Several chromatophore types possess motile organelles, which enable rapid changes in coloration. Recently, we described red fluorescence in a number of marine fish and argued that it may be used for private communication in an environment devoid of red. Here, we describe the discovery of a chromatophore in fishes that regulates the distribution of fluorescent pigments in parts of the skin. These cells have a dendritic shape and contain motile fluorescent particles. We show experimentally that the fluorescent particles can be aggregated or dispersed through hormonal and nervous control. This is the first description of a stable and natural cytoskeleton-related fluorescence control mechanism in vertebrate cells. Its nervous control supports suggestions that fluorescence could act as a context-dependent signal in some marine fish species and encourages further research in this field. The fluorescent substance is stable under different chemical conditions and shows no discernible bleaching under strong, constant illumination

Regulation of red fluorescent light emission in a cryptic marine fish

INTRODUCTION: Animal colouration is a trade-off between being seen by intended, intra- or inter-specific receivers while not being seen by the unintended. Many fishes solve this problem by adaptive colouration. Here, we investigate whether this also holds for fluorescent pigments. In those aquatic environments in which the ambient light is dominated by bluish light, red fluorescence can generate high-contrast signals. The marine, cryptic fish Tripterygion delaisi inhabits such environments and has a bright red-fluorescent iris that can be rapidly up- and down-regulated. Here, we described the physiological and cellular mechanism of this phenomenon using a neurostimulation treatment with KCl and histology. RESULTS: KCl-treatment revealed that eye fluorescence regulation is achieved through dispersal and aggregation of black-pigmented melanosomes within melanophores. Histology showed that globular, fluorescent iridophores on the anterior side of the iris are grouped and each group is encased by finger-like extensions of a single posterior melanophore. Together they form a so-called chromatophore unit. By dispersal and aggregation of melanosomes into and out of the peripheral membranous extensions of the melanophore, the fluorescent iridophores are covered or revealed on the anterior (outside) of the iris. CONCLUSION: T. delaisi possesses a well-developed mechanism to control the fluorescent emission from its eyes, which may be advantageous given its cryptic lifestyle. This is the first time chromatophore units are found to control fluorescent emission in marine teleost fishes. We expect other fluorescent fish species to use similar mechanisms in the iris or elsewhere in the body. In contrast to a previously described mechanism based on dendritic fluorescent chromatophores, chromatophore units control fluorescent emission through the cooperation between two chromatophore types: an emitting and an occluding type. The discovery of a second mechanism for fluorescence modulation strengthens our view that fluorescence is a relevant and adaptive component of fish colouration

Results of cell manipulation.

<p>Bars show results of neuronal and hormonal manipulation in terms of normalized fluorescent area in interradial membranes of <i>E. pellucida</i> before (white bars) and after treatment (shaded bars, respectively). Neuronal K⁺ stimulation significantly decreased fluorescent area (paired t-test, t = 10.5, df = 5, p<0.005). The Lidocain-treatment effectively inhibited this effect (paired t-test, t = 1.86, df = 4, p>0.13). Neurotransmitter-induced aggregation of fluorosomes (NA) was highly significant (paired t-test, t = 4.67, df = 9, p<0.001). Aggregation induced by MCH was significant (paired t-test, t = 5.19, df = 3, p<0.013) as well as α–MSH significantly induced dispersal in pre-aggregated cells (paired Wilcoxon, Z = 10.5, df = 5, p = 0.03). Bars include standard errors.</p

Aggregation of fluorescence.

<p>Classified according to melanophore index 5 (a) through melanophore index 1 (e, respectively). In a completely dispersed state (a) the nucleus becomes visible as there is only little cytoplasm one the apical and basal side of the nucleus in these flat cells. In the aggregated state (e), the nucleus is tightly packed with fluorosomes. Scale bar  = 100 µm.</p

Morphology of a fluorescent chromatophore.

<p>Confocal Laser scanning microscopy, excitation 510 nm. Fluorosomes are the only visible structures within the fluorescent cell, but in the dispersed state they show the cell outline as they fill the cytoplasm entirely. Scale bar  = 30 µm.</p

Distribution of erythrophores, melanophores and fluorescent chromatophores in the interradial membrane of a dorsal fin of <i>E. pellucida</i>.

<p><b>a</b>) Erythrophores (red) and melanophores (black) are visible in bright field microscopy. <b>b</b>) Fluorescent chromatophores appear in fluorescence microscopy. <b>c</b>) Overlay of a) and b). Note that erythrophores, melanophores and fluorescent chromatophores are spatially distributed and can be distinguished. Scale bar  = 400 µm.</p

Spectra of excitation and emission of the fluorescent pigment in fluorescent chromatophores.

<p>Fluorescence has a maximum at around 595 nm (dashed line) with an optimal excitation wavelength around 500 nm (solid line).</p