The fine structure of integumental erythrophores and the intracellular location of pteridine and carotenoid pigments in adult goldfish, Carassius auratus, were studied by means of cytochemistry, paper and thin-layer chromatography, ionophoresis, density-gradient centrifugal fractionation, and electron microscopy. The ultrastructure of erythrophores is characterized by large numbers of somewhat ellipsoidal pigment granules and a well-developed system of tubules which resembles endoplasmic reticulum. The combined morphological and biochemical approaches show that pteridine pigments of erythrophores are located characteristically in pigment granules and are the primary yellow pigments of these organelles. Accordingly, this organelle is considered to be the "pterinosome" which was originally found in swordtail erythrophores. Major pteridines obtainable from goldfish pterinosomes are sepiapterin, 7-hydroxybiopterin, isoxanthopterin, and 6-carboxyisoxanthopterin. Density-gradient fractions indicate that carotenoids are mostly associated with the endoplasmic reticulum. Both tyrosinase and possibly a tyrosinase inhibitor containing sulfhydryl groups are present in the pterinosome. The possible existence of a tyrosinase inhibitor is suggested by the marked increase of tyrosinase activity upon the addition of iodoacetamide or p-chloromercuribenzoic acid. In the light of their fine structure, pigmentary composition, and enzymatic properties, the erythrophores and pterinosomes are discussed with respect to their probable functions and their relationship to melanophores.
Classical light microscopic studies on pigmentation of Fundulus heteroclitus (killifish) indicated that there are three groups of light reflecting cells; one group on the surface of scales reflects white light, while two other deeper groups (the melaniridophores and the stratum argenteum) are iridescent. The results presented here show that: (1) The scale leucophores reflect white light by a Tyndall light-scattering mechanism, by virtue of the presence of randomly oriented organelles of "novel" morphology. (2) The iridophores of the melaniridophores contain stacks of irregularly-spaced, large reflecting platelets which function as an imperfect multiple thin layer interference system. (3) The stratum argenteum consists of a continuous layer(s) of iridophores with reflecting platelets which are so regularly packed as to approach an ideal multiple thin layer interference system. (4) In all three types of light reflecting cells, the dimensions and packing (orientation) of the reflecting organelles satisfactorily account for the chromogenic properties of the cells, including colors as viewed under transmitted, reflected, or polarized light. (5) The spacial relationships between these light reflecting cells and adjoining melanophores are different for each type of light reflecting cell. Furthermore, we propose to replace the term reflecting platelet with refractosome.
The physiological response and ultrastructure of the pigment cells of Trematomus bernacchii, an Antarctic teleost that lives under the sea ice north of the Ross Ice Shelf, were studied. In the integument, two types of epidermal chromatophores, melanophores and xanthophores, were found; in the dermis, typically three types of chromatophores--melanophores, xanthophores, and iridophores--were observed. The occurrence of epidermal xanthophore is reported for the first time in fish. Dermal melanophores and xanthophores have well-developed arrays of cytoplasmic microtubules. They responded rapidly to epinephrine and teleost melanin-concentrating hormone (MCH) with pigment aggregation and to theophylline with pigment dispersion. Total darkness elicited pigment aggregation in the majority of dermal xanthophores of isolated scales, whereas melanophores remained dispersed under both light and dark conditions. Pigment organelles of epidermal and dermal xanthophores that translocate during the pigmentary responses are carotenoid droplets of relatively large size. Dermal iridophores containing large reflecting platelets appeared to be immobile.
Extracts of brightly colored skins from nine amphibian species were analyzed chromatographically. In yellow skin in which xanthophores predominated, relatively large quantities of sepiapterin were found, while in red skin which was laden with erythrophores, three drosopterins were most prevalent. Frozen sections of skin indicated that pteridines were present within chromatophores, either alone or accompanied by carotenoids. It is concluded that sepiapterin and three drosopterins are utilized as pigments in amphibians and it is suggested that other less brightly colored pteridines also function in this respect. It no longer seems proper to make the tacit assumption that bright pigmentation of amphibians is due only to the presence of carotenoids.
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