Light modulates the melanophore response to alpha-MSH in Xenopus laevis: An analysis of the signal transduction crosstalk mechanisms involved

Melanin granule (melanosome) dispersion within Xenopus laevis melanophores is evoked either by light or alpha-MSH. We have previously demonstrated that the initial biochemical steps of light and alpha-MSH signaling are distinct, since the increase in cAMP observed in response to alpha-MSH was not seen after light exposure. cAMP concentrations in response to alpha-MSH were significantly lower in cells pre-exposed to light as compared to the levels in dark-adapted melanophores. Here we demonstrate the presence of an adenylyl cyclase (AC) in the Xenopus melanophore, similar to the mammalian type IX which is inhibited by Ca(2+)-calmodulin-activated phosphatase. This finding supports the hypothesis that the cyclase could be negatively modulated by a light-promoted Ca(2+) increase. In fact, the activity of calcineurin PP2B phosphatase was increased by light, which could result in AC IX inhibition, thus decreasing the response to alpha-MSH. St-Ht31, a disrupting agent of protein kinase A (PKA)-anchoring kinase A protein (AKAP) complex totally blocked the melanosome dispersing response to alpha-MSH, but did not impair the photo-response in Xenopus melanophores. Sequence comparison of a melanophore AKAP partial clone with GenBank sequences showed that the anchoring protein was a gravin-like adaptor previously sequenced from Xenopus non-pigmentary tissues. Co-immunoprecipitation of Xenopus AKAP and the catalytic subunit of PKA demonstrated that PKA is associated with AKAP and it is released in the presence of alpha-MSH. We conclude that in X laevis melanophores, AKAP12 (gravin-like) contains a site for binding the inactive PKA thus compartmentalizing PKA signaling and also possesses binding sites for PKC. Light diminishes alpha-MSH-induced increase of cAMP by increasing calcineurin (PP2B) activity, which in turn inhibits adenylyl cyclase type IX, and/or by activating PKC, which phosphorylates the gravin-like molecule, thus destabilizing its binding to the cell membrane. (C) 2009 Elsevier Inc. All rights reserved.

Development of kelp rockfish, Sebastes atrovirens (Jordan and Gilbert 1880), and brown rockfish, S. auriculatus (Girard 1854), from birth to pelagic juvenile stage, with notes on early larval development of black-and-yellow rockfish, S. chrysomelas (Jordan and Gilbert 1880), reared in the laboratory (Pisces: Sebastidae).

Larval kelp (Sebastes atrovirens), brown (S. auriculatus), and blackand-yellow (S. chrysomelas) rockfish were reared from known adults, to preflexion stage, nine days after birth for S. chrysomelas, to late postflexion stage for S. atrovirens, and to pelagic juvenile stage for S. auriculatus. Larval S. atrovirens and S. chrysomelas were about 4.6 mm body length (BL) and S. auriculatus about 5.2 mm BL at birth. Both S. atrovirens and S. auriculatus underwent notochord flexion at about 6–9 mm BL. Sebastes atrovirens transform to the pelagic juvenile stage at about 14–16 mm BL and S. auriculatus transformed at ca. 25 mm BL. Early larvae of all three species were characterized by melanistic pigment dorsally on the head, on the gut, on most of the ventral margin of the tail, and in a long series on the dorsal margin of the tail. Larval S. atrovirens and S. auriculatus developed a posterior bar on the tail during the flexion or postflexion stage. In S. atrovirens xanthic pigment resembled the melanistic pattern throughout larval development. Larval S. auriculatus lacked xanthophores except on the head until late preflexion stage, when a pattern much like the melanophore pattern gradually developed. Larval S. chrysomelas had extensive xanthic pigmentation dorsally, but none ventrally, in preflexion stage. All members of the Sebastes subgenus Pteropodus (S. atrovirens, S. auriculatus, S. carnatus, S. caurinus, S. chrysomelas, S. dalli, S. maliger, S. nebulosus, S. rastrelliger) are morphologically similar and all share the basic melanistic pigment pattern described here. Although the three species reared in this study can be distinguished on the basis of xanthic pigmentation, it seems unlikely that it will be possible to reliably identify field-collected larvae to species using traditional morphological and melanistic pigmentation characters. (PDF file contains 36 pages.)

Larval development of the southern sea garfish (Hyporhamphus melanochir) and the river garfish (H. regularis) (Beloniformes: Hemiramphidae) from South Australian waters

Larval development of the southern sea garfish (Hyporhamphus melanochir) and the river garfish (H. regularis) is described from specimens from South Australian waters. Larvae of H. melanochir and H. regularis have completed notochord flexion at hatching and are characterized by an elongate body with distinct rows of melanophores along the dorsal, lateral, and ventral surfaces; a small to moderate head; a heavily pigmented and long straight gut; a persistent pre-anal finfold; and an extended lower jaw. Fin formation occurs in the following sequence: caudal, dorsal and anal (almost simultaneously), pectoral, and pelvic. Despite the similarities between both species and among hemiramphid larvae in general, H. melanochir larvae are distinguishable from H. regularis by 1) having 58–61 vertebrae (vs. 51–54 for H. regularis); 2) having 12–15 melanophore pairs in longitudinal rows along the dorsal margin between the head and origin of the dorsal fin (vs. 19–22 for H. regularis); and 3) the absence of a large ventral pigment blotch anteriorly on the gut and isthmus (present in H. regularis). Both species can be distinguished from similar larvae of southern Australia (other hemiramphids and a scomberosocid) by differences in meristic counts and pigmentation.

Larval ontogeny of a percichthyid fish, Synagrops philippinensis (Günther) in Kagoshima Bay, southern Japan

The larval ontogeny of a developmental series (1.2-8.3mm body length, BL) of Synagrops philippinensis from Kagoshima Bay, southern Japan is described and illustrated. The yolk was completely absorbed in larva of ≥1.5 mm BL. Notochord flexion commenced at about 3.5mm BL and was completed by about 4.0-4.5mm BL. S. philippinensis larvae were distinguished from their congeners based on melanophore patterns, head spination and fin spines and rays. Larvae of 7.5-8.3 mm BL were characterized by anteriorly serrated pelvic spine, two anal spines, nine inner preopercular spines and no melanophore on lateral side of the caudal peduncle; 7.0 to 7.5mm BL larvae by the above characters except serration on pelvic spine; and yolk-sac, pre-flexion, flexing and post-flexion larvae up to 7.0mm BL by unique melanophores on lower lobe of pectoral finfold/fin.

Early developmental stages of two Secutor species (Family: Leiognathidae) collected from the Bak-khali river estuary of the Bay of Bengal, Bangladesh

Early developmental stages of two Secutor species, Secutor insidiator (Bloch) (11.9-36.0 mm standard length, SL) and Secutor ruconius (Hamilton-Buchanan) (14.0-33.0 mm SL) collected by ichthyoplankton net from the Bak-khali river estuary of the Bay of Bengal, Cox's Bazar, Bangladesh are described and illustrated. All of the fins with supporting spines and rays were present in the smallest collected sizes of both species. With growth of the specimens, significant changes in melanophore patterns were found. S. insidiator is similar to S. ruconius in having upward protracting mouth parts and body colouration, but can be distinguished easily by its more elongate body shape (body depth 38-47% of SL compared with 46-52% of SL in Secutor ruconius). Both the species occurred round the year from August 1998 to July 1999. The surface water temperature and salinity during the study period varied from 22.0-32) C and 10-37 ppt respectively.

Early developmental stages of Nandus nandus (Ham.)

In order to study the early developmental stages of Nandus nandus an experiment was conducted, where eggs and milt were obtained from the laboratory reared N nandus by stripping after 15 hours of 150 mg/kg body weight of carp PG extract injection. Then the eggs were fertilized in the laboratory and subsequent developmental stages were studied. First cleavage (two cell), four cell, eight cell, sixteen cell and multi cell stages were found 30, 50, 70, 105 and 160 minutes after fertilization respectively. Morula, early gastrula, middle gastrula, late gastrula and yolk plug stages were found 5, 8, 9, 11 and 13 hours after fertilization respectively. Hatching occurred within 20±2 hours after fertilization, and larvae were measured 1.60 mm in diameter. After one hour of hatching two melanophore bands were found at the caudal region of the body of the larvae. Eyes were first observed in l 0 hours, pectoral and pelvic fin buds appeared in 22 hours and well developed in 38 hours old larvae. Mouth cleft and brain lobes were visible when the larvae were 34 and 38 hours old respectively. Myomeres partially appeared in 16 hours, which were clearly visible in 74 hours old larvae. Larvae started wandering and searching for food after 56 hours of hatching. The yolk sac was completely absorbed when larvae became 62 hours old.

Effect of cage colour and light environment on the skin colour of Australian Snapper Pagrus auratus (Bloch & Schneider, 1801)

A two-factor experiment was performed to evaluate the effects of cage colour (black or white 0.5 m³ experiment cages) and light environment (natural sunlight or reduced level of natural sunlight) on the skin colour of darkened Australian snapper. Each treatment was replicated four times and each replicate cage was stocked with five snapper (mean weight=351 g). Snapper exposed to natural sunlight were held in experimental cages located in outdoor tanks. An approximately 70% reduction in natural sunlight (measured as PAR) was established by holding snapper in experimental cages that were housed inside a 'shade-house' enclosure. The skin colour of anaesthetized fish was measured at stocking and after a 2-, 7- and 14-day exposure using a digital chroma-meter (Minolta CR-10) that quantified skin colour according to the L*a*b* colour space. At the conclusion of the experiment, fish were killed in salt water ice slurry and post-mortem skin colour was quantified after 0.75, 6 and 22 h respectively. In addition to these trials, an ad hoc market appraisal of chilled snapper (mean weight=409 g) that had been held in either white or in black cages was conducted at two local fish markets. Irrespective of the sampling time, skin lightness (L*) was significantly affected by cage colour (P<0.05), with fish in white cages having much higher L* values (L*≈64) than fish held in black cages (L*≈49). However, the value of L* was not significantly affected by the light environment or the interaction between cage colour and the light environment. In general, the L* values of anaesthetized snapper were sustained post mortem, but there were linear reductions in the a* (red) and b* (yellow) skin colour values of chilled snapper over time. According to the commercial buyers interviewed, chilled snapper that had been reared for a short period of time in white cages could demand a premium of 10–50% above the prices paid for similar-sized snapper reared in black cages. Our results demonstrate that short-term use of white cages can reduce the dark skin colour of farmed snapper, potentially improving the profitability of snapper farming.

Cage colour and post-harvest K+ concentration affect skin colour of Australian snapper Pagrus auratus (Bloch & Schneider, 1801)

In an attempt to improve post-harvest skin colour in cultured Australian snapper Pagrus auratus, a two-factor experiment was carried out to investigate the effects of a short-term change in cage colour before harvest, followed by immersion in K⁺-enriched solutions of different concentrations. Snapper supplemented with 39 mg unesterified astaxanthin kg⁻¹ for 50 days were transferred to black (for 1 day) or white cages (for 1 or 7 days) before euthanasia by immersing fish in seawater ice slurries supplemented with 0, 150, 300, 450 or 600 mmol L⁻¹ K⁺ for 1 h. Each treatment was replicated with five snapper (mean weight=838 g) held individually within 0.2 m³ cages. L*, a* and b* skin colour values of all fish were measured after removal from K⁺ solutions at 0, 3, 6, 12, 24 and 48 h. After immersion in K⁺ solutions, fish were stored on ice. Both cage colour and K⁺ concentration significantly affected post-harvest skin colour (P<0.05), and there was no interaction between these factors at any of the measurement times (P>0.05). Conditioning dark-coloured snapper in white surroundings for 1 day was sufficient to significantly improve skin lightness (L*) after death. Although there was no difference between skin lightness values for fish held for either 1 or 7 days in white cages at measurement times up to 12 h, fish held in white cages for 7 days had significantly higher L* values (i.e. they were lighter) after 24 and 48 h of storage on ice than those held only in white cages for 1 day. K⁺ treatment also affected (improved) skin lightness post harvest although not until 24 and 48 h after removal of fish from solutions. Before this time, K⁺ treatment had no effect on skin lightness. Snapper killed by seawater ice slurry darkened (lower L*) markedly during the first 3 h of storage in contrast with all K⁺ treatments that prevented darkening. After 24 and 48 h of storage on ice, fish exposed to 450 and 600 mmol L⁻¹ K⁺ were significantly lighter than fish from seawater ice slurries. In addition, skin redness (a*) and yellowness (b*) were strongly dependent on K⁺ concentration. The initial decline in response to K⁺ was overcome by a return of a* and b* values with time, most likely instigated by a redispersal of erythrosomes in skin erythrophores. Fish killed with 0 mmol L⁻¹ K⁺ maintained the highest a* and b* values after death, but were associated with darker (lower L*) skin colouration. It is concluded that a combination of conditioning snapper in white surroundings for 1 day before harvest, followed by immersion in seawater ice slurries supplemented with 300–450 mmol L⁻¹ K⁺ improves skin pigmentation after >24 h of storage on ice.

Ontogenia da resposta endócrina em larvas de matrinxã Brycon amazonicus: ênfase nos eixos hipótise-tireóide e hipófise-tecido interrenal

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