Identification of protein components from the mature ovary of the sea urchin Evechinus chloroticus (Echinodermata: Echinoidea)

The gonads of sea urchins are a high value seafood product, with considerable research being undertaken worldwide on the development of sea urchin aquaculture. As the best prices are obtained for specific gonad attributes, research has also focused on the development of artificial diets that enhance gonad quality and quantity. Total protein has been used as a measure of gonad quality; yet no studies to date have applied proteomics technology to diet development. Here we use a MudPIT and 2-DE approach to describe the major proteins in mature ovaries of a New Zealand sea urchin Evechinus chloroticus. This tissue, which is a target seafood product, contained 138 proteins that were identified from the recently completed sea urchin genome (Strongylocentrotus purpuratus) with high confidence. The majority of these proteins had general functions, with only 12 related to ovarian reproductive function. Eighteen proteins were located on the 2-DE; four of these were directly identified from S. purpuratus protein sequences. In combination this paper shows that the genome resources of S. purpuratus can be used to identify proteins in sea urchins from different families; describes the proteome of E. chloroticus mature ovary; and, provides proteomic tools for analysis of gonads from other edible sea urchins.     

Carotenoids in the sea urchin Paracentrotus lividus: occurrence of 9'-cis-echinenone as the dominant carotenoid in gonad colour determination

Regular sampling of wild Paracentrotus lividus was carried out over a 12-month period to examine seasonal effects on the pigment profile and content of the gonads, especially in comparison to gonad colour. The major pigments detected in the gut wall were breakdown products of fucoxanthin, namely fucoxanthinol and amarouciaxanthin A. Lower levels of other dietary carotenoids (lutein and β-carotene) together with some carotenoids not found in the diet, namely isozeaxanthin and echinenone ( 20% total carotenoid) were also detected in the gut wall. The presence of echinenone in the gut wall demonstrates that this organ acts as a major site of carotenoid metabolism. Echinenone is the dominant carotenoid in the gonads, accounting for approx. 50–60% of the total pigment. Both all-trans and 9′-cis forms of echinenone were detected in both the gut wall and in the gonad, with levels of the 9′-cis form typically 10-fold greater than the all-trans form in the gonad. The detection of large levels of 9′-cis-echinenone in wild sea urchins is unexpected due to the absence of 9- or 9′-cis forms of carotenoids in the natural, algal, diet. Whilst echinenone clearly contributes towards gonad pigmentation, levels of this carotenoid, cannot be directly linked to a qualitative assessment of gonad colour in terms of market acceptability. Indeed, unacceptable gonad colouration can be seen with both very low and high levels of echinenone and total carotenoid. The presence of 9′-cis-echinenone as the major carotenoid contributing to the pigmentation/colour of the gonad is an important observation in terms of developing artificial diets for urchin cultivation.     

Testing for the Effects of Seasonal and Lunar Periodicity on the Reproduction of the Edible Sea Urchin <Emphasis Type="Italic">Tripneustes gratilla</Emphasis> (L) in Kenyan Coral Reef Lagoons

The annual and lunar reproductive cycle of the widely distributed edible sea urchin Tripneustes gratilla (L) was examined through measurements of gonad index, histological examination of gametogenesis, and induction of spawning with KCl injections. The population density and morphological characteristics of urchins at Diani, Kanamai, and Vipingo reef lagoons were also studied as well as the effects of seawater temperature and light on reproduction. Gonad growth started early during the northeast monsoon and reached a peak in June at the beginning of the southeast monsoon followed by a sharp decrease in gonad size of 50% in July and August towards the end of the southeast monsoons. Histological examination of gonads, revealed many different stages of gametogenesis with gametes present throughout the year, indicating continuous reproduction. There was a significant relationship between gonad index and lunar day with spawning occurring between lunar day 7 and 21, but spawning was not in perfect synchrony in the population. The population density of urchins at each reef is variable from year to year and was highest on average at Vipingo. Urchins at Kanamai had the lowest gonad indices, the largest jaws and smallest individuals an indication of food limitation. The gonads (roe) of T. gratilla at all three sites, were perpetually ‘runny’ an attribute that is not suitable for urchin fisheries. Studies to develop techniques to improve roe quality are recommended.     

Select light spectra affect gonad color in the sea urchin Lytechinus variegatus

It is suggested that gonad color in sea urchins depends upon the in vivo accumulation and metabolism of red and yellow carotenoid pigments. We hypothesized that differential light exposure could affect carotenoid deposition and, hence, gonad color in sea urchins. We therefore performed two experiments to determine whether light spectra affect the gonad color of Lytechinus variegatus. In the first experiment, urchins were fed a formulated feed supplemented with or without β-carotene and held beneath three lighting regimes designed to emit differing wavelengths of the visible spectrum. After 12 weeks, urchins were dissected and gonad color (CIE L*a*b*) was measured with a Pantone Capsure RM200. Actinic light significantly increased the value of a* (red) in gonad color. Color in the orange and yellow spectra in the gonads increased in individuals fed the β-carotene supplemented diet. In the second experiment, we cultured urchins for nine weeks under lamps specialized to emit UV radiation. All urchins in this experiment received diets supplemented with β-carotene. There was no significant difference in harvested gonad color between these treatments. These data suggest that light quality and dietary carotenoids affect carotenoid deposition in the gonads.     

Long-Term Changes in the Gonad Condition of the Sea Urchin Strongylocentrotus intermediusfrom Alekseev Bight (Amurskii Bay) under Polluted Conditions

The gonad condition of the sea urchin Strongylocentrotus intermediuscollected in August 1997 at two stations in Peter the Great Bay was examined. One of the stations was located in a polluted area (Alekseev Bight, Popov Island) and the other, in a relatively clean area (the Verkhovskii Islands). The results were compared with analogous data for 1984, 1985, and 1989. In 1997, the gonad condition of sea urchins inhabiting the two areas differed significantly. The mean value of the gonad index (GI) for sea urchins from Alekseev Bight was less than half and the maturity index was about twice that of sea urchins from the Verkhovskii Islands. The GI of sea urchins from Alekseev Bight decreased by a factor of 1.5 between 1984 and 1997. Pronounced histopathological changes were found in sea urchin gonads at this station: granular and hydropic dystrophy of oocytes, resorption and a sharp decrease in the number of gametes (in about 20% of the sea urchins, hardly any gametes were found in the gonads), changes in the morphology of accessory cells (hypertrophy, atrophy, and necrosis), and accumulation of lipofuscin in the cytoplasm of accessory cells and oocytes, in the hemal sinuses and mesentery. The suppressed gonad condition of the sea urchin S. intermediusin Alekseev Bight may be a consequence of the unfavorable environmental situation that formed in the bight in the 1980s–1990s. The main negative factor is anthropogenic pollution of Amurskii Bay.     

Changes of major carotenoids in gonads of sea urchins (Strongylocentrotus intermedius and S. nudus) at maturation

Changes in the carotenoid content in gonads of two sea urchins species were investigated during maturation. The content of echinenones and carotenes, the two major carotenoid fractions in gonads, is highest for Strongylocentrotus intermedius at the spawning gametogenic stage of gonad maturation for both sexes. For S. nudus, the content of these pigments is highest at stages of active gametogenesis and spawning for males and at the growth stage for females. A comparison of the carotenoid content dynamics during maturation of gonads for males, females and animals at the resting (sexual inactivity) stage was also carried out.     

Female ornamentation and egg carotenoids of six sympatric gobies

Belly colouration, gonad carotenoid concentration and skin transparency were quantified in gravid Gobiusculus flavescens, as well as in females of five sympatric gobies where belly ornamentation has not been described. Although G. flavescens females did, indeed, have far more colourful bellies than the other species, this could only in part be explained by a high concentration of total gonad carotenoids. Comparable, or occasionally higher, carotenoid levels were found in the gonads of other species. Instead, the unusual ornamentation of G. flavescens arises from a unique combination of carotenoid‐rich gonads and a highly transparent abdominal skin.     

The gonads of Aegla platensis Schmitt (Decapoda, Anomura, Aeglidae): a macroscopic and histological perspective

The aim of this study is to characterize the gonads of Aegla platensis, relating aspects of colour and size of ovaries, testes and vasa deferentia (VD) to histological observations. The ovaries are H‐shaped, extending from behind the stomach and ending at the 1st or 2nd abdominal somite. The male has a pair of testes from which the VD issue and extend over the pereon. For males and females, macroscopic categories of gonad development are defined. Stages of the male gonad are defined as types 1, 2 and 3. Type 1 includes the less developed gonads, and type 3 includes extremely developed testes and completely differentiated VD. The ovaries are classified into four stages according to their colour: I (white), II (yellow), III (orange) and IV (red), beginning with the least developed gonads. The number and size of oogonia and oocytes depend on the stage of the ovary and coincide with the different degrees of development as indicated by ovary colour. In males, the presence or absence of spermatozoa in the testes and VD is determined only for the more developed gonads. Neither spermathecae (female) nor spermatophores (male) were found. The lack of spermatophore in the present species is a rare characteristic among the anomurans.     

Carotenoid-binding proteins; accessories to carotenoid function

Understanding of the widespread biological importance of carotenoids is increasing. Accompanying this is the developing recognition that the interaction of carotenoids with other molecules, such as proteins, is also essential. Here the significance of carotenoid-protein interactions with respect to biological function is reviewed for three well characterised carotenoprotein complexes; crustacyanin, the orange carotenoid protein and glutathione-S-transferase P1. In addition a preliminary report is made on the recent partial purification of an echinenone-binding protein extracted from a New Zealand sea urchin, Evechinus chloroticus.     

The commercial potential of the common sea urchin Echinus esculentus from the west coast of Scotland

Sea urchin roe is a valuable food product. The commercial potential of Echinus esculentusL. (Echinodermata: Echinoidea) was assessed both by surveying field populations and gonad growth in response to artificial feeds. Urchins were collected from four locations on the Scottish west coast between June and August of 1998. The roe content was found to be of variable quality. A relatively low percentage of urchins (34.9%) had roe which was considered marketable both in terms of yield and colour. Urchins with a marketable yield of roe were found where algal growth or encrusting organisms were most prolific. Good roe colours and high roe yields were not necessarily coincident. The data suggest an urchin fishery is likely to be impractical economically. In tank-based trials large E. esculentus were fed either commercially available salmon feed, a pelletised urchin diet or the macroalgae Laminaria saccharina. Urchins fed the artificial diets increased in test diameter during the 24-week experiment. At the end of the trial urchins fed salmon feed had a significantly higher gonad and alimentary index than those in the other treatments. E. esculentusis able to successfully exploit a diet rich in animal-origin oils. The diets also influenced gonad colour. The proportion of urchins with marketable gonad colours was highest (85%) in the treatment receiving salmon feed. As this species appears robust in culture and as artificial diets promote somatic and gonadal growth, an investigation into the practicalities of a fully farmed approach to gonad production may be worthwhile.     

MYP基因在中间球海胆及杂交海胆生殖腺不同发育时期的转录表达差异

摘要: 应用实时荧光定量PCR技术对主要卵黄蛋白(Major yolk protein, MYP)基因在中间球海胆和杂交海胆(中间球海胆♀×光棘球海胆♂)的生殖腺不同发育时期的转录表达差异进行了分析。结果表明, MYP基因在海胆雌雄个体生殖腺中转录水平上的表达差异不明显; MYP基因在中间球海胆生殖腺发育的4个时期的表达呈快速下降的趋势, 而在杂交海胆呈缓慢下降的趋势。在分析的生殖腺4个发育期中, 中间球海胆雌雄个体 MYP 基因的表达量(以18S rRNA为参照)分别从44.55%和41.17%下降到9.59%和1.83%; 杂交海胆的雌雄个体分别从37.66%和36.66%下降到19.22%和12.55%。MYP基因的转录表达量差异与杂交后生殖腺产生的变异密切相关。     

Reproductive cycle and reproductive output of the sea urchin Loxechinus albus (Echinodermata: Echinoidea) from Beagle Channel, Tierra del Fuego, Argentina

Reproductive cycle, frequency and duration of spawning, energetic content of gonads, and reproductive output of the common green sea urchin Loxechinus albus were analyzed in the Beagle Channel (Tierra del Fuego) between May 2004 and May 2005. Gonad indices (GI, percentages of gonad mass in total body mass) were significantly higher in March, April, July, and August than in November and May, thus showing a negative correlation with the photoperiod. Highest GI values of mature individuals were observed in August, and spawning occurred from September to December. In females, the mass-specific energy content of gonads (ECG) was highest in spawned gonads and lowest in mature ones, while in males ECG values were higher in immature stage and lower in premature and mature stages. High ECG values can be explained by the abundance of nutritive phagocytes. Both ECG and total gonad energy content (TECG) were higher in females than in males. Mean reproductive output was 7.28% for females and 6.15% for males (expressed as the difference between mean GI of mature and spawned gonads) and 25.02 kJ for females and 19.26 kJ for males (expressed as the difference between mean TECG of mature and spawned gonads).     

Calcium-activated neutral proteinase in gametes of the sea urchinStrongylocentrotus droebachiensis

By Chromatographic separation of cytosol fraction of homogenates from gonads of females and males of green sea urchinStrongylocentrotus droebachiensis using an AcA-34 ultragel, protein fractions with mol. mass of 123 and 81 kDa are revealed, which have activity of calcium-activated proteinases. Dynamics of activity of the calcium-activated proteinases is studied during annual sexual cycles of the sea urchin. It is shown that the maximal activity of the studied enzyme in females and males of the sea urchin is present at the IV stage of gonad maturation, at the period of their trophic growth which is characterized by formation of stores of nutritient substances necessary for the subsequent development of embryos.     

Calcium-activated neutral proteinase in gametes of the sea urchinStrongylocentrotus droebachiensis

By Chromatographic separation of cytosol fraction of homogenates from gonads of females and males of green sea urchinStrongylocentrotus droebachiensis using an AcA-34 ultragel, protein fractions with mol. mass of 123 and 81 kDa are revealed, which have activity of calciumactivated proteinases. Dynamics of activity of the calcium-activated proteinases is studied during annual sexual cycles of the sea urchin. It is shown that the maximal activity of the studied enzyme in females and males of the sea urchin is present at the IV stage of gonad maturation, at the period of their trophic growth which is characterized by formation of stores of nutritient substances necessary for the subsequent development of embryos.     

Identification of key nutrients for gonadal development by comparative analysis of proximate composition and fatty/amino acid profile in tissues and eggs of Chinese sturgeon (Acipenser sinensis Gray, 1835)

The present study was conducted to characterize the proximate composition and fatty/amino acid profile in muscle, liver and gonads obtained from farmed male and female Chinese sturgeon (Acipenser sinensis) with gonad development at stage II, and to discern the key nutrients for gonads by comparing the nutrients between tissues and between fish at different gonad stages. Chinese sturgeon were reared in an outdoor circular cement pool at temperatures ranging from 8°C to 26°C, and fed with 70% artificial diets and 30% frozen forage fish at a feeding rate of 0.2%–1.5% body weight according to the water temperature. Three male and three female A. sinensis were sampled for analyses. Results showed that the lipid content in gonads was significantly higher than in other tissues in both males and females (p < .05). The protein content in male and female gonads was significantly lower than that in the muscle (p < .05). Total content of mono‐unsaturated fatty acids (MUFA) in females was significantly higher than in males (p < .05), while the total content of n‐6 PUFA in females was significantly lower than in males (p < .05). Palmitic acid (C16:0) and oleic acid (C18:1n9) were the major saturated fatty acid (SFA) and MUFA, respectively, in both females and males. The contents of C18:2n6, C20:4n6 (ARA), C18:3n3 and C22:6n3 (DHA) in male gonads were significantly higher than in females (p < .05). The contents of ARA, EPA and DHA in gonads were lower, possibly playing a pivotal role in gonad development. Comparing the nutrients of ovaries in broodstocks at stages III and IV and the eggs at stage IV, the present study suggests that the lipid and fatty acid profile in fish is vital to the gonad development and that proper improvement of lipid nutrition in the broodstock diet to provide sufficient energy and essential fatty acids would be beneficial for the gonad development of Acipenser sinensis.     

Fucoxanthin metabolites in egg yolks of laying hens

Feeding experiments were conducted with White leghorn laying hens fed a carotenoid depleted control diet (containing some zeaxanthin and lutein) or this diet supplemented with 15% seaweed meal of established carotenoid composition. Egg yolk colour was estimated by use of a Roche Yolk Colour Fan and by detailed quantitative and qualitative carotenoid analysis of individual eggs of three laying hens during 4 weeks. Identification of the carotenoids included HPLC, VIS, MS, ¹H NMR data and partial synthesis. The results confirmed that fucoxanthin, the major carotenoid in seaweed meal, is not transferred to the yolk. However, fucoxanthin gave rise to the metabolites fucoxanthinol, fucoxanthinol 3'-sulphate and paracentrone, that are ascribed to enzymatic modifications occurring in the hens. The difuranoid auroxanthin encountered in the egg yolk was ascribed to violaxanthin and/or its furanoid derivatives present in the seaweed meal. Colour of individual yolks varied considerably. The pigmentation level is discussed.     

Ultrastructure of the indifferent gonad in male and female pig embryos.

Pig embryos aged 24 days were obtained from artifically inseminated sows for ultrastructural study of the indifferent gonads. Sex was identified by chromosome analysis. The gonads are composed in both sexes of three different tissues: the surface epithelium, the gonadal blastema and the mesenchyme. The surface epithelial cells contained elongate mitochondria, granular endoplasmic reticulum, free polysomes, the Golgi complex, fine filaments and coated vesicles. The primitive cords were continuous with the surface epithelium and the interior of the gonad was occupied by blastema cells. They had prominent nucleoli, elongate mitochondria, granular endoplasmic reticulum, the Golgi complex, free polysomes, some lipid droplets and occasionally circular smooth membrane profiles resembling the agranular endoplasmic reticulum. Individual primordial germ cells were seen in all parts of the gonad. They were roundish with prominent nucleoli, globular mitochondria, granular endoplasmic reticulum, free polysomes, the Golgi complex, coated vesicles, lipid droplets and dense bodies. Degenerating cells and cells having pseudopods were also encountered. In comparison to the gonad at the age of 22 days, the primordium had grown into a longitudinal roundish protrusion and the number of primoridal germ cells had increased. Histological and ultrastructural observations showed that the pig gonads at the age of 24 days were similar in both sexes.     

The reproductive response of the sea urchins Paracentrotus lividus (G.) and Psammechinus miliaris (L.) to a hyperproteinated macrophytic diet

The sea urchins Paracentrotus lividus and Psammechinus miliaris are submitted to the same environmental conditions in the Bay of Brest. The relationship between seasonal changes in food source quality and their gonad production was investigated in reproducing experimentally these conditions. In a first stage two macroalgae (Palmaria palmata and Laminaria digitata) were tested. P. miliaris showed a stronger preference for P. palmata and over a year-long experiment both urchins progressively preferred P. palmata. Seasonal variations in the chemical composition of P. palmaria were observed in the Bay of Brest: total carbohydrates were important and the relative maximum (about 50%) was reached between February and August; the lipid level was low and had a relative maximum of about 1% in June and August. Total protein in P. palmaria was high compared to other seaweeds: the maximum value (25%) was observed in June, which was probably due to the maintenance of nitrogen nutrient in the bay.In the second stage of the study, seasonal changes in biochemical components of ingestion and absorption of the two sea urchins were followed in the laboratory using a monospecific diet of P. palmaria. The patterns of total carbohydrates and lipid absorption were very similar for both sea urchin species. Carbohydrates were absorbed strongly and uniformly, year round. Lipid absorption mimicked the lipid nutrient pattern in the food source. Only changes in protein absorption varied slightly between the two urchin species. Protein absorption was maximal for both species in February and June, but the quantity of absorbed protein was significantly higher in P. miliaris than in P. lividus during February. This increase was concomitant with protein storage in the sea urchin gonads, which peaked in February for P. miliaris and in June for P. lividus. P. lividus had a higher gonad production efficiency, based on gonad yield. The comparison between in situ data and the experimental results suggests that an algal diet more nitrogenous than the in situ algal food source would benefit the herbivorous P. lividus, rather than the more omnivorous species P. miliaris. Although P. milaris has been described as a species with large gonad production potential, P. lividus appears to be a more suitable species for echiniculture conditions.     

Gonadal growth and reproduction in the commercial sea urchin Paracentrotus lividus (Lamarck, 1816) (Echinodermata: Echinoidea) from southern Spain

The reproductive state of six populations of the commercial sea urchin P. lividus was investigated. Specimens were sampled monthly from six sites of southern Spain. The development of the gonad during the period of study was measured using histological methods and categorised by four stages for females and two for male specimens. A Gonadosomatic Index was also established. The six populations exhibited similar reproductive patterns, having mature gonads during almost all months while reconstitution and spent were present mainly in autumn. The population from Sancti Petri had the highest Gonadosomatic Index value. Higher food availability may explain this difference.