Gene expression levels of growth hormone receptor and insulin-like growth factor-I in gonads of maturing coho salmon (Oncorhynchus kisutch)

The expression levels of growth hormone receptor (GHR) and insulin-like growth factor (IGF-I) in gonads of maturing coho salmon were measured by real-time PCR. The GHR and IGF-I mRNAs showed different gene expression pattern between male and female. These results suggest that a sexual dimorphism may exist in the GHR and IGF-I mRNA expressions of coho salmon during gonadal maturation     

Spawning frequency of the Tsushima Current subpopulation of chub mackerel <Emphasis Type="Italic">Scomber japonicus</Emphasis> off Kyushu,Japan

Female Japanese chub mackerel Scomber japonicus of the Tsushima Current subpopulation were collected during the spawning season from March to May 2001. A total of 137 adult females were caught between midnight and daybreak. A considerable number of fish displayed new postovulatory follicles (POF), whereas there was no evidence of germinal vesicle breakdown or hydrated oocytes in any of the fish collected. This suggests a daily spawning synchronicity toward midnight. To estimate the spawning frequency (S), the female reproductive state was classified into four criteria based on the degenerative stage of the POFs and the developmental stage of the oocytes. To stage the POFs according to age and determine the stage duration, ovaries from S. japonicus were induced to spawn in the laboratory and were sampled 0–72 h after ovulation at appropriate intervals. The average S, which is evaluated from four different indices, was 16.9%, corresponding to the average female chub mackerel spawned every 5.9 days (8.5 times) during the 50 days.     

Purification and characterization of lipovitellin from Pacific saury Cololabis saira

ABSTRACT:   Lipovitellin (Lv), the major yolk protein derived from vitellogenin (Vg), was purified from vitellogenic ovaries of Pacific saury Cololabis saira using hydroxylapatite column chromatography followed by gel filtration. The apparent native mass of purified Lv was approximately 420 kDa, while the tertiary structure of Lv revealed by sodium dodecylsulfate–polyacrylamide gel electrophoresis was typical of teleost Lvs, consisting of a heavy chain (∼99 kDa) and a light chain (∼34 kDa). Western blot analysis using rabbit antiserum raised against Pacific saury Lv revealed a specific reaction with a polypeptide (∼194 kDa) that is present in serum from female Pacific saury but not in male serum, suggesting the approximately 194-kDa polypeptide to be the Vg monomer. This study describes the first step toward the development of specific immunoassays for Pacific saury Vg, which will be an effective tool for monitoring the reproductive development of this species.     

Atlantic salmon fed a nutrient package of surplus methionine,vitamin B12, folic acid and vitamin B6 improved growth and reduced the relative liver size,but when in excess growth reduced

Feeding plant‐based diet through smoltification of Atlantic salmon requires verification of the optimal level of 1C nutrients. Here, we fed Atlantic salmon plant‐based diets containing three different surplus amounts of the 1C nutrients; methionine, cobalamin (vitamin B12), pyridoxine (vitamin B6) and folic acid during 6 weeks in fresh water, through smoltification, followed by 3 months on‐growing period in salt water. The three diets were fed to fish dispersed in triplicate tanks throughout the experiment. Mean start body weight was 32 g. Dietary methionine levels in the diets were 6.7, 9.2 and 11.7 g/kg. Dietary B6 was 6.75, 8.45 and 11 mg/kg. Cobalamin was 0.16, 0.18 and 0.20 mg/kg. While dietary folic acid was 2.9, 4.8 and 6.3 mg/kg, diets are referred to as low, medium and high 1C diet. All other amino acids were similar between diets. The results showed no differences in growth or feed utilization in the fresh water period, but following the on‐growing salt water period, differences between diets occurred. The fish fed the medium 1C diet showed better growth, as compared to fish fed the low or high 1C diet (p = .009). The medium 1C fed fish showed a relative lower liver weight compared with fish fed low or high 1C diet (p = .025). Condition factor was better in fish fed the medium and high 1C diet as compared to those fed the low 1C diet (p = .0006). As expected, free methionine in liver, plasma and muscle increased by dietary methionine inclusion. Surplus vitamins only had minor effect on tissue concentrations. Based on these findings, we conclude that the micronutrient and methionine level presented in the medium 1C diet improved the growth, liver size and condition factor; however, more research is needed to evaluate the optimal requirement level for each of the 1C nutrients.     

Induced maturation and spawning: opportunities and applications for research on oogenesis

Broodstock management requires the ability to detect and regulate oocyte growth, acquisition of maturational competence, maturation of oocytes, and onset of ovarian atresia. Our research on temperate basses (genus Morone) has supported development of these capabilities. These investigations have revealed that accumulation of neutral lipid droplets and deposition of vitellogenin-derived yolk proteins in growing oocytes are independent processes with different sensitivities to changing day length and water temperature. In these fishes, completion of oocyte growth is marked by disappearance of vitellogenin from ovarian biopsy samples. Competence of females for induced spawning is predicted by the ability of biopsied follicles to initiate oocyte meiosis in vitro in response to insulin-like growth factor I. Cytoplasmic maturation of the oocytes is triggered by the maturation-inducing steroid hormone and can be monitored by evaluating degradation of the yolk proteins. Onset of ovarian atresia is indicated by the appearance of edema in the granulosa cell layer of biopsied follicles, and can be delayed for months by holding gravid females at abnormally low temperature (`cold banking'). These novel findings hold strong promise for application to other farmed fishes.     

Identification and primary structures of eel type I collagen proα1, proα2 and proα3

Many bony fish type I collagens have a characteristic third chain designated as α3(I). However, much less is known about the primary structure and distinction of the proα(I) chains. Their cDNAs were cloned by RT-PCR from the muscle tissue of Japanese eel, Anguilla japonica. Three cDNAs coding for the triple-helical domain of fibrillar collagen were identified as proα1(I), proα2(I) and proα3(I) chains by sequencing selected tryptic peptides isolated from eel type I collagen subunit chains, α1(I), α2(I) and α3(I). Eel proα3(I) had high amino acid sequence identity (81 %) to its proα1(I). The distribution of seven Cys residues in the C-propeptide of proα3(I) was identical to that of proα1(I). There was a third Cys residue at the 1,268th position from the N-terminus in proα1(I), though a supposed Cys residue at the 1,264th position in proα3(I) was replaced by a Ser residue. Similar replacement has been observed in the proα3(I) chains of trout and zebrafish. These combined results suggest that replacement of the Cys residue allows for the identification of fish collagen proα(I) previously not identified as proα3(I).     

Effects of thyroid hormone on gonadotropin-induced steroid production in medaka,Oryzias latipes,ovarian follicles

Blood and ovarian samples were collected at intervals of 4h prior to spawning time from medaka (Oryzias latipes) that were maturationally synchronized with artificial photoperiod (14h light: 10h dark). Plasma estradiol-17β (E₂) levels increased rapidly from 16h before spawning and peaked at 8h before spawning. Follicle-enclosed oocytes (ovarian follicles) at different stages of development were isolated from the ovaries and used to study the in vitro effects of thyroid hormone (triiodothyronine; T₃) on pregnant mare serum gonadotropin (GTH)-induced E₂ production. GTH at a concentration of 100 IU/ml stimulated E₂ production by ovarian follicles collected between 32 and 16h before spawning. At 32h before spawning, T₃ (5 ng/ml) administered along with GTH (100 IU/ml) resulted in a 3.5 fold increase in E₂ production, compared with GTH administered alone. These results suggest that T₃ can act on ovarian follicles directly to modulate GTH-stimulated E₂ production in the medaka.