Impaired female fertility in tubulointerstitial antigen-like 1-deficient mice

Tubulointerstitial nephritis antigen-like 1 (Tinagl1, also known as adrenocortical zonation factor 1 [AZ-1] or lipocalin 7) is a matricellular protein. Previously, we demonstrated that Tinagl1 expression was restricted to extraembryonic regions during the postimplantation period and detected marked expression in mouse Reichert’s membranes. In uteri, Tinagl1 is markedly expressed in the decidual endometrium during the postimplantation period, suggesting that it plays a physical and physiological role in embryo development and/or decidualization of the uterine endometrium during pregnancy. In the present study, in order to determine the role of Tinagl1 during embryonic development and pregnancy, we generated Tinagl1-deficient mice. Although Tinagl1^–/– embryos were not lethal during development to term, homologous matings of Tinagl1^–/– females and Tinagl1^–/– males showed impaired fertility during pregnancy, including failure to carry pregnancy to term and perinatal lethality. To examine ovarian function, ovulation was induced with equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG); the number of ovulated oocytes did not differ between Tinagl1^–/– and Tinagl1^flox/flox. In vitro fertilization followed by embryo culture also demonstrated the normal developmental potential of Tinagl1-null embryos during the preimplantation period. Our results demonstrate that Tinagl1 deficiency affects female mice and results in subfertility phenotypes, and they suggest that although the potential of Tinagl1^–/– oocytes is normal, Tinagl1 is related to fertility in adult females but is not essential for either fertilization or preimplantation development in vitro.     

Ligand-binding characteristics of feline insulin-binding immunoglobulin G

Polyclonal immunoglobulin (Ig) G autoantibodies against insulin have been identified in sera of healthy cats. We purified and fractionated insulin-binding IgGs from cat sera by affinity chromatography and analyzed affinity of insulin-binding IgGs for insulin and their epitopes. Following the passing of fraction A, which did not bind to insulin, insulin-binding IgGs were eluted into two fractions, B and C, by affinity chromatography using a column fixed with bovine insulin. Dissociation constant (KD) values between insulin-binding IgGs and insulin, determined by surface plasmon resonance analysis (Biacore™system), were 1.64e⁻⁴ M for fraction B (low affinity IgGs) and 2e⁻⁵ M for fraction C (high affinity IgGs). Epitope analysis was conducted using 16 peptide fragments synthesized in concord with the amino acid sequence of feline insulin by an enzyme-linked immunosorbent assay. Fractions B and C showed higher absorbance (affinity) of the peptide fragment of 10 amino acid residues at the carboxyl-terminal of the B chain (peptide No. 19), followed by peptide fragments of 6 to 15 amino acid residues of the B chain (peptide No. 8). Fraction C showed a higher absorbance to 7 to 16 amino acid residues of the B chain (peptide No. 5) compared with the absorbance of fraction B. Polyclonal insulin-binding IgGs may form a macromolecule complex with insulin through the multiple affinity sites of IgG molecules. Feline insulin-binding IgGs are multifocal and may be composed of multiple IgG components and insulin.     

Effect of lactoferrin on murine embryo development created from lipopolysaccharide-treated sperm

The effect of lactoferrin (LF) on embryo development was investigated by using lipopolysaccharide (LPS)-treated mouse sperm. For the development rate of the 2-cell stage embryo, the embryo derived from LPS- and LF-treated sperm showed similar survival rate to the control embryo. On day 12 after the embryo transfer into the recipient, the frequent abnormality was observed in the embryo derived from LPS-treated sperm, and the abnormality was tended to be inhibited in the embryo derived from LPS- and LF-treated sperm. These results imply that LF treatment on sperm contaminated with bacteria may facilitate the embryo development, which contribute to the improvement of infertility.     

Optimal dietary taurine level for growth of juvenile Japanese flounder Paralichthys olivaceus

The requirement for taurine in juvenile Japanese flounder Paralichthys olivaceus was determined by feeding diets containing various levels of taurine and cystine. Test diets supplemented with 0.5, 1.0 and 1.5% of taurine or with 0.5, 1.0 and 1.5% of L -cystine were prepared. The basal diet contained 55% protein from white fish meal. These diets were fed to juvenile Japanese flounder with an initial mean bodyweight of 0.9 g (total length (TL) 48 mm) for 5 weeks. Approximately 1.4% taurine content in the diet was required for optimum growth of juvenile flounder. A positive linear relationship was noted between the content of taurine accumulated in the muscle, liver and brain and the level of taurine in the diet. However, there was no increased taurine content in tissues of fish fed the cystine-supplemented diet. In contrast, the fish fed control and cystine-supplemented diets showed higher contents of cystathionine in the tissues. The concentration of cystathionine in tissues rapidly decreased with an increase of taurine in the diet. It was also observed that for each of the dietary groups, a trace amount of taurine was excreted. These results suggest that the taurine content in the diet affects the sulfur amino acid metabolism of juvenile Japanese flounder, and indicate that juvenile flounder are unable to biosynthesize taurine from cystine.     

Flesh quality of tilapia Oreochromis niloticus fed solely on raw Spirulina

ABSTRACT:   In order to simplify the food chain in a closed ecological recirculating aquaculture system, raw Spirulina platensis cultivated in 50-L photobioreactors were fed to tilapia Oreochromis niloticus as a uni-feed from the onset of exogenous feeding. The feasibility of using tilapia grown on raw Spirulina (body weight: 155.4 ± 3.9 g, standard length: 16.2 ± 0.2 cm; n  = 3) was investigated for 30 weeks for sashimi (sliced raw meat). Some of the rheological properties of the flesh were measured to clarify the obtained results of sensory evaluations of texture and fatness. The elastic modulus of flesh of the Spirulina -fed fish was significantly higher than that of the control fish (body weight: 168.9 ± 5.8 g, standard length: 17.8 ± 0.5 cm; n  = 3) that were fed commercial diets. In addition, the viscosity of the flesh of the Spirulina -fed fish was significantly lower than that of the control fish. There was no significant difference in the rupture strength between the two treatments. The rheological parameters in the muscle were found to coincide with the results of the sensory assessment. These results suggest that tilapia fed solely on raw Spirulina have a high flesh quality that is suitable for sashimi. The muscle lipid of the raw Spirulina -fed fish had lower levels of non-polar lipids such as triglycerides and partial glycerides, but higher levels of polar lipids such as phosphatidylcholine and phosphatidylethanolamine. The fatty acid profile of the raw Spirulina -fed fish muscle, especially in the polar lipid fraction, was characterized by an abundance of Σn-6 highly unsaturated fatty acids (20:3n-6, 20:4n-6, 22:4n-6, 22:5n-6).     

Effect of dietary supplementation with taurine, β-alanine andGABA on the growth of juvenile and fingerling Japanese flounder Paralichthysolivaceus

ABSTRACT:   Three diets supplemented with taurine, β-alanine andGABA and a control diet were fed to juvenile and fingerling Japaneseflounder to investigate the effects of the diets on growth and metabolicchanges of free amino acids in whole body and tissues. In experimentI, three diets supplemented with 1% each of taurine, β-alanineand GABA and a control diet were fed to juvenile Japanese flounderwith an initial mean body weight of 0.4 g for 4 weeks at20°C. In experiment II, the taurine-supplemented diet anda control diet were fed to fingerling Japanese flounder with an initialmean body weight of 15 g for 4 weeks at 20°C.Only supplementation of taurine in the diet of juvenile flounderimproved their growth performance in experiment I, but fingerlinggrowth performance of experiment II was not significantly relatedto taurine supplementation in the experimental diet. These resultssuggest that there is a greater requirement for taurine for thegrowth of juvenile Japanese flounder than fingerling Japanese flounder.     

Insulin-like Growth Factor 1 mRNA Expression in the Uterus of Streptozotocin-treated Diabetic Mice

Reproductive functions decline with the onset of diabetes in female mice. Diabetic mice have smaller uteri with an underdeveloped endometrium, suggesting diminished estrogen-induced growth. We aimed to clarify the changes in the estrous cycle and in insulin-like growth factor 1 (IGF1) expression in the uteri of streptozotocin (STZ)-treated diabetic mice, because IGF1 is one of the main growth factors involved in estrogen-induced uterine growth. ICR female mice were intraperitoneally administered STZ (10 mg/100 g BW), and blood glucose levels were determined. Mice with blood glucose levels > 200 mg/dl were classified as diabetic mice. The onset of diabetes was associated with acyclic estrous cycles. Diabetes was also induced with STZ in ovariectomized mice. Uterine Igf1 mRNA levels were reduced in ovariectomized STZ-treated diabetic mice. Estrogen is known to stimulate Igf1 mRNA expression in the uterus, but estrogen action was abolished in the uteri of STZ-treated diabetic mice. mRNA expressions of estrogen receptor α (ERα) and steroid hormone receptor coactivators (SRC-1/Ncoa1, SRC-2/Ncoa2, SRC-3/Ncoa3 and CBP/p300/Crebbp) were reduced in the uteri of ovariectomized STZ-treated diabetic mice. The present study demonstrates that diabetes induces a decline in female reproductive functions in mice. Igf1 expression in ovariectomized diabetic female mice was decreased, and decreased responsiveness to estrogen in the uteri of diabetic mice is probably associated with a reduction in ERα and steroid receptor coactivator mRNA expression.