Propionate tolerance test for determination of insulin secretion in a hyperglycemic Japanese Black steer

A propionate tolerance test (PTT) was used to determine the pathophysiology of a Japanese Black steer with hyperglycemia. In the hyperglycemic steer, a low insulin secretion was confirmed by a glucose tolerance test (GTT), so that the hyperglycemic steer was diagnosed as insulin-dependent diabetes mellitus. Although the plasma insulin concentration in the control cattle increased in response to propionate stimulation, a low insulin response to PTT was observed in the diabetic steer. The fact that both PTT and GTT determined that the diabetic steer had low insulin secretion suggests that the PTT might be an effective diagnostic tool for diabetes mellitus in cattle.     

Isolation and identification of F-spondin in the boar testis and its production during testis growth

F-spondin/vascular smooth muscle cell growth-promoting factor (VSGP), purified from the follicular fluid of adult bovine ovaries, has been identified as a promoter of neuronal differentiation and vascular smooth muscle growth. The objectives of the present study were (1) to clarify whether F-spondin is also produced in the testis, which is ontogenically equivalent to the ovary, and (2) to examine whether production of this protein changes with testicular growth. To isolate F-spondin from the testis, testicular homogenates obtained from 8-week-old boars were sequentially subjected to heparin-Sepharose chromatography, diethylaminoethyl (DEAE)-Sepharose chromatography, and reverse-phase high-performance liquid chromatography (RP-HPLC). The isolated protein had a molecular mass of approximately 110 kDa and was cross-reactive with anti-F-spondin antibody by Western blotting. The purified protein was further characterized by amino acid sequence analysis of its internal peptide. The sequence obtained was GEQCNIVPDN VD, and a homology search indicated that the purified protein is a homologue of rat, human, and bovine F-spondin. By fractionation of the same amounts of testis tissue obtained from 1-, 8-, 16-, and 40-week-old boars, we analyzed age-related production of F-spondin in the testis. Western blotting of the fractions obtained from RP-HPLC revealed the presence of a band at approximately 110 kDa, corresponding to F-spondin, in the testes obtained from boars between 1 and 16 weeks old, but this band was not detected at 40 weeks. These results clearly indicate that (1) the porcine testis produces F-spondin and that (2) production of this protein is evident in the immature porcine testis, but not the adult testis.     

Insulin and glucagon secretory patterns during propionate and arginine tolerance tests in Japanese black cattle with growth retardation

To evaluate the energy condition of cattle with growth retardation, propionate (PTT) and arginine tolerance tests (ATT) were carried out. The insulin/glucagon concentration ratio immediately before PTT or ATT in the cattle with growth retardation was lower than in the control. In the growth-retarded cattle, insulin-AUC(0-120 min) during PTT was lower than in the control, while glucagon-AUC(0-120 min) was the same as in the control. Insulin-AUC(0-120 min) during ATT in the cattle with growth retardation tended to be lower than in the control, whereas glucagon-AUC(0-120 min) was the same. Therefore, insulin-AUC(0-120 min)/glucagon-AUC(0-120 min) in the cattle with growth retardation was lower than in the control during both tolerance tests. The growth-retarded cattle showed lower insulin/glucagon ratio similar to that found in starved and lactating cattle, suggesting a lack of energy.     

H1foo is indispensable for meiotic maturation of the mouse oocyte

Oocyte-specific linker histone H1foo is localized in the oocyte nucleus, either diffusely or bound to chromatin, during the processes of meiotic maturation and fertilization. This expression pattern suggests that H1foo plays a key role in the control of gene expression and chromatin modification during oogenesis and early embryogenesis. To reveal the function of H1foo, we microinjected antisense morpholino oligonucleotides (MO) against H1foo into mouse germinal-vesicle stage oocytes. The rate of in vitro maturation of the antisense MO group was significantly lower than that of the control group. Eggs that failed to extrude a first polar body following injection of antisense MO arrested at metaphase I. Additionally, co-injection of in vitro synthesized H1foo mRNA along with antisense MO successfully rescued expression of H1foo and improved the in vitro maturation rate. There was no difference in the rate of parthenogenesis between the antisense MO and control groups. These results indicate that H1foo is essential for maturation of germinal vesicle-stage oocytes.     

A proteomic analysis of Nipponia nippon (ID#162)

We investigated the proteome of a female Crested Ibis (Nipponia nippon, ID#162) that died on March 10, 2010 at the Sado Japanese Crested Ibis Conservation Center. Protein preparations from the brain, trachea, liver, heart, lung, proventriculus, muscular stomach, small intestine, duodenum, ovary and neck muscle were subjected to in‐solution shotgun mass spectrometry (MS)/MS analyses using an LTQ Orbitrap XL mass spectrometer. A search of the National Center for Biotechnology Information Gallus gallus databases revealed 4253 GI (GenInfo Identifier) numbers with the sum of the same 11 tissues examined in the Crested Ibis. To interpret the obtained proteomics data, it was verified in detail with the data obtained from the brain of the Crested Ibis. It has been reported that drebrin A is specifically expressed in adult chicken brain. In the shotgun proteomic analyses of the Crested Ibis, we identified drebrin A as a brain‐specific protein. Furthermore, Western blotting analysis of the protein preparations from 10 tissues of the Crested Ibis and 150‐day‐old hens using anti‐drebrin antibodies showed intensive expression of approximately 110 kDa polypeptides of drebrin in both brains. We believe firmly that the present data will contribute to initial and fundamental steps toward understanding the Crested Ibis proteome.     

In vivo gonadotropic effects of recombinant Japanese eel follicle-stimulating hormone

Recombinant Japanese eel follicle-stimulating hormone (rjeFSH) produced by methylotrophic yeast was subjected to in vivo bioassay to assess its gonadotropic activities and availability to artificially induce maturation of this species. Intramuscular injections of rjeFSH into male eels at doses of 0.1 and 1.0 U/g body weight were repeated 3 times during 12 days. The rjeFSH significantly increased plasma 11-ketotestosterone levels, and induced both testicular growth and spermatogenesis in a dose-dependent manner. These results demonstrate that rjeFSH is effective in promoting male eel gamatogenesis in vivo. Instead of scarce native eel FSH, abundant recombinant eel FSH is now ready for future application to maturation induction of Japanese eel.     

Growth and ontogenetic migration of greeneye Chlorophthalmus albatrossis in Tosa Bay, Pacific Coast of South-Western Japan

ABSTRACT:   Growth and ontogenetic migration of greeneye Chlorophthalmus albatrossis were studied in the area from 100–400 m depths in Tosa Bay, from April 1996 to March 1997, using samples collected by an otter trawl net with a 1.3 mm mesh cod end. In the monthly changes of the standard length (SL) composition, four year classes were recognized, although the third- and fourth-year classes were not clearly separated in all the monthly samples. Individuals of 40–50 mm SL settled on the bottom of the 150–200 m depth zone from August to February. By May and June, they grew to 75–80 mm SL modal size, and migrated to 200–300 m depth. In one year after settlement, they grew to 125–130 mm SL, and migrated to 300–350 m depth. They grew to a maximum of 160 mm SL and were not sampled in the winter three years after settlement, suggesting either migration or mortality. Seasonal changes in the gonadosomatic index for combined samples of males and females showed that greeneye did not mature in Tosa Bay. It is suggested that the planktonic larvae are transported from the spawning grounds, the location of which remains unknown.     

Changes in plasma somatolactin levels during spawning migration of chum salmon (Oncorhynchus keta)

Plasma somatolactin (SL) concentrations were examined in chum salmon in relation to gonadal maturation; immature salmon in the Bering Sea at various stages of maturation, and mature salmon during upstream migration caught at the ocean, bay and river. Plasma SL concentrations as well as plasma prolactin (PRL) and growth hormone (GH) levels in the immature fish caught in the Bering Sea were maintained essentially at similar levels. Plasma SL in mature salmon increased significantly from the fish in the ocean to the fish in the river in both sexes. Although all the fish had fully developed gonads, females completed ovulation while still in the bay, whereas final spermeation in males was achieved after entry into the river. Thus, no clear correlation was seen between plasma SL levels and final gonadal maturation. On the other hand, plasma PRL concentrations in both male and female fish were higher in the fish in the river than those in the ocean and bay, and plasma GH levels were higher in both sexes in the fish in the bay and river than those in the ocean. Plasma levels of triglycerides, glucose, free fatty acids and ionized sodium and calcium were also examined. Significant-negative correlations were seen between plasma SL and plasma ionized calcium in mature male salmon, and between plasma SL and plasma triglycerides in mature female salmon. Although our findings do not rule out the possibility of the involvement of SL in final maturation, the results indicate that SL seems to be involved at least in energy and/or calcium metabolism during the spawning migration.