Immunohistochemical localization of three GnRH systems in brain and pituitary of Japanese flounder

ABSTRACT:   To clarify the possible roles of gonadotropin-releasing hormone (GnRH) in the reproduction of Japanese flounder Paralichthys olivaceus , localization of salmon GnRH (sGnRH), chicken GnRH-II (cGnRH-II), and sea bream GnRH (sbGnRH) immunoreactive (ir) cell bodies and fibers in the brain and pituitary were examined together with follicle stimulating hormone (FSH) and luteinizing hormone (LH)-ir cells in the pituitary by immunohistochemistry. sGnRH-ir cell bodies were localized in the ventromedial part of the rostral olfactory bulb and cGnRH-II-ir cell bodies were restricted to the midbrain tegmentum, while sbGnRH-ir cell bodies were evident in the preoptic area. sGnRH-ir fibers were distributed throughout the brain, especially abundant in the forebrain. cGnRH-II-ir fibers were also scattered in many areas of the brain with abundance in the midbrain, but sbGnRH-ir fibers were observed in the preoptic–hypothalamic area and innervated the pituitary. In the pituitary, neither sGnRH-ir fibers nor cGnRH-II-ir fibers were found, but sbGnRH-ir fibers were profuse in the neurohypophysis and invaded the proximal pars distalis, targeting FSH and LH cells. These results suggest that three GnRH systems can play different physiological roles in the brain of Japanese flounder. Among them, sbGnRH is considered to be involved in reproduction by stimulating gonadotropin secretion, while sGnRH and cGnRH-II can function as a neurotransmitter and/or neuromodulator within the brain in this species.     

Changes in brain and pituitary GnRH levels during ovarian maturation in wild female Japanese flounder

To elucidate the role of gonadotropin-releasing hormone (GnRH) in gonadal maturation in wild female Japanese flounder Paralichthys olivaceus, we monitored changes in the levels of seabream GnRH (sbGnRH) in the olfactory bulb, telencephalon, hypothalamus, and pituitary during ovarian development together with changes in plasma levels of testosterone (T), estradiol-17β (E₂), and 17α, 20β-dihydroxy-4-pregnen-3-one (DHP). Fish were caught offshore of the northern mainland of Japan in the Pacific Ocean at 3- to 4-week intervals between April and September by gill net. The netted fish were categorized into six groups based on ovarian stages: previtellogenic (April–early May), early yolk (April–late May), late yolk (late May–June), early spawning (June–August), late spawning (September), and termination (September) stages. The gonadosomatic index significantly increased from the previtellogenic to early spawning stages and decreased thereafter. In the olfactory bulb, no significant differences were observed in sbGnRH levels among the developmental stages. In contrast, sbGnRH levels in the telencephalon and hypothalamus were very high in the previtellogenic stage, lower in the early spawning stage, and relatively high in latter stages. sbGnRH levels in the pituitary were high in the previtellogenic stage and low in the early spawning stage. In addition, the relatively high levels of pituitary sbGnRH were found together with high plasma T, E₂, and DHP levels in fish in the late yolk stage. These results indicate that sbGnRH in the telencephalon, hypothalamus, and pituitary is involved in ovarian maturation and that sbGnRH may play an important role in the initiation of ovarian recrudescence in wild Japanese flounder.     

Yellowtail neuropeptide Y: molecular cloning,tissue distribution,and response to fasting

Neuropeptide Y (NPY) and cholecystokinin (CCK) play important roles in regulating appetite in vertebrates, including mammals and fish. Understanding the appetite mechanism is important in aquaculture to improve production performance. The yellowtail Seriola quinqueradiata is one of the most cultured fish in Japan, but little is known about its appetite hormones. In the present study, complementary DNA encoding for NPY was cloned in yellowtail and consists of 604 bp, in which deduced amino acid sequences show high identity to those of other teleosts. In tissue distribution, the npy and cck mRNAs were detected in all examined tissues (whole brain, telencephalon, optic tectum, hypothalamus, cerebellum, pituitary, retina, stomach, pyloric caeca, anterior intestine, liver, and kidney). In the fasting experiment, only npy mRNA expression in the hypothalamus responded to fasting, showing a significantly high value compared with that in control fish. The expression of cck mRNA in the examined tissues did not change with fasting. The npy mRNA expression in the hypothalamus might be involved in feeding regulation in yellowtail.     

Termite resistance and color of heartwood of hinoki (Chamaecyparis obtusa) trees in 5 half-sib families in a progeny test stand in Kyushu, Japan

Hinoki (Chamaecyparis obtusa) wood has been used as a structural material in Japan because of its superior mechanical properties, its excellent durability and the beautiful color of its heartwood. Variations of termite resistance and compositions of extractives among hinoki trees have been reported. However, genetic variation of termite resistance and the effect of heartwood color on termite resistance remain unknown. In this study, we report the characteristics of termite resistance and color indexes (L*, a* and b*) of heartwood of hinoki half-sib families in a progeny test stand. The survival days of termites and the mass loss of samples of hinoki heartwood differed significantly among hinoki families. Families with red-color heartwood had larger termite resistance than families with yellow-color heartwood. The termite resistances of individual samples from two families with yellow-color heartwood were as small as that of Pinus densiflora. Larger a* and smaller b* induced larger termite resistance of heartwood. The effect of DBH (diameter at breast height) of hinoki trees on termite resistance of hinoki families was small.     

Calming effect of orally administered γ‐aminobutyric acid in Shih Tzu dogs

The calming effects of γ‐aminobutyric acid (GABA) by oral administration were investigated in four adult Shih Tzu dogs. Three dosage levels (1, 2 and 4 mg/kg body weight) and non‐administration were tested by an increase and decrease method. Changes in activity (for 1.5 h) and urinary cortisol levels (pre‐administration, 3 and 7 h later) of dogs were monitored after administration. Without reference to dosage level, the mean times spent standing (P = 0.06), sitting (P < 0.05) and walking (P < 0.05) tended to decrease compared to non‐administration. A significant depression in the urinary cortisol level was observed at 7 h after administration (P < 0.05). These results indicate that orally administrated GABA exerts calming effects on dogs as well as humans.     

Socioeconomic Conditions Affecting Smallholder Timber Management in Gunungkidul District,Yogyakarta Special Region,Indonesia

Indonesia is one of the largest teak timber producers in the world. The Javanese State Forest Company has been a major producer of teak timber in Indonesia; however, log production decreased drastically due to severe illegal logging after the collapse of Suharto regime. In contrast, small-scale private forests (PFs) owned by local farmers have expanded and are expected to be a new source of teak timber. Long rotation is a critical factor in producing a larger diameter log with a higher heartwood proportion. However, harvest timing in PFs is traditionally decided based on individual farmers’ needs even if trees are still young and of small diameter. Therefore, traditional harvesting is an obstacle to producing high-quality teak timber. The objectives of this study are to (1) identify the household economies and PF management styles of local farmers, (2) characterize the local farmers who conduct traditional harvesting, and (3) suggest key considerations for PF policymaking. Key informant interviews and semi-structured interviews with local farmers were conducted in three villages in Gunungkidul district, Yogyakarta Special Region. The study identified the household economies, the ownership and management structures, and the traditional harvesting in PFs in the three villages, and reaffirmed diversity and complexity of PFs. It appears that PF management is influenced by topographic and socioeconomic conditions and differs widely across villages. Therefore, it is important to consider the diversity and complexity of PFs in PF policymaking.     

Rhodococcus equi virulence plasmids recovered from horses and their environment in Jeju, Korea: 90-kb type II and a new variant, 90-kb type V

Rhodococcus equi was isolated from fecal and soil samples from four native Jeju horse farms and six Thoroughbred farms in Jeju, Korea. The isolates were examined for the presence of virulence-associated 15-17-kDa antigens (VapA) by colony blotting, using the monoclonal antibody 10G5, and for the gene encoding VapA by PCR. R. equi was isolated from all 36 soil samples collected from the 10 farms with between 5.0 x 10(2) and 7.5 x 10(4) colony-forming units (cfu) per gram of soil, and from 37 of 40 fecal samples with between 5.0 x 10(1) and 1.1 x 10 (5) cfu per gram of feces. Virulent R. equi was isolated from seven farms and appeared in 2.0% of isolates (10 of 508). Of the 10 virulent isolates, four contained a 90-kb type II plasmid, which has been found in isolates from the Kiso native horses of Japan, and the other six contained a new variant, which did not display the EcoRI and EcoT22I digestion patterns of the 10 representative plasmids already reported (85-kb types I, II, III, and IV; 87-kb types I and II; 90-kb types I, II, III, and IV). We designated the new variant as the "90-kb type V" plasmid, because its EcoRI digestion pattern is similar to that of the 90-kb type II plasmid. This is the first report of the prevalence of virulent R. equi in Jeju, Korea. The same virulence plasmid type is found in both Korean and Japanese isolates, providing insight into the origin, ancestry, and dispersal of native horses in Korea and Japan.