Changes in the expression of toll-like receptor mRNAs during follicular growth and in response to lipopolysaccharide in the ovarian follicles of laying hens

The aim of this study was to determine the changes in the mRNA expression of Toll-like receptors (TLRs) in hen ovarian follicles during follicular growth and in response to lipopolysaccharide (LPS). White follicles and the fifth largest to largest follicles (WF and F(5)-F(1), respectively) were collected from laying hens. To examine the effects of LPS, the laying hens were treated intravenously with LPS (1 mg/kg BW) 0, 3, 6, 12 and 24 h before examination. Expressions of TLRs and IL-1beta in the theca and granulosa layers were examined by semi-quantitative RT-PCR. Immunocytochemistry was performed to identify immunoreactive TLR-4. The theca layer expressed TLR-2, TLR-4, TLR-5 and TLR-7, whereas the granulosa layer expressed only TLR-4 and TLR-5. The expression of TLR-4 and TLR-5 in the theca layer increased significantly during follicular growth. In the granulosa layer, the expression of TLR-5 increased, but that of TLR-4 was unchanged. Expression of TLR-4 increased significantly during the period of 6 to 12 h of LPS treatment in the theca layer and during the period of 12 to 24 h in the granulosa layer of F(3). Immunoreaction products for TLR-4 were observed in theca interna and granulosa layers of WF and F(5)-F(1), with the greater amount observed in the theca interna. LPS treatment significantly increased expression of IL-1beta in the theca layer after 3 h and in the granulosa layer during the period of 12 to 24 h. These results suggest that TLRs are expressed in ovarian follicles and that TLR-4 and TLR-5 expression increase with the growth of follicles. Enhanced expression of TLR-4 and IL-1beta by LPS in the theca and granulosa layers suggests possible roles of TLR in recognition of microorganisms.     

Effects of oral administration of colostrum whey in peripartum goat on antimicrobial peptides in postpartum milk

The present study was conducted to examine whether colostrum supplementation in peripartum goats increases the antimicrobial peptides in their milk. Goats were orally administered 2 ml of colostrum whey products (colostrum group) or water (control group) daily, from 2 weeks before until 2 weeks after kidding. Body weights of mothers and kids were measured. Blood, milk, and fecal samples were collected from the mothers, and blood samples were collected from the kids. Concentrations of milk antimicrobial peptides (beta‐defensin, cathelicidin, lactoferrin, S100A7, lactoperoxidase, and immunoglobulin A [IgA]) were determined. IgA and nutritional parameters (glucose, total cholesterol, triglyceride, ketone bodies, and non‐esterified fatty acids) were also determined in the blood of mothers and kids. Milk IgA and lactoferrin concentrations were higher in the colostrum group than in the control group. Conversely, lower milk concentrations of S100A7 were observed in the colostrum group than that in the control group. Plasma IgA concentrations were higher for kids from the colostrum group than for those from the control group. These results suggest that oral administration of colostrum in pregnant goats increases IgA concentration in postpartum milk, which can subsequently improve the health of their kids.     

Cloning and seasonal changes in ovarian expression of a TSH receptor in the channel catfish, Ictalurus punctatus

The cDNA encoding the thyroid stimulating hormone (TSH) receptor was isolated from the ventral aorta (thyroid follicles) and in both gonads of the channel catfish. The seasonal changes in ovarian expression of this receptor seem to be correlated to reproduction. TSH, but not LH or FSH, activated the recombinant TSH receptor expressed in COS cells. These results indicate that the isolated cDNA encodes a functional and highly specific TSH receptor. The seasonally regulated expression in the catfish ovary suggests a role for TSH in gametegenesis.     

Significance of local immunity in hen reproductive organs

The current paper describes aspects of local immunity in the ovary and oviduct, and the significance of immunity to reproductive functions in hens. The immunocompetent cell populations in the ovary and oviduct change with a positive correlation to sexual activity, and gonadal steroid is one of the key factors in the increase. Local immune responses mediated by major histocompatibility complex class II and T cell subsets occur in response to infection by Salmonella enteritidis, which may contaminate eggs. In the ovary, immunocompetent cells are also suggested to play roles in the regulation of ovarian functions. Macrophages and T cells are likely to enhance the regression of atretic follicles to maintain the ovarian tissue microenvironment. Autoantibodies to ovarian tissues appeared in the hens with low egg laying frequency, suggesting that the auto‐antibodies may be one of the factors in the decline of egg production. In the oviduct, local immunity possibly has a role in the selection of sperm, though the immunoreactions may also affect sperm survival leading to the decline in fertility. The concentration of yolk IgY, which plays a role in maternal immunity transmission, significantly decreases with the aging of birds, whereas it is significantly increased by estrogen. Therefore, the immune system plays significant roles not only in defense against infection, but also in the functions of reproductive organs. Investigations on the local immune system in the reproductive organs and factors affecting it are of importance for the production of sterile eggs and improvement of reproductive functions.     

Immunolocalization of sex steroid receptors in the epididymis and ductus deferens of immature and mature Japanese Quail, Coturnix Japonica

The goal of this study was to determine whether in the Japanese quail the male genital tract contains receptors for progesterone, androgen and estrogen (PR, AR and ER, respectively), which have significant roles in reproductive functions, and whether their localization changes during sexual maturation. The epididymis and ductus deferens (middle and ampulla regions) of immature (approximately 30-day-old) and mature male Japanese quail were collected and frozen sections of them were immunostained for PR, AR and ER. The immunoreaction products for AR and PR were found in the nuclei of epithelial cells in the efferent ductules, epididymal duct, and the middle and ampulla regions of the ductus deferens of mature and immature birds. In the mature birds, the epithelial cells of the efferent ductules, epididymal duct, and the middle and ampulla regions of the ductus deferens were positive for ER, although some of the cells in the ductus deferens were negative. The epithelial cells of the ductules in the epididymis stained positive for ER, but the immunoreactions were negligible in the ductus deferens of immature birds. These results suggest that the epididymis and ductus deferens in quail possesses PR, AR and ER receptors. Each receptor is expressed before sexual maturation, although enhancement of ER expression may occur during maturation.     

Rates of chemical weathering of porous rhyolites: 5-year measurements using the weight-loss method

To examine the rate of chemical weathering, field experiments using microweight-loss techniques were carried out. Two kinds of porous rhyolite forming lava domes in Kozu-shima Island were selected: (1) a younger rock (weathering period is 2.6 ka) and (2) an older rock (20 ka). Both rocks seem to have been very similar in their initial composition and structure. They have, however, different properties in the present depending on the difference in weathering period. Rock tablets of these rhyolites with a diameter of 3.5 cm and a thickness of about 1 cm were enclosed in a nylon mesh bag and placed in a soil–bedrock interface on a hillslope of central Japan for over 5 years. The younger rock has a lower weight loss by about 0.5%, and the older rock has a higher weight loss of 3% to 5%. These results show that older rock has a higher rate of chemical weathering than in younger rock. This supports Oguchi et al.'s (Oguchi, T.C., Hatta, T., Matsukura, Y., 1994. Changes in rock properties of porous rhyolite through 40,000 years in Kozu-shima Island, Japan. Geogr. Rev. Jpn. 67A, 775–793 (in Japanese, with English Abstr.); Oguchi, T.C., Hatta, T., Matsukura, Y., 1999. Weathering rates over 40,000 years based on changes in rock properties of porous rhyolite. Phys. Chem. Earth (A) 24, 861–870.) finding that the rate of change in chemical properties appears to accelerate with weathering time.     

Isolation and characterization of aminopeptidase (Jc-peptidase) from Japanese cedar pollen (Cryptomeria japonica)

An aminopeptidase, Jc-peptidase, was purified from Japanese cedar pollen by seven steps, including precipitation with ammonium sulfate, ion-exchange chromatography, gel filtration, hydrophobic interaction chromatography on phenyl-agarose, and high-performance liquid chromatography. Purified Jc-peptidease has a molecular weight of 42 kDa and hydrolyzes the synthetic substrates of L-phenylalanyl-4-methylcoumaryl-7-amide (Phe-MCA) with Km = 5 x 10(-5) M, Tyr-MCA with Km = 7 x 10(-4) M, Leu-MCA with Km = 1 x 10(-3) M, and Met-MCA with Km = 1 x 10(-3) M. Other MCA analogues such as Arg-MCA or Glu-MCA failed to serve as its substrates. The activity was inhibited in the presence of phebestin, [(2S,3R)-3-amino-2-hydroxy-4-phenylbutanoyl-L-valyl]-L-phenylalanine, with Ki = 4.7 x 10(-5) M, or bestatin, [(2S,3R)-3-amino-2-hydroxy-4-phenylbutanoyl]-L-leucine, with Ki = 1.1 x 10(-4) M. According to amino acid sequence analysis, the N-terminal amino group seems to be blocked. The physiological function of the aminopeptidase (Jc-peptidase) has not been clarified in vivo.     

Quality aspect of fish sauce prepared from underutilized fatty Japanese anchovy and rabbit fish

ABSTRACT:   To explore the potential utility of underutilized fish in the fish sauce industry, fish sauces were prepared from both raw and heat-induced meat of Japanese anchovy Engraulis japonica and rabbit fish Siganus fuscescens using wheat malt, and their quality aspects and sensory properties were assessed. Proximate composition of the fish meat represents protein as the major component (16.0–17.8%), other than moisture. Analyses of free amino acids, peptides including oligopeptides, and organic acids contents in fish sauces revealed suitability of both raw and heat-induced meat of the selected species in commercial fish sauce production. The variations in taste of raw meat fish sauces were species-specific, but the taste of fish sauces from heat-induced meat were similar.     

Developmental changes in pituitary–thyroid axis, and formation of gonads in leptocephali and glass eels of Anguilla spp.

SUMMARY: Pituitary, thyroid gland and gonads in leptocephali of Japanese eel Anguilla japonica (19.8–32.6 mm in total length), A. obscura (45.0 mm), and A. bicolor pacifica (49.5 mm) and those in glass eels of the Japanese eel were histologically and immunohistochemically examined in order to observe the developmental changes of these endocrine organs in the Anguillidae. The pituitary, consisting of adenohypophysis and neurohypophysis in Japanese eel leptocephali over 22.5 mm, did not contain thyroid stimulating hormone (TSH) immunoreactive cells. Such cells were, however, detectable in the more developed pituitaries of leptocephali of A. obscura and A. bicolor pacifica and in those of glass eels. Conversely, thyroxine (T₄)-immunoreactive thyroid follicles could be detected in all specimens, both leptocephalic and glass eel. Only in glass eels, gonads were found in the body cavity, and these gonads harbored one or two primordial germ cells (PGC) per cross-section. Our results indicate that thyroid hormones (TH) production started prior to TSH production, and that TSH and TH are both secreted during the metamorphosis from leptocephalus to glass eel. Therefore, it is plausible that the TSH–TH axis is involved in the metamorphosis from leptocephalus to glass eel, but not in the early growth from preleptocephalus to leptocephalus.