The existence of gonadotropin-releasing hormone-like peptides in the neural ganglia and ovary of the abalone, Haliotis asinina L.

Gonadotropin-releasing hormone (GnRH) is a neuropeptide that is conserved in both vertebrate and invertebrate species. In this study, we have demonstrated the presence and distribution of two isoforms of GnRH-like peptides in neural ganglia and ovary of reproductively mature female abalone, Haliotis asinina, using immunohistochemistry. We found significant immunoreactivities (ir) of anti-lamprey(l) GnRH-III and anti-tunicate(t) GnRH, but with variation of labeling intensity by each anti-GnRH type. lGnRH-III-ir was detected in numerous type 1 neurosecretory cells (NS1) throughout the cerebral and pleuropedal ganglia, whereas tGnRH-I-ir was detected in only a few NS1 cells in the dorsal region of cerebral and pleuropedal ganglia. In addition, a small number of type 2 neurosecretory cells (NS2) in cerebral ganglion showed lGnRH-III-ir. Long nerve fibers in the neuropil of ventral regions of the cerebral and pluropedal ganglia showed strong tGnRH-I-ir. In the ovary, lGnRH-III-ir was found primarily in oogonia and stage I oocytes, whereas tGnRH-ir was observed in stage I oocytes and some stage II oocytes. These results indicate that GnRH produced in neural ganglia may act in neural signaling. Alternatively, GnRH may also be synthesized locally in the ovary where it could induce oocyte development.     

Production and characterization of a monoclonal antibody against recombinant cathepsin L1 of Fasciola gigantica

Monoclonal antibodies (MoAbs) against a recombinant cathepsin L1 of Fasciola gigantica (rFgCatL1) were produced in vitro by fusion of BALB/c mice spleen cells immunized with rFgCatL1 and mouse myeloma cells. Reactivity and specificity of these MoAbs were evaluated by indirect ELISA and immunoblotting techniques. Seven MoAb clones were selected from the stable hybridoma clones, namely 1E10, 1F5, 3D11, 4B10, 4D3, 4E3 and 5E7. Clones 1E10, 1F5 and 3D11 were IgM, whereas clones 4B10, 4D3, 4E3 and 5E7 were IgG₁. All MoAbs had kappa light chain isotypes. All MoAbs reacted with rCatL1 at molecular weight (MW) 30 kDa and with the native CatL1 at MW 27 kDa in whole body (WB) extracts of metacercariae (Met), newly excysted juveniles (NEJ), 1, 3, 5-week-old juveniles (Ju), adult WB and adult excretory–secretory (ES) fractions, but not with adult tegumental antigens (TA). All of these MoAbs showed no cross-reactions with antigens of other parasites commonly found in ruminants and human, including Paramphistomum cervi, Eurytrema pancreaticum, Gigantocotyle explanatum, Schistosoma spindale, Schistosoma mansoni, Moniezia benedeni, Avitellina centripunctata, Trichuris sp., Haemonchus placei and Setaria labiato-papillosa. Localization of CatL1 in each developmental stages of F. gigantica by immunoperoxidase technique, using these MoAbs as probes, indicated that CatL1 was present at high concentration in the caecal epithelium and caecal lumen of metacercariae, NEJ, 1, 3, 5-week-old juveniles and adult fluke. This finding indicated that CatL1 is a copiously expressed parasite protein that is released into the ES, thus CatL1 and its MoAb could be a good candidate for immunodiagnosis of fasciolosis in ruminant and human.     

Changes in the levels of serotonin and dopamine in the central nervous system and ovary, and their possible roles in the ovarian development in the giant freshwater prawn, Macrobrachium rosenbergii

Serotonin or 5-hydroxytryptamine (5-HT) and dopamine (DA) are the two key neurotransmitters that control gonadal development in decapod crustaceans. This study investigated changes in the levels of 5-HT and DA in the CNS and ovary during different phases of the ovarian cycle of the freshwater prawn, Macrobrachium rosenbergii. The levels of 5-HT and DA were quantified by using High Performance Liquid Chromatography with electrochemical detection (HPLC-ECD). Moreover, changes of vitellogenin (Vg) concentrations in the hemolymph after treatment with 5-HT and DA (at doses of 2.5 × 10⁻⁶ and 2.5 × 10⁻⁷ mol per prawn) were also examined. 5-HT exhibited a gradual increase in concentration in the brain and thoracic ganglia from ovarian stage I (0.12 ± 0.01 nmol/mg, 0.22 ± 0.01 nmol/mg, respectively) to reach a maximum (0.66 ± 0.03 nmol/mg, 1.48 ± 0.03 nmol/mg, respectively) at ovarian stage IV. In contrast, DA in the brain and thoracic ganglia showed the highest concentrations at ovarian stage II (0.20 ± 0.01 nmol/mg, 1.27 ± 0.06 nmol/mg, respectively) and then decreased to the lowest concentrations (0.06 ± 0.01 nmol/mg, 0.28 ± 0.04 nmol/mg, respectively) at ovarian stage IV. The ovarian concentration of 5-HT was 0.53 ± 0.11 nmol/mg at ovarian stage I and gradually increased to 1.63 ± 0.16 nmol/mg at ovarian stage IV. In contrast, the concentration of DA was highest at ovarian stage I (29.05 ± 1.31 nmol/mg), and lowest at the ovarian stage IV (11.43 ± 0.74 nmol/mg). Injecting 5-HT into prawns significantly increased Vg concentration in the hemolymph at ovarian stage IV compared to control groups, and injecting DA into prawns had the opposite effect. The inverse relationship between 5-HT and DA levels in neural ganglia and ovaries, and their opposing effects on hemolymph Vg levels suggest that these two transmitters play opposite regulatory roles in controlling ovarian maturation and oocyte development in this species.     

The expression and distribution of a leptin receptor in the central nervous system,digestive organs,and gonads of the giant freshwater prawn, Macrobrachium rosenbergii

In the present study, the presence and distribution of leptin receptor (LEP-R) in central nervous system, digestive organs, gonads of giant freshwater prawn, Macrobrachium rosenbergii, were investigated with Western blot and immunohistochemistry. By Western blot a LEP-R with a molecular weight (MW) of 100?kDa was detected in the brain, thoracic ganglia, abdominal ganglia, hepatopancreas, all parts of the gastrointestinal tract, ovaries, and testes. In hepatopancreas and foregut, another intense positive band was detected at molecular weight of 30?kDa, which could be an isotype of LEP-R. By immunohistochemistry, LEP-R-ir was detected in the neurons, and neuropils in the brain, thoracic ganglia, and abdominal ganglia. In the gastrointestinal tract, there was intense LEP-R-ir in the apical part of the epithelial cells of the foregut, midgut, and hindgut. In addition, LEP-R-ir was found in the Restzellen(R)cells and Fibrillenzellen(F) cells in the hepatopancreas. In the ovary, LEP-R-ir was detected in early stage of oocytes and mature oocytes. Intense LEP-R-ir was observed in spermatogonia and spermatocytes of the small and orange claw male prawns. In addition, LEP-R was seen in the high epithelium of spermatic ducts from all male morphotypes. In summary, the detection of the LEP-R-ir suggests the existence of a LEP-R in several organs of M. rosenbergii. Through binding with leptin peptide, LEP-R may be an important signaling molecule that has critical functions in modulating and controlling food intake, energy expenditure, and reproduction in this prawn.     

Distribution of abalone egg-laying hormone-like peptide in the central nervous system and reproductive tract of the male mud crab, Scylla olivacea

The mud crab, Scylla olivacea, is a high value economic marine animal in Thailand. However, collection of these crabs from natural habitat for local consumption and export has caused rapid population decline. Hence, aquaculture of this species is required and to this measure understanding of endocrine control of their reproduction must be understood. Egg laying hormone (ELH) is a neuropeptide synthesized by the bag cells (neurons) in the abdominal ganglia of Aplysia gastropods. It plays a critical role in controlling egg production and laying in gastropods, and its possible homolog (ELH-like peptide) was reported in the neural and ovarian tissues of prawns and recently in female reproductive tract of the blue swimming crab, Portunus pelagicus. In this study, we have studied the histology of the male reproductive tract in Scylla olivacea which are comprised of anterior testis, posterior testis, early proximal spermatic duct (ePSD), proximal spermatic duct (PSD), middle spermatic duct (MSD) and distal spermatic duct (DSD), by immunohistochemistry, detected an abalone ELH- immunoreactivity (aELH-ir) in epithelium of ducts in posterior testis and epithelium of all parts of spermatic duct. Furthermore, we could detect aELH-ir in neurons of cluster 9, 11, olfactory neuropil (ON) in the brain and in the small neurons located between the third and the fourth thoracic neuropils (T3-T4) and between the fourth and the fifth thoracic neuropils (T4-T5) of thoracic ganglia. Thus, the presence of aELH in male S. olivacea was designated the role of female egg laying behavior in the male mud crab.