Cellular targets and pathways of yellow head virus infection in lymphoid organ of Penaeus monodon as studied by transmission electron microscopy

Negative-stained intact yellow head virus (YHV) was an enveloped bacilliform particle measuring 40-50 x 175-210 nmwith spike-like projections measuring 7-9 nm. The space between projections was 4-7 nm. YHV nucleocapsid was rod-shaped,measuring 35-40 x 160-200 nm, and the RNA genome had 40-50 turns in a helical structure. YHV infected both stromal matrixcells and haemocytes in the lymphoid tubule wall. The patterns of localisation of viral particles were similar in both cells. Thefully enveloped viral particles were detected at the cell membrane, endosome, rough endoplasmic reticulum, Golgi complexand secretory vesicles, and virions were exocytosed at the cell membrane. In the case of severe infection, unenveloped viralparticles could be detected in the cytoplasm, and they might be released by general breakdown and lysis of the highly infectedcells

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(I) GnRH-III and anti-tunicate(t) GnRH, but with variation of labeling intensity by each anti-GnRH type. IGnRH-III-ir was detected in numerous type1 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 type2 neurosecretory cells (NS2) in cerebral ganglion showed lGnRH-III-ir. Long nerve fibers in the neuropil of ventral regions of the cerebral and pleuropedal 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 oocytes development

Expression of the male reproduction-related gene (Mar-Mrr) in the spermatic duct of the giant freshwater prawn, Macrobrachium rosenbergii

Phosphorylated sperm proteins are crucial for sperm maturation and capacitation as a priori to their fertilization with eggs. In the freshwater prawn, Macrobrachium rosenbergii, a male reproduction-related protein (Mar-Mrr) was known to be expressed only in the spermatic ducts as a protein with putative phosphorylation and may be involved in sperm capacitation in this species. We investigated further the temporal and spatial expression of the Mar-Mrr gene using RT-PCR and in situ hybridization and the characteristics and fate of the protein using immunblotting and immunocytochemistry. The Mar-Mrr gene was first expressed in 4-week-old post larvae and the protein was produced in epithelial cells lining the spermatic ducts, at the highest level in the proximal region and decreased in the middle and distal parts. The native protein had a MW of 17 kDa and a high degree of serine/threonine phosphorylation. It was transferred from the epithelial cells to become a major protein at the anterior region of the sperm. We suggest that it is involved in sperm capacitation and fertilization in this open thelycal species and this is being investigated

The effects of biogenic amines, gonadotropin-releasing hormones and corazonin on spermatogenesis in sexually mature small giant freshwater prawns, Macrobrachium rosenbergii (De Man, 1879)

Neurotransmitters such as the serotonin (5-HT) and dopamine (DA), as well as the neurohormones gonadotropin-releasing hormones (GnRHs) and corazonin (Crz), are known to have various effects on decapod crustaceans, including ovarian maturation and spermatogenesis. The effects of these neurotransmitters and neurohormones on spermatogenesis in the small male freshwater prawns, Macrobrachium rosenbergii, have not been reported. So, we undertook histological and histochemical observations, as well as germ cell proliferation assays to examine the effects of 5-HT, DA, two exogenous GnRH isoforms (l-GnRH-III and oct-GnRH) and Crz. Ten experimental groups were injected with 5-HT and DA at 2.5 × 10−7 and 2.5 × 10−6 mol/prawn, and l-GnRH-III, oct-GnRH and Crz at 50 and 500 ng/gBW, at 4-day intervals from days 0 to 16. We found that prawns treated with 5-HT and GnRH isoforms exhibited significant increases in their testis-somatic index (TSI), seminiferous tubules at early maturation, i.e., stages I and III, with increased diameter of the tubules (DST), and germ cell proliferation, by days 4, 12 and 16, compared with saline control groups. In contrast, prawns treated with DA and Crz showed mostly seminiferous tubules at late maturation stages VIII and IX, and decreases of TSI, DST, and cell proliferation, by day 12, compared with saline control groups. By day 16 the Crz-treated prawns had died. These data indicate that 5-HT and GnRHs can stimulate spermatogenesis, while DA and Crz inhibit spermatogenesis. Consequently, hormonal treatment of male broodstocks in aquaculture with 5-HT and GnRHs could provide valuable tools to enhance reproduction by accelerating testicular maturation, leading to increased production of sperm