The sperm longevity and freezability in the modified BHSV extender of Thai Pradu-hangdum chicken

Pradu-hangdum is a distinctive Thai native pure breed chicken. To conserve the pure breed of this chicken species, the artificial insemination was invested. The key to the highly successful output of this technique is high quality of semen. Hence, the objective of the present study is to characterize the Pradu-hangdam sperms and their longevity and freezability in the BHSV extender. The semen of the 30 randomized male chicken were collected. The macroscopic and microscopic examination were used to determine the sperm characters. The results revealed that the Pradu-hangdam sperms contained normal a spiral-shaped head, mid-piece and tail and normal white cream color. The mean number of sperm concentration was 5.24×109 ± 1.54 sperms/mL. The mean volume was 0.22 ± 0.08 mL. The results of longevity in mean total motility of sperm was 85.20%, 56.00% and 36.33% storage for 1, 24, and 48 hours after semen collection in extender, respectively. The longevity of sperm storage in extender decreased significantly at 24 and 48 hours (P<0.05). The freezability of Pradu-hangdam semen significant decreased from 81.45% to 57.02% of motility (P<0.05) but in the range of acceptable result for insemination. In conclusion, this study provides the basic knowledge of sperm characters and their longevity which decreases in relation to the time after collection even though it was preserved and frozen in the BHSV of the acceptable data. Furthermore, the freezing technique and fertility rate should be a further study in the long-term preservation of Pradu-hangdam sperms

Distribution of Gaba in the nerve ganglia of haliotis asinina linnaeus

Gamma-aminobutyric acid (GABA) is a major neurotransmitter and effective settlement inducer in abalone aquaculture. This study aimed to explore the distribution of GABA within neural tissues of Haliotis asinina. Gamma-aminobutyric acid was found in neuronal cell type 1 of 3 major ganglia (i.e., cerebral, pleuropedal, and visceral ganglia) of both sexes. The distribution of GABA-immunoreactive (-ir) cells in the cerebral ganglion was concentrated mostly in the cortex region of the dorsal horn, whereas it was scattered throughout the pleuropedal ganglion, with more in the upper half. Gamma-aminobutyric acid-ir nerve fibers were found throughout the neuropils of the ganglia. The visceral ganglion had the least numbers of GABA-ir neurons compared with the other 2 ganglia. The cells were distributed mainly in the dorsal horn. We also observed GABA to be colocalized with 2 other neurotransmitters: serotonin (5-HT) and dopamine (DA). In the cerebral ganglion, fluorescence double staining of GABA and 5-HT, and GABA and DA showed immunoreactivity in separate cells and was also colocalized in the same cells. In the pleuropedal ganglion, the staining pattern was similar to the cerebral ganglion, but positive-staining cells were less numerous. In the visceral ganglion, GABA and DA, and GABA and 5-HT were colocalized in the same cell types. Overall, we found that GABAergic cells were most numerous in the cerebral ganglion of H. asinina. Further studies are required to determine the functions of these neurotransmitters in relation to their distribution

Molecular characterization and analysis of a truncated serotonin receptor gene expressed in neural and reproductive tissues of abalone

In molluscs, the neurotransmitter serotonin (5-HT) has been linked to a variety of biological roles including gamete maturation and spawning. The possible involvement of 5-HT in abalone gamete release was demonstrated by a dose-dependent increase in Haliotis rubra gonad contractile bioactivity following 5-HT stimulation. Physiological functions associated with 5-HT, are mediated through binding to 5-HT receptors. A cDNA encoding a putative 5-HT receptor consisting of 359 amino acids was isolated from the tropical abalone H. asinina, termed 5-HT1 ha. The 5-HT1 ha shares G-protein-coupled receptor motifs with metazoan 5-HT receptors, including predicted transmembrane domains, active sites for protein kinase action, and N-linked glycosylation sites. However, the third intracellular loop of 5-HT1 ha is relatively short, and only six transmembrane domains are predicted, implying a truncated receptor. Phylogenetic analysis with known 5-HT receptor genes suggests that 5-HT1 ha belongs to the type 1 5-HT receptor family. Expression analysis by RT-PCR showed that 5-HT1 ha&nbsp; mRNA was present in all tissues examined, including the neural ganglia and gonad tissues. Immunocytochemistry revealed the presence of 5-HT1 ha specifically within the soma of neuronal cells located in the outer cortex of both cerebral and pleuropedal ganglia. In ovarian and testicular tissues, 5-HT1 ha immunoreactivity was observed in epithelial cells of the outer capsule and connective tissue of the trabeculae to which the gamete follicles adhere. Whether this receptor transcript is translated to a functional protein needs to be verified, but if so, it could play a role in reproduction.<br /

Molecular analysis of two FMRFamide-encoding transcripts expressed during the development of the tropical abalone haliotis asinina

FMRFamide-related peptides (FaRPs) are involved in numerous neural functions across the animal kingdom and serve as important models for understanding the evolution of neuropeptides. Gastropod molluscs have proved to be particularly useful foci for such studies, but the developmental expression of FaRPs and the evolution of specific transcripts for different peptides are unclear within the molluscs. Here we show that FaRPs are encoded by two transcripts that appear to be splice variants of a single gene in the abalone, Haliotis asinina, which represents the basal vetigastropods. Has-FMRF1 comprises 1,438 nucleotides and encodes a precursor protein of 329 amino acids that can potentially produce two copies of FLRFamide, one copy each of TLAGDSFLRFamide, QFYRIamide, SDPDLDDVIRASLLAYSLDDSPNN, and SVATAPVEAKAVEAGNKDIE, and 13 copies of FMRFamide. The second 1,241-nucleotide transcript, Has-FMRF2, encodes a 206-amino acid precursor protein with single copies of FLRFamide and FMRFamide along with such extended forms as NFGEPFLRFamide, FDSYEDKALRFamide, and NGWLHFamide, in addition to SDPGEDMLKSILLRGAPSNNGLQY and DTUDETTUNDNAHSRQ. Both transcripts are present early in life and are expressed in different but overlapping patterns within the developing larval nervous system. Mass spectrometry and immunocytochemistry demonstrate that FaRPs are cleaved from larger precursors and localize to the developing nervous system. Our results confirm previous evidence that FaRPs are expressed early and potentially play many roles during molluscan development and suggest that the last common ancestor to living gastropods used alternative splicing of an FMRFamide gene to generate a diversity of FaRPs in spatially restricted patterns in the nervous system