Expression levels of taste‐related genes in palate and tongue tip,and involvement of transient receptor potential subfamily M member 5 (TRPM5) in taste sense in chickens

The elucidation of the mechanisms underlying the taste sense of chickens will contribute to improvements in poultry feeding, because the molecular mechanism of chickens’ taste sense defines the feeding behavior of chickens. Here we focused on the gene expressions in two different oral tissues of chickens – the palate, which contains many taste buds, and the tongue tip, which contains few taste buds. Using the quantitative real‐time polymerase chain reaction method, we found that the molecular markers for taste buds of chickens, that is α‐gustducin and vimentin, were expressed significantly highly in the palate compared to the tongue tip. Our analyses also revealed that transient receptor potential subfamily M member 5 (TRPM5), a cation channel involved in taste transduction in mammals, was also highly expressed in the palate compared to the tongue tip. Our findings demonstrated that the expression patterns of these genes were significantly correlated. We showed that the aversion to bitter solution was alleviated by a TRPM5 inhibitor in behavior of chickens. Taken together, our findings enabled us to develop a simple method for screening taste‐related genes in chickens. The use of this method demonstrated that TRPM5 was involved in chickens’ taste transduction, and that a TRPM5 inhibitor can alleviate chickens’ bitter taste perception of feed ingredients.     

Structure of bovine fungiform taste buds and their immunoreactivity for gustducin

The taste buds of bovine fungiform papillae were studied by light and electron microscopy using both histological and immunohistochemical methods. The taste buds existed in the epithelium of the apical region of the papillae. By electron microscopy, two types of taste cells, namely type I and type II cells, could be classified according to the presence of dense-cored vesicles, the cytoplasmic density and the cell shape. Type I cells were thin, had an electron-dense cytoplasm containing dense-cored vesicles, and possessed long thick apical processes in the taste pore. Type II cells were thick, had an electron-lucent cytoplasm containing many electron-lucent vesicles, rather than dense-cored vesicles, and possessed microvilli in the taste pore. Immunohistochemical staining with an antiserum against gustducin was investigated by both light and electron microscopy using the avidin-biotin complex (ABC) method. Some, but not all, of the type II cells exhibited gustducin immunoreactivity, whereas none of the type I cells showed any immunoreactivity.     

The effects of propofol, isoflurane and sevoflurane on vecuronium infusion rates for surgical muscle relaxation in dogs

OBJECTIVE: To compare the constant rate infusion (CRI) of vecuronium required to maintain a level of neuromuscular blockade adequate for major surgeries, e.g. thoracotomy or laparotomy, in dogs anaesthetized with a CRI of fentanyl and either propofol, isoflurane or sevoflurane. STUDY DESIGN: Prospective, randomized, cross-over study. ANIMALS: Thirteen male beagles (age, 9-22 months; body mass 6.3-11.3 kg). MATERIALS AND METHODS: Dogs were anaesthetized with propofol (24 mg kg(-1) hour(-1) IV CRI; group P), isoflurane (1.3% end-tidal concentration; group I) or sevoflurane (2.3% end-tidal concentration; group S) with fentanyl (5 microg kg(-1) hour(-1) IV, CRI). Sixty to seventy minutes after induction of anaesthesia, vecuronium was administered at a rate of 0.4, 0.3 and 0.2 mg kg(-1) hour(-1) in groups P, I and S respectively. To determine the degree of neuromuscular block, a peripheral nerve was stimulated electrically using the train-of-four (TO4) stimulus pattern. Evoked muscle contractions were evaluated using a neuromuscular monitoring device. Once the TO4 ratio reached 0, the continuous infusion rate was decreased and adjusted to maintain a TO4 count of 1. Continuous infusion was continued for 2 hours. The infusion rate of vecuronium was recorded 20, 40, 60, 80, 100 and 120 minutes after the start of infusion. RESULTS: The mean continuous infusion rates of vecuronium during stable infusion were 0.22 +/- 0.04 (mean +/- SD), 0.10 +/- 0.02 and 0.09 +/- 0.02 mg kg(-1) hour(-1) in groups P, I and S respectively. There were statistically significant differences between the rates in groups P and I and between the rates in groups P and S. Conclusions and clinical relevance In healthy dogs, the recommended maintenance infusion rate of vecuronium is 0.2 mg kg(-1) hour(-1) under CRI propofol-fentanyl anaesthesia and 0.1 mg kg(-1) hour(-1) during CRI fentanyl-isoflurane or sevoflurane anaesthesia.     

Comparison of feasibility and safety of epidural catheterization between cranial and caudal lumbar vertebral segments in dogs

To compare the technical difficulty and safety of epidural catheterization between cranial and caudal lumbar region, thirteen dogs were randomly assigned to a cranial lumbar group (group CraL, n=6) or a caudal lumbar group (group CauL, n=6) depending on different epidural sites, and one dog was used as a negative control without catheterization. After general anesthesia, an epidural catheter was advanced 10 cm cranially from the interspace of L1-L2 in group CraL or from lumbosacral space in group CauL. Dogs were euthanized and catheter position and tip location were confirmed by laminectomy. Spinal cord samples were examined by macro- and microscopic observations. Success rate, time taken for epidural space confirmation and catheter insertion were compared, and overall technical difficulty was evaluated subjectively. Epidural catheter was inserted successfully in all dogs. Time needed from needle skin puncture to catheter placement and saline injection was 226 ± 63 and 229 ± 26 sec in groups CraL and CauL without significant differences. Three dogs in group CraL suffered subcutaneous blood, but no spinal cord injuries were found. Subjective evaluation score of the overall technical difficulty was slightly but significantly higher in group CraL than in group CauL (P=0.009). Epidural catheterization in cranial lumbar region could be performed as feasible and safe as that at the caudal lumbar vertebral region in medium or large dogs.     

An astacin-like protease, alveolin, is released from cortical vesicles and induces changes in egg envelope proteins during fertilization of medaka (Oryzias latipes) eggs

Upon fertilization, two major protein groups, ZI-1,2 and ZI-3, composing the egg envelope are polymerized to insoluble forms via temporary formation of 132 kDa and 61–62 kDa intermediate proteins. We have shown that an astacin-like protease, `alveolin' is released from cortical vesicles into the perivitelline space after egg activation and induces initiation of the polymerization of egg envelope proteins. To clarify initial reactions of the polymerization, the primary structure of the 132 kDa protein was analyzed.     

Glycoprotein hormone receptor family in medaka (Oryzias latipes)

Four types of GPHR cDNAs have been cloned from ovary and testis of medaka (Oryzias latipes) and their gene constructions have been determined. Two of them are closely related to known fish receptors for FSH and LH, respectively. Changes in their mRNA levels were examined during the course of oogenesis. FSH receptor mRNA could not be detected from 20 h before ovulation, whereas LH mRNA remained 5 h before ovulation.     

Promoter analysis of two aromatase genes in the serial-sex changing gobiid fish, Trimma okinawae

Despite numerous endocrine studies on sex change in teleost, no general mechanism that mediates sex change has emerged. The gobiid fish, Trimma okinawae, can change sex in both directions repeatedly. This phenomenon of sex change in goby assigns it as an excellent animal model to elucidate the understanding mechanisms of sex change. In hermaphrodite fishes, estrogen plays a particularly important role in natural and experimentally induced sex change. To investigate the role of estrogen in the serial-sex changing fish T. okinawae, we cloned and analyzed the 5′-flanking regions of P450arom genes from goby genome DNA. Both regions have consensus sequences of TATA, CRE and ERE. Ad4 binding site was restricted in the region of P450aromA. These findings indicate that different regulators control the expression of the two P450arom genes.