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Digestibility of kamaboko-diet in totally isolated stomach of black rockfish Sebastes schlegeli by in vitro culture method
Abstracts
and five alternative polyadenylation mechanisms in the 5'UTR and 3'UTR of pdf mRNA, respectively. In QRT-PCR, total pdf mRNA level was shown to be fluctuated in a day. The analyses by in situ hybridization and immunocytochemistry clarified about 14 lateral neurons in the hemisphere expressing pdf mRNA and PDF peptide simultaneously. In the present study, we attempted to measure the expression levels of each alternative splicing mRNA variants (insertion/deletion) at 5'UTR by QRT-PCR. Also, we challenged to identify discriminately the cells expressing the splicing variants by in situ hybridization.
457
Comparing methylation status between summer and winter bees, although few methylated cytosines were found in winter bees, 4 cytosines were highly methylated in summer bees in the promoter region of one isoform of MAPK. Additionally, we showed that cytosines not only in transcribed regions of one isoform of MAPK but also in promoter region were methylated. These results suggested the difference of methylation level in MAPK promoter region results behavioral difference between summer and winter bees. doi:10.1016/j.cbpb.2008.09.040
doi:10.1016/j.cbpb.2008.09.038
38. Three-dimensional reconstruction of neural tracts and commissures in the bee brain
40. Responses to chemical stimuli from the totally isolated stomach of the black rock fish Sebastes schlegeli using an in vitro culture method
Hiroyuki Ai, Yuri Kusakabe, Tsunao Itoh, Div. of Biol. Dep. of Earth Sys. Sci., Fukuoka Univ. Fukuoka 814-0180, Japan
Makoto Igarashi, Minoru Kihara, Grad. Sch. Sci. Eng., Tokai Univ., Sapporo 005-8601, Japan
The neural mechanism of olfactory learning and memory of honeybee has been clarified, however that of the bee's instinctive behavior, like the waggle dance communication is still unclear. We have identified many interneurons which process the vibratory signal caused by waggle dance, by intracellular recording and staining. As a result, it has been found that the principle neurons related with the vibratory processing arborize in the posterior protocerebral lobe (PPL), dorsal lobe (DL) and suboesophageal ganglion (SEG). In these brain areas there are many neural tracts and commissures. In order to know which neural pathway our identified neurons run through, the neural tracts and commissures in the brain were dyed with fluorescence dextrans and three-dimensional images of these neural pathways were reconstructed from fluorescence images. It was found that our identified interneurons, DL-Int-1 and DL-Int-2, run through the medial antennocerebral tract and the maxillary supramedian commissure, respectively. Moreover, we identified three new neural pathways, named as dorsal lateral protocerebral lobe commissure (which connects among the optic lobe, bilateral PPL and central body), lateral protocerebral lobe descending tract (which connects between PPL and SEG and runs through the VNC) and ventral deutocerebral commissure (which connect between bilateral DLs).
The nutrient source for predatory fish is mostly protein, and the stomach is the primary digestive organ for dietary protein. However, the fish's environment makes study of digestive physiology difficult. We are thus developing a new experimental method to confirm gastric digestive enzyme output from the totally isolated, luminally perfused and externally batch-cultured black rockfish stomach. In this study, glass beads (5 mm O. D.) were inserted into the stomach as a basic physical stimulus at the start of the incubation. We then administered glycine to the internal perfusate as a dietary stimulus, or added histamine to the external culture solution as a vascular side stimulus after 120 min. We report stomach motility and the changes in pepsin-like enzyme activity in the internal perfusate during a 6 h incubation period.
doi:10.1016/j.cbpb.2008.09.039
39. Seasonal changes of methylation status in promoter region of a MAPK on honeybee genomic DNA Dai Hatakeyama a,b, Uli Mueller b, a Kagawa Sch. Pharm. Sci. Tokushima Bunri Univ., Kagawa 769-2193, Japan; b Department 8.3, Nat. Sci. Tech. III, Saarland Univ., Saarbruecken 66041, Germany Investigation of whole genomic sequence of honeybees was recently completed, and honeybees are focused as suitable model animals for understanding detailed mechanisms of DNA methylation. Using antibody against methylated cytosine, we performed an immunohistochemical, immunoblotting-based and ELISA-based experiments to quantify total methylation levels in whole genomic DNA, and showed that the methylation level in total genomic DNA in honeybee brain is significantly lower than that in mammalian and Drosophila brain. We focused on genes regulated by the transcription factor CREB and found the CRE sequences in the promoter region of 2 isoforms of MAPKs. Bisulfite sequence technique showed that the promoter regions of MAPKs had at least 2 or 4 methylated cytosines and that the methylation occurred at sequences of CpA and CpC.
doi:10.1016/j.cbpb.2008.09.041
41. Digestibility of kamaboko-diet in totally isolated stomach of black rockfish Sebastes schlegeli by in vitro culture method Tomohiro Suzuki a, Fumiaki Itou a, Makoto Igarashi b, Minoru Kihara a,b, Department Marine Biol. Sci., Tokai Univ., Sapporo 005-8601, Japan; b Grad. Sch. Sci. Eng., Tokai Univ., Sapporo 005-8601, Japan a
This is the first study to measure diet digestibility in the totally isolated and cultured stomach. We prepared commercial kamaboko gel in 5 mm cubes as a pseudo-diet, and then inserted the pseudodiets into the isolated stomach. The stomach was then cultured for 6 h at 20 °C and the luminal perfusate collected every 90 min. We measured the difference in dry weights of these diets before and after the incubation. Moreover, we compared enzyme activities in the gastric perfusate of the kamaboko group with those of a group given glass beads. Kamaboko cube dry weight decreased up to 10% after 6 h incubation. Pepsin-like enzyme activity was always higher in the kamaboko group. These results indicate that the totally isolated and cultured stomach can digest a pseudo-diet of kamaboko. doi:10.1016/j.cbpb.2008.09.042
42. Bioresources characteristics of chum and pink salmon considered from their changes in rhodopsin–porphyropsin ratio Eiichi Hasegawa, Fisheries Research Agency, National Salmon Resources Center, Sapporo 062-0922, Japan
and five alternative polyadenylation mechanisms in the 5'UTR and 3'UTR of pdf mRNA, respectively. In QRT-PCR, total pdf mRNA level was shown to be fluctuated in a day. The analyses by in situ hybridization and immunocytochemistry clarified about 14 lateral neurons in the hemisphere expressing pdf mRNA and PDF peptide simultaneously. In the present study, we attempted to measure the expression levels of each alternative splicing mRNA variants (insertion/deletion) at 5'UTR by QRT-PCR. Also, we challenged to identify discriminately the cells expressing the splicing variants by in situ hybridization.
457
Comparing methylation status between summer and winter bees, although few methylated cytosines were found in winter bees, 4 cytosines were highly methylated in summer bees in the promoter region of one isoform of MAPK. Additionally, we showed that cytosines not only in transcribed regions of one isoform of MAPK but also in promoter region were methylated. These results suggested the difference of methylation level in MAPK promoter region results behavioral difference between summer and winter bees. doi:10.1016/j.cbpb.2008.09.040
doi:10.1016/j.cbpb.2008.09.038
38. Three-dimensional reconstruction of neural tracts and commissures in the bee brain
40. Responses to chemical stimuli from the totally isolated stomach of the black rock fish Sebastes schlegeli using an in vitro culture method
Hiroyuki Ai, Yuri Kusakabe, Tsunao Itoh, Div. of Biol. Dep. of Earth Sys. Sci., Fukuoka Univ. Fukuoka 814-0180, Japan
Makoto Igarashi, Minoru Kihara, Grad. Sch. Sci. Eng., Tokai Univ., Sapporo 005-8601, Japan
The neural mechanism of olfactory learning and memory of honeybee has been clarified, however that of the bee's instinctive behavior, like the waggle dance communication is still unclear. We have identified many interneurons which process the vibratory signal caused by waggle dance, by intracellular recording and staining. As a result, it has been found that the principle neurons related with the vibratory processing arborize in the posterior protocerebral lobe (PPL), dorsal lobe (DL) and suboesophageal ganglion (SEG). In these brain areas there are many neural tracts and commissures. In order to know which neural pathway our identified neurons run through, the neural tracts and commissures in the brain were dyed with fluorescence dextrans and three-dimensional images of these neural pathways were reconstructed from fluorescence images. It was found that our identified interneurons, DL-Int-1 and DL-Int-2, run through the medial antennocerebral tract and the maxillary supramedian commissure, respectively. Moreover, we identified three new neural pathways, named as dorsal lateral protocerebral lobe commissure (which connects among the optic lobe, bilateral PPL and central body), lateral protocerebral lobe descending tract (which connects between PPL and SEG and runs through the VNC) and ventral deutocerebral commissure (which connect between bilateral DLs).
The nutrient source for predatory fish is mostly protein, and the stomach is the primary digestive organ for dietary protein. However, the fish's environment makes study of digestive physiology difficult. We are thus developing a new experimental method to confirm gastric digestive enzyme output from the totally isolated, luminally perfused and externally batch-cultured black rockfish stomach. In this study, glass beads (5 mm O. D.) were inserted into the stomach as a basic physical stimulus at the start of the incubation. We then administered glycine to the internal perfusate as a dietary stimulus, or added histamine to the external culture solution as a vascular side stimulus after 120 min. We report stomach motility and the changes in pepsin-like enzyme activity in the internal perfusate during a 6 h incubation period.
doi:10.1016/j.cbpb.2008.09.039
39. Seasonal changes of methylation status in promoter region of a MAPK on honeybee genomic DNA Dai Hatakeyama a,b, Uli Mueller b, a Kagawa Sch. Pharm. Sci. Tokushima Bunri Univ., Kagawa 769-2193, Japan; b Department 8.3, Nat. Sci. Tech. III, Saarland Univ., Saarbruecken 66041, Germany Investigation of whole genomic sequence of honeybees was recently completed, and honeybees are focused as suitable model animals for understanding detailed mechanisms of DNA methylation. Using antibody against methylated cytosine, we performed an immunohistochemical, immunoblotting-based and ELISA-based experiments to quantify total methylation levels in whole genomic DNA, and showed that the methylation level in total genomic DNA in honeybee brain is significantly lower than that in mammalian and Drosophila brain. We focused on genes regulated by the transcription factor CREB and found the CRE sequences in the promoter region of 2 isoforms of MAPKs. Bisulfite sequence technique showed that the promoter regions of MAPKs had at least 2 or 4 methylated cytosines and that the methylation occurred at sequences of CpA and CpC.
doi:10.1016/j.cbpb.2008.09.041
41. Digestibility of kamaboko-diet in totally isolated stomach of black rockfish Sebastes schlegeli by in vitro culture method Tomohiro Suzuki a, Fumiaki Itou a, Makoto Igarashi b, Minoru Kihara a,b, Department Marine Biol. Sci., Tokai Univ., Sapporo 005-8601, Japan; b Grad. Sch. Sci. Eng., Tokai Univ., Sapporo 005-8601, Japan a
This is the first study to measure diet digestibility in the totally isolated and cultured stomach. We prepared commercial kamaboko gel in 5 mm cubes as a pseudo-diet, and then inserted the pseudodiets into the isolated stomach. The stomach was then cultured for 6 h at 20 °C and the luminal perfusate collected every 90 min. We measured the difference in dry weights of these diets before and after the incubation. Moreover, we compared enzyme activities in the gastric perfusate of the kamaboko group with those of a group given glass beads. Kamaboko cube dry weight decreased up to 10% after 6 h incubation. Pepsin-like enzyme activity was always higher in the kamaboko group. These results indicate that the totally isolated and cultured stomach can digest a pseudo-diet of kamaboko. doi:10.1016/j.cbpb.2008.09.042
42. Bioresources characteristics of chum and pink salmon considered from their changes in rhodopsin–porphyropsin ratio Eiichi Hasegawa, Fisheries Research Agency, National Salmon Resources Center, Sapporo 062-0922, Japan