菜採 佐野 - Academia.edu (original) (raw)

Papers by 菜採 佐野

Journal of Fish Diseases, Mar 1, 1992

A method of in situ hybridization with biotinylatcd probes is described for the specific detectio... more A method of in situ hybridization with biotinylatcd probes is described for the specific detection of the Herpesvirus cvprini {CHV) genome, both in CHV infeeted eell eultures and tissue speeimens prepared from earp fry infeeted with the same virus. From moleeularly eloned Pstl-fragments of CHV DNA. two fragments were selected as probes for in situ hybridization which had no deteetable homology, cither with DNA of fathead minnow (FHM) eells used for virus propagation, or with the DNA of several fish herpesviruses. The method of in situ hybridization was based on a system using streptavidinbiotinylated alkaline phosphatase for the detection of hybridization.

Aquaculture, 1995

To assess cytotoxic activity of carp (Cyprinus carpio), head kidney leukocytes were examined with... more To assess cytotoxic activity of carp (Cyprinus carpio), head kidney leukocytes were examined with special reference to the effects of rearing water temperature and of assay temperature. Leukocytes as effector cells were separated from head kidney using Histopaque 1077 and cytotoxic activities were measured by the release of 51Cr from target cells which were K562, human chronic myelogenous leukemia cells. Cytotoxic activity of leukocytes from carp kept at higher temperature (25 °C) was lower at lower assay temperature (10 °C) than at higher assay temperature (25 °C). However, activity from carp acclimated to low temperature (10 °C) increased on 10 °C assay and decreased on 25 °C assay. Cellular composition of effector cells changed in connection with the above phenomenon. In carp acclimated to 10 °C, small lymphocytes decreased while the others, e.g. large lymphocytes, granulocytes and macrophages, increased. Accommodation of carp natural killer-like cells to environmental temperatures may be regulated by changing the cellular composition of effector cells.

CABI eBooks, 2017

The geographical distribution; economic impacts; diagnosis of the infection; pathology; pathophys... more The geographical distribution; economic impacts; diagnosis of the infection; pathology; pathophysiology; and the protective and control strategies against Oncorhynchus masou virus and Cyprinid herpesvirus are discussed.

Plankton and Benthos Research, Feb 28, 2020

Corbicula leana and C. fluminea are hermaphroditic and ovoviviparous freshwater clams. Although t... more Corbicula leana and C. fluminea are hermaphroditic and ovoviviparous freshwater clams. Although they are considered to reproduce using self-fertilization, the possibility of outcrossing was suggested due to lineage discordance between mitochondrial and genomic DNA. In these species, outcrossing means egg parasitism by the spermatozoa from one or more other clams, because they reproduce by androgenesis in which only the nucleus of spermatozoa is transmitted to the progeny. Moreover, the presence of males in these species was reported in the previous study, and they were estimated to reproduce by egg parasitism of hermaphrodites. In this study, we investigated the paternity of juveniles in the brood pouches of six hermaphrodites by comparing the genotypes of the brooded juveniles, brooding clams, and neighboring adult clams using two microsatellite DNA markers. Brooded juveniles showed either identical genotypes to the parental clam or different genotypes from their parent in one clam with brooding. The genotypes of brooded juveniles were identical to those of neighboring hermaphrodites and males. These results indicate that androgenetic Corbicula reproduce not only by self-fertilization but also by egg parasitism, with outcrossing among hermaphrodites and from males to hermaphrodites.

Journal of Fish Diseases, 1993

Heipcnirits cvprini (CHV) genome was traced in carp, Cvprinus carpio L., after iLUtc infection b\... more Heipcnirits cvprini (CHV) genome was traced in carp, Cvprinus carpio L., after iLUtc infection b\ the method of in \itu hybridization with biotinylated probes, Xhc viral Tnd spinal ner\'cs. However, ut this stage, viral antigens were nttt detected and the \irus wds ner\es ind is issdei ited with the induelion ind reeunencc of p ipillom is

Veterinary Immunology and Immunopathology, Jul 1, 1995

A new fluorochromasia method using a fluorescence microplate reader has been established to assay... more A new fluorochromasia method using a fluorescence microplate reader has been established to assay spontaneous cytotoxic activity of carp leucocytes. This method is characterized by using propidium iodide (PI) for staining dead target cells and a fluorescence microplate reader for measurement of the fluorescence of PI. K562 as target cells were prepared in 96-well flat-bottom microplates, and carp leucocytes were added as effector cells. After 2.5 h incubation, PI was added to each well. After an additional 1.5 h incubation, fluorescence of each well was measured. Correlation between this method and 51Cr-release assay was obtained. The results demonstrated that this new fluorochromasia method can be used to assay cytotoxic activity of carp leucocytes.

Journal of Shellfish Research, 2015

ABSTRACT The vasa orthologs have been used as a specific germ line molecular marker in many anima... more ABSTRACT The vasa orthologs have been used as a specific germ line molecular marker in many animal species. In this study, the expression of the vasa ortholog (povlg1) in adult and juvenile pearl oysters (Pinctada fucata) was observed by in situ hybridization. The in situ hybridization with povlg1 made it possible to detect the immature germ cells, which could not be detected by hematoxylin and eosin staining. During the reproductive season (May to July), spermatogonia, spermatocytes, oogonia, and oocytes showed povlg1 expression in the gonads of mature adult pearl oysters. By contrast, in spent pearl oysters in the nonreproductive season (September and October), only small ovoid (8.9×5.2 mm) povlg1-positive cells were observed in the base of acini. In juvenile (1 mo old) pearl oysters, a clump of germ cells first formed from several cells that were distributed symmetrically and lateral to the visceral mass. These cells then migrated to the ventrolateral periphery of the visceral mass in 2-mo-old oysters. The cells then migrated posteriorly along the periphery of the visceral mass with increasing cell numbers and size in 4-mo-old oysters. This observation of immature germ cell distribution and migration provides useful information for the control of gametogenesis and about pearl quality.

Veterinary Immunology and Immunopathology, 1995

A new fluorochromasia method using a fluorescence microplate reader has been established to assay... more A new fluorochromasia method using a fluorescence microplate reader has been established to assay spontaneous cytotoxic activity of carp leucocytes. This method is characterized by using propidium iodide (PI) for staining dead target cells and a fluorescence microplate reader for measurement of the fluorescence of PI. K562 as target cells were prepared in 96-well flat-bottom microplates, and carp leucocytes were added as effector cells. After 2.5 h incubation, PI was added to each well. After an additional 1.5 h incubation, fluorescence of each well was measured. Correlation between this method and 51Cr-release assay was obtained. The results demonstrated that this new fluorochromasia method can be used to assay cytotoxic activity of carp leucocytes.

Aquaculture, 1995

To assess cytotoxic activity of carp (Cyprinus carpio), head kidney leukocytes were examined with... more To assess cytotoxic activity of carp (Cyprinus carpio), head kidney leukocytes were examined with special reference to the effects of rearing water temperature and of assay temperature. Leukocytes as effector cells were separated from head kidney using Histopaque 1077 and cytotoxic activities were measured by the release of 51Cr from target cells which were K562, human chronic myelogenous leukemia cells. Cytotoxic activity of leukocytes from carp kept at higher temperature (25 °C) was lower at lower assay temperature (10 °C) than at higher assay temperature (25 °C). However, activity from carp acclimated to low temperature (10 °C) increased on 10 °C assay and decreased on 25 °C assay. Cellular composition of effector cells changed in connection with the above phenomenon. In carp acclimated to 10 °C, small lymphocytes decreased while the others, e.g. large lymphocytes, granulocytes and macrophages, increased. Accommodation of carp natural killer-like cells to environmental temperatures may be regulated by changing the cellular composition of effector cells.

Fish & Shellfish Immunology, 2014

Protective efficacies of three antigenic proteins (3-hydroxyacyl-CoA dehydrogenase (HCD), ATP syn... more Protective efficacies of three antigenic proteins (3-hydroxyacyl-CoA dehydrogenase (HCD), ATP synthase beta subunit (atpD), and glutamate dehydrogenase (gdhA)) against Flavobacterium psychrophilum were investigated in ayu (Plecoglossus altivelis). Recombinant proteins of HCD, atpD, and gdhA were expressed in Escherichia coli BL21 cells. Ayu were then vaccinated with inactivated cells via the intraperitoneal route. Compared with the empty BL21- and PBS-injected groups, the vaccinated group had a significantly longer survival time after challenge with F. psychrophilum. The antibody titers against each recombinant protein were significantly higher in serum from vaccinated fish, compared with serum from control fish. Results of indirect immunofluorescence assays using serum indicated that the HCD, atpD, and gdhA proteins are located on the surface of F. psychrophilum. These results suggest that these three surface proteins are protective antigens and are good candidates for development of vaccines against bacterial cold-water disease in ayu.

Zoological Science, 2008

In many bivalve species, paternal and maternal mitochondrial DNA (mtDNA) from sperm and eggs is t... more In many bivalve species, paternal and maternal mitochondrial DNA (mtDNA) from sperm and eggs is transmitted to the offspring. This phenomenon is known as doubly uniparental inheritance (DUI). In these species, sperm mtDNA (M type) is inherited by the male gonad of the offspring. Egg mtDNA (F type) is inherited by both male and female somatic cells and female gonadal cells. In Mytilidae, sperm mitochondria are distributed in the cytoplasm of differentiating male germ cells because they are transmitted to the male gonad. In the present study, we investigated maternal inheritance of mtDNA in the Pacific oyster, Crassostrea gigas. Sequence analysis of two mitochondrial non-coding regions revealed an identical sequence pattern in the gametes and adductor muscle samples taken from six males and five females. To observe whether sperm mitochondria were specifically located in the cytoplasm of differentiating germ cells, their distribution was recorded in C. gigas fertilized eggs by vital staining with MitoTracker Green. Although the 1D blastomere was identified in the cytoplasm of differentiating germ cells, sperm mitochondria were located at the 1D blastomere in only 32% of eggs during the 8-cell stage. Thus, in C. gigas, sperm mitochondria do not specifically locate in the germ cell region at the 1D blastomere. We suggest that the distribution of sperm mitochondria is not associated with germ cell formation in C. gigas. Furthermore, as evidenced by the mtDNA sequences of two non-coding regions, we conclude that mitochondrial DNA is maternally inherited in this species.

Development, Growth & Differentiation, 2011

Doubly uniparental inheritance (DUI) of mitochondrial (mt) DNA has been reported in the blue muss... more Doubly uniparental inheritance (DUI) of mitochondrial (mt) DNA has been reported in the blue mussel Mytilus galloprovincialis. In DUI, males inherit both paternal (M type) and maternal (F type) mtDNA. Here we investigated changes in M type mtDNA copy numbers and mitochondrial mass in testicular cells by real-time polymerase chain reaction and flow cytometry. The ratios of M type mtDNA copy numbers to nuclear DNA content were not different between haploid (1n), diploid (2n) and tetraploid (4n) spermatogenic cells. The mitochondrial mass decreased gradually during spermatogenesis. These results suggest that mtDNA and mitochondrial mass are maintained during spermatogenesis. We then traced M type mtDNA in larvae after fertilization. M type mtDNA was maintained up to 24 h after fertilization in the male-biased crosses, but decreased significantly in femalebiased crosses (predicted by Mito Tracker staining pattern). These results are strikingly different from those reported for mammals and fish, where it is well known that the mitochondria and mtDNA are reduced during spermatogenesis and that sperm mitochondria and mtDNA are eliminated soon after fertilization. Thus, the M type mtDNA copy number is maintained during spermatogenesis and in the development of male larvae to sustain the DUI system in the blue mussel.

Journal of Fish Diseases, Mar 1, 1992

A method of in situ hybridization with biotinylatcd probes is described for the specific detectio... more A method of in situ hybridization with biotinylatcd probes is described for the specific detection of the Herpesvirus cvprini {CHV) genome, both in CHV infeeted eell eultures and tissue speeimens prepared from earp fry infeeted with the same virus. From moleeularly eloned Pstl-fragments of CHV DNA. two fragments were selected as probes for in situ hybridization which had no deteetable homology, cither with DNA of fathead minnow (FHM) eells used for virus propagation, or with the DNA of several fish herpesviruses. The method of in situ hybridization was based on a system using streptavidinbiotinylated alkaline phosphatase for the detection of hybridization.

Aquaculture, 1995

To assess cytotoxic activity of carp (Cyprinus carpio), head kidney leukocytes were examined with... more To assess cytotoxic activity of carp (Cyprinus carpio), head kidney leukocytes were examined with special reference to the effects of rearing water temperature and of assay temperature. Leukocytes as effector cells were separated from head kidney using Histopaque 1077 and cytotoxic activities were measured by the release of 51Cr from target cells which were K562, human chronic myelogenous leukemia cells. Cytotoxic activity of leukocytes from carp kept at higher temperature (25 °C) was lower at lower assay temperature (10 °C) than at higher assay temperature (25 °C). However, activity from carp acclimated to low temperature (10 °C) increased on 10 °C assay and decreased on 25 °C assay. Cellular composition of effector cells changed in connection with the above phenomenon. In carp acclimated to 10 °C, small lymphocytes decreased while the others, e.g. large lymphocytes, granulocytes and macrophages, increased. Accommodation of carp natural killer-like cells to environmental temperatures may be regulated by changing the cellular composition of effector cells.

CABI eBooks, 2017

The geographical distribution; economic impacts; diagnosis of the infection; pathology; pathophys... more The geographical distribution; economic impacts; diagnosis of the infection; pathology; pathophysiology; and the protective and control strategies against Oncorhynchus masou virus and Cyprinid herpesvirus are discussed.

Plankton and Benthos Research, Feb 28, 2020

Corbicula leana and C. fluminea are hermaphroditic and ovoviviparous freshwater clams. Although t... more Corbicula leana and C. fluminea are hermaphroditic and ovoviviparous freshwater clams. Although they are considered to reproduce using self-fertilization, the possibility of outcrossing was suggested due to lineage discordance between mitochondrial and genomic DNA. In these species, outcrossing means egg parasitism by the spermatozoa from one or more other clams, because they reproduce by androgenesis in which only the nucleus of spermatozoa is transmitted to the progeny. Moreover, the presence of males in these species was reported in the previous study, and they were estimated to reproduce by egg parasitism of hermaphrodites. In this study, we investigated the paternity of juveniles in the brood pouches of six hermaphrodites by comparing the genotypes of the brooded juveniles, brooding clams, and neighboring adult clams using two microsatellite DNA markers. Brooded juveniles showed either identical genotypes to the parental clam or different genotypes from their parent in one clam with brooding. The genotypes of brooded juveniles were identical to those of neighboring hermaphrodites and males. These results indicate that androgenetic Corbicula reproduce not only by self-fertilization but also by egg parasitism, with outcrossing among hermaphrodites and from males to hermaphrodites.

Journal of Fish Diseases, 1993

Heipcnirits cvprini (CHV) genome was traced in carp, Cvprinus carpio L., after iLUtc infection b\... more Heipcnirits cvprini (CHV) genome was traced in carp, Cvprinus carpio L., after iLUtc infection b\ the method of in \itu hybridization with biotinylated probes, Xhc viral Tnd spinal ner\'cs. However, ut this stage, viral antigens were nttt detected and the \irus wds ner\es ind is issdei ited with the induelion ind reeunencc of p ipillom is

Veterinary Immunology and Immunopathology, Jul 1, 1995

A new fluorochromasia method using a fluorescence microplate reader has been established to assay... more A new fluorochromasia method using a fluorescence microplate reader has been established to assay spontaneous cytotoxic activity of carp leucocytes. This method is characterized by using propidium iodide (PI) for staining dead target cells and a fluorescence microplate reader for measurement of the fluorescence of PI. K562 as target cells were prepared in 96-well flat-bottom microplates, and carp leucocytes were added as effector cells. After 2.5 h incubation, PI was added to each well. After an additional 1.5 h incubation, fluorescence of each well was measured. Correlation between this method and 51Cr-release assay was obtained. The results demonstrated that this new fluorochromasia method can be used to assay cytotoxic activity of carp leucocytes.

Journal of Shellfish Research, 2015

ABSTRACT The vasa orthologs have been used as a specific germ line molecular marker in many anima... more ABSTRACT The vasa orthologs have been used as a specific germ line molecular marker in many animal species. In this study, the expression of the vasa ortholog (povlg1) in adult and juvenile pearl oysters (Pinctada fucata) was observed by in situ hybridization. The in situ hybridization with povlg1 made it possible to detect the immature germ cells, which could not be detected by hematoxylin and eosin staining. During the reproductive season (May to July), spermatogonia, spermatocytes, oogonia, and oocytes showed povlg1 expression in the gonads of mature adult pearl oysters. By contrast, in spent pearl oysters in the nonreproductive season (September and October), only small ovoid (8.9×5.2 mm) povlg1-positive cells were observed in the base of acini. In juvenile (1 mo old) pearl oysters, a clump of germ cells first formed from several cells that were distributed symmetrically and lateral to the visceral mass. These cells then migrated to the ventrolateral periphery of the visceral mass in 2-mo-old oysters. The cells then migrated posteriorly along the periphery of the visceral mass with increasing cell numbers and size in 4-mo-old oysters. This observation of immature germ cell distribution and migration provides useful information for the control of gametogenesis and about pearl quality.

Veterinary Immunology and Immunopathology, 1995

A new fluorochromasia method using a fluorescence microplate reader has been established to assay... more A new fluorochromasia method using a fluorescence microplate reader has been established to assay spontaneous cytotoxic activity of carp leucocytes. This method is characterized by using propidium iodide (PI) for staining dead target cells and a fluorescence microplate reader for measurement of the fluorescence of PI. K562 as target cells were prepared in 96-well flat-bottom microplates, and carp leucocytes were added as effector cells. After 2.5 h incubation, PI was added to each well. After an additional 1.5 h incubation, fluorescence of each well was measured. Correlation between this method and 51Cr-release assay was obtained. The results demonstrated that this new fluorochromasia method can be used to assay cytotoxic activity of carp leucocytes.

Aquaculture, 1995

To assess cytotoxic activity of carp (Cyprinus carpio), head kidney leukocytes were examined with... more To assess cytotoxic activity of carp (Cyprinus carpio), head kidney leukocytes were examined with special reference to the effects of rearing water temperature and of assay temperature. Leukocytes as effector cells were separated from head kidney using Histopaque 1077 and cytotoxic activities were measured by the release of 51Cr from target cells which were K562, human chronic myelogenous leukemia cells. Cytotoxic activity of leukocytes from carp kept at higher temperature (25 °C) was lower at lower assay temperature (10 °C) than at higher assay temperature (25 °C). However, activity from carp acclimated to low temperature (10 °C) increased on 10 °C assay and decreased on 25 °C assay. Cellular composition of effector cells changed in connection with the above phenomenon. In carp acclimated to 10 °C, small lymphocytes decreased while the others, e.g. large lymphocytes, granulocytes and macrophages, increased. Accommodation of carp natural killer-like cells to environmental temperatures may be regulated by changing the cellular composition of effector cells.

Fish & Shellfish Immunology, 2014

Protective efficacies of three antigenic proteins (3-hydroxyacyl-CoA dehydrogenase (HCD), ATP syn... more Protective efficacies of three antigenic proteins (3-hydroxyacyl-CoA dehydrogenase (HCD), ATP synthase beta subunit (atpD), and glutamate dehydrogenase (gdhA)) against Flavobacterium psychrophilum were investigated in ayu (Plecoglossus altivelis). Recombinant proteins of HCD, atpD, and gdhA were expressed in Escherichia coli BL21 cells. Ayu were then vaccinated with inactivated cells via the intraperitoneal route. Compared with the empty BL21- and PBS-injected groups, the vaccinated group had a significantly longer survival time after challenge with F. psychrophilum. The antibody titers against each recombinant protein were significantly higher in serum from vaccinated fish, compared with serum from control fish. Results of indirect immunofluorescence assays using serum indicated that the HCD, atpD, and gdhA proteins are located on the surface of F. psychrophilum. These results suggest that these three surface proteins are protective antigens and are good candidates for development of vaccines against bacterial cold-water disease in ayu.

Zoological Science, 2008

In many bivalve species, paternal and maternal mitochondrial DNA (mtDNA) from sperm and eggs is t... more In many bivalve species, paternal and maternal mitochondrial DNA (mtDNA) from sperm and eggs is transmitted to the offspring. This phenomenon is known as doubly uniparental inheritance (DUI). In these species, sperm mtDNA (M type) is inherited by the male gonad of the offspring. Egg mtDNA (F type) is inherited by both male and female somatic cells and female gonadal cells. In Mytilidae, sperm mitochondria are distributed in the cytoplasm of differentiating male germ cells because they are transmitted to the male gonad. In the present study, we investigated maternal inheritance of mtDNA in the Pacific oyster, Crassostrea gigas. Sequence analysis of two mitochondrial non-coding regions revealed an identical sequence pattern in the gametes and adductor muscle samples taken from six males and five females. To observe whether sperm mitochondria were specifically located in the cytoplasm of differentiating germ cells, their distribution was recorded in C. gigas fertilized eggs by vital staining with MitoTracker Green. Although the 1D blastomere was identified in the cytoplasm of differentiating germ cells, sperm mitochondria were located at the 1D blastomere in only 32% of eggs during the 8-cell stage. Thus, in C. gigas, sperm mitochondria do not specifically locate in the germ cell region at the 1D blastomere. We suggest that the distribution of sperm mitochondria is not associated with germ cell formation in C. gigas. Furthermore, as evidenced by the mtDNA sequences of two non-coding regions, we conclude that mitochondrial DNA is maternally inherited in this species.

Development, Growth & Differentiation, 2011

Doubly uniparental inheritance (DUI) of mitochondrial (mt) DNA has been reported in the blue muss... more Doubly uniparental inheritance (DUI) of mitochondrial (mt) DNA has been reported in the blue mussel Mytilus galloprovincialis. In DUI, males inherit both paternal (M type) and maternal (F type) mtDNA. Here we investigated changes in M type mtDNA copy numbers and mitochondrial mass in testicular cells by real-time polymerase chain reaction and flow cytometry. The ratios of M type mtDNA copy numbers to nuclear DNA content were not different between haploid (1n), diploid (2n) and tetraploid (4n) spermatogenic cells. The mitochondrial mass decreased gradually during spermatogenesis. These results suggest that mtDNA and mitochondrial mass are maintained during spermatogenesis. We then traced M type mtDNA in larvae after fertilization. M type mtDNA was maintained up to 24 h after fertilization in the male-biased crosses, but decreased significantly in femalebiased crosses (predicted by Mito Tracker staining pattern). These results are strikingly different from those reported for mammals and fish, where it is well known that the mitochondria and mtDNA are reduced during spermatogenesis and that sperm mitochondria and mtDNA are eliminated soon after fertilization. Thus, the M type mtDNA copy number is maintained during spermatogenesis and in the development of male larvae to sustain the DUI system in the blue mussel.