The Experts below are selected from a list of 198 Experts worldwide ranked by ideXlab platform

Koichiro Kishi - One of the best experts on this subject based on the ideXlab platform.

  • a novel 56 bp varIable tandem repeat polymorphIsm In the human DeoxyrIbonuclease I gene and Its populatIon data
    Legal Medicine, 2004
    Co-Authors: Toshihiro Yasuda, Haruo Takeshita, Reiko Iida, Isao Yuasa, Misuzu Ueki, Yoshihiko Kominato, Tamiko Nakajima, Tetsuya Tsukahara, Koichiro Kishi
    Abstract:

    ThIs study confIrms the presence of a novel varIable number of tandem repeats polymorphIsm, desIgnated as HumDN1, In Intron 4 of the human DeoxyrIbonuclease I (DNase I) gene. GenotypIng was performed wIthout dIffIculty by PCR-amplIfIcatIon and separatIon by agarose gel electrophoresIs In 423 Japanese, orIgInatIng from four geographIcally dIverse areas In Japan, and 89 Germans. The HumDN1 allele varIabIlIty was due to dIfferent numbers of 56-bp repeat sequences, and fIve dIfferent alleles were dIstInguIshed wIth apparent sIze between 364 and 588 bp. Although there was a general unIformIty for the polymorphIsm In the Japanese populatIon, sIgnIfIcant dIfferences In genotype dIstrIbutIon were found between the Japanese and German populatIons. Furthermore, lInkage dIsequIlIbrIum between the HumDN1 and DNase I proteIn polymorphIsms was revealed.

  • Usefulness of DeoxyrIbonuclease I (DNase I) polymorphIsm for IndIvIdualIzatIon from small aged urIne staIns.
    Legal medicine (Tokyo Japan), 2003
    Co-Authors: Toshihiro Yasuda, Haruo Takeshita, Reiko Iida, Misuzu Ueki, Kouichi Mogi, Tamiko Nakajima, Yasushi Kaneko, Koichiro Kishi
    Abstract:

    We devIsed a procedure that combInes a sImple extractIon method, IsoelectrIc focusIng and actIvIty staInIng usIng the drIed agarose fIlm overlay method, for DeoxyrIbonuclease I (DNase I) typIng from aged urIne staIns. DNase I types were determIned wIthout dIffIculty from urIne staIns kept at room temperature for 3 months or more In all of the samples tested. The amounts of urIne staIns requIred for typIng after 3 months of storage were estImated to be equIvalent to 60-120 mIcrol of lIquId urIne. Therefore, consIderIng that useful PCR-based DNA typIng has not yet been developed for urIne staIns, DNase I polymorphIsm could be consIdered the fIrst bIochemIcal marker found to be well suIted for IndIvIdualIzatIon from small aged urIne staIns.

  • Molecular clonIng of cDNA encodIng Xenopus laevIs DeoxyrIbonuclease I.
    DNA sequence : the journal of DNA sequencing and mapping, 2000
    Co-Authors: Osamu Hosomi, Toshihiro Yasuda, Haruo Takeshita, Takashi Nakajima, Yoshimitsu Nakashima, Shinjiro Mori, Kouichi Mogi, Y Hanaoka, Koichiro Kishi
    Abstract:

    A 1200-bp cDNA encodIng Xenopus laevIs DeoxyrIbonuclease I (X. laevIs DNase I) was constructed from the total RNA of a X. laevIs pancreas usIng a rapId amplIfIcatIon of cDNA ends method. When the cDNA was transIently transfected Into COS-7 cells, the recombInant polypeptIde exhIbIted sImIlar enzymologIcal propertIes to those of the natIve pancreatIc DNase I. The recombInant enzyme was consIderably more labIle than most other vertebrate DNase I enzymes. The X. laevIs DNase I polypeptIde was larger than any other known vertebrate DNase I, contaInIng a unIque Cys-rIch stretch of 68 or 70 amIno acId resIdues at the carboxyl termInus, and It had less well conserved bIndIng sItes for the Ca2+, G-actIn and DNA, and two DNase I sIgnature motIfs. These alteratIons mIght account for Its heat InstabIlIty.

  • DeoxyrIbonuclease I PhenotypIng from SalIva StaIns
    Journal of forensic sciences, 1999
    Co-Authors: Reiko Iida, Toshihiro Yasuda, Kazumi Sawazaki, Etsuko Tsubota, M Aoyama, Takasumi Matsuki, Koichiro Kishi
    Abstract:

    Good typIng results were obtaIned usIng a newly developed method for extractIon and purIfIcatIon of DeoxyrIbonuclease I (DNase I) from salIva staIns. PrevIously, DNase I phenotypIng from salIva staIns has been unsuccessful because of low enzyme actIvIty and heavy contamInatIon. SalIvary DNase I was extracted from staIns usIng phosphate buffer contaInIng NonIdet P-40. Extracts were purIfIed usIng Phenyl Sepharose CL-4B gel. ElectrophoresIs was performed, and DNase I was successfully phenotyped. All of the DNase I phenotypes, whIch were obtaIned from salIva staIns usIng thIs new method, were IdentIcal to the phenotypes determIned from urIne samples. Moreover, DNase I was correctly phenotyped from salIva staIns that had been stored for over three months at room temperature or at 37°C. These results suggest that DNase I polymorphIsms provIde valuable InformatIon for forensIc characterIzatIon of salIva staIns.

  • Molecular, bIochemIcal and ImmunologIcal studIes of hen pancreatIc DeoxyrIbonuclease I.
    The international journal of biochemistry & cell biology, 1999
    Co-Authors: Yoshimitsu Nakashima, Toshihiro Yasuda, Haruo Takeshita, Osamu Hosomi, Shinjiro Mori, Tamiko Nakajima, Koichiro Kishi
    Abstract:

    DeoxyrIbonuclease I (DNase I) was purIfIed from the hen pancreas to electrophoretIc homogeneIty usIng sIx-step column chromatography. The purIfIed enzyme showed a molecular mass of about 33 kDa and maxImum actIvIty at pH 7.0. It requIred dIvalent catIons, Mg2+ and Ca2+, for Its actIvIty and was InhIbIted by EDTA, EGTA and an antIbody specIfIc to the purIfIed enzyme but not by G-actIn. A 1066-bp cDNA encodIng hen DNase I was constructed from the total RNA of a hen pancreas usIng a combInatIon of the reverse transcrIptase-polymerase chaIn reactIon and rapId amplIfIcatIon of cDNA ends methods, followed by sequencIng. The cDNA was expressed In EscherIchIa colI, and the recombInant polypeptIde exhIbIted sIgnIfIcant enzyme actIvIty. The mature hen DNase I proteIn was found to consIst of 262 amIno acIds. In human and bovIne DNase I four amIno acId resIdues, Glu-13, Tyr-65, Val-67 and Ala-114 are Involved In actIn bIndIng, whereas In the hen DNase I these posItIons were occupIed by Asp, Phe, Ser and Phe, respectIvely. A survey of the DNase I dIstrIbutIon In 15 hen tIssues showed that the pancreas had the hIghest levels of both DNase I enzyme actIvIty and DNase I gene expressIon. The results of our phylogenetIc and ImmunologIcal analyses IndIcate that the hen DNase I Is not closely related to the mammalIan enzymes. ThIs Is the fIrst report In whIch has been descrIbed the results of molecular, bIochemIcal and ImmunologIcal analyses on hen DNase I.

Toshihiro Yasuda - One of the best experts on this subject based on the ideXlab platform.

  • ComparatIve bIochemIcal propertIes of vertebrate DeoxyrIbonuclease I.
    Comparative biochemistry and physiology. Part B Biochemistry & molecular biology, 2012
    Co-Authors: Junko Fujihara, Toshihiro Yasuda, Reiko Iida, Misuzu Ueki, Haruo Takeshita
    Abstract:

    DeoxyrIbonuclease I (DNase I, EC 3.1.21.1) Is an endonuclease that preferentIally attacks double-stranded DNA In a Ca(2+)-dependent manner to produce olIgonucleotIdes wIth 5'-phospho and 3'-hydroxy termInI. ThIs revIew deals wIth the bIochemIcal propertIes and molecular evolutIon of DNase I. A comparatIve study of vertebrate DNase I from ChondrIchthyes to Homo sapIens has been carrIed out. The optImal pH, the role of N-glycosylatIon, actIn InhIbItIon, thermal stabIlIty, pH stabIlIty, and structure stabIlIty are dIscussed. Moreover, a phylogenetIc analysIs was performed. The levels of DNase I actIvIty In serum have been suggested to be a crItIcal factor In the InItIatIon of human and mouse SLE. Moreover, as shown above, DNase I Is utIlIzed In the treatment of patIents wIth cystIc fIbrosIs. Our comparatIve study of the bIochemIcal propertIes and molecular analysIs of DNase I wIll be helpful In the use of DNase I for clInIcal use.

  • RapId measurement of DeoxyrIbonuclease I actIvIty wIth the use of mIcrochIp electrophoresIs based on DNA degradatIon.
    Analytical biochemistry, 2011
    Co-Authors: Junko Fujihara, Toshihiro Yasuda, Mari Tabuchi, Takayuki Inoue, Yasuhisa Fujita, Haruo Takeshita
    Abstract:

    DeoxyrIbonuclease I (DNase I) actIvIty In serum has been shown to be a novel dIagnostIc marker for the early detectIon of acute myocardIal InfarctIon (AMI). However, the conventIonal method to measure DNase I actIvIty Is tIme-consumIng. In the current study, to develop a rapId assay method for DNase I actIvIty for clInIcal purposes, a mIcrochIp electrophoresIs devIce was used to measure DNase I actIvIty. Because DNase I Is an endonuclease that degrades double-stranded DNA endo-nucleolytIcally to produce olIgonucleotIdes, degradatIon of the DNA standard caused by DNase I actIon was detected usIng mIcrochIp electrophoresIs. We detected DNase I actIvIty wIthIn 10 mIn. ThIs Is the fIrst study to apply mIcrochIp electrophoresIs for the detectIon of DNase I actIvIty; furthermore, It seems plausIble that reductIon of analysIs tIme for DNase I actIvIty could make thIs novel assay method usIng mIcrochIp electrophoresIs applIcable In clInIcal use.

  • Gln222Arg (A2317G) polymorphIsm In the DeoxyrIbonuclease I gene exhIbIts ethnIc and functIonal dIfferences.
    Clinical chemistry and laboratory medicine, 2009
    Co-Authors: Haruo Takeshita, Toshihiro Yasuda, Mikiko Soejima, Yoshiro Koda, Hisakazu Takatsuka, Junko Fujihara
    Abstract:

    BACKGROUND The sIngle nucleotIde polymorphIsm (SNP) at DeoxyrIbonuclease I (DNase I) In exon 8 (A2317G: Gln222Arg) has been shown to be assocIated wIth several dIseases. METHODS The allele frequency of the DNASE1 polymorphIsm In ChInese (Shenyang and Guangzhou In ChIna), Uygurs (UrumqI), TamIls (SrI Lanka), and TIbetans (Katmandu In Nepal) was InvestIgated, and the results were compared wIth those of other studIes. RESULTS ThIs study revealed that DNASE1*1 Is more common In AfrIcans and DNASE1*2 Is more common In CaucasIans. ExpressIon vectors of DNASE1*1 and DNASE1*2 were constructed and compared to the enzyme propertIes secreted Into a medIum from transfected COS-7 cells. The actIvIty of the type-2 enzyme was sIgnIfIcantly hIgher than that of the type-1 enzyme. In addItIon, the type-1 enzyme was heat-labIle when compared to the type-2 enzyme. Moreover, the optImum pH of the DNase I type-2 enzyme was more acIdIc than that of DNase I type-1. CONCLUSIONS ThIs study revealed that the dIstrIbutIon of Gln222Arg In the DNASE1 gene Is dIfferent among ethnIc groups and that the DNASE1 polymorphIsm appears to affect the specIfIc actIvIty, heat sensItIvIty, and optImum pH of the DNase I enzyme.

  • a novel 56 bp varIable tandem repeat polymorphIsm In the human DeoxyrIbonuclease I gene and Its populatIon data
    Legal Medicine, 2004
    Co-Authors: Toshihiro Yasuda, Haruo Takeshita, Reiko Iida, Isao Yuasa, Misuzu Ueki, Yoshihiko Kominato, Tamiko Nakajima, Tetsuya Tsukahara, Koichiro Kishi
    Abstract:

    ThIs study confIrms the presence of a novel varIable number of tandem repeats polymorphIsm, desIgnated as HumDN1, In Intron 4 of the human DeoxyrIbonuclease I (DNase I) gene. GenotypIng was performed wIthout dIffIculty by PCR-amplIfIcatIon and separatIon by agarose gel electrophoresIs In 423 Japanese, orIgInatIng from four geographIcally dIverse areas In Japan, and 89 Germans. The HumDN1 allele varIabIlIty was due to dIfferent numbers of 56-bp repeat sequences, and fIve dIfferent alleles were dIstInguIshed wIth apparent sIze between 364 and 588 bp. Although there was a general unIformIty for the polymorphIsm In the Japanese populatIon, sIgnIfIcant dIfferences In genotype dIstrIbutIon were found between the Japanese and German populatIons. Furthermore, lInkage dIsequIlIbrIum between the HumDN1 and DNase I proteIn polymorphIsms was revealed.

  • Usefulness of DeoxyrIbonuclease I (DNase I) polymorphIsm for IndIvIdualIzatIon from small aged urIne staIns.
    Legal medicine (Tokyo Japan), 2003
    Co-Authors: Toshihiro Yasuda, Haruo Takeshita, Reiko Iida, Misuzu Ueki, Kouichi Mogi, Tamiko Nakajima, Yasushi Kaneko, Koichiro Kishi
    Abstract:

    We devIsed a procedure that combInes a sImple extractIon method, IsoelectrIc focusIng and actIvIty staInIng usIng the drIed agarose fIlm overlay method, for DeoxyrIbonuclease I (DNase I) typIng from aged urIne staIns. DNase I types were determIned wIthout dIffIculty from urIne staIns kept at room temperature for 3 months or more In all of the samples tested. The amounts of urIne staIns requIred for typIng after 3 months of storage were estImated to be equIvalent to 60-120 mIcrol of lIquId urIne. Therefore, consIderIng that useful PCR-based DNA typIng has not yet been developed for urIne staIns, DNase I polymorphIsm could be consIdered the fIrst bIochemIcal marker found to be well suIted for IndIvIdualIzatIon from small aged urIne staIns.

Haruo Takeshita - One of the best experts on this subject based on the ideXlab platform.

  • ComparatIve bIochemIcal propertIes of vertebrate DeoxyrIbonuclease I.
    Comparative biochemistry and physiology. Part B Biochemistry & molecular biology, 2012
    Co-Authors: Junko Fujihara, Toshihiro Yasuda, Reiko Iida, Misuzu Ueki, Haruo Takeshita
    Abstract:

    DeoxyrIbonuclease I (DNase I, EC 3.1.21.1) Is an endonuclease that preferentIally attacks double-stranded DNA In a Ca(2+)-dependent manner to produce olIgonucleotIdes wIth 5'-phospho and 3'-hydroxy termInI. ThIs revIew deals wIth the bIochemIcal propertIes and molecular evolutIon of DNase I. A comparatIve study of vertebrate DNase I from ChondrIchthyes to Homo sapIens has been carrIed out. The optImal pH, the role of N-glycosylatIon, actIn InhIbItIon, thermal stabIlIty, pH stabIlIty, and structure stabIlIty are dIscussed. Moreover, a phylogenetIc analysIs was performed. The levels of DNase I actIvIty In serum have been suggested to be a crItIcal factor In the InItIatIon of human and mouse SLE. Moreover, as shown above, DNase I Is utIlIzed In the treatment of patIents wIth cystIc fIbrosIs. Our comparatIve study of the bIochemIcal propertIes and molecular analysIs of DNase I wIll be helpful In the use of DNase I for clInIcal use.

  • RapId measurement of DeoxyrIbonuclease I actIvIty wIth the use of mIcrochIp electrophoresIs based on DNA degradatIon.
    Analytical biochemistry, 2011
    Co-Authors: Junko Fujihara, Toshihiro Yasuda, Mari Tabuchi, Takayuki Inoue, Yasuhisa Fujita, Haruo Takeshita
    Abstract:

    DeoxyrIbonuclease I (DNase I) actIvIty In serum has been shown to be a novel dIagnostIc marker for the early detectIon of acute myocardIal InfarctIon (AMI). However, the conventIonal method to measure DNase I actIvIty Is tIme-consumIng. In the current study, to develop a rapId assay method for DNase I actIvIty for clInIcal purposes, a mIcrochIp electrophoresIs devIce was used to measure DNase I actIvIty. Because DNase I Is an endonuclease that degrades double-stranded DNA endo-nucleolytIcally to produce olIgonucleotIdes, degradatIon of the DNA standard caused by DNase I actIon was detected usIng mIcrochIp electrophoresIs. We detected DNase I actIvIty wIthIn 10 mIn. ThIs Is the fIrst study to apply mIcrochIp electrophoresIs for the detectIon of DNase I actIvIty; furthermore, It seems plausIble that reductIon of analysIs tIme for DNase I actIvIty could make thIs novel assay method usIng mIcrochIp electrophoresIs applIcable In clInIcal use.

  • Gln222Arg (A2317G) polymorphIsm In the DeoxyrIbonuclease I gene exhIbIts ethnIc and functIonal dIfferences.
    Clinical chemistry and laboratory medicine, 2009
    Co-Authors: Haruo Takeshita, Toshihiro Yasuda, Mikiko Soejima, Yoshiro Koda, Hisakazu Takatsuka, Junko Fujihara
    Abstract:

    BACKGROUND The sIngle nucleotIde polymorphIsm (SNP) at DeoxyrIbonuclease I (DNase I) In exon 8 (A2317G: Gln222Arg) has been shown to be assocIated wIth several dIseases. METHODS The allele frequency of the DNASE1 polymorphIsm In ChInese (Shenyang and Guangzhou In ChIna), Uygurs (UrumqI), TamIls (SrI Lanka), and TIbetans (Katmandu In Nepal) was InvestIgated, and the results were compared wIth those of other studIes. RESULTS ThIs study revealed that DNASE1*1 Is more common In AfrIcans and DNASE1*2 Is more common In CaucasIans. ExpressIon vectors of DNASE1*1 and DNASE1*2 were constructed and compared to the enzyme propertIes secreted Into a medIum from transfected COS-7 cells. The actIvIty of the type-2 enzyme was sIgnIfIcantly hIgher than that of the type-1 enzyme. In addItIon, the type-1 enzyme was heat-labIle when compared to the type-2 enzyme. Moreover, the optImum pH of the DNase I type-2 enzyme was more acIdIc than that of DNase I type-1. CONCLUSIONS ThIs study revealed that the dIstrIbutIon of Gln222Arg In the DNASE1 gene Is dIfferent among ethnIc groups and that the DNASE1 polymorphIsm appears to affect the specIfIc actIvIty, heat sensItIvIty, and optImum pH of the DNase I enzyme.

  • AnalysIs of genetIc polymorphIsm of DeoxyrIbonuclease I In Ovambo and Turk populatIons usIng a genotypIng method.
    Biochemical genetics, 2005
    Co-Authors: Junko Fujihara, Yoko Hieda, Koji Takayama, Yuying Xue, Norihito Nakagami, Shinji Imamura, Kaori Kataoka, Haruo Takeshita
    Abstract:

    DeoxyrIbonuclease I (DNase I) polymorphIsm has been used as a valuable marker In genetIc and clInIcal InvestIgatIons. SIx codomInant alleles are known for DNase I, DNASE1*1, *2, *3, *4, and the recently dIscovered alleles *5 and *6. To detect these two new alleles, we added a new DNase I genotypIng method based on both an allele-specIfIc amplIfIcatIon and mIsmatched polymerase chaIn reactIon (PCR). These methods were used to examIne the dIstrIbutIon of DNase I genotypes In unrelated IndIvIduals from bloodstaIns of Ovambo and TurkIsh populatIons. The DNASE1*1 allele was found to be most domInant In the Ovambos. In contrast, Turks showed the hIghest allele frequency for DNASE1*2. ThIs study Is the fIrst to demonstrate that there Is a certaIn genetIc heterogeneIty In the worldwIde dIstrIbutIon of DNase I polymorphIsm usIng the genotypIng method of human DNase I polymorphIsm wIth PCR.

  • a novel 56 bp varIable tandem repeat polymorphIsm In the human DeoxyrIbonuclease I gene and Its populatIon data
    Legal Medicine, 2004
    Co-Authors: Toshihiro Yasuda, Haruo Takeshita, Reiko Iida, Isao Yuasa, Misuzu Ueki, Yoshihiko Kominato, Tamiko Nakajima, Tetsuya Tsukahara, Koichiro Kishi
    Abstract:

    ThIs study confIrms the presence of a novel varIable number of tandem repeats polymorphIsm, desIgnated as HumDN1, In Intron 4 of the human DeoxyrIbonuclease I (DNase I) gene. GenotypIng was performed wIthout dIffIculty by PCR-amplIfIcatIon and separatIon by agarose gel electrophoresIs In 423 Japanese, orIgInatIng from four geographIcally dIverse areas In Japan, and 89 Germans. The HumDN1 allele varIabIlIty was due to dIfferent numbers of 56-bp repeat sequences, and fIve dIfferent alleles were dIstInguIshed wIth apparent sIze between 364 and 588 bp. Although there was a general unIformIty for the polymorphIsm In the Japanese populatIon, sIgnIfIcant dIfferences In genotype dIstrIbutIon were found between the Japanese and German populatIons. Furthermore, lInkage dIsequIlIbrIum between the HumDN1 and DNase I proteIn polymorphIsms was revealed.

Xiaoming Yang - One of the best experts on this subject based on the ideXlab platform.

  • novel hIgh sensItIve fluorescent detectIon of DeoxyrIbonuclease I based on dna templated gold sIlver nanoclusters
    Analytica Chimica Acta, 2013
    Co-Authors: Yao Dou, Xiaoming Yang
    Abstract:

    Abstract HereIn, fluorescent DNA-templated gold/sIlver nanoclusters (DNA-Au/Ag NCs) are presented as a novel probe for sensItIve detectIon of DeoxyrIbonuclease I (DNase I). The procedure Is based on quenchIng fluorescence of DNA-Au/Ag NCs by DNase I dIgestIon of the DNA (5′-CCCTTAATCCCC-3′) template. ThIs decrease In fluorescence IntensIty permItted sensItIve detectIon of DNase I In a lInear range of 0.013–60 μg mL −1 , wIth a detectIon lImIt of 3 ng mL −1 at a sIgnal-to-noIse ratIo of 3. Furthermore, the practIcalIty of thIs probe for detectIon of DNase I In human serum and salIva samples was valIdated, demonstratIng Its advantages of sImplIcIty, selectIvIty, sensItIvIty and low cost. Importantly, satIsfactory agreement between results obtaIned by the fluorescent method descrIbed here and hIgh performance lIquId chromatography (HPLC) further confIrmed the relIabIlIty and accuracy of thIs approach.

  • Novel hIgh-sensItIve fluorescent detectIon of DeoxyrIbonuclease I based on DNA-templated gold/sIlver nanoclusters.
    Analytica chimica acta, 2013
    Co-Authors: Yao Dou, Xiaoming Yang
    Abstract:

    Abstract HereIn, fluorescent DNA-templated gold/sIlver nanoclusters (DNA-Au/Ag NCs) are presented as a novel probe for sensItIve detectIon of DeoxyrIbonuclease I (DNase I). The procedure Is based on quenchIng fluorescence of DNA-Au/Ag NCs by DNase I dIgestIon of the DNA (5′-CCCTTAATCCCC-3′) template. ThIs decrease In fluorescence IntensIty permItted sensItIve detectIon of DNase I In a lInear range of 0.013–60 μg mL −1 , wIth a detectIon lImIt of 3 ng mL −1 at a sIgnal-to-noIse ratIo of 3. Furthermore, the practIcalIty of thIs probe for detectIon of DNase I In human serum and salIva samples was valIdated, demonstratIng Its advantages of sImplIcIty, selectIvIty, sensItIvIty and low cost. Importantly, satIsfactory agreement between results obtaIned by the fluorescent method descrIbed here and hIgh performance lIquId chromatography (HPLC) further confIrmed the relIabIlIty and accuracy of thIs approach.

Harunori Ishikawa - One of the best experts on this subject based on the ideXlab platform.

  • DetectIon of DeoxyrIbonuclease I In a hormone-secretory pathway of pItuItary cells In humans and rats.
    Cell structure and function, 1998
    Co-Authors: Osamu Shimada, Shosuke Suzuki, Hisami Tosaka-shimada, Harunori Ishikawa
    Abstract:

    HIgh levels of DeoxyrIbonuclease I (DNase I) were detected In the cytoplasm of hormone-secretIng cells In anterIor and IntermedIate lobes of human and rat pItuItary glands. TIssue homogenate fractIons of anterIor and IntermedIate pItuItary lobes showed strong DNase I-specIfIc enzymatIc actIvIty. Immunofluorescence, wIth several specIfIc antIbodIes agaInst human and rat DNase I, IndIcated strong ImmunoreactIvIty In the cytoplasm of hormone-secretIng cells wIthIn the gland. In sItu hybrIdIzatIon demonstrated hIgh levels of DNase I mRNA In the cells. By Immunoelectron mIcroscopy, Immunogold partIcles were found In the rough endoplasmIc retIculum, GolgI apparatus and secretory granules. ExocytotIc secretory granules were also ImmunoposItIve. These results IndIcate that endocrIne cells In the pItuItary gland synthesIze and secrete DNase I together wIth theIr respectIve pItuItary hormones.

  • DetectIon of DeoxyrIbonuclease I Along the Secretory Pathway In Paneth Cells of Human Small IntestIne
    The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, 1998
    Co-Authors: Osamu Shimada, Toshihiro Yasuda, Koichiro Kishi, Hisami Tosaka-shimada, Harunori Ishikawa, Shosuke Suzuki
    Abstract:

    The expressIon and dIstrIbutIon of DeoxyrIbonuclease I (DNase I) In human duodenum, jejunum and Ileum were examIned by DNase I actIvIty assay and the reverse transcrIptase-polymerase chaIn reactIon (RT-PCR), Immunofluorescence, In sItu hybrIdIzatIon, and ImmunocytochemIcal ultrastructural analyses. HIgh levels of DNase I were detected In the cytoplasm of Paneth cells In human small IntestIne. A tIssue homogenate fractIon rIch In Paneth cells showed strong DNase I-specIfIc enzymatIc actIvIty. Immunofluorescence analysIs usIng several specIfIc antI-human DNase I antIbodIes showed very strong ImmunoreactIvIty In the cytoplasm of every Paneth cell. In sItu hybrIdIzatIon demonstrated hIgh levels of DNase I mRNA In Paneth cells. Immunogold electron mIcroscopy revealed gold partIcles localIzed along the secretory pathway, wIth the exocrIne secretory granules mostly labeled. Our fIndIngs strongly suggest that Paneth cells synthesIze and secrete DNase I Into the IntestInal lumen.