Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

DOI： 10.1016/j.placenta.2023.08.057

researchmap

Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

DOI： 10.1016/j.jri.2023.103883

researchmap

Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

DOI： 10.1016/j.jri.2022.103729

researchmap

Language：Japanese   Publisher：(公社)日本産科婦人科学会  

researchmap

Language：Japanese   Publisher：(公社)日本産科婦人科学会  

researchmap

Web of Science

researchmap

Language：English   Publisher：W B SAUNDERS CO LTD  

Web of Science

researchmap

Language：Japanese   Publisher：(一社)日本サイトメトリー学会  

researchmap

Language：Japanese   Publisher：(一社)日本内分泌外科学会  

researchmap

Language：Japanese   Publisher：(公社)日本臨床細胞学会  

researchmap

Language：Japanese   Publisher：(一社)日本内分泌外科学会  

researchmap

Language：Japanese   Publisher：(一社)日本サイトメトリー学会  

researchmap

Language：Japanese   Publisher：日本生殖免疫学会  

researchmap

Language：Japanese   Publisher：(一社)日本腎臓学会  

researchmap

Language：Japanese   Publisher：(公社)日本産科婦人科学会  

researchmap

Language：Japanese   Publisher：(公社)日本産科婦人科学会  

J-GLOBAL

researchmap

Language：Japanese   Publisher：(公社)日本産科婦人科学会  

CiNii Books

researchmap

Other Link： https://projects.repo.nii.ac.jp/?action=repository_uri&item_id=456675

Language：Japanese   Publisher：日本生殖免疫学会  

researchmap

Language：Japanese   Publisher：(株)メディカルレビュー社  

胎盤の網羅的マイクロRNA(miRNA)発現解析から、妊娠高血圧腎症(PE)の胎盤においてmiRNAの発現異常が起きており、異常miRNAの標的分子である胎盤特異的エストラジオール(E2)合成酵素(HSD17B1)の調節不全が引き起こされていることが明らかとなった。さらに、妊婦血液中でHSD17B1が検出可能であり、血漿HSD17B1値はPEの新たな予知マーカーになる可能性が示された。胎盤のトランスクリプトームにより、周産期医療のための新しい予知・診断ツールの開発がさらに進展することが期待される。(著者抄録)

researchmap

Language：Japanese   Publisher：日本産婦人科・新生児血液学会  

妊娠高血圧腎症の胎盤でmicroRNA(miRNA)の発現異常が起きており、異常を示しているmiRNAの標的分子である胎盤特異的estradiol合成酵素、17β-水酸化ステロイド脱水素酵素(HSD17B1)の調節不全が引き起こされていることが明らかとなった。妊婦血液中でHSD17B1が検出可能であり、血漿HSD17B1は、妊娠高血圧腎症の新たな予知マーカーになる可能性が示された。採血というルーチン検査によって胎盤由来のmiRNAおよびその標的分子の情報を得ることができ、周産期医療のための新しい予知・診断ツールとしての開発が期待される。(著者抄録)

researchmap

Language：Japanese   Publisher：(株)メディカルドゥ  

第19番染色体上でmicroRNA(miRNA)がクラスターを形成している領域に由来し,胎盤において特異的に発現しているmiRNAが同定された。妊娠高血圧症候群の胎盤において発現異常を認めるmiRNAにより引き起こされる胎盤機能異常がはじめて明らかにされた。妊娠期間中,この胎盤特異的miRNAがエクソソームを介して胎盤より放出され,母体血液中に移行して循環している。採血というルーチン検査によって胎盤由来のmiRNA情報を得ることができ,周産期医療のための新しい予知・診断ツールとして臨床応用が期待される。(著者抄録)

researchmap

Language：Japanese   Publisher：日本産婦人科・新生児血液学会  

researchmap

Language：Japanese   Publisher：(一社)日本解剖学会  

researchmap

Language：Japanese   Publisher：(一社)日本外科学会  

researchmap

Language：Japanese   Publisher：(公社)日本産科婦人科学会  

CiNii Books

researchmap

Other Link： https://projects.repo.nii.ac.jp/?action=repository_uri&item_id=448229

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：W B SAUNDERS CO LTD  

Web of Science

researchmap

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：ELSEVIER IRELAND LTD  

DOI： 10.1016/j.jri.2010.08.051

Web of Science

researchmap

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：W B SAUNDERS CO LTD  

Web of Science

researchmap

Language：Japanese   Publisher：医学書院  

DOI： 10.11477/mf.1542101381

CiNii Books

J-GLOBAL

researchmap

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：W B SAUNDERS CO LTD  

Web of Science

researchmap

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：W B SAUNDERS CO LTD  

Web of Science

researchmap

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：W B SAUNDERS CO LTD  

Web of Science

researchmap

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：W B SAUNDERS CO LTD  

Web of Science

researchmap

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：W B SAUNDERS CO LTD  

Web of Science

researchmap

Language：Japanese   Publisher：(公社)日本産科婦人科学会  

CiNii Books

researchmap

Other Link： https://projects.repo.nii.ac.jp/?action=repository_uri&item_id=437971

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：W B SAUNDERS CO LTD  

Web of Science

researchmap

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：W B SAUNDERS CO LTD  

Web of Science

researchmap

Language：English   Publisher：W B SAUNDERS CO LTD  

The purpose of the present study was to localize glucose-6-phosphatase (G6Pase) activity in the human placenta at various gestational stages and to compare them to pre-eclamptic placenta activity. Ultrastructural enzyme-histochemical analysis of G6Pase was performed using cerium and lead as capturing agents. Precipitates indicative of G6Pase activity were observed in the endoplasmic reticulum and the nuclear envelope of the syncytiotrophoblasts in near-term placenta obtained from women with normal pregnancies. In placenta taken from women with severe pre-eclampsia, the localization pattern, enzyme activity intensity, and morphology of the endoplasmic reticulum did not differ from normal pregnancies. Stringent control experiments were performed also to ensure specific detection of G6Pase activity. The results indicate that cytochemically detectable G6Pase is present in the human placenta. This enzyme may play significant roles in carbohydrate metabolism in the human placenta. (C) 1999 W. B. Saunders Company Ltd.

DOI： 10.1053/plac.1998.0346

Web of Science

researchmap

researchmap

Language：English   Publisher：JAPAN SOC HISTOCHEMISTRY & CYTOCHEMISTRY  

To improve the sensitivity of in situ hybridization (ISH) carried out using oligonucleotide probes without radionuclides, we employed a; new method entailing amplification of ISH signals by catalyzed reporter deposition. This method is based on biotin-tyramine-catalyzed signal amplification (BT-CSA) and was used to detect expression of a relatively scarce polynucleotide. kinesin 3A mRNA, in the central nervous system. Strong signals were detected in the hippocampus and in the granular cell layer of the cerebellum, but expression of kinesin 3A mRNA was not detected in Purkinje cells. ISH performed without the use of radionuclides has the advantage of being comparatively easy to perform, but has the disadvantage of low sensitivity. We showed that the low sensitivity can be overcome by using BT-CSA. Our method also had the advantage of enabling visualization of ISH signals via enzymatic reaction with horseradish peroxidase or by fluorescent staining.

DOI： 10.1267/ahc.32.275

Web of Science

researchmap

researchmap

Language：English   Publisher：HISTOCHEMICAL SOC INC  

We demonstrate a fluorescent ultrasmall immunogold probe, FluoroNanogold (FNG), to be a versatile reporter system for immunocytochemical labeling of ultrathin cryosections. FNG-labeled molecules in the same ultrathin cryosections can be resolved by two imaging techniques (i.e., fluorescence and electron microscopy). Lactoferrin, a marker protein for the specific granules in human neutrophils, was employed as the target for FNG immunolabeling. The spatial resolution of the fluorescence signal from FNG-labeled specific granules was compatible with that of silver-enhanced gold signal from the same granules in electron microscopy. Our results confirm that FNG can be used as a probe for high-resolution correlation between immunofluorescence and electron microscopy.

Web of Science

researchmap

Language：English   Publisher：HISTOCHEMICAL SOC INC  

This report describes the subcellular distribution of 5'-nucleotidase (5'-NT) in rat photoreceptor cells and pigment epithelial cells processed by rapid-freeze enzyme cytochemistry. There was a striking difference in the ultrastructural localization of 5'-NT activity between rod outer segments after freeze-substitution fixation and conventional fixation. By rapid-freezing enzyme cytochemistry, 5'-NT activity was localized in the extradiscal space of intact nonvacuolated discs, whereas by conventional cytochemistry it was shown in the intradiscal space of artifactual vacuolated discs. In the freeze-substituted retinal cells, an appreciable difference in functional 5'-NT molecules was also found. The soluble 5'-NT on the cytoplasmic side of the disc membrane was vital in the rod outer segments, whereas the membrane-bound ecto-5'-NT on the exoplasmic (external) surface of the apical process was active in the pigment epithelial cells. Rapid-freezing enzyme cytochemistry should be worth employing as a method to reveal the fine localization of enzyme activity at the level of cell ultrastructures, which are poorly preserved by conventional fixation, and should provide information approximate to that in living cells.

DOI： 10.1177/002215549804600914

Web of Science

researchmap

Language：English   Publisher：WILEY-LISS  

In this article, we review the immunocytochemical literature With respect to a comparison between conventional colloidal gold and ultrasmall gold particles as immunoprobes. We discuss the relative advantages and disadvantages of each of these types of particles for immunocytochemical applications. We present results from our own laboratories, in which we compared these immunoprobes in selected experimental situations. In addition, we discuss our work on the use of a fluorescently labeled ultrasmall immunoprobe for correlative microscopy. Microsc. Res. Tech. 42:13-23, 1998. (C) 1998 Wiley-Liss, Inc.

DOI： 10.1002/(SICI)1097-0029(19980701)42:1<13::AID-JEMT3>3.3.CO;2-M

Web of Science

PubMed

researchmap

Language：English   Publisher：ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD  

Adeno-associated virus (AAV) vectors, derived from a non-pathogenic parvovirus, are considered to be an appropriate gene transfer vehicle for both dividing and non-dividing cells. In the present study, we investigated whether the rat heart could be efficiently transduced with AAV vectors. Rat cardiac myocytes (CM) were infected with AAV-lacZ vector containing beta-galactosidase (beta-gal) gene in vitro, and the expression of beta-gal in CM was evaluated by X-gal staining and beta-gal ELISA, With increasing multiplicities of Infection (MOI), more than 60% of CM were stained positively with X-gal, and the beta-gal expression increased to 31.1 +/- 4.6 ng/mg protein in a MOI-dependent manner (MOI: 10(4) to 10(6) particles/cell). The beta-gal expression was also increased in an incubation period-dependent manner (1-24 h). beta-gal expression was maximal at day 3 and then gradually decreased with time, However, beta-gal expression at day 14 was almost the same level as that at day 1 (45.5 +/- 5.9 v 55.2 +/- 6.2 ng/mg protein). Excised rat right ventricular papillary muscles were also infected with AAV-lacZ ex vivo, When the beta-gal expression was evaluated by X-gal staining, more than 80% of CM in the papillary muscles were stained positively, indicating efficient gene transfer into CM using AAV vectors. These findings suggest that AAV vectors are promising for cardiac gene therapy, (C) 1998 Academic Press.

DOI： 10.1006/jmcc.1998.0697

Web of Science

researchmap

Language：English   Publisher：WILEY-LISS  

Human scalp hair is important as a diagnostic clue to many diseases, in medical jurisprudential investigations, and also as a subject of cosmetic treatments. While many ultrastructural studies of the human hair root including the hair follicle have been reported, few studies have been done on the human hair shaft. We report here the ultrastructure of human scalp hair shafts prepared by a rapid-freezing technique followed by freeze-substitution fixation that allows the observation of fine cell structures.
Healthy scalp hair shafts from Japanese females 12-13 years of age were rapid-frozen and then freeze-substituted in OsO4-acetone. In addition, this technique was applied to the study of some changes of the hair shafts (i.e., hair damaged by thioglycolic acid cold permanent waving and white hair).
By this method, the hair shaft was rapid-frozen throughout without appreciable ice damage although the hair shaft was nearly 100 mu m in diameter. The rapid-freezing technique resulted in excellent preservation of the ultrastructure of the hair shafts: lamellar structures in the cuticle and fine fibrous ultrastructures in the cortex were observed without chemical treatments. Thioglycolic acid treatment affected the ultrastructure of both the cuticle and the cortex. Except for the absence of melanin granules, no significant differences in the ultrastructure were observed between white hair and black hair.
The rapid-freezing technique followed by freeze-substitution fixation appears to be the most reliable approach for the morphological evaluation of fully keratinized cells and tissues. Anat. Rec. 251:406-413, 1998. (C) 1998 Wiley-Liss, Inc.

DOI： 10.1002/(SICI)1097-0185(199807)251:3<406::AID-AR17>3.0.CO;2-S

Web of Science

researchmap

Language：English   Publisher：UNIV BASQUE COUNTRY PRESS  

To examine the presence and distribution of Six family gene products in a variety of tissues at various developmental stages and in various cell types, we prepared specific antibodies against recombinant Six gene products. The distribution of Six2 and Six4 was examined by immunostaining in the developing mouse embryo. Production of Six2 was detected at E8.5 mainly in the mesenchyme, while Six4 was present in nuclei of neuronal cells in the peripheral region of the mantle layer of developing brain and spinal cord and in various ganglia at E10.5 and E11.5. Specific DNA binding activities of the Six proteins were analyzed by gel retardation super-shift assays using nuclear extracts from different rat tissues and cell lines. Six5 was the dominant isoform observed in the adult kidney, liver and lung but not in the brain. Six4 was not detected in all tested adult tissues, however, it was present in embryonic (FD21) lung nuclear extracts. In contrast, Six4 was detected in a variety of cultured cell lines, including HeLa, 3T3, MDCK and C2C12. Our results suggest that Six4 plays a specific role in the differentiation or maturation of neuronal cells, while Six5 is an adult type Six gene isoform product and is distributed in the kidney, liver and lung.

Web of Science

researchmap

Language：English   Publisher：SPANDIDOS PUBL LTD  

We describe a predictive marker (CD95) for the responsiveness to tretinoin (RA) in acute promyelocytic leukemia (APL). Functional CD95 expression during RA treatment have been observed only in those patients who responded to RA. Expression of CD95 (Fas antigen), which plays a major role in apoptosis, was determined by fluorescence activated cell sorter (FACS) analysis. APL cases in which no enhancement of CD95 expression was observed showed no response to RA and did not obtain complete remission. We propose that CD95 can predict the clinical response to RA probably due to differentiation.

Web of Science

researchmap

Language：English   Publisher：HISTOCHEMICAL SOC INC  

We describe a new freeze-fracture cytochemical technique consisting of combined immunocytochemistry and enzyme cytochemistry. This technique reveals the relationship between molecules in biological membranes by double labeling with two different cytochemical markers (i.e., immunogold probes and cerium). In this method, antigens were detected with specific primary antibodies and appropriate secondary immunoprobes, Subsequently, alkaline phosphatase activity was detected with cerium as the capture agent on the same replicas. Octyl-glucoside (OG) digestion before the cytochemical reactions was crucial to the success of this combined method. OG is an efficient detergent and OG digestion can preserve both immunocytochemical antigenicity and enzyme activity on replicas. As an initial examination, we applied this technique to the study of glycosyl-phosphatidyl-inositol-anchored proteins and adhesion molecules in human neutrophils. The method described here should serve as a unique additional approach for the study of topology and dynamics of molecules in biomembranes.

DOI： 10.1177/002215549804600103

Web of Science

researchmap

Language：English   Publisher：JAPAN SOC HISTOCHEMISTRY & CYTOCHEMISTRY  

Although many hair proteins have been investigated biochemically, little information is available on their subcellular distribution in human hair. We report here on the immunoelectron microscopic technique for defining the ultrastructural localization of hair proteins, especially hair keratins, in human scalp hair shafts: a combination of a rapid-freezing, freeze-substitution fixation without chemical fixatives, and subsequent immunocytochemistry (i.e., rapid-freezing immunocytochemistry). The hair shafts were rapid-frozen, freeze-substituted in acetone without chemical fixatives, and then embedded in LR Whiteresin. Subsequently, ultrathin-sectioned samples were stained for hair keratins by an immunogold technique. Rapid-freezing followed by freeze-substitution without chemical fixatives well-preserved not only the fine structure of the hair shafts but also the antigenicity of hair keratins for immunocytochemistry. In the cortex, hair keratins were present mainly on the macrofibrils. In the cuticle, they were also located primarily in the endocuticle, which did not show the fibrous structure like the macrofibrils did. Rapid-freezing immunocytochemistry appears to be the most viable approach for revealing the macromolecular architecture of human hair, which is a completely keratinized tissue and one of the most delicate tissues in preparation for transmission electron microscopy.

DOI： 10.1267/ahc.31.237

Web of Science

researchmap

Language：English   Publisher：OXFORD UNIV PRESS  

Cyclic guanosine monophosphate (cGMP)-phosphodiesterase (PDE) in the outer segment of vertebrate retinal rod cells is one of key enzymes mediating phototransduction. We report here on light-induced PDE activity in intact rat retinal rod cells processed by rapid-freezing enzyme cytochemistry, a new morphological technique that is a combination of rapid-freezing, freeze-substitution fixation, and subsequent enzyme cytochemistry. This technique quickly immobilizes and preserves both enzyme activity and the cell ultrastructure in a state approximating living conditions; consequently, it has proved useful for cytochemical detection of light-induced PDE activity. Using this technique we observed that the catalytic site of PDE molecules in rapid-frozen outer segments was predominantly located in the extradiscal spaces; PDE activity was significantly greater in light than in darkness; and illumination elicited marked increases in PDE activity in dark-adapted cells. Light-induced PDE activity was first cytochemically detected after 3 s of illumination and reached a peak within 5 s, at which time it was in virtually the same as that seen in fully light-adapted cells. In addition, cytochemical guanosine triphosphatase (GTPase) activity in dark-adapted cells, as well as corresponding PDE activity, increased in a time-dependent manner with illumination duration; this acceleration in GTPase activity closely paralleled the PDE activity. Thus, our results suggest, in part, the existence of reciprocal regulation of PDE and activated transducin a subunit, thereby modulating light adaptation in rod cells.

Web of Science

researchmap

Language：English   Publisher：JAPAN SOC HISTOCHEMISTRY & CYTOCHEMISTRY  

The purpose of this study was to characterize the ultrastructural and enzyme-histochemical features of Hofbauer cells in the placental villi of second-trimester abortions. Hofbauer cells in the abortive placenta had more numerous cytoplasmic processes and more prominent phagosomes as compared to gestational age-matched normal control placenta. Acid phosphatase activity was demonstrated in both the lysosomes and on phagosomal membranes of these cells. Our results indicate that Hofbauer cells in second-trimester abortions are activated or stimulated in situ. These activated Hofbauer cells might play important roles in the pathophysiology of preterm parturition.

DOI： 10.1267/ahc.31.447

Web of Science

researchmap

researchmap

Language：English  

In this mini-review, we describe new enzyme cytochemical methodology combined with cryotechniques, which we have developed in order to reveal the ultrastructural localization of enzymes in biological membranes. This combined approach has been shown to be a valuable tool to study topology, dynamics, and function of enzymes in a more lifelike state.

Scopus

PubMed

researchmap

researchmap

researchmap

Language：English   Publisher：ELSEVIER SCIENCE BV  

Objectives: Recombinant viral vectors based on the nonpathogenic parvovirus, adeno-associated virus (AAV), have a number of attractive features for gene therapy, including the ability to transduce non-dividing cells and its long-term transgene expression. In this study, an AAV vector containing bacterial beta-galactosidase gene (lacZ) was used to transduce cultured rat vascular smooth muscle cells (VSMC) in vitro and rat thoracic aortas ex vivo. Methods: VSMC were transduced with AAV-lacZ at multiplicities of infection (MOI) ranging from 5.0 x 10(5) to 1.0 x 10(7). Expression of beta-galactosidase (beta-gal) in VSMC was evaluated by X-gal staining and a beta-gal ELISA method. Excised rat aortas were incubated with medium containing AAV-lacZ. Expression of beta-gal in the aortic segments was evaluated by X-gal staining. Results: With increasing MOI, up to 50% of cultured VSMC were positive by X-gal staining and the beta-gal expression increased up-to 15 ng/mg protein. The expression gradually decreased during the culture but was detectable for at least 1 month. In the ex vivo study, AAV vectors transduced endothelial and adventitial cells in rat aortic segments, while no expression was seen in medial VSMC. Conclusions: AAV vectors can efficiently transduce rat VSMC in vitro. AAV-mediated ex vivo gene transfer into the normal aorta resulted in efficient gene transfer into endothelial and adventitial cells but not into medial VSMC. These findings suggest that AAV-based vectors are promising for use in cardiovascular gene therapy. (C) 1997 Elsevier Science B.V.

DOI： 10.1016/S0008-6363(97)00163-6

Web of Science

researchmap

Language：English   Publisher：JAPAN SOC HISTOCHEMISTRY & CYTOCHEMISTRY  

During the past decade, freeze-fracture cytochemistry, the combination of freeze-fracture electron microscopy with cytochemistry, has greatly contributed to the investigation of the macromolecular architecture of biological membranes. The application of enzyme cytochemical labeling technique to freeze-fracture cytochemistry (i.e., freeze-fracture enzyme cytochemistry) has not received considerable attention although by enzyme cytochemical labeling technique many enzyme molecules can be labeled with metal cations and enzyme cytochemistry has been of great utility in cell biology studies. In this study, we report freeze-fracture enzyme cytochemistry: ''label-fracture method'' and ''fracture-label method''. Freeze-fracture enzyme cytochemistry was applied to the study of acid phosphatase, 5'-nucleotidase in the proximal tubular epithelium of the rat kidney, ecto-adenosine triphosphatase in human neutrophils and alkaline phosphatase in rat neutrophils, and was shown to be a technique that visualized these enzyme activities on replicas. Cerium as the capture agent was a useful enzyme cytochemical probe in this technique. Lead was applicable for labeling of plasma membrane-associated enzyme molecules. This technique leads to new applications that may extend the usefulness of freeze-fracture cytochemistry for the analysis of biomembrane structure.

DOI： 10.1267/ahc.30.77

Web of Science

researchmap

Language：English   Publisher：OXFORD UNIV PRESS  

We show a new freeze-fracture enzyme cytochemistry technique, i.e. fracture-labelling by enzyme cytochemistry. Freeze-fracture replication is carried out first and subsequently enzyme molecules in the split-membrane halves of cellular membranes are labelled with enzyme cytochemical markers. A replica-digestion treatment before the cytochemical reactions is a key step in this method. Triton X-100, saponin, and ultrasonication provided adequate cleaning of the replicas with good preservation of enzyme activity. Enzyme cytochemical fracture-labelling was applied to the study of alkaline phosphatase (ALPase)-positive granules in unstimulated human neutrophils. ALPase activity was found primarily on the exoplasmic membrane halves of intracellular small granules. Using X-ray microanalysis, we confirmed that the electron-dense deposit on replicas was the reaction product demonstrating ALPase activity. The results obtained by this method should provide unique information for the understanding of structure and function of biological membranes.

Web of Science

researchmap

researchmap

Language：English   Publisher：JAPAN SOC HISTOCHEMISTRY & CYTOCHEMISTRY  

The fusion of lysosomes to phagosomes was observed under high voltage electron microscopy, in 4 mu m thick rat retinal sections with the aid of acid phosphatase cytochemistry. The study of thick sections facilitates the observation of the moment of fusion in stereo view from two tilted pictures. From this study, the contents of the lysosome pored into the phagosome through the orifice. shortly after the collision of the two organelles. The hydrolytic enzymes such as acid phosphatase spread in a sheet under the limiting membrane of the phagosome to finally form a balloon of the reaction product. In some case the ballooning appeared to be doubled. The outer skin of the reaction product may be the result of a wrapping mechanism of phagolysosomes.

DOI： 10.1267/ahc.30.675

Web of Science

researchmap

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：AMER SOC CELL BIOLOGY  

Web of Science

researchmap

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：AMER SOC CELL BIOLOGY  

Web of Science

researchmap

Language：English   Publisher：Elsevier B.V.  

We identified five cDNA clones of the Sir gene family which are expressed in retina. They are Six2 Six3α and Six3β (which are derived from alternative splicing forms), Six5, and AREC3/Six4. All of these Six family genes possess extensive sequence similarity among each other in the so-homologous region (Six domain and homeodomain) but differ greatly in structure in some other regions. The amino acid sequence similarity of the so-homologous region to the previously identified AREC3/Six4 is 70.1% for Six2, 57.3% for Six3α and Six3β, and 70.3% for Six5. The expression of these genes was observed in inner and outer nuclear layer, ganglion cell layer, and pigment epithelium of mouse retina by in situ hybridization. The so-homologous region of each Six family protein has specific DNA binding activity. Six5 and Six2 bind to the same sequence as does AREC3/Six4, while Six3 does not. These observations suggest that some of the Six family genes can regulate the same target genes.

DOI： 10.1016/0014-5793(96)00899-X

Scopus

PubMed

researchmap

Language：English   Publisher：JAPANESE SOC ELECTRON MICRO BUS CENTER ACAD SOC JAPAN  

We present a novel freeze-fracture cytochemistry method based upon enzyme cytochemistry, By this method, freeze-fractured membranes are labeled with cerium as an enzyme cytochemical marker on replicas. The cerium capture method was suitable for freeze-fracture enzyme cytochemistry because the cerium phosphate reaction product is stable after replica-cleaning. As a model system, acid phosphatase, which is a well-known lysosomal marker, was detected on freeze-fractured membranes of lysosomes in the proximal tubular epithelium of the rat kidney, This technique should be a useful addition for analyzing the ultrastructure of biological membranes.

Web of Science

researchmap

researchmap

researchmap

researchmap

Language：English   Publisher：AMER PHYSIOLOGICAL SOC  

RACTK1 cDNA encodes a pH-sensitive K+ channel from the renal cortical collecting tubule cells, which is only localized in the kidney by Northern blots (Suzuki, M., K. Takahashi, M. Ikeda, H. Hayakawa, A. Ogawa, Y. Kawaguchi, and O. Sakai. Nature Lend. 367: 642-645, 1994). We further investigated the localization of the channel by immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR). Immunohistochemical analysis showed that the signal was detected in various epithelia, including the luminal membrane of the cortical collecting tubules, artery of the kidney, and pancreatic ducts. Interestingly, the signal was also detected in the coronary vascular smooth muscle and cerebral artery. These tissue distributions were confirmed by localization of mRNA determined by RT-PCR. These findings suggest that RACTK1 encoding pH-sensitive K+ channels is widely distributed in epithelia as well as in vascular smooth muscle cells.

Web of Science

researchmap

Language：English   Publisher：ELSEVIER SCIENCE BV  

In human neutrophils, alkaline phosphatase (AlkPase), a low-affinity receptor for IgG (FcRIIIB), and complement decay accelerating factor (DAF) are glycosyl-phosphatidylinositol (GPI)-anchored proteins. Varying greatly in biological function these three integral membrane proteins exhibit regulated cell surface expression in neutrophils. Defined by their common membrane-linkage motif, AlkPase, FcRIIIB, and DAF can be released from the lipid bilayer by the action of phosphatidylinositol-specific phospholipase C and are relatively resistant to low temperature extraction with Triton X-100 (TX-100). In this study we show that neutrophil AlkPase, FcRIII, and DAF display differential extractibility; they are relatively insensitive to TX-100 solubilization at 4 degrees C, but are readily extracted with TX-100 at 37 degrees C or by the detergent octyl glucoside at 4 degrees C. The differential extractibility of these GPI-anchored proteins is the same in unstimulated cells, where these proteins exist primarily in an intracellular pool, and stimulated cells, where they are expressed principally at the cell surface. However, no differential extraction effect is observed with two neutrophil transmembrane proteins, complement receptor 1 (CD35, CR1) and MHC Class I in either stimulated or unstimulated cells.

Web of Science

researchmap

DOI： 10.11410/kenbikyo1950.30.69

CiNii Books

researchmap

Language：Japanese  

CiNii Books

researchmap

Language：English   Publisher：HISTOCHEMICAL SOC INC  

We present a new application for the use of small immunogold particles (approximate to 1.4-nm diameter) for ultrastructural immunocytochemistry. These small gold particles have been used on ultra-thin cryosections in conjunction with a silver enhancement procedure that does not degrade ultrastructural detail. We have used the human neutrophil as a model system, in which known protein markers of two different cytoplasmic granules were localized, in the development of this procedure. The 1.4-nm immunogold particles coupled with silver enhancement yield intense labeling for localization of lactoferrin, a marker for the specific granules, and myeloperoxidase, a marker for the azurophil granules. Double labeling in which one antigen was visualized with 1.4-nm gold and silver enhancement and a second antigen was detected with colloidal gold-IgG on the same ultra-thin cryosection was successfully achieved. We also show that 1.4-nm diameter immunogold particles penetrate into cryosectioned neutrophils to a greater extent than 5-nm or 10-nm immunogold probes. These results show that small immunogold particles, along with silver enhancement, are a useful addition to the immunolabeling methods available for use with ultra-thin cryosections.

Web of Science

researchmap

Language：English   Publishing type：Article, review, commentary, editorial, etc. (scientific journal)   Publisher：HISTOCHEMICAL SOC INC  

One of the more routine steps associated with cryosectioning of ultra-thin and semi-thin sections for immunocytochemistry is the transfer of sections to EM grids or coverslips. In a study of the granules of human neutrophils, we have found that the composition of the transfer medium was crucial to the success of these experiments.

Web of Science

researchmap

researchmap

Language：English   Publisher：EDITIONS PHYSIQUE  

Web of Science

researchmap

Language：English   Publisher：HISTOCHEMICAL SOC INC  

We combined immunocytochemistry and enzyme cytochemistry to localize two different proteins on the same ultra-thin cryosection. In this method the immunocytochemical localization is visualized with colloidal gold probes and the enzyme cytochemical detection is achieved with cerium as the capture agent. The immunocytochemistry is conducted first so that any potential adverse effects of the enzyme cytochemical procedure will not alter the antibody binding properties of the cryosections.

Web of Science

researchmap

researchmap

Language：English   Publisher：JAPAN SOC HISTOCHEMISTRY & CYTOCHEMISTRY  

The precise establishment for the subcellular localization of guanylate cyclase would very much aid in clarifying a number of important involvements of cGMP metabolism on biological phenomena. Therefore, as reported, attempts were made to develop a cytochemical method for guanylate cyclase (GCase), using lead citrate as capture agent. Lead citrate acts as an inhibitor of GCase but 21.3% of its activity remains reserved in the tissue. The optimum pH examined on cytochemical sections was between 7.5 to 8.5, the manganese ion working as activator. In the rat retina given immersion fixation, the GCase activity was defined in the space of the disc membranes on the rod outer segment. However, after application of rapid freeze substitution fixation on the retina, the enzyme activity appeared at the cytoplasmic side of the disc membranes, showing good correlation with the biochemical estimations.

DOI： 10.1267/ahc.25.181

Web of Science

researchmap

researchmap

Language：English   Publisher：JAPAN SOC HISTOCHEMISTRY & CYTOCHEMISTRY  

It is of increasing interest to observe the precise localization of cyclic 3',5'-nucleotide phosphodiesterase (PDEase) activity in rod outer segments at the ultrastructural level. The cytochemical method of PDEase localization at the ultrastructural level was developed by Florendo et al.. However, there were two major difficulties in the use of snake venom as an exogenous 5'-Nucleotidase (5'-Nase): 1) the marked ultrastructural damage due to snake venom, and 2) the inadequate penetration of exogenous 5'-Nase into tissues. Therefore, two improvements for the demonstration of PDEase activity were carried out by the case of: 1) a purified 5'-Nase as exogenous 5'-Nase for a better preservation of cell morphology, and 2) 40 mum sections made with the use of a freezing microtome for a better penetration of 5'-Nase into the tissues.
PDEase activity using purified 5'-Nase was observed along the disc in the rod outer segment, and no retinal detachment was observed between the outer segments and pigment epithelium, the ultrastructure being excellently preserved, as compared with the use of snake venom. By applying this improved method to freeze-substitution technique, the catalytic site of PDEase was localized on the cytoplasmic surface of the disc membranes. This new technique would be useful in detecting the precise localization of PDEase activity.

DOI： 10.1267/ahc.25.697

Web of Science

researchmap

Language：English   Publisher：JAPAN SOC HISTOCHEMISTRY & CYTOCHEMISTRY  

Since cell function is supported by many constituent enzymes in organized array, it is important that the site of enzyme activities in combination with fine ultrastructures be studied.
In the retina, the cGMP metabolizing enzymes included in phosphodiesterase are estimated to play a key role in action potential formation. Therefore, the site and arrangement of these enzymes in the rod outer segment are of histochemical interest and have yet to be clarified. The new tool, freeze substitution enzyme histochemistry developed in our laboratory enables us to visualize the exact activity and also the sites of enzymes as they are in the living state. With this method, the active sites of guanylate cyclase and phosphodiesterase have been demonstrated, but before having been able to specify the precise active sites at the ultrastructured level, the contrast obtained by rapid freeze substitution enzyme histochemistry on the biological membranes had to be increased. This contrast enhancement has been successfully achieved applying higher concentrations of fixatives in combination with tannic acid. Other attempts were the extension of fixation time as well as the application of higher temperatures. Potassium ferrocyanide osmium post-fixation also appeared effective. With the contrast enhancement achieved by various combinations in pretreatment, the reaction products of guanylate cyclase and phosphodiesterase were demonstrated at the cytoplasmic side of the disc membranes, which result is in good agreement with the biochemical estimations. Rapid freeze substitution enzyme histochemistry may be useful in detecting the precise locations of substances in living conditions.

DOI： 10.1267/ahc.24.121

Web of Science

researchmap

researchmap

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：HISTOCHEMICAL SOC INC  

Web of Science

researchmap

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：HISTOCHEMICAL SOC INC  

Web of Science

researchmap

researchmap

researchmap

researchmap

researchmap

Language：English   Publisher：JAPAN SOC HISTOCHEMISTRY & CYTOCHEMISTRY  

DOI： 10.1267/ahc.22.139

Web of Science

researchmap

researchmap

researchmap

researchmap

researchmap

Event date： 2021.10

researchmap

Event date： 2018.9

researchmap