巻号一覧
23 巻 (1990)
- 6 号 p. 753-
- 5 号 p. 601-
- 4 号 p. 427-
- 3 号 p. 261-
- 2 号 p. 139-
- 1 号 p. 1-
23 巻, 3 号
選択された号の論文の13件中1～13を表示しています
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YOHEI HOSOKAWA, YASUNARI TSUCHIHASHI, SHOJI MITSUFUJI, KAZUHIKO TOKITA ...1990 年 23 巻 3 号 p. 261-270
発行日: 1990年
公開日: 2009/10/28
ジャーナル フリーBromodeoxyuridine (BrdU) immunohistochemistry has recently been introduced for the visualization of DNA-synthesizing nuclei. To determine the optimal conditions for detecting BrdU-incorporated nuclear DNA in formalin- or ethanol-fixed, paraffin-embedded human gastric tissues, we tested several different pretreatment procedures of hydrolysis with HCl and enzymatic digestion with actinase before the immunohistochemical staining. Three cases of surgically resected stomachs were perfused with artificial blood containing BrdU and fixed in formalin, ethanol or with their combination for different durations. The results of BrdU-immunohistochemical stainings were checked and the labeling indices were compared. As for hydrolysis, treatment with 2N HCl at 25°C for 90min was sufficient to detect the immunoreactivity in the stomach irrespective of fixative conditions tested. However, to obtain the most suitable stainings an additional enzymatic digestion was needed, and the optimal condition with actinase was different according to the fixation time in formalin- and/or ethanol-fixed tissues. There are also a few differences in the demonstrability of nuclear BrdU among various kinds of cells, i.e. normal epithelial cells, immunoblasts in the lymphoid follicle and cancer cells. Our method could extend the range of application of BrdU immunohistochemistry for cell kinetic studies using human surgically resected organs.抄録全体を表示PDF形式でダウンロード (7158K) -
MALINI LALORAYA, G. PRADEEP KUMAR, MANISH NIVSARKAR, MANMOHAN M. LALOR ...1990 年 23 巻 3 号 p. 271-279
発行日: 1990年
公開日: 2009/10/28
ジャーナル フリーThe distribution of the enzyme Cu-Zn superoxide dismutase in the ovary and uterus during early pregnancy has been dealt with in this histochemical study. Cu-Zn superoxide dismutase is found to be present in membrana granulosa of Graafian follicles and ovulated follicles, corpora lutea and blood vessels of the ovary. This presence has been attributed a functional significance in the process of luteal steroidogenesis. The marked low levels of uterine Cu-Zn superoxide dismutase and its noticeable absence in the endometrium, the site of blastocyst attachment, at 5.00 A.M. on Day 5 of pregnancy is evident. The possible involvement of these lowered levels in implantation is discussed.抄録全体を表示PDF形式でダウンロード (8275K) -
HIROKAZU KITAMURA1990 年 23 巻 3 号 p. 281-294
発行日: 1990年
公開日: 2009/10/28
ジャーナル フリーA couple of new sensitive methods have been established for the histochemical detection of acidic glycoconjugates in electron microscopy. These are combined staining methods consisting of a high or low iron diamine preembedding and thiocarbohydrazide silver proteinate postembedding sequence (HID or LID-TCH-SP) followed by a physical development (PD) procedure. The HID or LID-TCH-SP-PD methods have been applied to a series of acidic glycoconjugate-containing ultrastructures from different organs of the rat and mouse such as the aorta, kidney, pancreas and spleen, in comparision with the HID or LID-TCH-SP techniques. The new methods are signigicantly more sensitive than the HID or LID-TCH-SP technique. Their specificity is sufficient.抄録全体を表示PDF形式でダウンロード (13307K) -
ITS APPLICATION TO THE DETECTION OF mRNAS FOR MYELIN PROTEINS (PLP AND MBP) IN MOUSE BRAIN SECTIONSCHIYO SHIOTA, KATSUHIKO MIKOSHIBA1990 年 23 巻 3 号 p. 297-306
発行日: 1990年
公開日: 2009/10/28
ジャーナル フリーIn situ hybridization is a method for identifying specific DNA or RNA sequences in cells or tissues. It was originally developed for the detection of specific DNA sequences in chromosomes (2, 6), then extended to the detection of specific endogenous mRNA in cells (5). Various kinds of nucleotide sequences, such as complementary DNA (cDNA), antisense RNA and synthetic oligonucleotide that have been labeled with radioactive or otherwise detectable molecules, could be acceptable as probes for in situ RNA hybridization, as well as Northern hybridization. Among these probes, the radioactive cDNA probe is the most widely used for in situ hybridization. With a rapidly increasing number of cDNAs corresponding to molecules of interest, in situ hybridization has been extensively used in various fields such as neurobiology, endocrinology, developmental biology, etc. The detection of mRNAs for myelin basic protein (MBP) and myelin proteolipid protein (PLP) (7, 12, 13) is a good example of successful application of in situ hybridization. Here we describe our evaluation of some technical conditions which are generally used in in situ hybridization. We also show the distribution of MBP and PLP mRNAs in the mouse brain sections revealed by the relatively simple but effective method of in situ hybridization.抄録全体を表示PDF形式でダウンロード (11733K) -
MITSUHIRO KAWATA, KAZUNARI YURI, KENZO KUMAMOTO1990 年 23 巻 3 号 p. 307-325
発行日: 1990年
公開日: 2009/10/28
ジャーナル フリーIn situ hybridization histochemistry with synthetic oligodeoxynucleotide probes was applied to investigate the localization of tissue mRNAs. Specific probes for mRNAs to vasopressin- and oxytocin-preprohormone were designed by using the computer-based homology search program. The probes were labeled with 3H- deoxycytidine triphosphate (dCTP) or 35S-deoxyadenosine triphosphate (dATP) by terminal deoxynucleotidyl transferase. The specificity of the tailed probe was again confirmed by the homology search. Issues for the properties of isotopes and data analysis of the hybridizatioin signals in the autoradiography were discussed from the methodological point of view.抄録全体を表示PDF形式でダウンロード (11807K) -
TAKEHIKO KOJI, PAUL K. NAKANE1990 年 23 巻 3 号 p. 327-334
発行日: 1990年
公開日: 2009/10/28
ジャーナル フリーTo analyze the expression of specific mRNA at the level of individual cells, non-radioactive in situ hybridization has been a most powerful technique. In the process of in situ hybridization, the use of formamide is usually required in order to reduce the melting temperature (Tm) of nucleic acids. However, formamide is an expensive and unstable reagent, and more importantly, formamide in itself has some deteriorative effects such as nonspecific staining and morphological damage on the results of in situ hybridization. In this study, we examined the use of a mix-ture (Nm) of nucleotides (AMP, GMP, UMP, CMP) as an alternative to formamide in our non-radioactive hybridization system with thymine-thymine (T-T) dimerized DNA probe. When the effects of Nm on re-annealing of denatured pBR 322 DNA were investigated by electrophoretic patterns on agarose gel, it was confirmed that Nm (about 20-200mg/ml) reduced the Tm of pBR 322 DNA. On dotblot hybridization using Nm, we obtained sensitive and specific results similar to that of formamide. Finally, on frozen sections of rat pituitary glands fixed by perfusion of 4% paraformaldehyde in phosphate buffered saline (pH 7.4), prolactin mRNA was successfully localized in situ using Nm instead of formamide.抄録全体を表示PDF形式でダウンロード (7535K) -
DETECTION OF CARCINOEMBRYONIC ANTIGEN (CEA) mRNA AND ONCOGENE TRANSCRIPTS IN MW-FIXED COLORECTAL CARCINOMAS USING SULFONATED PROBESMASAHIKO MUROTANI, HIDEKI MORIMOTO, TAKUSHI MONDEN, MASAHIKO HIGASHIYA ...1990 年 23 巻 3 号 p. 335-349
発行日: 1990年
公開日: 2009/10/28
ジャーナル フリーMicrowave (MW) energy has been reported to permit rapid fixation and good conservation of tissue antigens as well as of tissue morphology. In this paper, we examined the applicability of MW-fixed tissue specimens to in situ hybridization. The preliminary analysis by Northern blot hybridization revealed that no degradation of mRNA occurred following MW irradiation to tissue samples. To determine the optimal conditions for in situ hybridization of MW-fixed tissue sections, we first investigated the in situ localization of carcinoembryonic antigen (CEA) mRNA in MW-fixed colorectal carcinomas using a sulfonated cDNA probe. The results indicated that pretreatment of sections with HCl could be omitted and the concentration of proteinase K for tissue digestion could be reduced in MW-fixed specimens. Thus, these specimens provided stronger CEA mRNA signals when compared with routinely fixed ones. On the basis of this result, we examined oncogene expression in 26 colorectal cancer specimens fixed by MW irradiation, and successfully detected Ha-ras mRNA in 6 (23%), Ki-ras mRNA in 7 (27%), and c- myc mRNA in 12 (46%) specimens. The results of in situ hybridization for the on-cogene mRNAs agreed well with those obtained by the immunohistochemical analysis of oncogene products using the specific antibodies to p21ras and p62myc. Our observations indicated the usefulness of rapid MW fixation for the preservation of mRNA in tissue preparations. MW-fixed tissue samples are applicable to in situ hybridization as well as to the immunohistochemical analysis of the expression of various genes.抄録全体を表示PDF形式でダウンロード (19202K) -
SUMIHARE NOJI, NAOKI TAKAHASHI, TSUTOMU NOHNO, EIKI KOYAMA, TOMOICHIRO ...1990 年 23 巻 3 号 p. 353-366
発行日: 1990年
公開日: 2009/10/28
ジャーナル フリーImproved procedures for a simplified in situ hybridization using paraffinembedded tissues and riboprobes were described. We found that when tissues were fixed with 4% paraformaldehyde and embedded in paraffin, harsh pretreatments so far employed, such as proteolytic, heat, or HCl treatment each damaging cell morphology to some degree, are not essential to expose mRNAs on the surface of the sections subjected. Typical results are shown for the detection of NGF and EGF mRNAs in mouse submandibular glands. Furthermore, we demonstrated that our method is also applicable to tissues decalcified with 5% EDTA, as shown in the detection of elastin mRNAs in the calcified tissues in the mouse oral region. We applied our method to determine expression patterns of a new mouse homeobox-containing gene, designated Hox-3.5. Hox-2.1 and Hox-1.3 in embryos. The Hox-3.5 gene is expressed intensively in the rhombencephalon and spinal cord of 12 day embryos, and in the three (at least) cervical ganglion near the occipital bone of 13 day embryos. In the 14 day old embryos, expression of the Hox-3.5 gene is spatially limited at least to the epithelium of both lung and intestine, derived from endoderm. These results suggest that spatial expression of the Hox-3.5 gene may play an important role in formation of ectodermal tissues in 12 day embryos and then in generation of the digestive and respiratory tubes from endoderm in 14 day embryos.抄録全体を表示PDF形式でダウンロード (14709K) -
YOSHIO WAKAMATSU, HISATO KONDOH1990 年 23 巻 3 号 p. 367-374
発行日: 1990年
公開日: 2009/10/28
ジャーナル フリーThe in situ hybridization technique allows us to identify expressing cells, to assess expression levels semi-quantitatively, and hence to reveal the spatiotemporal order of expression of various genes in developing embryos. We report here the conditions for detection of embryonic N-myc transcript in paraffin sections in the form of standard protocols and technical notes.抄録全体を表示PDF形式でダウンロード (3094K) -
TADAHISA KITAMURA, SACHIHIKO WATANABE1990 年 23 巻 3 号 p. 375-386
発行日: 1990年
公開日: 2009/10/28
ジャーナル フリーIntermediate filaments of the CNS, neurofilament and glial filament, vary in amount with cell types and locations and with normal and pathological conditions. In order to establish whether these differences depend on the amount of their cytoplasmic messenger RNA (mRNA), we tried to detect the mRNA of the peptides constituting these filaments by Northern blot analysis and by in situ hybridization. We found mRNAs of both peptides in the perinuclear zone of nerve cells or glial cells of the rat brain. Thus, intermediate filament peptides are synthesized in the perikaryal cytoplasm, and not in the processes where the filaments exist. The neurons with thicker and longer axons tended to have larger amounts of NF-L mRNA in the cytoplasm. In the retrograde degeneration of rat facial nucleus, the amount of NF-L mRNA decreased, indicating a decline of its synthesis in the stage of axon regeneration. GFAP mRNA increased in the area of the damaged facial nucleus. The GFAP gene of the protoplasmic astroglia located in these areas may be induced by the brain damage, and this may lead to the formation of gliosis in the gray matter.抄録全体を表示PDF形式でダウンロード (12567K) -
COMPARISON IN PRIMARY AND METASTASIZED TUMORS AND IN THE ESTABLISHED CELL LINENAOTOSHI KANDA, YASUHIDE HAYASHI, RYOJI HANADA, KEIKO YAMAMOTO, EIZO A ...1990 年 23 巻 3 号 p. 389-399
発行日: 1990年
公開日: 2009/10/28
ジャーナル フリーAmplification of oncogene N-myc and the adjacent DNA fragments, clone 8 and G21 was observed in the primary neuroblastoma from a female patient at stage III and in the metastatic tumors from the same patient. The three DNA loci were amplified to the same degree in all tumor specimens, including an established cell line (SCMC-N3) of the primary tumor. Neither visible double minutes (DMs) nor homogeneously staining regions (HSRs) were recognized in the primary tumor by light microscope. However, numerous DMs were found in the established cell line with elevated N-myc expression, and in situ hybridization showed them to be the site of amplification for the N-myc, clone 8 and G21. These results suggest that the DNA amplification occurred in the primary tumor prior to metastasis and, probably, in the small DMs or precursors of DMs which were undetactable by light microscope.抄録全体を表示PDF形式でダウンロード (4276K) -
SAKAN MAEDA, SOHEI KITAZAWA, MITSUZO HORIO, KAZUHIRO SATONAKA, TAKAHIR ...1990 年 23 巻 3 号 p. 401-409
発行日: 1990年
公開日: 2009/10/28
ジャーナル フリーPDF形式でダウンロード (5562K) -
JOHJI INAZAWA, TATSUO ABE1990 年 23 巻 3 号 p. 411-422
発行日: 1990年
公開日: 2009/10/28
ジャーナル フリーThe chromosomal localization of four newly isolated carcinoembryonic antigen (CEA) family genes was determined by the combined use of the Southern blot hybridization of hybrid cells and cell sorted chromosome DNAs, and in situ hybridization. CEA, one of the most widely used human tumor markers, is a member of a gene family comprising over 10 closely-related genes and constitutes a subfamily within the immunoglobulin (Ig) superfamily. Recent success of analysis at molecular level revealed that nonspecific cross-reacting antigen (NCA) and pregnancy specific β1-glycoprotein. (PSβG) also belong to the CEA gene family. We mapped CEA, NCA, and two different PSβG genes to chromosome 19 at band q13.2. These results suggest that almost all CEA family genes are located within a limited region on chromosome 19 in a manner similar to the major histocompatibility (MHC) genes, which also belong to the Ig superfamily, and which are localized within bands p21.1-p21.3 on chromosome 6.抄録全体を表示PDF形式でダウンロード (3897K)
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