PCR In Situ Amplification and Catalyzed Signal Amplification. Approaches of Higher Sensitive, Non-Radioactive In Situ Hybridization.

Tani, Yoichi

doi:10.1267/ahc.32.261

Cited by 14 publications

(8 citation statements)

References 49 publications

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“…This is also in good agreement with the fact that a low expression of mRNA such as Srr mRNA (copies of Srr mRNA: GAPDH mRNA = 1:1000) (Fig. 1) is detected as a single, tiny-dotted signal when detected by in situ hybridization with TSA (Tani, 1999;Suzuki et al, 2003;Jiang et al, 2004;Yoshikawa et al, 2004;Dobashi et al, 2006).…”

Section: O P R O V E T H E V a L I D I T Y O F R E S U L T S F R O M supporting

confidence: 87%

The serine racemase mRNA is expressed in both neurons and glial cells of the rat retina

Takayasu

Yoshikawa

Watanabe

et al. 2008

Archives of Histology and Cytology

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IntroductionD-Serine acts as an obligatory and endogenous coagonist for the N-methyl-D-aspartate (NMDA) receptorrelated glycine site in the mammalian brain (Hashimoto et al., 1993;Matsui et al., 1995;Schell et al., 1995;Hashimoto and Oka, 1997;Danysz and Parsons, 1998). D-Serine is synthesized by serine racemase (Srr), which converts L-serine to D-serine. The regional profile of Srr closely resembles those of endogenous D-serine and NMDA receptors, which are highest in the forebrain and lowest in the hindbrain (Yoshikawa et al., 2004). Previous immunohistochemical studies have demonstrated that D-serine and Srr were localized exclusively in glial fibrillary acidic protein (GFAP)-positive astrocytes (Hashimoto and Oka, 1997;Schell et al., 1997; Wolosker et al., 1999;Mustafa et al., 2004). However, recent studies using immunohistochemistry, Western blot, and in situ hybridization have demonstrated that D-serine (Yasuda et al., 2001;Puyal et al., 2006) and Srr (Kartvelishvily et al., 2006;Yoshikawa et al., 2006Yoshikawa et al., , 2007 are signifi cantly expressed in neurons of tissue sections, and of primary cultures of the rat brain as compared with astrocytes.Compared with the brain, the retina contains fewer cell types, many of which have recognizable morphologies and consistent positions, making any investigation of the mechanisms for the central nervous system more The serine racemase mRNA is expressed in both neurons and glial cells of the rat retina

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Section: O P R O V E T H E V a L I D I T Y O F R E S U L T S F R O M supporting

confidence: 87%

The serine racemase mRNA is expressed in both neurons and glial cells of the rat retina

Takayasu

Yoshikawa

Watanabe

et al. 2008

Archives of Histology and Cytology

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“…In situ hybridization using nonradioactive probes includes the CSA method by which the detection sensitivity is amplified using tyramine, a phenol derivative. This method has been used in immunohistochemistry and immunoblotting, and its application to ISH has allowed the detection of viral DNA that had been at undetectable levels by the usual methods, and also of minute amounts of mRNA 12. By applying ISH and the CSA method for 20 patients with IPMT, the results were almost the same as those of immunohistochemical staining, confirming the COX‐2 expression at the mRNA level.…”

Section: Discussionmentioning

confidence: 76%

“…In situ hybridization was performed in 20 patients with IPMT by the catalyzed signal amplification (CSA) method12, 13 using Gen Point System (DAKO Japan, Kyoto, Japan). Formalin fixed paraffinized embedded sections were deparaffinized and hydrophilized in 0.1% diethylpyrocarbonate.…”

Section: Methodsmentioning

confidence: 99%

Immunohistochemical analysis and in situ hybridization of cyclooxygenase‐2 expression in intraductal papillary‐mucinous tumors of the pancreas

Niijima

Yamaguchi

Ishihara

et al. 2002

Cancer

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BACKGROUND The objective of this study was to investigate COX‐2 expression in intraductal papillary‐mucinous tumor of the pancreas (IPMT) using immunohistochemical staining (IH) and in situ hybridization (ISH). METHODS Immunohistochemical staining of COX‐2 was performed using samples from 42 patients with IPMT (hyperplasia, 10; adenoma, 13; noninvasive adenocarcinoma, 13; invasive adenocarcinoma, 6) and from 10 patients with ductal pancreatic adenocarcinoma, 10 with chronic pancreatitis, and 6 normal pancreatic tissues as controls. Also, COX‐2 was determined in five patients with IPMT noninvasive adenocarcinoma, in whom all histologic types, hyperplasia, adenoma, and adenocarcinoma were observed in the same excised specimens. Furthermore, IH of proliferating cell nuclear antigen (PCNA) was performed, and the labeling index (LI) was calculated to investigate the correlation with COX‐2. To confirm COX‐2 mRNA, the authors performed ISH in 20 IPMT patients. RESULTS COX‐2 was positive in 0%, 0%, and 10% of pancreatic duct epithelial cells from normal pancreatic tissue, chronic pancreatitis, and IPMT hyperplasia, respectively. Whereas it was positive in 69%, 77%, 67%, and 80% of IPMT adenoma, IPMT noninvasive adenocarcinoma, IPMT invasive adenocarcinoma, and ductal pancreatic adenocarcinoma, respectively, showing significant differences between IPMT hyperplasia and IPMT adenoma or IPMT adenocarcinoma (noninvasive and invasive adenocarcinoma). In the same patient, COX‐2 was negative in the hyperplasia region but positive in adenoma and adenocarcinoma regions, showing results reflecting the progression of the disease. In the COX‐2 negative group, PCNA‐LI was 19.2 ± 17.9%, and 33.5 ± 15.7% in the positive group, a significant difference. On ISH, COX‐2 mRNA was expressed in three of four and seven of eight COX‐2 positive patients with IPMT adenoma and adenocarcinoma, respectively. CONCLUSIONS COX‐2 was highly expressed in adenoma and adenocarcinoma in IPMT, showing a relation to the histologic grade of IPMT. Cancer 2002;94:1565–73. © 2002 American Cancer Society. DOI 10.1002/cncr.10358

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“…Compared with PCR in situ amplification, CSA is simpler and faster and does not require thermal cycling or the specialized equipment. 29 In a preliminary experiment, we failed to obtain any signal using RNA probes to detect ETBR mRNA (data not shown), but we obtained satisfactory signals with PNA probes using the CSA method. The results of our trial clearly demonstrate the advantages of PNA probes and the CSA method.…”

Section: Discussionmentioning

confidence: 80%

Enhanced expression of endothelin B receptor at protein and gene levels in human cirrhotic liver

Yokomori¹,

Oda²

2001

Gastroenterology

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PCR In Situ Amplification and Catalyzed Signal Amplification. Approaches of Higher Sensitive, Non-Radioactive In Situ Hybridization.

Cited by 14 publications

References 49 publications

The serine racemase mRNA is expressed in both neurons and glial cells of the rat retina

The serine racemase mRNA is expressed in both neurons and glial cells of the rat retina

Immunohistochemical analysis and in situ hybridization of cyclooxygenase‐2 expression in intraductal papillary‐mucinous tumors of the pancreas

Enhanced expression of endothelin B receptor at protein and gene levels in human cirrhotic liver

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