Tags

Type your tag names separated by a space and hit enter

Paraquat-induced gene expression in rat lung tissues using a differential display reverse transcription-polymerase chain reaction.
Arch Toxicol. 2002 Sep; 76(9):530-7.AT

Abstract

Increased formation of reactive oxygen species is a cause of paraquat (PQ)-induced injury and also provides a link between the signaling pathways and transcriptional events that regulate the expression of a large number of genes. However, the molecular mechanisms involved in PQ-induced injury remain unclear. To investigate the changes in gene expression at the onset of PQ injury, we used the differential display-polymerase chain reaction (PCR) method. Rats were treated intraperitoneally with 20 mg/kg PQ, and after 3 h the lungs were immediately excised. Samples of mRNA from normal and treated rats were used to prepare radiolabeled cDNAs, which were electrophoresed. Then the transcription levels were compared. We isolated 26 fragments of cDNA that were potentially affected by PQ, and determined their nucleotide sequences. Six clones of interest were selected and analyzed further. The reverse transcript-PCR based on their sequence information confirmed the differential expression for five clones: four clones were up-regulated and one was down-regulated. We were particularly interested in two genes that had homology with the known gene: TATA box-binding protein-associated factor, RNA polymerase II, B, 150 kDa (TAFIIB), and a candidate gene for lipodystrophy, Lpin2. Both genes were significantly up-regulated within 3 h of PQ intake and the stimulation continued during our 24-h observation period. In addition, up-regulation of Lpin2 was observed in the lungs, but not in the liver and kidneys. In situ hybridization using lung sections showed that the expression of both genes was strongly visualized in Clara cells and in alveolar macrophages. These findings suggest a stimulation of transcription levels and changes in lipid metabolism in Clara cells and in macrophages in the lungs, which result in their playing a crucial role at the onset of PQ-driven pulmonary injury.

Authors+Show Affiliations

Department of Legal Medicine, Kawasaki Medical School, 577 Matsushima, Kurashiki 701-0192, Japan. tomita@bcc.kawasaki-m.ac.jpNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

12242611

Citation

Tomita, Masafumi, et al. "Paraquat-induced Gene Expression in Rat Lung Tissues Using a Differential Display Reverse Transcription-polymerase Chain Reaction." Archives of Toxicology, vol. 76, no. 9, 2002, pp. 530-7.
Tomita M, Nohno T, Okuyama T, et al. Paraquat-induced gene expression in rat lung tissues using a differential display reverse transcription-polymerase chain reaction. Arch Toxicol. 2002;76(9):530-7.
Tomita, M., Nohno, T., Okuyama, T., Nishimatsu, S., & Adachi, J. (2002). Paraquat-induced gene expression in rat lung tissues using a differential display reverse transcription-polymerase chain reaction. Archives of Toxicology, 76(9), 530-7.
Tomita M, et al. Paraquat-induced Gene Expression in Rat Lung Tissues Using a Differential Display Reverse Transcription-polymerase Chain Reaction. Arch Toxicol. 2002;76(9):530-7. PubMed PMID: 12242611.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Paraquat-induced gene expression in rat lung tissues using a differential display reverse transcription-polymerase chain reaction. AU - Tomita,Masafumi, AU - Nohno,Tsutomu, AU - Okuyama,Toshiko, AU - Nishimatsu,Shin-ichiro, AU - Adachi,Junko, Y1 - 2002/07/25/ PY - 2002/05/13/received PY - 2002/06/10/accepted PY - 2002/9/21/pubmed PY - 2003/4/17/medline PY - 2002/9/21/entrez SP - 530 EP - 7 JF - Archives of toxicology JO - Arch Toxicol VL - 76 IS - 9 N2 - Increased formation of reactive oxygen species is a cause of paraquat (PQ)-induced injury and also provides a link between the signaling pathways and transcriptional events that regulate the expression of a large number of genes. However, the molecular mechanisms involved in PQ-induced injury remain unclear. To investigate the changes in gene expression at the onset of PQ injury, we used the differential display-polymerase chain reaction (PCR) method. Rats were treated intraperitoneally with 20 mg/kg PQ, and after 3 h the lungs were immediately excised. Samples of mRNA from normal and treated rats were used to prepare radiolabeled cDNAs, which were electrophoresed. Then the transcription levels were compared. We isolated 26 fragments of cDNA that were potentially affected by PQ, and determined their nucleotide sequences. Six clones of interest were selected and analyzed further. The reverse transcript-PCR based on their sequence information confirmed the differential expression for five clones: four clones were up-regulated and one was down-regulated. We were particularly interested in two genes that had homology with the known gene: TATA box-binding protein-associated factor, RNA polymerase II, B, 150 kDa (TAFIIB), and a candidate gene for lipodystrophy, Lpin2. Both genes were significantly up-regulated within 3 h of PQ intake and the stimulation continued during our 24-h observation period. In addition, up-regulation of Lpin2 was observed in the lungs, but not in the liver and kidneys. In situ hybridization using lung sections showed that the expression of both genes was strongly visualized in Clara cells and in alveolar macrophages. These findings suggest a stimulation of transcription levels and changes in lipid metabolism in Clara cells and in macrophages in the lungs, which result in their playing a crucial role at the onset of PQ-driven pulmonary injury. SN - 0340-5761 UR - https://www.unboundmedicine.com/medline/citation/12242611/Paraquat_induced_gene_expression_in_rat_lung_tissues_using_a_differential_display_reverse_transcription_polymerase_chain_reaction_ L2 - https://doi.org/10.1007/s00204-002-0379-x DB - PRIME DP - Unbound Medicine ER -