Gene transfer by adenovirus in smooth muscle cells.
Abstract: We report adenovirus-mediated gene transfer into airway smooth muscle cells in cultured cells and organ-cultured tracheal segments. Incubation of cultured rat tracheal myocytes with virus (5 x 10(8) pfu/ml) for 6 h resulted in beta-galactosidase expression in 94.8 +/- 2.5% of cells (n = 4). Following incubation of thin (less than 200 microns diameter) equine trachealis muscle segments with virus in organ culture (5 x 10(8)-5 x 10(10) pfu/ml) the average expression of the Lac Z gene was approximately 19 +/- 10% (n = 9). Expression was markedly improved, however, in segments from neonatal rats (13-21 days). In two experiments in which the mucosa and serosa were removed, nearly all cells expressed beta-galactosidase, whereas in a third experiment in which the tissue was not dissected, about 40% of cells were stained. Viral infection had no effect on tension development of strips following organ culture. In vitro gene transfer may provide a useful method to alter protein expression and examine the effect of this alteration on excitation/contraction coupling in smooth muscle.
Publication Date: 1996-08-01 PubMed ID: 8897661DOI: 10.1016/0034-5687(96)00016-3Google Scholar: Lookup
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- Journal Article
- Research Support
- U.S. Gov't
- P.H.S.
Summary
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The research involves utilizing adenovirus-mediated gene transfer into smooth muscle cells found in airways using both cell cultures and organ-cultured tracheal segments. Its results contribute to our understanding of gene manipulation in airway smooth muscle cells.
Gene Transfer
- The researchers have managed to successfully transfer genes using adenovirus, a group of common viruses, to airway smooth muscle cells. These cells play a crucial role in respiratory functioning.
- The main technique involved incubating cultured rat tracheal myocytes, a type of muscle cell, with the virus. The most successful results led to beta-galactosidase expression in over 94% of cells. This enzyme is often used as a marker in laboratories, suggesting that the gene transfer was successful.
Organ-culture Experimentation
- The research paper also speaks about experiments on organ-cultured tracheal segments. For these experiments, equine trachealis muscle segments were used. The average expression of a particular gene, Lac Z, was noticed in around 19% of the scenarios.
- The expression rate was significantly better in neonatal rat segments, which is an intriguing finding suggesting that younger, developing tissues may be more receptive to such techniques.
Viral Infection and Tension Development
- It is valuable to note that the viral infection involved in the gene transfer did not have a discernible impact on the tension development in these tissues post-culture. This is promising, indicating that such gene transfer techniques might not interfere significantly with cell functioning and physiology.
Implications
- The in vitro gene transfer illustrated in this research paper can be a beneficial method to study the effect of altered protein expression on excitation and contraction in smooth muscle cells.
- This kind of research can influence our understanding of smooth muscle cells’ behavior, particularly those in the respiratory system, and may have implications in studying or treating certain conditions related to muscle behavior or response.
Cite This Article
APA
Yu MF, Ewaskiewicz JI, Adda S, Bailey K, Harris V, Sosnoski D, Tomasic M, Wilson J, Kotlikoff MI.
(1996).
Gene transfer by adenovirus in smooth muscle cells.
Respir Physiol, 105(1-2), 155-162.
https://doi.org/10.1016/0034-5687(96)00016-3 Publication
Researcher Affiliations
- Department of Animal Biology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia 19104-6046, USA.
MeSH Terms
- Adenoviridae / genetics
- Animals
- Cells, Cultured
- Gene Expression Regulation / genetics
- Gene Transfer Techniques
- Histocytochemistry
- Horses
- Lac Operon / genetics
- Muscle Contraction / physiology
- Muscle, Smooth / metabolism
- Rats
- Trachea / metabolism
- Transfection / genetics
- beta-Galactosidase / genetics
- beta-Galactosidase / metabolism
Grant Funding
- HL41084 / NHLBI NIH HHS
- HL45239 / NHLBI NIH HHS
- T32HL07027 / NHLBI NIH HHS
Citations
This article has been cited 1 times.- Levine JA, Jensen MD, Eberhardt NL, O'Brien T. Adipocyte macrophage colony-stimulating factor is a mediator of adipose tissue growth. J Clin Invest 1998 Apr 15;101(8):1557-64.
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