Abstract
Synthesis of a number of rat liver proteins, including albumin, fibrinogen, apolipoprotein Al, and transferrin, is elevated in the nephrotic syndrome (NS). Increased synthesis of these proteins is regulated at the transcriptional level and occurs in the context of increased mRNA encoding each protein. Changes in albumin, fibrinogen, apolipoprotein Al, and transferrin mRNA levels in total cellular RNA isolated from the livers of normal rats and rats with passive Heymann nephritis were measured using a kinetically monitored, reverse transcriptase-initiated PCR (kRT-PCR) assay. The kRT-PCR assay rapidly quantitated changes in rat liver mRNA levels with an accuracy comparable to that of more labor-intensive mRNA quantitation methods. The relative levels of β-actin, apolipoprotein Al, fibrinogen, and albumin mRNAs were very similar in total cellular RNA isolated from rat liver versus H4C3 hepatocytes in culture, suggesting that the H4C3 hepatocyte is an appropriate model for studying expression of genes encoding proteins secreted by the liver. Taken together, the results demonstrate the feasibility of using the kRT-PCR assay for isolation and characterization of a soluble factor responsible for elevated synthesis of hepatocyte mRNAs associated with the nephrotic syndrome.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 437-443 |
| Number of pages | 7 |
| Journal | Methods: A Companion to Methods in Enzymology |
| Volume | 13 |
| Issue number | 4 |
| DOIs | |
| State | Published - Dec 1997 |
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ASJC Scopus subject areas
- Molecular Biology
- Biochemistry, Genetics and Molecular Biology(all)
Cite this
Rat liver transcript profiling in normal and disease states using a kinetic polymerase chain reaction assay. / Kang, John J.; Kaysen, George; Jones, Hardin; Holland, Michael J.
In: Methods: A Companion to Methods in Enzymology, Vol. 13, No. 4, 12.1997, p. 437-443.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Rat liver transcript profiling in normal and disease states using a kinetic polymerase chain reaction assay
AU - Kang, John J.
AU - Kaysen, George
AU - Jones, Hardin
AU - Holland, Michael J.
PY - 1997/12
Y1 - 1997/12
N2 - Synthesis of a number of rat liver proteins, including albumin, fibrinogen, apolipoprotein Al, and transferrin, is elevated in the nephrotic syndrome (NS). Increased synthesis of these proteins is regulated at the transcriptional level and occurs in the context of increased mRNA encoding each protein. Changes in albumin, fibrinogen, apolipoprotein Al, and transferrin mRNA levels in total cellular RNA isolated from the livers of normal rats and rats with passive Heymann nephritis were measured using a kinetically monitored, reverse transcriptase-initiated PCR (kRT-PCR) assay. The kRT-PCR assay rapidly quantitated changes in rat liver mRNA levels with an accuracy comparable to that of more labor-intensive mRNA quantitation methods. The relative levels of β-actin, apolipoprotein Al, fibrinogen, and albumin mRNAs were very similar in total cellular RNA isolated from rat liver versus H4C3 hepatocytes in culture, suggesting that the H4C3 hepatocyte is an appropriate model for studying expression of genes encoding proteins secreted by the liver. Taken together, the results demonstrate the feasibility of using the kRT-PCR assay for isolation and characterization of a soluble factor responsible for elevated synthesis of hepatocyte mRNAs associated with the nephrotic syndrome.
AB - Synthesis of a number of rat liver proteins, including albumin, fibrinogen, apolipoprotein Al, and transferrin, is elevated in the nephrotic syndrome (NS). Increased synthesis of these proteins is regulated at the transcriptional level and occurs in the context of increased mRNA encoding each protein. Changes in albumin, fibrinogen, apolipoprotein Al, and transferrin mRNA levels in total cellular RNA isolated from the livers of normal rats and rats with passive Heymann nephritis were measured using a kinetically monitored, reverse transcriptase-initiated PCR (kRT-PCR) assay. The kRT-PCR assay rapidly quantitated changes in rat liver mRNA levels with an accuracy comparable to that of more labor-intensive mRNA quantitation methods. The relative levels of β-actin, apolipoprotein Al, fibrinogen, and albumin mRNAs were very similar in total cellular RNA isolated from rat liver versus H4C3 hepatocytes in culture, suggesting that the H4C3 hepatocyte is an appropriate model for studying expression of genes encoding proteins secreted by the liver. Taken together, the results demonstrate the feasibility of using the kRT-PCR assay for isolation and characterization of a soluble factor responsible for elevated synthesis of hepatocyte mRNAs associated with the nephrotic syndrome.
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UR - http://www.scopus.com/inward/citedby.url?scp=0031414739&partnerID=8YFLogxK
U2 - 10.1006/meth.1997.0549
DO - 10.1006/meth.1997.0549
M3 - Article
C2 - 9480787
AN - SCOPUS:0031414739
VL - 13
SP - 437
EP - 443
JO - Methods
JF - Methods
SN - 0076-6879
IS - 4
ER -