The Effect of Reduced Aldosterone Levels on 11β-HSD Isoform Expression in Normal and Hypertensive Rat Kidney Tissue Using q-PCR
Location
CSU Ballroom
Start Date
28-4-2009 10:00 AM
End Date
28-4-2009 12:00 PM
Student's Major
Chemistry and Geology
Student's College
Science, Engineering and Technology
Mentor's Name
Theresa Salerno
Mentor's Department
Chemistry and Geology
Mentor's College
Science, Engineering and Technology
Description
11β-Hydroxysteroid Dehydrogenase (11β-HSD) exists in two isoforms, 11β-HSDl and 11β-HSD2. These two enzymes regulate levels of glucocorticoids ; 11β-HSDl converts inactive cortisone to active cortisol, while 11β-HSD2 catalyzes the opposite reaction. Since cortisol and aldosterone both bind to the mineralocortocoid receptor (MR), increases in cortisol can result in hypertension. The goals of this experiment were to measure the effect of decreased aldosterone levels on the levels of the 11β-HSD isoenzymes. Specifically, we assessed whether there is compensation by the 11β-HSD isoenzymes to account for the observations that blood pressures were not lowered by the decreased aldosterone levels. The specific question addressed was whether there was an upregulation or downregulation of messenger RNA expressions for either isoenzyme and whether the subsequent effect on cortisol levels resulted in hypertension. Our research focused on the experimental design and the development of methodology; design of primers and probes, RNA isolation and quantification, reverse transcription of RNA to cDNA, and real time polymerase chain reaction (qPCR). Kidney tissues were obtained and RNAs were successfully extracted from normotensive control Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHR), which underwent surgical destruction of the adrenal glands or a sham surgery. A real time PGR method was optimized and then used to compare mRNA levels of the two isoenzymes relative to mRNA levels of a housekeeping gene.
The Effect of Reduced Aldosterone Levels on 11β-HSD Isoform Expression in Normal and Hypertensive Rat Kidney Tissue Using q-PCR
CSU Ballroom
11β-Hydroxysteroid Dehydrogenase (11β-HSD) exists in two isoforms, 11β-HSDl and 11β-HSD2. These two enzymes regulate levels of glucocorticoids ; 11β-HSDl converts inactive cortisone to active cortisol, while 11β-HSD2 catalyzes the opposite reaction. Since cortisol and aldosterone both bind to the mineralocortocoid receptor (MR), increases in cortisol can result in hypertension. The goals of this experiment were to measure the effect of decreased aldosterone levels on the levels of the 11β-HSD isoenzymes. Specifically, we assessed whether there is compensation by the 11β-HSD isoenzymes to account for the observations that blood pressures were not lowered by the decreased aldosterone levels. The specific question addressed was whether there was an upregulation or downregulation of messenger RNA expressions for either isoenzyme and whether the subsequent effect on cortisol levels resulted in hypertension. Our research focused on the experimental design and the development of methodology; design of primers and probes, RNA isolation and quantification, reverse transcription of RNA to cDNA, and real time polymerase chain reaction (qPCR). Kidney tissues were obtained and RNAs were successfully extracted from normotensive control Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHR), which underwent surgical destruction of the adrenal glands or a sham surgery. A real time PGR method was optimized and then used to compare mRNA levels of the two isoenzymes relative to mRNA levels of a housekeeping gene.
Recommended Citation
Dittrich, Kristina and Linet Nyarobi. "The Effect of Reduced Aldosterone Levels on 11β-HSD Isoform Expression in Normal and Hypertensive Rat Kidney Tissue Using q-PCR." Undergraduate Research Symposium, Mankato, MN, April 28, 2009.
https://cornerstone.lib.mnsu.edu/urs/2009/poster-session-C/23
