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.

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Apr 28th, 10:00 AM Apr 28th, 12:00 PM

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