Anagha Rama Varma

Centennial High School, Ellicott City, MD 21042

The Effect of Lisinopril on the Desiccation Resistance of Male Drosophila melanogaster

The Effect of Lisinopril on the Desiccation Resistance of Male Drosophila melanogaster
Anagha Rama Varma
Centennial High School, Ellicott City, MD
Teacher and/or Mentor: Mrs. Michelle Bagley/Dr. Jeff Leips; Department of Biological Sciences;
University of Maryland, Baltimore County

Lisinopril, a commonly prescribed drug to treat high blood pressure, has been found to have
various positive side effects in humans, including the reduction of frailty. Recent work has
discovered that these same effects can be observed in Drosophila melanogaster, also known as
the common fruit fly. As the genetic pathway connected to RAS in humans is conserved in
Drosophila, understanding the effects of Lisinopril on phenotypic traits in Drosophila can help to
gain a better understanding of how the drug works in humans and may help to reveal an
overlooked genetic effect in humans. Since Lisinopril has been found to alter gene expression in
the midgut of Drosophila, which maintains water-solute balance in the organism, it may be
important to the mechanism that helps maintain water-solute balance. If so, desiccation is an
ideal phenotypic trait to examine in order to better understand the consequences of Lisinoprils
impact on gene expression. This study will examine the desiccation resistance of Lisinopril-
treated Drosophila melanogaster and compare it with that of untreated (control) flies.
 ACKNOWLEDGMENTS

I would like to thank Dr. Jeff Leips of the Department of Biological Sciences, University

of Maryland, Baltimore Country for mentoring me, allowing me to use the lab facilities, and

providing valuable comments on this project. I would also like to thank Ms. Sara Ohadi-

Hamadani for her help. I must also thank Mrs. Michelle Bagley of Centennial High School for all

of her help and encouragement with this project.

 TABLE OF CONTENTS

List of figures...1

Introduction..2

Methods and Materials.2

Results..4

Conclusion.......8

References9
List of Figures

Figure 1 Average Desiccation Resistance of Ral-73 of Drosophila melanogaster 5

Figure 2 Average Desiccation Resistance of Ral-304 of Drosophila melanogaster 6

Figure 3 Average Desiccation Resistance of Ral-229 of Drosophila melanogaster 7

Figure 4 Average Desiccation Resistance of Ral-229 (aged 5 weeks) of Drosophila

melanogaster

8
 INTRODUCTION

Lisinopril is a drug approved by the FDA for use in treating high blood pressure in

humans. It works by restricting the angiotensin converting enzyme (ACE) in the human renin-

angiotensin system (RAS), a critical regulator of sodium balance (Harrison-Bernard) which

converts the hormone angiotensin type 1 (Ang I) to angiotensin type 2 (Ang II). Ang II is a

vasoconstrictor; restricting ACE lowers the production of Ang II, and thus lowers blood pressure

(Abadir; Abadir et. al.; Krcobic; Lisinopril; Renin-angiotensin system Homo sapiens). In

addition to treatment of high blood pressure, the drug Lisinopril is associated with many positive

side effects, such as improved speed and reduced frailty. Frailty is defined as syndrome in which

three or more of the following trends occur: unintentional weight loss, fatigue, weakness,

decreased walking speed, and less physical activity (Fried et. al.; Xue). Lisinopril has been

shown to increase weight retention among older adults (Schellenbaum et. al.) and improve

walking speed among older adults with exercise (Buford et. al.). It has been shown that

Lisinopril-treated Drosophila melanogaster experience effects similar to those observed in

humans despite lacking RAS. D. melanogaster does contain a genetic pathway similar to the one

associated with RAS in humans, in that they possess genes homologous to ACE, such as the

angiotensin converting enzyme gene (ANCE) and angiotensin converting enzyme related

(ACER) (Fournier et. al.). As Lisinopril also appears to impact expression of genes related to

water-solute balance regulation in Drosophila, and likewise operates on RAS in humans, which

is related to maintenance of water-solute balance in humans, it is possible that Lisinoprils effects

may have a genetic link that is not yet fully understood. Additionally, Lisinopril has been found

to impact individuals differently. As such, this study will compare its effects on the desiccation
resistance of more than one genotype, in order to compare any genetically based differences that

may occur.

METHODS AND MATERIALS

General Methods

The first experiment was conducted with the purpose of determining the desiccation of

resistance of flies not exposed to Lisinopril. To conduct this experiment, experimental flies were

collected from the populations of three genetic lines that are subsets of the Drosophila Genetic

Reference Panel (Mackay et. al): lines Ral-229, Ral-304, and Ral-73. Lines 229, 304, and 73

were selected because they were found to respond differently to the drug, having varying degrees

of improvement in lifespan and strength. By using these lines, genetically based differences in

the impact of Lisinopril on desiccation resistance might be observed. These lines were first

mated and then expanded; line 229 was expanded to about 60 vials, whereas 304 and 73 were

expanded to about 80 vials each. After one week, all adult flies were cleared from the vials, and

the emerging flies were mated. These adults were then cleared, and males were collected from

the emerging flies. About eighty males were collected per line. These flies were then maintained

in six cages, separated by treatment and line. There were about forty flies in each cage, and these

were maintained using regular food. Once the caged flies were one week old, the first trial was

conducted; half of the flies of each line were subjected to the desiccation assay and the rest were

maintained in control vials, each of which were empty save for a cotton ball at the bottom. A

second trial was then conducted with a new batch of male flies, aged to five weeks.
Desiccation Assay

A standard method was used to tests desiccation resistance (Ngheim, Gibbs, Rose &

Bradley; Folk, Han & Bradley). Sixty vials were prepared in total; thirty vials were control vials,

empty save for a cotton ball, and thirty were experimental (desiccation) vials. The experimental

vials contained 4 grams anhydrous Drierite at the bottomthis desiccant was then covered by a

cotton ball. The vials were sealed with Parafilm, to prevent the entrance of moisture. Once the

flies were at the proper age, which depended on the trial, they were transferred to the vials from

the cages. In order to transfer the flies, the pre-prepared vials were placed in ice. Once cold, the

vials were opened, and the flies were transferred into them with an aspirator. Four flies were

placed in each vial. Once they were within, the Parafilm was resealed, and the filled vial was

removed from the ice and labeled with line number and treatment. The vials were observed, and

the number dead in each vial was recorded at hourly intervals over a twenty-four hour period.

RESULTS

Overall, it was found that the different genotypes varied considerably with regards to

desiccation resistance. Line 304 appeared to have the greatest ability to resist desiccation while

aged one week, surviving exposure to desiccant for an average of 22.9 hours. Compared to one

week old line 73, which survived desiccation death for an average of 11.4 hours, and one week

old line 229, which survived on average for 14.8 hours, line 304 performed far better. The

following graphs show the average desiccation resistance for the three lines.
Figure 1

Average Desiccation Resistance of Ral-73 Drosophila melanogaster

1
Treatment

0 5 10 15 20 25

Average Duration of Life (Hours)

As one can see from Figure 1 above, the average duration of life for line 73 flies exposed to

desiccant was approximately 11.4 hours, whereas the control flies lived on average for about

19.5 hours. Meanwhile, flies of line 304 lived on average up to 22.9 hours when exposed to

desiccant, whereas control flies lived 24 hours, as can be observed in Figure 2 below.
Figure 2

Average Desiccation Resistance of Ral-304 Drosophila melanogaster

1
Treatment

22.2 22.4 22.6 22.8 23 23.2 23.4 23.6 23.8 24 24.2

Average Duration of Life (Hours)

One week old 229 flies, however, resisted desiccation death on average for 14.8 hours, and

control flies lived for an average of 22.0 hours, as can be seen in Figure 3 below.
Figure 3

Average Desiccation Resistance of Ral-229 Drosophila melanogaster

1
Treatment
1

0 5 10 15 20 25

Average Duration of Life (Hours)

Finally, 229 flies that were five weeks of age seemed survived desiccation for an average of 11.7

hours, and control flies survived for about 13.8 hours on average, as shown in Figure 4.
Figure 4

Average Desiccation Resistance of Ral-229 Drosophila melanogaster (aged 5 weeks)

11.66
Treatment

13.84

10.5 11 11.5 12 12.5 13 13.5 14 14.5

Average Duration of Life (Hours)

Thus, it is seen that there is much variation between the genotypes in regards to their ability to

resist desiccation. Categorical analysis conducted with SAS software confirmed that there is a

statistically significant link between genotype and desiccation resistance.

CONCLUSION

A statistically significant link between genotype and desiccation resistance of male

Drosophila melanogaster has been established. Although a second trial could not be conducted

with lines 73 and 304 for the first experiment, this will be carried out in future. However, the
next step will be to conduct an experiment comparing the desiccation resistance of flies exposed

to Lisinopril with that of flies not exposed to the drug.

References

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Discusses renin-angiotensin system and how it is affected and how it affects the body

as aging occurs.

Abadir, P. M., Foster, D. B., Crow, M., Cooke, C. A., Rucker, J. J., Jain, A., . . . Watson, J. D.

(2011). Identification and characterization of a functional mitochondrial angiotensin

system. PNAS, 108(36), 14849-14854. http://dx.doi.org/10.1073/pnas.1101507108

Discusses a mitochondrial renin-angiotensin system and provides general background

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Buford, T. W., Manini, T. M., Hsu, F.-C., Cesari, M., Anton, S. D., Nayfield, S., . . . LIFE

Research Group. (2012). Angiotensin-converting enzyme inhibitor use by older adults is

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Geriatrics Society, 60(7), 1244-1252. http://dx.doi.org/10.1111/j.1532-5415.2012.04045

Folk, D. G., Han, C., & Bradley, T. J. (n.d.). Water acquisition and partitioning in Drosophila

melanogaster: Effects of selection for desiccation-resistance. Journal of Experimental

Biology, 204, 3323-3331. Retrieved from

http://jeb.biologists.org/content/204/19/3323.full

Fournier, D., Luft, F. C., Bader, M., Ganten, D., & Andrade-Navarro, M. A. (2012). Emergence

and evolution of the renin-angiotensin-aldosterone system. Journal of Molecular

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0894-z

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