Ivermectin Effects on Ivermectin Effects on Caenorhabditis elegans Caenorhabditis elegans

(C.elegans)(C.elegans)

Moyosore Olagoke

Ivermectin C.elegans

Background InformationBackground Information

1.Anti-parasitic MedicationStrongyloides stercoralis

Onchocerciasis volvulus Pubic lice infestation Scabies Wuchereria bancrofti

2 Mechanism of action Bind to GABA Channel Hyperpolarization by ↑Cl Permeability

http://www.pnas.org/cgi/content/full/97/6/2674/F2

C95H146O28

Background InformationBackground Information

Phylum NematodaPhylum Nematoda

Feed on OP50 bacteriaFeed on OP50 bacteria

PseudocoelomatesPseudocoelomates

Thrashing motion (contraction)Thrashing motion (contraction)

Female deposit 100,000 or more egg/dayFemale deposit 100,000 or more egg/day

Most individuals are HermaphroditesMost individuals are Hermaphrodites

Life Cycle : >12hoursLife Cycle : >12hours

Caenorhabditis elegans

ResearchResearch

Purpose :To determine the effect of Purpose :To determine the effect of Ivermectin on Ivermectin on wild type ( wild type (C.elegansC.elegans))

Plan to extend strainPlan to extend strain

ComplicationsComplications Method of introductionMethod of introduction Drug TestDrug Test Resistance strainResistance strain

Ivermectin response to OP50 Ivermectin response to OP50

bacteriabacteria Ivermectin has no effect on OP-50 Ivermectin has no effect on OP-50

bacteria bacteria

Avermectin is isolated from strepto Avermectins

Drug IntroductionDrug Introduction

Ivermectin→ Agar

Sample calculation

Stock 1- 0.1g/1000цl in 100% DMSO

Stock 2- 10цl of stock 1 in 9.99ml 1%DMSO

15цl of stock 2 in 8цl Agar

10цg/цl x 15цl= 150цg

150 цg/8цl = 62.5цg/цl

Wild type Wild type C.elegansC.elegans

Ivermectin: Wildtype Ivermectin: Wildtype wormworm

62.5цg/цl

Fewer numbers

No apparent movement

Worm appears transparent

18.75цg/цlSame conclusionMovement is perceived (~3 worms)

ComparisonComparison

IvermectinControl

Touch insensitiveTouch insensitive=Tapping=Tapping

Future AnalysisFuture Analysis

Growth Assay: Growth Assay: Count the number of gravid Count the number of gravid adult adult

Reflects Pharyngeal functionReflects Pharyngeal function

Mechanosensation: Mechanosensation: Touch sensitivity Touch sensitivity

Neurological ResponseNeurological Response

Motility assay: Motility assay: Number of worm still Number of worm still thrashingthrashing

LocomotionLocomotion

ReferencesReferences

American Society of, Health-System Pharmacists Inc.American Society of, Health-System Pharmacists Inc. Patient Drug Information From MedMasterPatient Drug Information From MedMaster. .

Maryland: 2007.Maryland: 2007.

Jamison, Dean T., and Joel G. Breman. Jamison, Dean T., and Joel G. Breman. Disease Control Priorities in Developing CountriesDisease Control Priorities in Developing Countries. 2nd. New York: . 2nd. New York:

Oxford University Press, 1993.Oxford University Press, 1993.

John Keane and Leon Avery John Keane and Leon Avery Mechanosensory inputs influence Caenorhabditis elegansMechanosensory inputs influence Caenorhabditis elegans pharyngeal activity via ivermectin sensitivity genes.pharyngeal activity via ivermectin sensitivity genes. Genetics. 2003 May; 164(1): 153–162. PMCID: PMC1462566Genetics. 2003 May; 164(1): 153–162. PMCID: PMC1462566

Griffiths, Anthony J.F., and Jeffery H. Miller. Griffiths, Anthony J.F., and Jeffery H. Miller. HumanGeneticsHumanGenetics. . 7th ed. New York: W.H. Freeman,1996.7th ed. New York: W.H. Freeman,1996.

Hart, Anne C. "Behavior." Hart, Anne C. "Behavior." Worm BookWorm Book July 3 2006. 04 April 2008July 3 2006. 04 April 2008<http://www.ncbi.nlm.nih.gov/books/bv.fcgi?rid=wormbook.<http://www.ncbi.nlm.nih.gov/books/bv.fcgi?rid=wormbook.

capter.7133capter.7133>.>.