The Role of The Role of Estrogens in the Estrogens in the Hippocampal-Hippocampal-

Neuropeptide YNeuropeptide YDamaly FerreiraDamaly Ferreira

Jana VeliJana Velisskovkovaa

Albert Einstein College of Albert Einstein College of MedicineMedicine

KeywordsKeywords

Estrogen- female sex hormones that are secreted by Estrogen- female sex hormones that are secreted by the ovary and responsible for typical female the ovary and responsible for typical female characteristics.characteristics.Seizure- a sudden burst of electric activity in the Seizure- a sudden burst of electric activity in the brainbrainEpilepsy- a seizure disorderEpilepsy- a seizure disorderHippocampus- a part of the brain affected by estrogen Hippocampus- a part of the brain affected by estrogen and is associated with transferring information into and is associated with transferring information into memorymemoryNPY- a powerful inhibitory system in the hippocampal NPY- a powerful inhibitory system in the hippocampal hilar region implicated in the regulation of seizure hilar region implicated in the regulation of seizure activity.activity.Status Epilepticus- an episode or continuous seizure Status Epilepticus- an episode or continuous seizure of 30 minutes or moreof 30 minutes or more

IntroductionIntroduction

Objective: In our experiment, our goal is Objective: In our experiment, our goal is to figure out the role estrogens play in to figure out the role estrogens play in the Hippocampus- Neuropeptide Y the Hippocampus- Neuropeptide Y region (NPY).region (NPY).Hypothesis: Estrogens are accountable Hypothesis: Estrogens are accountable for the actions produced in the NPY.for the actions produced in the NPY.Literature Review: In past studies, there Literature Review: In past studies, there have been human and animal studies have been human and animal studies that have created a lot of controversial that have created a lot of controversial data that debate the role of estrogens data that debate the role of estrogens with seizures.with seizures.

Materials and MethodsMaterials and MethodsOvariectomyOvariectomyTreat Sprague Dawley rats with PPTTreat Sprague Dawley rats with PPT PPTPPT

4,4,4-(4-propy-[1H]-pyrazole-1,3,5-triyl)trisphenol4,4,4-(4-propy-[1H]-pyrazole-1,3,5-triyl)trisphenolER beta and ER alpha agonistER beta and ER alpha agonistC24H22N2O3 C24H22N2O3 H2O H2O

PerfusionPerfusionBrain is taken out of skull Brain is taken out of skull Brain is immersed into 2-methylbutane, Brain is immersed into 2-methylbutane, then stored into storage freezerthen stored into storage freezerWashed with PBS (saline solution)Washed with PBS (saline solution)

Materials and Methods Materials and Methods con’tcon’t

ImmunohistochemistryImmunohistochemistry50 50 µl of Hµl of H22OO2 + 2 + 5 ml of PBS5 ml of PBS

Wash wells with PBS (5 minutes each)Wash wells with PBS (5 minutes each)

160 µl of BSA + 5 ml of PBS + 20 µl 160 µl of BSA + 5 ml of PBS + 20 µl Triton into wells for 90 minutesTriton into wells for 90 minutes

Apply NPY antibody + 5 ml PBS for 90 Apply NPY antibody + 5 ml PBS for 90 minutesminutes

Put 200 µl of solution in each wellPut 200 µl of solution in each well

Take out NPY antibody solution and Take out NPY antibody solution and wash 3 times with PBSwash 3 times with PBS

Materials and Methods Materials and Methods con’tcon’t

Immunohistochemistry con’tImmunohistochemistry con’t100 µl of BSA + 5 ml PBS + 25 µl anti-100 µl of BSA + 5 ml PBS + 25 µl anti-rabbit in wells for 1 hrrabbit in wells for 1 hr

Wash 3 times with PBSWash 3 times with PBS

Apply AB in wells and settle for 1 hrApply AB in wells and settle for 1 hr

Wash 3 times with PBSWash 3 times with PBS

DABDAB

Expected ResultsExpected Results

The PPT used, will hopefully detect The PPT used, will hopefully detect which Estrogen Receptor was triggered.which Estrogen Receptor was triggered.

In the maps of the slides, we will In the maps of the slides, we will hopefully see whether the Estrogen hopefully see whether the Estrogen created any damage to the hippocampal created any damage to the hippocampal area, or whether the NPY protected the area, or whether the NPY protected the hippocampal area from any damage.hippocampal area from any damage.

ReferencesReferencesImmunohistochemistry con’tImmunohistochemistry con’tAre the role of Estrogens in Seizures and Epilepsy: the bad Are the role of Estrogens in Seizures and Epilepsy: the bad guys or the good guys? 2005, Elsevier Ltd, Bronx, NY.guys or the good guys? 2005, Elsevier Ltd, Bronx, NY.Carol Camfield, M.D 2004. What is epilepsy? Carol Camfield, M.D 2004. What is epilepsy? http://www.epilepsy.com/101/ep101_epilepsy.htmlhttp://www.epilepsy.com/101/ep101_epilepsy.html, 2 Aug 2006 , 2 Aug 2006 Centre for Cancer Education [on-line] 1997. Centre for Cancer Education [on-line] 1997. http://cancerweb.ncl.ac.uk/cgi-bin/omd?http://cancerweb.ncl.ac.uk/cgi-bin/omd?query=seizure&action=Search+OMD, 3 Aug 2006.query=seizure&action=Search+OMD, 3 Aug 2006.Centre for Cancer Education [on-line] 1998. Centre for Cancer Education [on-line] 1998. http://cancerweb.ncl.ac.uk/cgi-bin/omd?query=status+epileptichttp://cancerweb.ncl.ac.uk/cgi-bin/omd?query=status+epilepticusus, 2 Aug 2006. , 2 Aug 2006. The American Heritage® Dictionary of the English Language The American Heritage® Dictionary of the English Language [on-line] 2000. [on-line] 2000. http://dictionary.reference.com/browse/estrogenhttp://dictionary.reference.com/browse/estrogen, 2 Aug 2006., 2 Aug 2006. Veliskova, J., Velisek, L., Galanopoulou, A.S. and Sperber, E.F.: Veliskova, J., Velisek, L., Galanopoulou, A.S. and Sperber, E.F.: Neuroprotective effects of estrogens on hippocampal cells in Neuroprotective effects of estrogens on hippocampal cells in adult female rats following status epilepticus. Epilepsia, 41 adult female rats following status epilepticus. Epilepsia, 41 Suppl.6:S30-35,(2000). Suppl.6:S30-35,(2000).

AcknowledgementsAcknowledgements

Jana Veliskova, M.D., Ph.D. Jana Veliskova, M.D., Ph.D. Hong Wong Hong Wong ZunjuZunjuTamara DennisTamara DennisDr. Sat BhattacharyaDr. Sat BhattacharyaHarlem Children SocietyHarlem Children SocietyMSKCCMSKCCThe AudienceThe Audience