The Perilipin Homologue, Lipid Storage Droplet 2, Regulates Sleep Homestasis and Prevents Learning Impairments Following Sleep

Loss Maggie Johnson, Biol. ! ", #lass $%$$ Fri. http://maggieseworld.weebly.com/ I. Introduction &. Insu''i(ient sleep results in glu(ose intoleran(e, insulin resistan(e and o)esit* B. Sleep deprivation results in (ognitive impairments and death #. Sleep homeostasis restores vital )iologi(al 'un(tions +. Identi'*ing pathwa*s that regulate sleep homeostasis through (ontrol groups and environmental o)servations with humans, 'lies and rats ,. Man* )ehaviors in'luen(ed )* intera(tions )etween genes and environment F. La(- o' 'ood availa)ilit* in(reases duration o' wa-ing .. #omparison )etween wa-ing indu(ed )* starvation vs. wa-ing indu(ed )* sleep deprivation H. .oal% Providing geneti( eviden(e that lipid meta)olism pla*s role in regulating sleep homeostasis II. Materials and Methods &. /0da* old mutant 'lies B. Sleep Re(ording i. !" mm glass tu)es ii. 0minute )ins #. Sleep +eprivation
i. Sleep nulli'*ing apparatus 1S2&P3

ii. +eprived o' sleep )etween 4 and 5 hours +. Starvation

i. 6 agar solution ii. 5" um etomo7ir ,. Photosensitivit* i. Lightened and dar-ened (ham)ers F. 8uinine9Humidit* Sensitivit* i. : (m (*lindri(al tu)e ii. Filter paper wetted with ;uinine .. 8P#R H. Trigl*(eride Measurements I. <il Red < Staining i. #<5 ii. :6 para'ormaldeh*de in phosphate )u''ered saline iii. Rinsed in !$6 isopropanol J. Blue Food +*e III. Results &. Flies show large homeostati( response a'ter 4 h o' sleep deprivation B. &PS signi'i(antl* disrupted )* sleep deprivation )ut un(hanged with starvation #. Flies have large sleep re)ound a'ter 4 h o' sleep deprivation )ut do not have a homeostati( response a'ter 4 h o' starvation IV. Discussion &. Starvation in(reases wa-ing not (ompensated )* sleep re)ound and does not result in learning de'i(its B. Brummer has e7aggerated sleep re)ound #. Lsd5 withstands negative e''e(ts o' 5 h o' sleep deprivation +. Lipid handling relevant to response to sleep loss ,. Sleep regulator* pathwa*s and lipid meta)olism ver* (onne(ted F. &m*lase should )e evaluated

.. &denosine ma* pla* a role

Maggie Johnson, Biol. ! ", Fri. $%$$ Title: The Perilipin Homologue, Lipid Storage Droplet 2, Regulates leep Homeostasis and !re"ents #earning Impairments $ollowing leep #oss

Introduction: Humans, 'lies and rats have shown with (on(lusive eviden(e that sleep deprivation (an result in 'ar0rea(hing ph*siologi(al damage, in(luding glu(ose intoleran(e, insulin resistan(e, o)esit*, (ognitive impairments and death. Sleep homeostasis, the in(rease o' sleep o)served 'ollowing sleep loss, helps reinstate vital )iologi(al 'un(tions lost during sleep deprivation. Man* )ehaviors are shaped )* inter(ommuni(ation )etween genes and the environment, su(h as starvation, whi(h in'luen(es the response to prolonged wa-ing. +uring indu(ed starvation, 'lies and rats e7perien(e in(reased duration o' wa-ing )ut do not have an adverse rea(tion as the* do with sleep deprivation. Reason %or Research: The genes )rummer 1)mm3 and Lipid storage droplet 5 1Lsd53 ma* assist with sleep regulation. Lipid meta)olism ma* prote(t against the ph*siologi(al damage prevalent with sleep deprivation. Materials and Methods: /0da* old 'lies were deposited in !" mm glass tu)es with regular la) 'ood and o)served with the Tri-ineti(s a(tivit*0monitoring s*stem, whi(h are (onne(ted to (a)les 'or data su)mission. The 'lies were deprived o' sleep using the sleep0nulli'*ing apparatus 1S2&P3 )etween 4 to 5 hours. Sleep homeostasis was determined individuall* using minutes gained divided )* minutes lost during the re(over* pro(ess. Flies were also deposited into 'ull* dar-ened tu)es with 6 agar solution 'or the same amount o' time as the 'lies )eing sleep deprived. &'ter the starvation period 'lies returned to their normal eating pro(ess to determine sleep homeostasis. The 'lies utili=ed lighted and dar-ened tu)es that were (overed with 'ilter paper. The paper in the lighted tu)es was dampened with M 8uinine h*dro(hloride solution whi(h helped show the humidit* on the paper. Flies were weighed and homogeni=ed to withdraw the lipids and the lipid levels were measured using a (urve o' -nown trigl*(eride ad>a(ent to it. The <il Red < staining was used as a stain to re(ord the data on Histo)ond slides and then were divided in :6 para'ormaldeh*de in

phosphate )u''ered saline. ?hen the 'lies were released to eat the normal 'ood again it was o)serva)le )* tainting the 'ood with )lue d*e whi(h (ould )e seen in the a)domen o' the 'l*. Results: ?a-ing indu(ed )* starvation does not initiate a sleep re)ound. Flies )egan to die with sleep deprivation at around $ hours, even as a group. ?ith starvation methods in the agar solution, there was immediate and reo((urring wa-ing )ut when the* went )a(- onto regular la) 'ood there was no sleep re)ound. Discussion: Starvation was (hosen in this stud* )e(ause it is naturall* o((urring with man* spe(ies and it (ould )e easil* studied in a (ontrol group. Starvation (auses (onsistent wa-ing periods )ut it doesn@t have negative neurologi(al or ph*si(al e''e(ts as is shown with sleep deprivation. )mm and Lsd5 were assigned )e(ause o' their a)ilit* to alter the )od*@s response to starvation. &nimals and mammals have learned to adapt through evolutionar* measures when wa-ing has )een prevalent in their environments. &onclusion: Proper lipid handling ma* )e a)le to aid with sleep loss, displa*ing the )ond )etween lipid meta)olism and (onsistent sleep. The meta)olism is adversel* a''e(ted )* sleep loss, whi(h has alread* )een proven in man* prior studies. The stud* aims to e7amine whi(h lipid is )est 'or sleep homeostasis )ut it has not )een resear(hed well enough at this point. Studies monitoring (ognitive )ehavior 'ollowing sleep deprivation (an help to isolate genes that attri)ute to (onsistent wa-ing periods.

Bi)liograph*% http%99www.plos)iolog*.org9arti(le9in'o6/&doi 65F $. /4 65F>ournal.p)io. $$$:!!