Jennifer Lee Intraneuronally injected amyloid beta inhibits long-term potentiation in rat hippocampal slices -By: Izumi Nomura

!ajime "a#echi and Nobuo $ato Biol %&'%L- L(& "he article I)*e chosen to re*ie+ Intraneuronally injected amyloid beta inhibits longterm potentiation in rat hippocampal slices is about the effect that the protein amyloid beta ,-./ has on someone)s ability to form long term potentiation0 "his is significant because long term potentiation is directly related to one)s ability to form long term memories and is something that is interrupted by -lzheimer)s disease0 It has been pro*en in recent studies that e1tracellular amyloid beta buildup on the brain)s surface is a trait related to -lzheimer)s disease and is li#ely the reason long term potentiation is inhibited in these patients0 ,-lzheimer)s disease/ is associated +ith intracellular neurofibrillary tangles and e1tracellular amyloid beta ,-./ pla2ues in the regions of the brain that are responsible for learning and memory ,3eddy/0 "he function of the protein -. isn)t entirely understand but it)s belie*ed to play *arious roles in the body0 4ne possible function of the protein is its in*ol*ement in neuron plasticity0 5hen the protein turns pathogenic is +hen there are e1cessi*e amounts of it or +hen its structure has been altered to be a slightly longer and stic#ier form of the peptide ,6enetics/0 "he reason +hy this longer stic#ier protein creates problems is because it clumps together and forms large masses that can hinder synaptic processes and cause ner*e damage0 "he same problem of aggregation occurs +hen there is too much of the protein being produced0 "hese problems occur +hen the amyloid beta precursor protein is mutated and can no longer control the amount or e1act type of protein that it creates in the form of the amyloid beta protein0 "he e1cess protein has been obser*ed to induce e1citoto1icity in the ner*e cells of the brain and as a result cell damage and death0 71citoto1icity in this case is +hen the e1cess amounts of -. build up in the membrane of the mitochondria of these ner*e cells leads to an influ1 of calcium ions0 "his leads to o*erstimulation of the ner*e cells and the damage that pre*ents -lzheimer)s patients from being able to form long term memories05hile it has been consistently pro*en that e1tracellular -. inhibits long term potentiation no studies ha*e been done to see the effect of intracellular buildup of the protein0 "hat is the purpose of the article I)*e chosen0 Its goal is to in*estigate if there is a connection bet+een high intracellular -. le*els and difficulty in forming long term potentiation0 Because no research has been done to determine the effects of intracellular amyloid beta the research article I chose to re*ie+ is using rat hippocampal slices to in*estigate this topic0 "he researchers hypothesize that intraneuronal buildup of the protein is a precursor to -lzheimer)s disease and leads to the clumps of -. on the brain that are consistently related to the inability to form long term potentiation0 8re*ious e1periments ha*e also supported that there may be significance in the intracellular protein buildup as the follo+ing citation indicates0 In the brain -. peptides e1ist both +ithin defined deposits ,pla2ues and *ascular/ and as a di*erse array of other forms including soluble membrane-associated and intracellular species that might play far more significant roles in the production of dementia than the molecules se2uestered in e1tracellular pla2ues ,3oher/0 "his statement sho+s that researchers ha*e long thought that there is more to the disease than the protein pla2ue that is found on the surface of the brain0 It also points out that +hile any aggregation of -. is significant that the buildup on the inside of the cell could ha*e e*en more of an impact on the disease than the popularly studied e1tracellular clumps0 Being able to find a relationship bet+een intracellular -. and the de*elopment of

-lzheimer)s disease could help future researchers in finding a possible treatment or cure for the disease0 "he only current connection bet+een the protein and -lzheimer)s is because of the e1tracellular buildup0 "his means that if the intracellular protein buildup led to e1cessi*e amounts of e1tracellular protein this research topic +ould be e1tremely *aluable to research in the field of not only -lzheimer)s disease but other forms of dementia as +ell0 In order to test their hypothesis the researchers used hippocampal slices of a rat)s brain0 "hey sliced the hippocampus into portions that +ere 9((:m thic# and #ept them at a regulated temperature in a constant medium composed of mostly Na;l as +ell as other compounds +hich contribute to normal body chemistry0 "he researchers then diluted three different types of the -. and diffused them into the cytoplasm of the brain cells0 "he particular brain cells +hich +ere selected to be obser*ed are #no+n as pyramidal brain cells because they are in*ol*ed in neuroplasticity and therefore are directly related to impaired -lzheimer)s brain function0 In order to test +hether the injected amyloid beta has an effect on long term potentiation of the brain cells the researchers used a method called theta burst stimulation0 "heta burst stimulation is described as a form of repetiti*e transcranial magnetic stimulation0 5hen applied to motor corte1 it leads to after-effects on corticospinal and corticocortical e1citability that may reflect L"8<L"=-li#e synaptic effects ,=i Lazarro >i/0 Because this method is able to simulate long term potentiation li#e synaptic action it)s *ery useful in this research0 -fter the stimulation is applied to the brain cells the researchers measure the e1citatory postsynaptic current ,78?;/0 "he magnitude of this current is able to determine ho+ effecti*ely long term potentiation is ta#ing place because a larger magnitude means more effecti*e L"80 "he results of this research led to se*eral different conclusions0 "he particular data that I thin# is interesting and is most rele*ant to the hypothesis is the one stating that intracellular concentrations of -. +ill impair long term potentiation0 "his conclusion +as reached by the researchers +hen they e1amined the differing 78?; measurements of the pyramidal cells injected +ith -. protein and compare them to the measurements of the cells +ithout the intracellular protein injections0 "his particular conclusion is only true +hen the cells are injected +ith the specific type of amyloid beta protein -.-0 "he figure that goes along +ith this result consists of a set of si1 different graphs0 "he first t+o graphs - and B represent respecti*ely the magnitude of 78?; measured after theta burst stimulation is used on cells +ithout injected protein the control and the other graph ,B/ representing the 7?8; magnitudes +ith the injected -.- o*er a consistent period of time0 In graph - the magnitudes of the 7?8; readings are as a +hole higher than those found in graph B0 It appears that the a*erage of the graph - results are about &&@ pico-amps +hile the a*erage of graph B)s 78?; measurements are about A@( pico-amps0 "his significant decrease in 78?; readings sho+s that the presence of -. in the graph B readings led to a decrease in the ability of the neurons to undergo long term potentiation0 It also seems li#e the readings in graph B decrease more rapidly after the theta burst than the data in graph -0 "he ne1t graph in this figure graph ; is a representation of the different 7?8; data +hich compares cells injected +ith -.- *ersus cells injected +ith the same peptide +ith the re*ersed order of amino acids -.-0 In this graph the cells injected +ith the -. ha*e o*erall lo+er percent of baseline 78?; for e*ery time recorded after theta burst stimulation than any of the data collected from cells injected +ith the re*erse peptide0 -lso recorded on this graph is the percent of baseline 78?; +hen cells are injected +ith no amyloid beta protein0 "hese readings are almost identical to those ta#en from cells injected +ith -.-

0 "his indicates that +hile -.- inhibits long term potentiation the re*erse peptide of this protein has little effect on this particular action of the cells0 6raphs = 7 and B of this figure demonstrate that se*eral different inhibitors injected along +ith the amyloid beta can re*erse the inhibitory effects that the protein has on long term potentiation0 "he inhibitors used in this research are ?B&(%@C( and D8780 "he final graph B sho+s an o*erall summary of all of the results0 It)s a graph +hich sho+s the different a*erage percent baseline 78?; measurements for cells injected +ith the follo+ing: no -. ,control/ 3-. ,re*erse/ -. ?BE -. ,?B&(%@C(/ and D8E -. ,D878/0 Bor the final graph the statistical analysis -N4>- +as used o*er the four e1perimental groups just described0 =unnett)s test +as then used to identify that L"8 inhibition +as larger in the cells injected +ith -.-0 -ll the cells sho+ a percent of about A@( e1cept for the cell only injected +ith amyloid beta0 "his indicates the ability of D878 and ?B&(%@C( in counteracting the long term potentiation inhibitory actions by -.0 4ne of the +ea#nesses +ith this research effort is the difference bet+een the results using a rat)s brain and that of a human0 5hile rats are e1cellent laboratory animals it)s undeniable that their brains function differently than that of humans especially +hen discussing memory and learning li#e this research is primarily doing0 "here aren)t possibilities to test the human brain in a similar +ay but more could be learned about -lzheimer)s through obser*ation of the disease in a person0 - second +ea#ness of this particular e1periment is that there +as *ery little similar research done on this topic before0 "his is the first recorded research done on the effects of intracellular -. +hich leads you to be at least a little s#eptical of the results0 5ithout further research it)s hard to belie*e that these single results can be too credible0 5hile this research had its faults there +ere se*eral reasons +hy it +as able to ma#e a strong statement0 "he methods it used for instance theta burst stimulation and 78?; are both highly accepted in the scientific community and lea*e little room for interpretation of the results0 "his leads to *ery e1act data +hich is hard to read in different +ays +hen a proper control is used0 -nother strength of the e1periment is the ample amount of funding a*ailable for research in the field of -lzheimer)s0 ;urrently because -lzheimer)s is becoming a rising issue +ith the increase in lifespan there is an e1cess of money being poured into research about it0 "his means that in*estigating -lzheimer)s +as a *ery useful and interesting topic for the researchers to choose and is something that +ill be highly funded and +ill be of great use to a lot of similar research in the near future0 I belie*e an interesting follo+ up to this e1periment +ould be to research possible treatments and solutions to -lzheimer)s disease and the amyloid beta buildup that is li#ely its cause0 4ne possible solution to the e1cess amyloid beta is to use mesenchymal stem cells +hich are belie*ed to brea# do+n the -. protein0 In *itro results indicate that ,mesenchymal stem cells/ might be useful *ehicles for deli*ering anti--. acti*ity ,!abisch/0 I thin# this +ould be an interesting follo+ up e1periment for these researchers because they could use the same e1perimental techni2ues in*ol*ing rat hippocampal brain slices to test +hat type of effect these mesenchymal stem cells ha*e on e1cess -. on the brain)s surface0 It)s also an interesting research topic because these stem cells can be easily obtained from umbilical cords +hich +ould other+ise ha*e been thro+n a+ay after becoming unattached0

5or#s ;ited:

=i Lazarro > 8ilato B =ileone D0 &((C0 "he physiological basis of the effects of intermittent theta burst stimulation of the human motor corte10 "he Journal of 8hysiology FInternetG0 Fcited &(A% 4ct A&G0 -*ailable from: http:<<jp0physoc0org<content< @CH<AH<%C'A0fullItarget-' 6enetics !ome 3eference0 &(A% ?ep &%0 -88 FInternetG0 J0?0 National Library of DedicineK FDay &(A& cited &(A% 4ct A&G0 -*ailable from: http:<<ghr0nlm0nih0go*<gene<-88 !abisch ! ?chmid B0 &(A(0 7fficient processing of -lzheimerLs disease amyloid-Beta peptides by neuroectodermally con*erted mesenchymal stem cells0 ?tem ;ells and =e*elopment FInternetG0 cited &(A% 4ct AAG AM,@/: H&M-H%%0 -*ailable from: http:<< online0liebertpub0com0librarylin#0uncc0edu<doi<abs<A(0A(CMscd0&((M0((9@ 3eddy !00 &((M0 -myloid beta mitochondrial structural and functional dynamics in -lzheimerLs disease0 71perimental Neurology FInternetG0 F&(A% cited &(A% 4ct AAG >olume &AC ,issue &/: ,p&CH-&M&/0 -*ailable from: http:+++0sciencedirect0com0librarylin#0uncc0edu< science<article<pii<?((A99CCH(M((A%(' 3oher - 7sh ; $o#john "0 &((M0 -myloid beta peptides in human plasma and tissues and their significance for -lzheimerLs disease0 -lzheimerLs and =ementia FInternetG0 F&(A% cited &(A% 4ct AAG @,A/: AC-&M0 -*ailable from: http:<< +++0sciencedirect0com0librarylin#0uncc0edu<science<article<pii<?A@@&@&H((C(&CMH%