Aw ni ce aw ka taa Nature.com kan. I bɛ baara kɛ ni navigatɛri min ye, o cogoya dan ye CSS dɛmɛni ye. Walasa ka ɲɛtaa ɲuman sɔrɔ, an b’a ɲini i fɛ i ka baara kɛ ni i ka navigatɛri kura ye (walima i ka Compatibility Mode bali Internet Explorer kɔnɔ). O waati kɔnɔna na, walasa ka dɛmɛn sabati, an bɛ site web jira k’a sɔrɔ an ma styling walima JavaScript kɛ.
Asidi propionic (PPA) bɛ kɛ ka mitokondriyaw ka baarakɛbaliya jɔyɔrɔ sɛgɛsɛgɛ farikolojidɛsɛbanaw la i n’a fɔ autizmu spectre disorder. A dɔnna ko PPA bɛ mitokondriyaw ka biyojenizi, farikoloɲɛnajɛ ani fɛnw jiginni tiɲɛ. Nka, PPA nɔ minnu bɛ mitokondriyaw ka fanga la, u farali ani u farali ɲɔgɔn kan, olu bɛ to gɛlɛya la k’a sababu kɛ nin fɛɛrɛ ninnu waati gɛlɛn ye. Yan, an bɛ baara kɛ ni ja hakɛ jateminɛ fɛɛrɛw ye minnu bɛ ɲɔgɔn dafa walasa ka sɛgɛsɛgɛli kɛ cogo min na PPA bɛ nɔ bila mitokondriyaw ka ultrastructure, morphologie ani dynamics la SH-SY5Y seliluw la minnu bɛ i n’a fɔ senfagabana. PPA (5 mM) ye dɔ bɔ kosɛbɛ mitokondriyaw yɔrɔ la (p < 0,01), Feret cɛmancɛ ni a lamini na (p < 0,05), ani yɔrɔ 2 la (p < 0,01). Mitokondriyaw ka ko kɛlenw ɲininikɛla sɛgɛsɛgɛli y’a jira ko dɔ farala kosɛbɛ (p < 0,05) fisiyɔn ni fusili ko kɛlenw na, o la, mitokondriyaw ka rezo dafalen mara degun cogoyaw la. Ka fara o kan, cMYC (p < 0,0001), NRF1 (p < 0,01), TFAM (p < 0,05), STOML2 (p < 0,0001) ani OPA1 (p < 0,05) ka ARNm jirali dɔgɔyara kosɛbɛ. 01). O bɛ mitokondriyaw cogoya, biyojɛnɛsi ani u fanga labɛncogo kura jira walasa ka baara sabati degun cogoyaw la. An ka kunnafoniw bɛ hakilina kura di PPA nɔfɛkow kan mitokondriyaw ka fanga kan ani ka ja lajɛ fɛɛrɛw nafa jira walasa ka sariyasunba gɛlɛnw sɛgɛsɛgɛ minnu bɛ mitokondriyaw ka degun jaabiw la.
Mitokondriyaw ye mɔgɔ jɔnjɔnw ye selilɛriw ka baara suguya caman na ka tɛmɛ u jɔyɔrɔ danma kan fanga sɔrɔli ni biyosintɛzi la. Mitokondriyaw ka farikoloɲɛnajɛ ye kalisiyɔmu taamasiɲɛw ɲɛnabɔbaga jɔnjɔn ye, farikoloɲɛnajɛ ni redox homeostasis, funu taamasiɲɛw, epijeni caman cili, selilɛriw caya, danfara ani selilɛri saya porogaramulen Kɛrɛnkɛrɛnnenya la, mitokondriyaw ka farikoloɲɛnajɛ nafa ka bon kosɛbɛ senfagabana yiriwali, u ka ɲɛnamaya ani u ka baara la, wa a sen bɛ kosɛbɛ senfagabana jiracogo suguya caman na2,3,4.
San tan tɛmɛnenw kɔnɔ, farikoloɲɛnajɛ cogoya bɔra kɛnɛ kan i n’a fɔ farikolojidɛsɛ, danfara, kɔgɔli ani plastiki ɲɛnabɔbaga cɛmancɛ5,6. Kɔsa in na, mitokondriyaw cogoya ni u fanga kɛra mitozi yɔrɔ nafamaw ye kɛrɛnkɛrɛnnenya la, o ye wale ye min bɛ mitokondriyaw kɛnɛmanw ka kulu mara selilu kɔnɔ. Mitokondriyaw ka fanga bɛ labɛn ni sira gɛlɛnw ye minnu bɛ tali kɛ ɲɔgɔn na, k’a ta mitokondriyaw ka biyojenizi ni biyoenergitiki la ka se mitokondriyaw ka faranfasi, fusion, transport ani clearance ma7,8. Nin jɛ-ka-baara fɛɛrɛ ninnu dɔ tiɲɛni bɛ mitokondriyaw ka rezo kɛnɛmanw marali tiɲɛ, wa a bɛ na ni baarakɛcogo kɔlɔlɔ juguw ye farikolojidɛsɛ la9,10. Tiɲɛ na, mitokondriyaw ka fanga dɔgɔyali bɛ ye hakiliɲagami, farikolojidɛsɛ ani hakiliɲagami bana caman na, autizmu spectrum disorders (ASD)11,12 fana sen bɛ o la.
ASD ye farikolojidɛsɛ ye min tɛ kelen ye, a ka jamu ni a ka jamu cogoya ka gɛlɛn. ASD ciyɛntacogo tɛ sɔsɔ, nka a sababu min bɛ sɔrɔ molekiyɔmu kɔnɔ, o ma faamuya ka ɲɛ. Kunnafoni dalajɛli ka bɔ misaliw la minnu bɛ kɛ ka kɔn kɛnɛyaso ɲɛ, kɛnɛyaso kalanw, ani kunnafonidilanw molekiyɔmu caman na, olu bɛ dalilu caman di mitokondriyaw baarakɛbaliya kan ASD13,14. An ye ADN metilasi sɛgɛsɛgɛli kɛ ka tɛmɛn jamu bɛɛ kan ASD banabagatɔw ka kulu dɔ la ani ka jamu metilasi danfaralenw dɔn minnu lajɛlen don mitokondriyaw farikoloɲɛnajɛ siraw kan O kɔfɛ, an ye diferential methylation (metilasi danfaralen) fɔ mitokondriyaw ka biyojɛnɛsi ni u ka dinamiki cɛmancɛ-labɛnbagaw la, o min ni ADNm kopi hakɛ cayalenba ani sugunɛ farikoloɲɛnajɛ cogoya caman caman cili bɛ tali kɛ ɲɔgɔn na ASD16 kɔnɔ. An ka kunnafoniw bɛ dalilu caman di minnu b’a jira ko mitokondriyaw ka fanga ni omɛostasi jɔyɔrɔ ka bon ASD banakisɛfagalanw na. O la, ka fɛɛrɛ faamuyali ɲɛ ka ɲɛsin jɛɲɔgɔnya min bɛ mitokondriyaw ka fanga, u cogoya ani u baarakɛcogo cɛ, o ye laɲini jɔnjɔn ye ɲininiw na minnu bɛ senna hakilibanaw kan minnu bɛ sɔrɔ mitokondriyaw ka baarakɛbaliya filanan fɛ.
Tuma caman na, baara bɛ kɛ ni molekiyɔmu fɛɛrɛw ye walasa ka jamu kɛrɛnkɛrɛnnenw jɔyɔrɔ sɛgɛsɛgɛ mitokondriyaw ka degun jaabiw la. Nka, o fɛɛrɛ in bɛ se ka dantigɛ ni mitotiki kɔlɔsili fɛɛrɛw ye minnu bɛ kɛ yɔrɔ caman na ani minnu bɛ kɛ waati kɔnɔ. Ka fara o kan, mitokondriyaw ka jɛnɛya danfara jirali ye baarakɛcogo caman yeli jiralan ye min tɛ ɲɛ, kɛrɛnkɛrɛnnenya la, a ka c’a la, jɛnɛya hakɛ danmadɔ dɔrɔn de bɛ sɛgɛsɛgɛ. O la, fɛɛrɛ tilennen caman fɔra walasa ka mitokondriyaw ka baara ni biyoenerjetiki kalan17. Mitokondriyaw cogoya bɛ tali kɛ mitokondriyaw ka fanga la kosɛbɛ. Mitokondriyaw cogoya, u ni ɲɔgɔn cɛ ani u jɔcogo nafa ka bon fanga sɔrɔli la ani mitokondriyaw ni selilɛriw ka ɲɛnamaya la5,18. Ka fara o kan, mitozi yɔrɔ suguyaw sinsinnen bɛ mitokondriyaw cogoya caman yeli kan, minnu bɛ se ka kɛ mitokondriyaw ka baarakɛbaliya laban nafamaw ye ani ka kɛ basigi ye fɛɛrɛko kalan nataw la.
Mitokondriyaw cogoya bɛ se ka kɔlɔsi k’a ɲɛsin u yɛrɛ ma ni transmission electron microscopy (TEM) ye, o bɛ kɛ sababu ye ka seliluw ka ultrastructure sɛgɛsɛgɛli caman kɛ. TEM bɛ mitokondriyaw kirisita cogoya, u cogoya ani u jɔcogo jira k’a ɲɛsin mitokondriyaw kelen-kelen bɛɛ ɲɛnabɔli ma, sanni a ka da jɛnɛya sɛbɛnni dɔrɔn kan, farikolojɔlifɛnw jirali walima mitokondriyaw ka baarakɛcogo paramɛtiriw kan selilɛriw kɔnɔ17,19, Ka fara o kan, TEM bɛ mitokondriyaw ni farikolo yɔrɔ wɛrɛw cɛ jɛɲɔgɔnyaw kalanni nɔgɔya, i n’a fɔ endoplasmic reticulum ani autophagosomes, minnu jɔyɔrɔ ka bon mitokondriyaw ka baara la ani homeostasis la21, O la, o bɛ TEM kɛ daminɛ ɲuman ye mitokondriyaw ka baarakɛbaliya kalanni na ka sɔrɔ ka sinsin sira kɛrɛnkɛrɛnnenw walima jamu kɛrɛnkɛrɛnnenw kan. Ni mitokondriyaw ka baara bɛ ka caya ka taa a fɛ ni hakiliɲagami ye, mago jɛlen bɛ yen ka se ka mitokondriyaw cogoya ni u fanga sɛgɛsɛgɛ k’a ɲɛsin u yɛrɛ ma ani ka kɛɲɛ ni hakɛ ye senfagabana modɛliw kɔnɔ in vitro.
Nin barokun in kɔnɔ, an bɛ mitokondriyaw ka fanga sɛgɛsɛgɛ mitokondriyaw ka baarakɛbaliya senfagabana modɛli kɔnɔ autizmu spektrum bana la. An ye propionyl-CoA carboxylase beta (PCCB) metilasi danfaralen fɔ ka tɛmɛ ASD15 kɔnɔ, o ye mitokondriyaw ka propionyl-CoA karɔkisilazi anzimu PCC ka yɔrɔ fitinin dɔ ye. A dɔnna ko PCC ka baarakɛcogo jugu bɛ kɛ sababu ye ka poropiyɔni bɔlenw dalajɛ ni baga ye, asidi poropiyɔni (PPA)23,24,25. A jirala ko PPA bɛ senfagabana farikoloɲɛnajɛ tiɲɛ ani ka kɛwale caman Changer in vivo ani a ye baganw ka misali sabatilen ye walasa ka senfagabana yiriwali fɛɛrɛw sɛgɛsɛgɛ minnu bɛ ASD26,27,28 la. Ka fara o kan, a fɔra ko PPA bɛ mitokondriyaw ka seko ni dɔnko tiɲɛ, biyojenizi ani ninakili in vitro ani a kɛra baarakɛminɛn ye kosɛbɛ walasa ka mitokondriyaw ka baarakɛbaliya misali kɛ senfagabana kɔnɔ29, Nka, PPA ka mitokondriyaw ka baarakɛbaliya nɔ min bɛ mitokondriyaw cogoya ni u fanga la, o ma faamuya kosɛbɛ hali bi.
Nin kalan in bɛ baara kɛ ni ja lajɛcogo dafalenw ye walasa ka PPA nɔw hakɛ jateminɛ mitokondriyaw cogoya, u fanga ani u baara la SH-SY5Y selilɛriw kɔnɔ. Fɔlɔ, an ye TEM fɛɛrɛ dɔ labɛn walasa ka fɛn caman yeli ye mitokondriyaw cogoya ni ultrastructure17,31,32. Ni an y’a jateminɛ ko mitokondriyaw33 ka fanga ka bon, an ye baara kɛ fana ni mitokondriyaw ka ko kɛlenw sigiyɔrɔmako (MEL) sɛgɛsɛgɛli ye walasa ka fɛn caman sɛgɛsɛgɛ fɛn caman Changements dans le balance de fission et fusion events, mitokondriyaw hakɛ ani hakɛ PPA degun kɔnɔ. A laban na, an y’a sɛgɛsɛgɛ ni mitokondriyaw cogoya n’u ka fanga bɛ tali kɛ fɛn caman Changements dans la expression de gènes sur les biogénèses, fission et fusion. Ni an y’u bɛɛ lajɛlen ta, an ka kunnafoniw b’a jira ko gɛlɛya min bɛ mitokondriyaw ka fanga ɲɛnabɔcogo fɛɛrɛw ka gɛlɛya ɲɛfɔli la. An bɛ TEM nafa jira mitokondriyaw cogoya kalanni na i n’a fɔ mitozi labanyɔrɔ min bɛ se ka suman SH-SY5Y selilɛriw kɔnɔ. Ka fara o kan, an b’a jira ko TEM kunnafoniw bɛ kunnafoni nafamaw di ni u farala ɲɔgɔn kan ni ja lajɛ fɛɛrɛw ye minnu fana bɛ ko kɛlenw minɛ minnu bɛ fanga sɔrɔ farikoloɲɛnajɛ degunw jaabi la. Molecules (molecules) ɲɛnabɔli fɛɛrɛw minnu bɛ senfagabana selilɛri mitosi dɛmɛ, olu taamasiɲɛ wɛrɛw bɛ se ka kɛ sababu ye ka hakilina nafamaw di farikoloɲɛnajɛ ni senfagabana banaw mitokondriyaw yɔrɔ kan.
Walasa ka mitokondriyaw degun bila mɔgɔ la, SH-SY5Y selilɛriw furakɛra ni PPA ye ni 3 mM ani 5 mM sodium propionate (NaP) ye. Sani TEM ka kɛ, sɛgɛsɛgɛli kɛlenw tun bɛ kɛ ni fɛnɲɛnamafagalanw labɛnni ye ni kiriyɔzɛni ye ni degunba ni nɛnɛ ye (Ja 1a). An ye mitokondriyaw ja sɛgɛsɛgɛli pipeline otomatiki dɔ labɛn walasa ka mitokondriyaw jamanadenw cogoya 8 suman biologique replicate saba kɔnɔ. An y’a ye ko PPA furakɛli ye fɛn naani Changé kosɛbɛ: yɔrɔ 2, yɔrɔ, lamini, ani Feret cɛmancɛ (Ja 1b–e). Yɔrɔ 2 dɔgɔyara kosɛbɛ ni PPA furakɛli ye ni milimɛtɛrɛ 3 ni milimɛtɛrɛ 5 ye (p = 0,0183 ani p = 0,002, o cogo kelen na) (Ja 1b), ka sɔrɔ yɔrɔ (p = 0,003), lamini (p = 0,0106) ani Feret cɛmancɛ bɛɛ dɔgɔyara kosɛbɛ. Dɔgɔyaliba kɛra (p = 0,0172) 5 mM furakɛli kulu la ni i ye a suma ni kɔrɔsili kulu ye (Ja 1c–e). Dɔgɔyaliba kɛra yɔrɔ ni lamini na, o y’a jira ko selilu minnu furakɛra ni 5 mM PPA ye, olu ka mitokondriyaw ka dɔgɔn, u lamininen don kosɛbɛ, wa ko o mitokondriyaw janya ka dɔgɔn ka tɛmɛ selilu kɔrɔsilenw ta kan. O fana bɛ Bɛn ni Feret cɛmancɛ dɔgɔyaliba ye, o ye paramɛtiri yɛrɛmahɔrɔnyalen ye min b’a Jira ko fɛnɲɛnamafagalanw dakunw ni ɲɔgɔn cɛ janyaba bɛ Dɔgɔya. Yɛlɛma minnu kɛra kirisitaw ka ultrastructure la, olu kɔlɔsira: kirisitaw ma ye kosɛbɛ PPA degun nɔfɛ (Ja 1a, panɛli B). Nka ja bɛɛ ma cristae ka ultrastructure jira ka jɛya, o la, o fɛn caman sɛgɛsɛgɛli ma kɛ. Nin TEM kunnafoniw bɛ se ka ko saba jira minnu bɛ se ka kɛ: (1) PPA bɛ farali bonya walima ka fusili bali, o bɛ kɛ sababu ye ka mitokondriyaw minnu bɛ yen, olu bonya ka dɔgɔya; (2) biyojɛnɛsi min bɛ bonya, o bɛ mitokondriyaw kura, minnu ka dɔgɔn walima (3) a bɛ o fɛɛrɛ fila bɛɛ bila ɲɔgɔn na. Hali ni nin cogoya ninnu tɛ se ka faranfasi ni TEM ye, fɛn caman b’a jira ko fɛn caman b’a jira ko fɛn caman bɛ yeli kɛ mitokondriyaw ka omɛostasi ni u ka fanga la PPA degun kɔrɔ. O kɔfɛ, an ye fɛn wɛrɛw sɛgɛsɛgɛ walasa ka nin fanga ninnu ni fɛɛrɛ minnu bɛ se ka kɛ u kɔnɔ, olu jira ka taa a fɛ.
Asidi propionic (PPA) bɛ mitokondriyaw cogoya sɛmɛntiya kokura. (a) Representative transmission electron microscopy (TEM) ja minnu b’a jira ko mitokondriyaw hakɛ bɛ dɔgɔya ani ko mitokondriyaw bɛ dɔgɔya ani ka kɛ lamini ye ni PPA furakɛli cayara 0 mM (min ma furakɛ), 3 mM ani 5 mM. Tile bilenmanw bɛ mitokondriyaw jira. (b–e) SH-SY5Y selilu minnu furakɛra ni PPA ye lɛrɛ 24 kɔnɔ, olu labɛnna TEM kama ani u jaabiw sɛgɛsɛgɛra ni Fiji/ImageJ ye. Paramɛtiri 8 la, naani ye danfaraba jira kunbɛnni (min ma furakɛ, 0 mM PPA) ni furakɛli kɛlen (3 mM ani 5 mM PPA) selilɛriw cɛ. (b) Mara 2, (c) Yɔrɔ, (d) Lamini, (e) Feret cɛmancɛ. Sɛgɛsɛgɛli min kɛra sira kelen fɛ (kɔlɔsili ni furakɛli) ani Dunnett ka sɛgɛsɛgɛli caman kɛli kɛra walasa ka danfarabaw dɔn (p < 0,05). Donanw yɔrɔw bɛ mitokondriyaw hakɛ danmadɔ jira seli kelen-kelen bɛɛ la, ani filiyɔrɔw bɛ cɛmancɛ ± SEM jira. Kunnafoni minnu jiralen don, olu bɛ n = 3 jira, a dɔgɔyalenba ye selilu 24 ye seginkanni kelen na; ja 266 de sɛgɛsɛgɛra u bɛɛ lajɛlen na ; * b’a jira ko p < 0,05, ** b’a jira ko p < 0,01.
Walasa ka dɔ fara a kan ka mitokondriyaw ka fanga jaabi cogo min na PPA la, an ye mitokondriyaw nɔgɔ ni tetramethylrhodamine ethyl ester (TMRE) ye ani ka baara kɛ ni waati tɛmɛnen mikroskɔpi ni MEL sɛgɛsɛgɛli ye walasa ka mitokondriyaw sigiyɔrɔmali ni u hakɛ jateminɛ sanga 24 kɔfɛ sanga 24 kɔfɛ PPA milimɛtɛrɛ 3 ni 5 la. Fisiyɔn ni fusili ko kɛlenw furakɛli. (Jiracogo 2a). MEL sɛgɛsɛgɛli kɔfɛ, mitokondriyaw sɛgɛsɛgɛra ka taa a fɛ walasa ka mitokondriyaw jɔcogo hakɛ n’u hakɛ danmadɔ jateminɛ. An ye dɔ farali fitinin ye nka a ka bon kosɛbɛ fisiyɔn ko kɛlenw hakɛ la minnu bɛ kɛ 3 mM [4,9 ± 0,3 (p < 0,05)] ni i y’a suma ni farali [5,6 ± 0,3 (p < 0,05) )] ani fusili [5,4 ± 0,5 (p < 0,05)] ani fusili [5,4 ± 0,5 (p < 0,05)] 0,05)] <0,05)] ko kɛlenw cayara kosɛbɛ 5 mM la ni i ye a suma ni kɔrɔsili ye (Ja 3b). Mitokondriyaw hakɛ cayara kosɛbɛ 3 [32,6 ± 2,1 (p < 0,05)] ani 5 mM [34,1 ± 2,2 (p < 0,05)] (Ja 3c) , ka sɔrɔ mitokondriyaw sigicogo kelen-kelen bɛɛ hakɛ danmadɔ ma Changé (Ja 3c). 3d). Ni an y’o bɛɛ lajɛlen ta, o b’a jira ko mitokondriyaw ka fanga sɛgɛsɛgɛli kokura bɛ kɛ jaabi musakabɔlan ye min bɛ mitokondriyaw ka rezo dafalen mara ka ɲɛ. Fisiyɔn ko kɛlenw hakɛ cayalenba 3 mM PPA la, o b’a jira ko mitokondriyaw hakɛ cayalenba dɔ bɛ bɔ mitokondriyaw fiyɛli la, nka n’i y’a jateminɛ ko mitokondriyaw hakɛ danmadɔ bɛ to a jɔyɔrɔba la, biyojenizi tɛ se ka ban i n’a fɔ jaabi musaka wɛrɛ. Nka, nin kunnafoniw bɛ bɛn ni mitokondriyaw jɔcogo misɛnninw ye minnu ye bulama ye, minnu kɔlɔsira TEM fɛ, wa u bɛ fɛn caman fana jira mitokondriyaw ka fanga la minnu bɛ sɔrɔ PPA fɛ.
Asidi propiyɔni (PPA) bɛ mitokondriyaw labɛncogo kura fangama bila walasa ka rezo dafalen mara. SH-SY5Y seliluw ladonna, ka furakɛ ni 3 ani 5 mM PPA ye sanga 24 kɔnɔ ka sɔrɔ ka nɔgɔ ni TMRE ni Hoechst 33342 ye o kɔfɛ MEL sɛgɛsɛgɛli kɛra. (a) Waati tɛmɛnen mikoroskɔpi ja jiralenw minnu bɛ kulɛri ni fanga maksimali jateminɛ binarilenw jira waati 2 (t2) la cogoya kelen-kelen bɛɛ la. Mara sugandilen minnu jiralen don ja fila kelen-kelen bɛɛ la, olu bɛ bonya ka jira 3D la waati saba danfaralenw na (t1-t3) walasa ka fanga jira waati kɔnɔ; fusion ko kɛlenw bɛ jira ni jiri ye; fisiyɔn ko kɛlenw bɛ jira ni jiri ye. A jiralen bɛ ni ɲɛ bilenman ye. (b) Ko minnu bɛ kɛ ni fanga ye, olu hakɛ cɛmancɛ la cogoya kelen-kelen bɛɛ la. (c) Mitokondriyaw ka sigida hakɛ dantɛmɛnen selilɛri kelen kɔnɔ. (d) Mitokondriyaw sigicogo kelen-kelen bɛɛ hakɛ dantɛmɛnen (μm3) seli kelen kɔnɔ. Kunnafoni minnu jiralen don, olu ye n = 15 selilu jira furakɛli kulu kelen-kelen bɛɛ la. Misaliw jiralen bɛ cɛmancɛ ± SEM jira, sɛgɛsɛgɛli kɛcogo = 10 μm, * p < 0,05.
Asidi propionic (PPA) bɛ kɛ sababu ye ka jamu minnu bɛ tali kɛ mitokondriyaw ka fanga la, olu ka sɛbɛnnikɛcogo damatɛmɛ. SH-SY5Y seliluw furakɛra ni 3 ani 5 mM PPA ye fo lɛri 24 kɔnɔ. Jɛnɛya hakɛ danfaralen kɛra ni RT-qPCR ye, ka a normalize ka kɛ B2M ye. Mitokondriyaw ka biyojenizi jɛnɛya (a) cMYC, (b) TFAM, (c) NRF1 ani (d) NFE2L2. Mitokondriyaw ka fusili ni farali jamu (e) STOML2, (f) OPA1, (g) MFN1, (h) MFN2 ani (i) DRP1. Danfaraba minnu bɛ u ni ɲɔgɔn cɛ (p < 0,05) olu sɛgɛsɛgɛra ni sira kelen ANOVA ye (kɔlɔsili ni furakɛli) ani Dunnett ka sɛgɛsɛgɛli caman kɛli ni ɲɔgɔn cɛ : * b’a jira ko p < 0,05, ** b’a jira ko p < 0,01, ani **** b’a jira ko p < 0,0001. Barw bɛ jiracogo cɛmancɛ jira ± SEM. Kunnafoni minnu jiralen don, olu bɛ n = 3 (STOML2, OPA1, TFAM), n = 4 (cMYC, NRF1, NFE2L2), ani n = 5 (MFN1, MFN2, DRP1) biologique replicates jira.
Kunnafoni minnu bɔra TEM ni MEL sɛgɛsɛgɛliw la ɲɔgɔn fɛ, olu b’a jira ko PPA bɛ mitokondriyaw cogoya ni u fanga caman sɛmɛntiya. Nka, o ja lajɛcogo ninnu tɛ hakilina di fɛɛrɛw kan minnu bɛ o wale ninnu ɲɛminɛ. O la, an ye mitokondriyaw ka fanga, biyojɛnɛsi ani mitozi ɲɛnabɔbaga jɔnjɔn 9 ka ARNm jiracogo sɛgɛsɛgɛ PPA furakɛli jaabi la. An ye selilɛri miyɛloma onkojɛni (cMYC), nukilila ninakili facteur (NRF1), mitokondriyali transkripsiyɔn facteur 1 (TFAM), NFE2-like transcription factor BZIP (NFE2L2), gastrin-like protein 2 (STOML2), optique nerve atrophie 1 (OPA1 ), Mitofusin 1 (MFN1), Mitofusin 2 (MFN2) ani dinamini-related protein 1 (DRP1) furakɛli kɔfɛ lɛri 24 kɔnɔ ni 3 mM ani 5 mM PPA ye. An ye 3 mM (p = 0,0053, p = 0,0415 ani p < 0,0001, o cogo kelen na) ani 5 mM (p = 0,0031, p = 0,0233, p < 0,0001) PPA furakɛli kɔlɔsi. (Jaa 3a–c) la. ARNm jirali dɔgɔyali tun bɛ bɔ a tata hakɛ la : cMYC, NRF1 ani TFAM jirali dɔgɔyara ni siɲɛ 5,7, 2,6 ani 1,9 ye 3 mM la, ani siɲɛ 11,2, 3 ani 2,2 ni 5 mM ye. O ni ɲɔgɔn cɛ, cɛmancɛ redox biyojenizi jɛnɛya NFE2L2 ma Changé PPA hakɛ si la, hali ni o ɲɔgɔnna dɔ kɔlɔsira ka bɔ a tata hakɛ la, n’o ye ka dɔ bɔ a jiracogo la (Ja 3d).
An ye klasiki jɛnɛya minnu sen bɛ fisiyɔn ni fusili ɲɛnabɔli la, olu fana jiracogo sɛgɛsɛgɛ. A bɛ miiri ko STOML2 sen bɛ fusili, mitofaji ani biyojenizi la, wa a jirali dɔgɔyara kosɛbɛ (p < 0,0001) ni 3 mM (2,4-yɔrɔ caman yeli) ani 5 mM (2,8-zeli yeli) PPA ye (Ja 1). 3d). O cogo kelen na, OPA1 fusion jeninida jirali dɔgɔyara ni 3 mM (1,6 yeli yeli) ani 5 mM (1,9 siɲɛ caman yeli) PPA (p = 0,006 ani p = 0,0024, o cogo kelen na) (Ja 3f). Nka, an ma danfaraba sɔrɔ fusion gènes MFN1, MFN2 walima fission gène DRP1 jirali la 24-h PPA degun kɔnɔ (Ja 3g–i). Ka fara o kan, an y’a ye ko fusion ni fission proteine ​​naani (OPA1, MFN1, MFN2 ani DRP1) hakɛ ma Changé cogo kelen na (Ja 4a–d). A nafa ka bon k’a dɔn ko nin kunnafoniw bɛ waati kelen jira, wa a bɛ se ka kɛ ko u tɛ fɛn caman yeli jira farikolojɔli dumuniw jiracogo walima u baara hakɛ la PPA degun fɔlɔw la. Nka, ni dɔ bɔra kosɛbɛ cMYC, NRF1, TFAM, STOML2 ani OPA1 jirali la, o b’a jira ko mitokondriyaw ka farikoloɲɛnajɛ, u ka biyojenizi, ani u ka baarakɛcogo bɛrɛbɛrɛ sɛbɛnnikɛcogo la. Ka fara o kan, nin kunnafoniw bɛ jatebɔ fɛɛrɛw nafa jira walasa ka fɛn labanw sɛgɛsɛgɛli kɛ tigitigi mitokondriyaw ka baarakɛcogo la.
Fusion ni fission factor proteine ​​hakɛ ma Changé asidi propionic (PPA) furakɛli kɔfɛ. SH-SY5Y seliluw furakɛra ni 3 ani 5 mM PPA ye fo lɛri 24 kɔnɔ. Protéines hakɛ jateminɛna ni Western blot analysis ye, ani expression hakɛw kɛra normal ye ka se proteine ​​totale ma. Protéines expression moyenne ani représentant Western blots de target et total proteine ​​jiralen don. a – OPA1, b – MFN1, c – MFN2, d – DRP1. Barw bɛ cɛmancɛ ± SEM jira, wa kunnafoniw jiralenw bɛ n = 3 biologique replicates jira. Sanga caman (p < 0,05) kɛra ni ɲɔgɔndan sɛgɛsɛgɛli ye min bɛ kɛ sira kelen fɛ ani Dunnett ka sɛgɛsɛgɛli. Jeli ni blot fɔlɔ jiralen don ja S1 na.
Mitokondriyaw ka baarakɛbaliya bɛ tali kɛ sigida caman banaw la, k’a ta farikoloɲɛnajɛ, dusukun ni farikolo banaw la ka se hakilibanaw ma1,10. Bana caman minnu bɛ mɔgɔ farikolo tiɲɛ ani minnu bɛ mɔgɔ farikolo tiɲɛ, olu bɛ tali kɛ mitokondriyaw ka baarakɛbaliya la, o b’a jira ko o farikolo yɔrɔw nafa ka bon hakili ka ɲɛnamaya bɛɛ kɔnɔ. O bana ninnu ye Parkinson ka bana, Alzheimer bana ani ASD3,4,18 ye. Nka, ka se ka kunsɛmɛnasumaya sɔrɔ walasa ka nin bana ninnu sɛgɛsɛgɛ, o ka gɛlɛn, kɛrɛnkɛrɛnnenya la, fɛɛrɛko siratigɛ la, o b’a to selilɛriw ka misali sigicogo bɛ kɛ fɛɛrɛ wɛrɛ wajibiyalen ye. Nin kalan in kɔnɔ, an bɛ baara kɛ ni selilɛriw ka misali kɛcogo ye min bɛ baara kɛ ni SH-SY5Y selilɛriw ye minnu furakɛra ni PPA ye walasa ka mitokondriyaw ka baarakɛbaliya min kɔlɔsira senfagabana banaw la, kɛrɛnkɛrɛnnenya la autizmu spektrum banaw la, o lajɛ kokura. Ni aw bɛ baara kɛ ni nin PPA misali in ye walasa ka mitokondriyaw ka fanga sɛgɛsɛgɛ senfagabana kɔnɔ, o bɛ se ka kɛ sababu ye ka hakilina di ASD sababu kan.
An ye sɛgɛsɛgɛli kɛ walasa ka baara kɛ ni TEM ye walasa ka fɛn caman yeli ye mitokondriyaw cogoya la. A nafa ka bon k’a dɔn ko TEM ka kan ka kɛ ka ɲɛ walasa a ka se ka baara kɛ ka ɲɛ. Cryo-specimens labɛnni bɛ kɛ sababu ye ka senfagabana sigicogo lakana ka ɲɛ ni selilu yɔrɔw sirili ye waati kelen na ani ka dɔ bɔ fɛn dilannenw dilanni na Ka bɛn o ma, an y’a kɔlɔsi ko SH-SY5Y selilu minnu bɛ i n’a fɔ senfagabana, olu kɔnɔna na, farikolo yɔrɔw ma tiɲɛ ani mitokondriyaw janyalen don (Ja 1a). O b’a jira ko nafa min bɛ kiriyɔzɛni labɛnni fɛɛrɛw la walasa ka mitokondriyaw cogoya kalan senfagabana selilɛriw misaliw la. Hali ni hakɛ sumani nafa ka bon kosɛbɛ TEM kunnafoniw sɛgɛsɛgɛli kuntilenna na, bɛnkan tɛ yen hali bi, fɛn kɛrɛnkɛrɛnnen minnu ka kan ka suman walasa ka mitokondriyaw cogoya caman sɛmɛntiya. Ka da kalan caman kan minnu ye mitokondriyaw cogoya sɛgɛsɛgɛ hakɛ la17,31,32, an ye mitokondriyaw ja sɛgɛsɛgɛli pibiliki otomatiki dɔ labɛn min bɛ cogoya dantigɛli 8 suman, n’olu ye : yɔrɔ, yɔrɔ2, ɲɛda hakɛ, lamini, sɛrɛkili, degɛrɛ , Feret cɛmancɛ. ani laminikogo.
Olu cɛma, PPA ye yɔrɔ 2, yɔrɔ, lamini, ani Feret cɛmancɛ dɔgɔya kosɛbɛ (Ja 1b–e). O y’a jira ko mitokondriyaw kɛra fitinin ye ani ka kɛ bulama ye, o bɛ bɛn ni ɲinini tɛmɛnenw ye minnu y’a jira ko mitokondriyaw yɔrɔ dɔgɔyara sanga 72 kɔfɛ PPA30 ka mitokondriyaw degun kɔfɛ. Nin cogoya ninnu bɛ se ka mitokondriyaw farali jira, o ye fɛɛrɛ wajibiyalen ye walasa ka fɛn tiɲɛnenw bɔ mitokondriyaw ka rezo la walasa k’u tiɲɛni sabati mitofaji fɛ35,36, Faan wɛrɛ fɛ, mitokondriyaw hakɛ danmadɔ dɔgɔyali bɛ se ka kɛ sababu ye ka biyojenizi caya, o min bɛ na ni mitokondriyaw misɛnninw bɔli ye minnu bɛ bange. Fisiyɔn walima biyojenizi cayalenba bɛ jaabi musaka jira walasa ka mitosis mara mitokondriyaw degunw kama. Nka, mitokondriyaw ka bonya dɔgɔyali, fusion (fusion) tiɲɛni, walima bana wɛrɛw tɛ se ka bɔ kɛnɛ kan.
Hali n’a y’a sɔrɔ ja minnu bɛ dilan ni TEM ye minnu ka bon kosɛbɛ, olu bɛ sira Di fɛnɲɛnɛma cogoyaw dɔnni ma mitokondriyaw kelen-kelen bɛɛ hakɛ la, o fɛɛrɛ in bɛ ja fila-fila-jatebɔ Kɛ waati kelen na. Walasa ka jaabi fangamaw kalan farikoloɲɛnajɛ degunw na, an ye mitokondriyaw nɔgɔ ni TMRE ye ani ka baara kɛ ni waati tɛmɛnen mikroskɔpi ye ni MEL sɛgɛsɛgɛli ye, o min bɛ sira di fɛn caman yeli ye 3D la, fɛn minnu bɛ yeli kɛ mitokondriyaw ka rezo kɔnɔ waati kɔnɔ An ye fɛn misɛnninw ye nka fɛn nafamaw yeli ye mitokondriyaw ka fanga la PPA degun kɔrɔ (Ja 2). Ni 3 mM ye, fisiyɔn ko kɛlenw hakɛ cayara kosɛbɛ, ka sɔrɔ fusili ko kɛlenw tora i n’a fɔ kɔrɔsili la. Dɔ farala fisiyɔn ni fusiliko fila bɛɛ hakɛ kan, o kɔlɔsira 5 mM PPA la, nka o fɛn caman ɲɔgɔnna yeli tun bɛ bɛn ɲɔgɔn ma, o b’a jira ko fisiyɔn ni fusili kinetiki bɛ se balannako ma ni u hakɛ ka ca (Ja 2b). Mitokondriyaw hakɛ danmadɔ ma Changé PPA 3 ni 5 mM fila bɛɛ la, o b’a jira ko mitokondriyaw ka rezo dafalen maralen don (Ja 2d). O b’a jira ko mitokondriyaw ka rezo fangamaw bɛ se ka farikoloɲɛnajɛ degun nɔgɔmanw jaabi walasa ka homeostasis mara ka ɲɛ k’a sɔrɔ u ma kɛ sababu ye ka rezow tigɛtigɛ. Ni 3 mM PPA ye, fiɲɛbɔli caya bɛ se ka kɛ sababu ye ka wuli ka taa balannako kura la, nka fɛnɲɛnɛma-ko kura jugumanba de ka kan ka kɛ ka ɲɛsin degun ma min bɛ sɔrɔ PPA hakɛ caman fɛ.
Mitokondriyaw hakɛ cayara PPA degun hakɛ fila bɛɛ la, nka mitokondriyaw hakɛ danmadɔ ma Changé kosɛbɛ (Ja 2c). O bɛ se ka kɛ sababu ye ka biyojenizi caya walima ka tila ka caya; nka, ni dɔ bɔra kosɛbɛ mitokondriyaw hakɛ danmadɔ la, a ka ca a la, biyosintɛzi bɛ bonya. Nka, kunnafoniw minnu bɛ ja 2 kɔnɔ, olu bɛ musakabɔcogo fila sɔrɔli sinsin: dɔ farali farali ko kɛlenw hakɛ kan, min bɛ bɛn mitokondriyaw farali wulicogo ma, ani ko kɛlenw hakɛ caya, min bɛ bɛn mitokondriyaw ka biyojenizi ma. A laban na, degun nɔgɔmanw musakabɔcogo fangama bɛ se ka kɛ walew ye minnu bɛ kɛ waati kelen na, minnu bɛ kɛ ni fission, fusion, biogenesis ani mitophagie ye. Hali ni sɛbɛnnikɛla tɛmɛnenw y’a jira ko PPA bɛ mitozi30,39 ani mitofaji29 bonya, an bɛ daliluw di mitokondriyaw ka faranfasi ni fusili dinamiki labɛnni kura kan PPA jaabi la. Nin kunnafoniw bɛ fɛnɲɛnɛmafagalanw sinsin minnu kɔlɔsira TEM fɛ, wa u bɛ hakilina wɛrɛw di fɛɛrɛw kan minnu bɛ tali kɛ PPA ka mitokondriyaw ka baarakɛbaliya la.
Ikomi TEM ni MEL sɛgɛsɛgɛli si ma dalilu tilennenw di jamu ɲɛnabɔli fɛɛrɛw kan minnu bɛ fɛnw cogoya caman yeli kɔlɔsilenw kɔnɔ, an ye jamu minnu sen bɛ mitokondriyaw ka farikoloɲɛnajɛ la, biyojenizi la, ani u ka fɛnɲɛnamafagalanw na, olu ka ARN jiracogo sɛgɛsɛgɛ. cMYC proto-oncogene ye sɛbɛnnikɛlan ye min bɛ mitokondriyaw, glycolyse, amino acides ani fatty acids metabolism labɛnni na40. Ka fara o kan, a dɔnna ko cMYC bɛ mitokondriyaw ka jamu 600 ɲɔgɔn jiracogo ɲɛnabɔ minnu sen bɛ mitokondriyaw ka sɛbɛnni na, u bamanankan na, ani u lajɛcogo gɛlɛnw na, NRF1 ni TFAM41 fana sen bɛ o la. NRF1 ni TFAM ye mitosis ɲɛmɔgɔba fila ye, minnu bɛ baara kɛ PGC-1α duguma walasa ka ADNmt caman ɲɔgɔnnabɔli baara. O sira in bɛ baara kɛ ni cAMP ni AMPK taamasiɲɛw ye, wa a bɛ fanga musaka ni farikoloɲɛnajɛ degunw jateminɛ. An ye NFE2L2 fana sɛgɛsɛgɛ, n’o ye mitokondriyaw ka biyojenizi ɲɛnabɔbaga ye min bɛ wele ko redox, walasa k’a dɔn ni PPA nɔw bɛ se ka sɔrɔ oksidan degun fɛ.
Hali ni NFE2L2 jirali ma Changé, an ye dɔgɔyali sɔrɔ min bɛ bɔ a tata hakɛ la tuma bɛɛ cMYC, NRF1 ani TFAM jirali la furakɛli kɔfɛ lɛrɛ 24 kɔnɔ ni 3 mM ani 5 mM PPA ye (Ja 3a–c). cMYC jirali jigincogo fɔra ka tɛmɛ ko o ye jaabi ye mitokondriyaw ka degun na42, wa a kɔfɛ, cMYC jirali dɔgɔyali bɛ se ka kɛ sababu ye ka mitokondriyaw ka baarakɛbaliya bila mitokondriyaw ka farikoloɲɛnajɛ la, ka rezow jɛɲɔgɔnya, ani membrane polarisation A nafa ka bon kosɛbɛ, cMYC fana sen bɛ mitokondriyaw ka faranfasi ni fusili labɛnni na42,43, wa a dɔnna ko a bɛ dɔ fara DRP1 fosforilasi ni mitokondriyaw sigiyɔrɔmali kan selilu tilali waati44, ka fara mitokondriyaw cogoya caman sɛgɛsɛgɛli cɛmancɛ kan senfagabana jirisunw kɔnɔ Tiɲɛ na, fibroblaste minnu tɛ cMYC sɔrɔ, olu bɛ mitokondriyaw hakɛ dɔgɔyali jira, o bɛ bɛn fɛn caman ma minnu bɛ sɔrɔ PPA43 degun fɛ. Nin kunnafoniw bɛ jɛɲɔgɔnya nafama dɔ jira, nka min ma jɛya fɔlɔ, cMYC ni mitokondriyaw ka fanga cɛ, o bɛ laɲini nafama di PPA degunw ka sɛgɛsɛgɛli nataw ma.
NRF1 ni TFAM dɔgɔyali bɛ bɛn cMYC jɔyɔrɔ ma i n’a fɔ sɛbɛnnikɛlan nafama. Nin kunnafoniw fana bɛ bɛn ni sɛgɛsɛgɛli tɛmɛnenw ye hadamadenw ka kɔnɔbara kansɛribanakisɛw kan minnu y’a jira ko PPA ye NRF1 mRNA jirali dɔgɔya sanga 22 kɔnɔ, o min ni ATP dɔgɔyali bɛ tali kɛ ɲɔgɔn na ani ka ROS46 caya. Nin sɛbɛnnikɛla ninnu fana y’a jira ko TFAM jirali cayara sanga 8,5 kɔnɔ nka a seginna a daminɛ hakɛ la sanga 22 kɔnɔ. O ni Kim ni a ɲɔgɔnnaw ta tɛ kelen ye. (2019) y’a jira ko TFAM mRNA jirali dɔgɔyara kosɛbɛ PPA degun lɛrɛ 4 kɔfɛ SH-SY5Y selilɛriw kɔnɔ; nka, sanga 72 kɔfɛ, TFAM furakisɛ jirali cayara kosɛbɛ ani ADNmt kopi hakɛ cayara kosɛbɛ. O la, an ye min kɔlɔsi sanga 24 kɔfɛ, o dɔgɔyali min kɛra mitokondriyaw ka biyojenizi jɛnɛya la, o tɛ a bɔ a la ko mitokondriyaw hakɛ cayali bɛ tali kɛ biyojenizi baarakɛcogo la waati fɔlɔw la. Nin ɲinini tɛmɛnenw y’a jira ko PPA bɛ PGC-1α mRNA ni farikolojɔlifɛnw wuli kosɛbɛ SH-SY5Y selilɛriw kɔnɔ sanga 4 ni miniti 30 kɔnɔ, ka sɔrɔ asidi propiyɔni bɛ mitokondriyaw ka biyojeni sabati misiden sugunɛbilenniw la PGC-1α fɛ sanga 12 ni miniti 39 kɔnɔ. A nafa ka bon kosɛbɛ, PGC-1α tɛ NRF1 ni TFAM ka sɛbɛnnikɛlan tilennen dɔrɔn ye, nka a jirala fana ko a bɛ MFN2 ni DRP1 ka baara ɲɛnabɔ ni fisiyɔn ni fusili ɲɛnabɔli ye Ni an y’u bɛɛ lajɛlen ta, o bɛ fɛɛrɛw jɛlenw jira minnu bɛ mitokondriyaw ka jaabiw musakabɔcogo ɲɛnabɔ minnu bɛ sɔrɔ PPA fɛ. Ka fara o kan, an ka kunnafoniw b’a jira ko biyojɛnɛsi ni farikoloɲɛnajɛ kɛcogo sɛbɛnniko ɲɛnabɔli bɛrɛbɛnbaliyaba bɛ PPA degun kɔrɔ.
STOML2, OPA1, MFN1, MFN2 ani DRP1 jamu bɛ mitokondriyaw ka faranfasi, fusili ani dinamiki cɛmancɛ ɲɛnabɔbagaw cɛma37,48,49. Jɛnkulu caman wɛrɛ bɛ yen minnu bɛ mitokondriyaw ka fanga la, nka, a jirala ka tɛmɛn ko STOML2, OPA1 ani MFN2 bɛ metilasi danfara la ASD kuluw kɔnɔ,16 ani yɛrɛmahɔrɔnya kalan caman ye fɛn dɔw jira minnu ye fɛn caman Changements dans les transcriptions facteurs de réponse de l’affaires de mitokondrienne50, 52. OPA1 ni STOML2 fila bɛɛ jirali dɔgɔyara kosɛbɛ ni 3 mM ni 5 mM PPA furakɛli ye (Ja 3e, f). OPA1 ye mitokondriyaw ka ɲɔgɔn sɔrɔli ɲɛnabɔbaga klasiki dɔ ye ni jɛɲɔgɔnya tilennen ye ni MFN1 ni 2 ye, wa a jɔyɔrɔ ka bon cristae remodeling ni mitokondriyaw cogoya la53. STOML2 jɔyɔrɔ tigitigi min bɛ mitokondriyaw ka fanga la, o ma jɛya hali bi, nka daliluw b’a jira ko a jɔyɔrɔ ka bon mitokondriyaw ka ɲɔgɔn sɔrɔli la, biyojenizi la, ani mitofaji la.
STOML2 sen bɛ mitokondriyaw ninakili jɛɲɔgɔnya sabatili la ani ninakili cakɛda complexes (finakili cakɛda complexes) dilancogo la54,55 wa a jirala ko a bɛ kansɛribanakisɛw ka farikoloɲɛnajɛ cogoyaw Changer kosɛbɛ Kɔlɔsiliw y’a jira ko STOML2 bɛ mitokondriyaw ka seko ni dɔnko sabati ni BAN ni karidiyolipini ka jɛɲɔgɔnya ye 55, 57, 58. Ka fara o kan, sɛgɛsɛgɛli yɛrɛmahɔrɔnyalenw y’a jira ko STOML2 ni PINK1 ka jɛɲɔgɔnya bɛ mitofaji labɛn Kɛrɛnkɛrɛnnenya la, a fɔra ko STOML2 bɛ jɛɲɔgɔnya kɛ ni MFN2 ye k’a ɲɛsin a ma ani k’a sabati ani fana a jɔyɔrɔ ka bon OPA1 isofɔmu janw sabatili la, a kɛtɔ ka poroteyizi bali min bɛ OPA1 tiɲɛni ɲɛnabɔ53,61, STOML2 jirali dɔgɔyali min kɔlɔsira PPA waleyaliw la, o bɛ se ka kɛ sababu ye ka nin fusili furakisɛ ninnu kɛ ka tiɲɛ ka tɛmɛ siraw fɛ minnu bɛ tali kɛ ubiquitin ni proteasome la48. Hali ni STOML2 ni OPA1 jɔyɔrɔ tigitigi ma jɛya PPA jaabi fangama la, nin fusili jamu ninnu jirali dɔgɔyali (Jaa 3) o bɛ se ka balansi tiɲɛ farali ni farali cɛ ani ka kɛ sababu ye ka mitokondriyaw hakɛ dɔgɔya (Jaa 3). 1).
Faan wɛrɛ fɛ, OPA1 furakisɛ jirali ma Changé lɛrɛ 24 kɔfɛ, ka sɔrɔ MFN1, MFN2 walima DRP1 ka ARNm ni farikolojɔlifɛnw hakɛ ma Changé kosɛbɛ PPA furakɛli kɔfɛ (Ja 3g-i, Jaa 4). O bɛ se k’a jira ko fɛn ma Changé (Yɛlɛma) o fɛn ninnu labɛnni na minnu bɛ mitokondriyaw ka fusili ni u farali la. Nka, a ka kan ka kɔlɔsi ko nin jamu naani kelen-kelen bɛɛ fana bɛ labɛn ni fɛn caman sɛgɛsɛgɛli kɔfɛ (PTM) ye minnu bɛ farikolojɔli baara kɔlɔsi. OPA1 bɛ ni ɲɔgɔndan suguya 8 ye minnu bɛ tigɛtigɛ ni poroteyolitiki ye mitokondriyaw la walasa ka isofɔmu fila danfaralenw dilan 63 . Balannako min bɛ isofɔmu janw ni isofɔmu surunw cɛ, o laban bɛ OPA1 jɔyɔrɔ jira mitokondriyaw ka ɲɔgɔn sɔrɔli la ani mitokondriyaw ka rezo ladonni na64. DRP1 baara bɛ ɲɛnabɔ ni kalisiyɔmu/kalmodulini-dependant proteine ​​kinase II (CaMKII) fosforilasi ye, ka sɔrɔ DRP1 tiɲɛni bɛ ɲɛnabɔ ni ubiquitination ni SUMOylation ye A laban na, DRP1 ni MFN1/2 fila bɛɛ ye GTPasew ye, o la, baara bɛ se ka nɔ bila GTP bɔli hakɛ la mitokondriyaw la 66 . O la, hali ni nin farikolojɔlifɛn ninnu jirali bɛ to a cogo la, a bɛ se ka kɛ ko o tɛ farikolojɔli baara walima u sigiyɔrɔma caman jira67,68. Tiɲɛ na, PTM poroteyini minnu bɛ yen, olu ka teli ka kɛ lafasali sira fɔlɔ ye min ka baara ye ka degun jugumanw jaabiw cɛsiri. Ni farikoloɲɛnajɛ degun danmadɔ bɛ an ka misali la, a ka c’a la, PTM bɛ dɔ fara fusion ni fission protéines ka baara kan walasa ka mitokondriyaw dafalen segin a cogo kɔrɔ la k’a sɔrɔ a ma kɛ ni o gènes baarakɛcogo wɛrɛw ye ARNm walima proteine ​​hakɛ la.
Ni an y’u bɛɛ lajɛlen ta, sanfɛ kunnafoniw bɛ mitokondriyaw cogoya ɲɛnabɔli gɛlɛn ni waati-dafalen jira ani gɛlɛya minnu bɛ o fɛɛrɛw ɲɛfɔli la. Walasa ka jamu jiracogo sɛgɛsɛgɛ, a ka kan fɔlɔ ka jamu laɲini kɛrɛnkɛrɛnnenw dɔn sira kɔnɔ. Nka, an ka kunnafoniw b’a jira ko jamu minnu bɛ sira kelen kan, olu tɛ jaabi cogo kelen na degun kelen na. Tiɲɛ na, kalan tɛmɛnenw y’a jira ko jamu suguya wɛrɛw bɛ sira kelen na, olu bɛ se ka waati jaabi cogoya wɛrɛw jira30,46. Ka fara o kan, sɛbɛnni kɔfɛ fɛɛrɛ gɛlɛnw bɛ yen minnu bɛ jɛɲɔgɔnya tiɲɛ sɛbɛnni ni jeninida baara cɛ. Protéomic kalanw bɛ se ka hakilina di PTMw ni proteine ​​baara nɔfɛkow kan, nka u bɛ gɛlɛyaw fana lase u ma i n’a fɔ fɛɛrɛw minnu tɛ baara kɛ kosɛbɛ, taamasiyɛn ni mankan hakɛ caman, ani ɲɛfɔli jugu.
O hukumu kɔnɔ, mitokondriyaw cogoya kalanni ni TEM ni MEL ye, o bɛ se ka kɛ sababu ye ka ɲininkali jɔnjɔnw ɲɛnabɔ jɛɲɔgɔnya min bɛ mitokondriyaw ka fanga ni baarakɛcogo cɛ ani o bɛ nɔ bila bana na cogo min na. Min nafa ka bon kosɛbɛ, o ye ko TEM bɛ fɛɛrɛ tilennen di walasa ka mitokondriyaw cogoya suman i n’a fɔ mitokondriyaw ka baarakɛbaliya ni u ka fanga labanyɔrɔ min bɛ ɲɔgɔn sɔrɔ51. MEL fana bɛ fɛɛrɛ tilennen dɔ di walasa ka fisiyɔn ni fusili ko kɛlenw ye selilu sigida saba kɔnɔ, o bɛ sira di mitokondriyaw sɛgɛsɛgɛli fangama hakɛ dɔnni ma hali ni fɛn ma Changements dans la expression de gènes Yan, an bɛ mitokondriyaw jateminɛ fɛɛrɛw nafa jira mitokondriyaw bana filananw na. A ka ca a la, nin bana ninnu bɛ sɔrɔ farikoloɲɛnajɛ degun nɔgɔman basigilenw fɛ minnu bɛ sɔrɔ mitokondriyaw ka rezow labɛncogo kura nɔgɔlen fɛ sanni ka mitokondriyaw tiɲɛni jugumanba sɔrɔ. Nka, mitokondriyaw ka musaka min ka kan ka kɛ walasa ka mitosis mara degun basigilen kɔnɔ, o bɛ na ni baarakɛcogo kɔlɔlɔbaw ye. Hakilila-sɛnɛko siratigɛ la, nin musakabɔcogo ninnu faamuyali ɲuman bɛ se ka kunnafoni nafamaw di farikolo-ɲɛnajɛ-bana (pleiotropic neuropathologie) kan min bɛ tali kɛ mitokondriyaw ka baarakɛbaliya la.
A laban na, an ka kunnafoniw b’a jira ko jatebɔ fɛɛrɛw nafa ka bon walasa ka baarakɛnafolo kɔlɔlɔw faamuya, minnu bɛ sɔrɔ jɛɲɔgɔnya gɛlɛnw na minnu bɛ sɔrɔ jeninida jiracogo, farikolojɔlifɛnw caman cili, ani farikolojɔli baara cɛ minnu bɛ senfagabana mitokondriyaw ka fanga kɔlɔsi. An ye PPA kɛ ka mitokondriyaw ka baarakɛbaliya misali kɛ senfagabana selilɛri modɛli kɔnɔ walasa ka hakilina sɔrɔ ASD mitokondriyaw yɔrɔ kan. SH-SY5Y selilu minnu furakɛra ni PPA ye, olu ye fɛn caman jira mitokondriyaw cogoya la: mitokondriyaw kɛra fitininw ye ani u kɛra bulama ye, ani kirisitaw ma ɲɛfɔ ka ɲɛ ni u kɔlɔsira TEM fɛ. MEL sɛgɛsɛgɛli b’a jira ko nin fɛn caman Changements bɛ kɛ ɲɔgɔn fɛ ni dɔ farali ye fission ni fusion ko kɛlenw kan walasa ka mitokondriyaw ka rezo mara ka kɛɲɛ ni farikoloɲɛnajɛ degun nɔgɔmanw ye. Ka fara o kan, PPA bɛ mitokondriyaw ka farikoloɲɛnajɛ ni omɛostazi ka sɛbɛnnikɛcogo tiɲɛ kosɛbɛ. An y’a jira ko cMYC, NRF1, TFAM, STOML2, ani OPA1 ye mitokondriyaw ɲɛnabɔbaga jɔnjɔnw ye minnu tiɲɛna PPA degun fɛ, wa u bɛ se ka jɔyɔrɔba ta PPA ka fɛn caman cili la mitokondriyaw cogoya n’u baarakɛcogo la. Kalan nataw ka kan ka kɛ walasa ka PPA ka waati caman yeli dɔn ka ɲɛ jamu jiracogo ni farikolojɔli baara la, sigida la, ani bamanankan bamanankan kɔfɛ fɛn caman caman cili la. An ka kunnafoniw b’a jira ko sariyasunba minnu bɛ mitokondriyaw ka degun jaabi cɛmancɛ la, olu ka gɛlɛya n’u ka ɲɔgɔndɛmɛ jira, wa u b’a jira ko TEM ni ja lajɛ fɛɛrɛ wɛrɛw nafa ka bon fɛɛrɛko kalanw na minnu ɲɛsinnen don kosɛbɛ.
SH-SY5Y selilɛri (ECACC, 94030304-1VL) sanna Sigma-Aldrich fɛ. SH-SY5Y seliluw falenna Dulbecco ka Eagle’s medium/F-12 balofɛnw ɲagaminen caman na (DMEM/F-12) ani L-glutamine (SC09411, ScienCell) la, santimɛtɛrɛ 25 fiyɛli la, minnu farala 20% denso kɔnɔ misiw ka joli la (FBS) (10493106, ThermoFisher Scientific) ani 1% penisilini-sɛrɛpitomisini (P4333-20ML, Sigma-Aldrich) ni a bɛ kɛ 37 °C la, 5% CO2. Bugunw kɛra subculture ye fo ka se 80% confluence ma ni 0,05% tripsin-EDTA (15400054, ThermoFisher Scientific) ye, ka u kɛ centrifuge la garamu 300 la ka u bila plati la ni density ye selilu 7 × 105 ɲɔgɔn/ml. Kɔrɔbɔli bɛɛ kɛra SH-SY5Y selilɛriw kan minnu ma danfara bɔ tɛmɛsira 19–22 cɛ. PPA bɛ kɛ i n’a fɔ NaP. Aw bɛ NaP mugu (CAS No. 137-40-6, kemikaliya formula C3H5NaO2, P5436-100G, Sigma-Aldrich) kɛ MilliQ ji sumalen na fo ka se M 1 ma, ka a mara 4 °C la. Furakɛli don na, aw bɛ nin furaji in kɛ ni PPA M 1 ye ka se milimɛtɛrɛ 3 ma ani PPA mililitiri 5 kɛ ji la min tɛ serum ye (DMEM/F-12 ni L-glutamine ye). Furakɛli hakɛ minnu kɛra kɛlɛli bɛɛ la, olu tun ye PPA tɛ (0 mM, kɔrɔsili), 3 mM, ani 5 mM PPA. Kɔrɔbɔliw kɛra biologique replication saba la dɔgɔmannin na.
SH-SY5Y seliluw kɛra kisɛ ye santimɛtɛrɛ 25 fiyɛliw kɔnɔ ni hakɛ ye 5,5 × 105 selilu/ml ye ka u falen fo lɛri 24. PPA furakɛli farala fiyɛlilan kan sani a ka kɛ lɛri 24 kɔnɔ. Aw bɛ selilɛriw lajɛ ka kɛɲɛ ni baganw ka farikolo yɔrɔw sɛnɛcogo nɔgɔmanw ye (a ɲɛfɔlen don sanfɛ). Aw bɛ selilɛri kurukuruni kɛ glutaraldehyde 2,5% 100 μl kɔnɔ, 1× PBS kɔnɔ ka a mara 4 °C la fo ka se a baara ma. SH-SY5Y seliluw kɛra santirifugu ye waati kunkurunnin kɔnɔ walasa ka seliluw kɛ pellet ye ani ka glutaraldehyde 2,5% bɔ, 1× PBS jilama. Aw bɛ segin ka bɔgɔmugu kɛ agarozi jeli 4% kɔnɔ min labɛnna ji dilanlen na (agarozi ni bɔgɔ hakɛ hakɛ ye 1:1 ye). Agarose yɔrɔw tun bɛ bila gris kan plati flakɛlenw kan ka u datugu ni 1-hexadecene ye ka sɔrɔ ka u bila nɛnɛ na ni tansiyɔnba ye. Aw bɛ sɛgɛsɛgɛli kɛlenw bila asetɔni jalen 100% la -90°C la sanga 24 kɔnɔ. O kɔfɛ, aw bɛ funteni kɔrɔta ka se -80°C ma, ka sɔrɔ ka osiyɔmu tetorokisidi 1% ni glutaraldehyde 0,1% jilama fara a kan. A sɛgɛsɛgɛli kɛra -80°C la sanga 24 kɔnɔ. O kɔfɛ, funteni cayara dɔɔni dɔɔni ka se so funteni ma tile damadɔ kɔnɔ: ka bɔ – 80 °C la ka se – 50 °C ma sanga 24 kɔnɔ, ka se – 30 °C ma sanga 24 kɔnɔ, ka se – 10 °C ma sanga 24 kɔnɔ ani laban na ka se so funteni ma. goniyahakɛ.
Kɔrɔyɔzɛni labɛnni kɔfɛ, sɛgɛsɛgɛli kɛlenw kɛra ni resin ye ani ka yɔrɔ finmanw (∼100 nm) dilan ni Leica Reichert UltracutS ultramicrotome (Leica Microsystems) ye. Yɔrɔw nɔgɔlen don ni uranili asetati 2% ni sitɛrɛti plɔmu ye. Misaliw kɔlɔsira ni FEI Tecnai 20 transmission electron microscope (ThermoFisher (kɔrɔlen FEI), Eindhoven, Pays-Bas) ye min bɛ baara kɛ 200 kV (Lab6 transmetteur) ani Gatan CCD kamera (Gatan, Angletɛri) min bɛ ni Tridiem fanga filɛri ye.
Fɛɛrɛko segin-ka-bɔnye kelen-kelen bɛɛ la, seli kelen ja 24 ni kɔ sɔrɔla, o bɛɛ lajɛlen ye ja 266 ye. Ja bɛɛ sɛgɛsɛgɛra ni Region of Interest (ROI) makɔrɔ ye ani Mitochondria macro ye. Mitokondriyaw makɔrɔ sinsinnen bɛ fɛɛrɛw kan minnu bɔra kɛnɛ kan17,31,32 ani a bɛ sira di TEM jaw baaracogo ma min bɛ kɛ ni batch tilancɛ ye Fidji/ImageJ69. Kuma surun na : ja bɛ wuli ka wuli ka wuli ni rolling ball kɔkanna bɔli ye (60 pixel radius) ani FFT bandpass filter (ka baara kɛ ni 60 ani 8 pixel sanfɛla ni duguma dan ye) ani vertical line suppression ni orientation tolerance ye 5% ye. Ja min baaralen don, o bɛ dantigɛ a yɛrɛma ni entropi maksimali algorisimu ye ani mask binaire bɛ dilan. Ja yɔrɔ minnu bɛ tali kɛ ROI sugandilenw bolo la TEM ja raw kɔnɔ, olu bɔra, ka mitokondriyaw taamasiɲɛw jira ani ka joli-sira-funu ni yɔrɔ wɛrɛw bɔ minnu ni ɲɔgɔn cɛ ka bon. ROI bɔlen kelen-kelen bɛɛ la, fɛnɲɛnamafagalan fila minnu ka bon ni pixel 600 ye, olu sɛgɛsɛgɛra, ani fɛnɲɛnamafagalanw yɔrɔ, u lamini, aksidan belebelew ni u fitininw, Feret cɛmancɛ, u laminicogo, ani u laminicogo suman ni Fidji/ImageJ ka sumanikɛcogo jɔlenw ye. Merrill, Flippo, ani Strack (2017) kɔfɛ, yɔrɔ 2, fɛnɲɛnamafagalanw ɲɛda hakɛ (aksidanba ni aksidan fitinin hakɛ), ani cogoya hakɛ (FF) jatebɔ kɛra ka bɔ nin kunnafoniw na, yɔrɔ min na FF = lamini 2/4pi x yɔrɔ. Paramɛtiri formula ɲɛfɔli bɛ se ka sɔrɔ Merrill, Flippo ani Strack (2017) kɔnɔ. Makɔrɔ minnu kofɔlen don, olu bɛ sɔrɔ GitHub kan (aw ye kunnafoni sɔrɔcogo sɛbɛn lajɛ). Ni an y’a jateminɛ, fɛnɲɛnamafagalan 5600 ɲɔgɔn sɛgɛsɛgɛra PPA furakɛli kelen kɔnɔ, o bɛɛ lajɛlen kɛra fɛnɲɛnamafagalan 17000 ɲɔgɔn ye (kunnafoni ma jira).
SH-SH5Y seliluw bilala sɛnɛfɛnw na minnu bɛ so 8 kɔnɔ (ThermoFisher, #155411) walasa ka a to u ka nɔrɔ su fɛ, o kɔfɛ, u bilala ni TMRE 1:1000 (ThermoFisher, #T669) ani Hoechst 33342 1:200 (Sigma-Aldrich, H6024) ye. kalali. Jaw sɔrɔla ni laser 405 nm ani 561 nm ye miniti 10 lamini na, ani ja raw sɔrɔla i n’a fɔ z-stacks minnu kɔnɔ ja mikorografu 10 bɛ sɔrɔ ni az taamasiɲɛ ye 0,2 μm ye ja karamɔgɔ ni ɲɔgɔn cɛ waati 12 nataw la Jaw lajɛra ni Carl Zeiss LSM780 ELYRA PS.1 super-resolution platform ye (Carl Zeiss, Oberkochen, Alimanjamana) ni LCI Plan Apochromate 100x/1.4 Oil DIC M27 lens ye. Jaw sɛgɛsɛgɛra ImageJ kɔnɔ ni pibiliki ye min ɲɛfɔra ka tɛmɛ ani ImageJ plugin ye walasa ka fusion ni fission ko kɛlenw suman, mitokondriyaw jɔcogo hakɛ cɛmancɛ, ani mitokondriyaw hakɛ cɛmancɛ seli kelen kɔnɔ MEL makɔrɔw bɛ sɔrɔ GitHub kan (aw ye kunnafoni sɔrɔcogo sɛbɛn lajɛ).
SH-SY5Y seliluw falenna kɔlɔn wɔɔrɔ kɔnɔ ni u hakɛ ye 0,3 × 106 selilu/mL ye sanga 24 kɔnɔ ka sɔrɔ ka furakɛli kɛ. ARN bɔra ni Quick-RNATM Miniprep protocole ye (ZR R1055, Zymo Research) ni fɛn dɔw sɛmɛntiyalen don: aw bɛ ARN lysis tampon 300 μl fara kɔlɔn kelen-kelen bɛɛ kan sani a ka bɔ ani ka sɛgɛsɛgɛli kɛ sɛgɛsɛgɛli kelen-kelen bɛɛ kan i n’a fɔ fɛɛrɛ laban ni DNase/RNase elution 30 μl ye. -ji hɔrɔnyalen. Sɛgɛsɛgɛli bɛɛ kɛra u hakɛ ni u jogo la ni NanoDrop ND-1000 UV-Vis Spectrophotometer ye. Protéines totales ka bɔ cellules lysates la, o sɔrɔla ni 200 μl RIPA lysis buffer ye, ani proteine ​​hakɛ jateminɛna ni Bradford protein assay ye70.
ADNc labɛnni kɛra ni TetroTM cDNA Synthesis Kit (BIO-65043, Meridian Bioscience) ye ka kɛɲɛ ni a dilabaga ka laadilikanw ye ni fɛn dɔw sɛmɛntiyalen don. ADNc kɛra 20-μl waleyali la ni ARN bɛɛ lajɛlen 0,7 fo 1 μg ye. Primerw sugandira ka bɔ sɛbɛn 42, 71, 72, 73, 74, 75, 76, 77, 78 la minnu bɔra ka tɛmɛ (Taabolo S1) ani sonda minnu bɛ taa ɲɔgɔn fɛ, olu dilannen don ni PrimerQuest baarakɛminɛn ye min bɔra Integrated DNA Technologies fɛ. Nafa sɔrɔli jɛnɛya bɛɛ kɛra normalize ye ka kɛɲɛ ni nukilila B2M jamu ye. STOML2, NRF1, NFE2L2, TFAM, cMYC ani OPA1 ka jeninida jiracogo jateminɛna ni RT-qPCR ye. Master mixture tun ye LUNA Taq polymerase (M3003L, New England Biolabs), 10 μM ɲɛfɛla ni kɔfɛ primers, cDNA, ani PCR-grade ji ye walasa ka 10 μL hakɛ laban sɔrɔ farikoloɲɛnajɛ kelen-kelen bɛɛ la. Tilali ni farali jamu (DRP1, MFN1/2) jirali suman ni TaqMan multiplex assays ye. Luna Universal Probe qPCR Master Mix (M3004S, New England Biolabs) kɛra ka kɛɲɛ ni a dilabaga ka laadilikanw ye ni fɛn misɛnninw ye. Multiplex RT-qPCR master mix kɔnɔ, 1X LUNA Taq polimerazi, 10 μM ɲɛfɛla ni kɔfɛla, 10 μM sonda, cDNA, ani PCR-grade ji bɛ sɔrɔ, o bɛ na ni 20 μL hakɛ laban ye farikoloɲɛnajɛ kelen-kelen bɛɛ la. RT-qPCR kɛra ni Rotor-Gene Q 6-plex ye (QIAGEN RG—seriyali nimɔrɔ: R0618110). Bisi bolicogo jiralen bɛ jatebɔsen S1 kɔnɔ. ADNc sɛgɛsɛgɛli bɛɛ kɛra ka caya siɲɛ saba ani ka kurufe jɔnjɔn dɔ dilan ni dilancogo tan ye min bɛ tugu ɲɔgɔn na. Outliers minnu bɛ samples saba kɔnɔ ni cycle threshold standard deviation (Ct) >0,5 ye, olu bɔra sɛgɛsɛgɛli la walasa ka kunnafoniw lasegincogo sabati30,72. Jɛnɛya jiracogo danfaralen jatebɔ kɛra ni 2-ΔΔCt79 fɛɛrɛ ye.
Protéines (60 μg) ɲagamina ni Laemmli doni tampon ye ni 2:1 hakɛ ye ka boli 12% protein gel kan min kulɛri tɛ (Bio-Rad #1610184). Protéines (poroteyini) bilala PVDF (polyvinylidene fluoride) membrane (#170-84156, Bio-Rad) kɔnɔ ni Trans-Blot Turbo system (#170-4155, Bio-Rad) ye. Aw bɛ membrane datugu ka a bila ni farikolojidɛsɛ fura fɔlɔ bɛnnenw ye (OPA1, MFN1, MFN2, ani DRP1) (a dilanna 1:1000) fo lɛri 48, o kɔfɛ, a bɛ tobi ni banakisɛfagalan filananw ye (1:10.000) fo lɛri 1. O kɔfɛ, membranes ja kɛra ni Clarity Western ECL Substrate (#170-5061, Bio-Rad) ye, ka sɛbɛn ni Bio-Rad ChemiDoc MP system ye. ImageLab version 6.1 kɛra ka Western blot sɛgɛsɛgɛli kɛ. Jeli ni blot fɔlɔ jiralen don ja S1 na. Banakisɛfagalanw kunnafoniw bɛ sɔrɔ jatebɔsen S2 kɔnɔ.
Kunnafoni kuluw bɛ jira i n’a fɔ hakɛ danmadɔ ni fili hakɛ dantigɛlen (SEM) min bɛ sɔrɔ misali yɛrɛmahɔrɔnyalen saba la dɔgɔmannin na. Kunnafoni kuluw sɛgɛsɛgɛra ni u bɛ cogo la ni Shapiro-Wilks ka sɛgɛsɛgɛli ye (fo n’a fɔra cogo wɛrɛ la) ka sɔrɔ ka Gauss ka jatebɔ ni danfaralanw jateminɛ minnu bɛ bɛn ɲɔgɔn ma ani ka taa ɲɛ ni sɛgɛsɛgɛliw ye. Ka fara kunnafonidilanw sɛgɛsɛgɛli kan ni Fisher ka MEL LSD ye (p < 0,05), sira kelen ANOVA (furakɛli ni kɔrɔsili hakɛ danmadɔ), ani Dunnett ka sɛgɛsɛgɛli caman kɛli ye walasa ka nafa dɔn (p < 0,05). P hakɛ nafamaw jiralen don ja in kɔnɔ i n’a fɔ *p < 0,05, **p < 0,01, ***p < 0,001, ****p < 0,0001. Jateminɛw ni jatebɔw bɛɛ kɛra ka dilan ni GraphPad Prism 9.4.0 ye.
Fidji/ImageJ makɔrɔ minnu bɛ kɛ TEM ja sɛgɛsɛgɛli kama, olu bɛ sɔrɔ foroba la GitHub kan: https://github.com/caaja/TEMMitoMacro. Mitokondriyaw ka ko kɛlenw ɲininikɛla (MEL) makɔrɔ bɛ sɔrɔ foroba la GitHub kan: https://github.com/rensutheart/MEL-Fiji-Plugin.
Meiliana A., Devi NM ani Vijaya A. Mitokondriyaw: farikoloɲɛnajɛ, omɛostasi, degun, kɔrɔya ani epijenitiki ɲɛnabɔbaga ŋanaw. Indonezikan na. Biomedical Science (Biyomedical Science) ye. J. 13, 221-241 (2021) Kalanjɛ ni sɛbɛnni: kalanjɛ ni sɛbɛnni gafew.
Ben-Shachar, D. Mitokondriyaw ka baarakɛbaliya fan caman fɛ hakiliɲagami bana na, complexe I i n’a fɔ bana laɲini min bɛ se ka kɛ. Sizofreni (Schizophrénie) bana. nafolomafɛn. 187, 3–10 (2017) Ɲɛjirali dɔw.
Bose, A. ani Beal, MF Mitokondriyaw ka baarakɛbaliya Parkinson ka bana na. J. Neurochimie (sɔgɔsɔgɔninjɛ). 139, 216–231 (2016) Kalanjɛ ni sɛbɛnni: kalanjɛ ni sɛbɛnni gafew.
Sharma VK, Singh TG ani Mehta V. Mitokondriyaw degun: binkanni laɲiniw Alzheimer bana na. Mitokondriyaw 59, 48–57 (2021) Kalanjɛ ni sɛbɛnni: kalanjɛ ni sɛbɛnni gafew.
Belenguer P., Duarte JMN, Shook PF ani Ferreira GK Mitokondriyaw ni hakili: biyoɛnerji ani fɛn wɛrɛw. Neurotoxins (banakisɛfagalanw). nafolomafɛn. 36, ɲɛ 219–238 (2019) kɔnɔ.
Rangaraju, V. ani a ɲɔgɔnnaw. Mitokondriyaw pleiotropique : mitokondriyaw nɔ min bɛ senfagabana yiriwali ni banaw la. J. Hakilila-jɔ-sɛnɛ. 39, 8200–8208 (2019) Kalanjɛ ni sɛbɛnni: kalanjɛ ni sɛbɛnni gafew.
Cardaño-Ramos, C. ani Morais, V. A. Mitokondriyaw ka biyojɛnɛsi senfagabana kɔnɔ: cogo ani yɔrɔ min. diɲɛnatigɛko. J. Mohr ye baara kɛ. dɔnniya ye. 22, san 13059 (2021 sàn).
Yu, R., Lendahl, U., Nister, M. ani Zhao, J. Baganw ka mitokondriyaw ka baarakɛcogo ɲɛnabɔli: sababuw ni gɛlɛyaw. ɲɛfɛla. endocrine (endocrine) ye. (Lausanne) 11, 374 (2020) Bamako, Mali.
Khacho, M. ani Slack, RS Mitokondriyaw ka fanga sɔrɔcogo ɲɛnabɔli la: k’a ta hakili la min bɛ ka yiriwa ka taa a bila balikuw ka hakili la. yiriwali. dinamikɛ ye. 247, 47–53 (2018) Kalanjɛ ni sɛbɛnni: kalanjɛ ni sɛbɛnni gafew.