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AChE mRNA stability in mammalian skeletal muscle, studied in vitro
Zajc-Kreft, Katarina (Author), Kreft, Samo (Author), Grubič, Zoran (Author)

URLURL - Presentation file, Visit http://www.dlib.si/details/URN:NBN:SI:doc-8VD9L6LE This link opens in a new window

Abstract
Tako elektromehanska aktivnost kot tudi denervacija povzročita padec ravni acetilholinesteraze (AChE) mRNA v hitri podganji skeletni mišici. Povečana hitrost razgradnje je eden od možnih mehanizmov za zmanjšanje ravni AChE mRNA v denervirani mišici. Da bi to preverili, smo iz podganje m. SM izolirali deproteneizirano RNA in jo izpostavili delovanju mišičnih subceličnih frakcij,pripravljenih iz kontralaterarne m. SM. Po določenih časovnih intervalih, smo z neradioaktivno Northern blot analizo določili količino nerazgrajene AchE mRNA. Vsebnost AChE mRNA v mišici ostane nespremenjena tekom prvih 5 ur po denervaciji, potem pa strmo pade v naslednjih 13 urah. Nadaljnje padanje je veliko počasnejše. Daljši transkript (3.5kb) je manj stabilen kot krajši (2.3 kb). V denervirani mišici se je znižala tidi vsebnostalfa aktinske kisline mRNA, in to s podobno hitrostjo kot AChE mRNA, kar nakazuje, da AChE mRNA v takih razmerah ni prizadeta soecifično. AChE mRNAse je razgrajevala z vsemi testiranimi subceličnimi frakcijami. Postmitohondrijska in postpolisomalna frakcija sta imeli večjo kapaciteto razgradnje kot polisomalna frakcija. Ugotovili smo, da je uporabljen pristop primeren za proučevanje kapacitete razgradnje specifičnih mRNA v odrasli skeletni mišici. Naši preliminarni rezultati kažejo, da je za padec AChE mRNA po denervaciji vsaj deloma odgovorna povečana nagnjenost k razgradnji te molekule.

Language:English
Keywords:acetilholinesteraza, podganja skeletna mišica, RNA, denervacija
Work type:Not categorized (r6)
Tipology:1.01 - Original Scientific Article
Organization:FFA - Faculty of Pharmacy
Year:1998
Publisher:Slovensko kemijsko društvo
Number of pages:str. 35-44
Numbering:Let. 45, št. 1
UDC:577.3
ISSN on article:1318-0207
COBISS.SI-ID:255857 Link is opened in a new window
Views:732
Downloads:116
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Record is a part of a journal

Title:Acta chimica slovenica
Shortened title:Acta chim. slov.
Publisher:Slovensko kemijsko društvo, =Slovenian Chemical Society
ISSN:1318-0207
COBISS.SI-ID:14086149 This link opens in a new window

Secondary language

Language:English
Title:In vitro določanje stabilnosti AChE mRNA v skeletni mišici sesalcev
Abstract:
Acetylcholinesterase (AChE) mRNA in fast ret skeletal muscle is downregulated by both, electromechanical activity and denervation. One candidate mechanism that could explain decreased level of AChE mRNA in the denervated muscle is increased rate of its degradation. In order to test this possibility, total deproteined RNA was isolated from rat . SM and exposed to subcellular musculafractions prepared from contralateral m SM. After selected time intervals, we determined remaining AChE mRNA by nonradioactive Northern blot analysis. AChE mRNA remained at the same level during first 5 hours after denervation and abruptly fell after subsequent 13h. Further decrease in the transcript level proceed at much slower rate. Longer transcript (3.5kb) was more affected than the shorte (2.3kb) one. The level of alpha-actin mRNA was also decreased in the denervated muscle, and the rate of its disappearance wassimilar to that of AChE mRNA, suggesting that AChE mRNA is not specifically affected under such conditions. Degradations of AChE mRNA was observed in all subcellular fractions studied. Postmitochondrial and postpolysomal fractions exibited higher rate than polysomal fraction. We find experimental approach demonstraded here suitable for studies of degradation capacities of the specific mRNAs in the adult skeletal muscle. Our preliminary results suggests, that fall of AChE mRNA after denervation at least partly results from increased degradation of transcripts under such conditions.


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