Specific gene knockdown mediated by the antisense oligodeoxynucleotides (AODNs) strategy emerged as a rapid and effective tool for probing gene role in plant cells, particularly tip-growing pollen tubes. Here, we describe the protocol for the successful employment of AODN technique in growing tobacco pollen tubes, covering AODN design, application, and analysis of the results. We also discuss the advantages and drawbacks of this method.
Snížená exprese ubikviterní NRl podjednotky NMDA receptoru v některých oblastech mozku osob trpících schizofrenií je dávána do souvislosti s jednonukleotidovými polymorfizmy (SNP) v genu pro tuto podjednotku (GRINl). Také neurovývojový (animální) model psychóze podobného chovaní SOdenního potkana (kmen Wistar, 50. PND), založený na intrakraniální infuzi kyseliny chinolinové (QUIN; 250 nmol/0,25 pL NaCl do každé z postranních mozkových komor) neonatálním potkanům (12. PND), charakterizuje pokles exprese proteinu ubikviterní NRl podjednotky v obou hipokampech, který provází porucha ve zpracování sensorických informací (deficit v prepulzní inhibici úlekové reakce) pozorovaná také u osob trpících schizofrenií. Protože však změnu v proteinu NRl podjednotky provázely 1 zmeny v expresi většiny NR2 podjednotek, nebylo možné prokázat přímý vztah mezi sníženou expresí NRl a změnami v prepulzní inhibici akustického úleku. Proto jsme do obou hipokampů potkana 50. PND infundovali antisense oligodeoxynukleotid (aODN) specifický pro NRl podjednotku. Za 24 hod. po infuzi jsme prokázali snížení exprese proteinu hipokampální NRl, ale prepulzní inhibice akustického úleku zůstala nezměněna. V práci se navrhuje další postup při vymezování úlohy NMDA-Rl v patofyziologii schizofrenie.
Decreased expression of ubiquitous NMDA-NR1 subunit in some schizophrenic brain regions has been related to several single nuc- leotide polymorphisms (SNP) in the subunit gene ( GRIN1 ). Also neurodevelopmental (animal) model of schizophrenia-like behaviour of 50-day-old Wistar rat males, which were neonatally treated with quinolinic acid (QUIN; 250 nmol/0.25 μL NaCl into each later al cerebral ventricle on postnatal day 12), exhibited a decrease in the hippocampal subunit NR1 protein together with a deficit in the sen- sorimotor gating (deficit in the prepulse inhibition of startle reaction) accompanying schizophrenia. Because changes in the su bunit NR1 protein went with changed expression in most of NR2 subunits, it was impossible to reveal a direct relation between the dec reased expression of NR1 protein and changed prepulse inhibition of the acoustic startle. Therefore, we infused bilaterally a single d ose of antisense oligodeoxynucleotide (aODN) specific for NR1 subunit into ventral hippocampi of 50-day-old rat males. Twenty four hou rs later we found decreased hippocampal levels of NR1 protein, but prepulse inhibition of the acoustic startle remained unchanged. We discuss further study course delimitating the role of NMDA-R1 subunit in pathophysiology of schizophrenia.
- MeSH
- Oligodeoxyribonucleotides, Antisense genetics MeSH
- Research Support as Topic MeSH
- Financing, Organized MeSH
- Hippocampus physiology drug effects MeSH
- Disease Models, Animal MeSH
- Rats, Wistar physiology genetics MeSH
- Receptors, N-Methyl-D-Aspartate genetics drug effects MeSH
- Reflex, Acoustic physiology genetics MeSH
- Schizophrenia etiology genetics MeSH
- Reflex, Startle genetics drug effects MeSH
- Blotting, Western methods utilization MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- MeSH
- Oligodeoxyribonucleotides, Antisense physiology genetics MeSH
- Financing, Government MeSH
- Genetic Therapy MeSH
- Humans MeSH
- RNA, Small Interfering physiology genetics MeSH
- MicroRNAs physiology genetics MeSH
- Gene Expression Regulation genetics immunology MeSH
- RNA Interference immunology MeSH
- Check Tag
- Humans MeSH
