PMID- 10355826
OWN - NLM
STAT- MEDLINE
DCOM- 19990715
LR  - 20061115
IS  - 1087-2906 (Print)
IS  - 1087-2906 (Linking)
VI  - 9
IP  - 2
DP  - 1999 Apr
TI  - Differential knockdown of delta-opioid receptor subtypes in the rat brain by 
      antisense oligodeoxynucleotides targeting mRNA.
PG  - 203-11
AB  - Two antisense oligodeoxynucleotides (A-ODN), targeting delta-opioid receptor mRNA 
      (DOR) and two mismatch ODN sequences (mODN) were continuously infused for 24 days 
      into the lateral brain ventricles of Wistar rats. The density of delta-opioid 
      receptors in rat brain homogenates was measured by saturation binding experiments 
      using four selective ligands, two agonists ([D-Ala2, Glu4]-deltorphin and DPDPE) 
      and two antagonists (Dmt-Tic-OH and naltrindole), and by immunoblotting SDS 
      solubilized receptor protein. In brain membranes of mODN or saline-infused rats, 
      the rank order of delta-opioid receptor density, calculated by Bmax values of the 
      four delta-opioid receptor ligands, was: [D-Ala2, Glu4]deltorphin approximately 
      Dmt-Tic-OH approximately naltrindole (86-118 fmo/mg protein) > DPDPE (73.6+/-6.3 
      fmol/mg protein). At the end of the 24 day infusion of A-ODN targeting DOR 
      nucleotide sequence 280299 (A-ODN280-299), the Bmax of DPDPE (62.4+/-3.2 fmol/mg 
      protein) was significantly higher than that of Dmt-Tic-OH (31.5+/-3.9 fmol/mg 
      protein). Moreover, both the Kd value for DPDPE saturation binding and the Ki 
      value for Dmt-Tic-OH displacement by DPDPE were halved. In contrast, an A-ODN 
      treatment targeting exon 3 (A-ODN741-760) decreased the specific binding of 
      [D-Ala2, Glu4]deltorphin and Dmt-Tic-OH significantly less (67%-81%) than the 
      binding of DPDPE (53%), without changes in DPDPE Ki and KD values. No A-ODN 
      treatment modified the specific binding of the micro-opioid agonist DAMGO and of 
      the k-selective opioid receptor ligand U69593. On the Western blot of solubilized 
      striatum proteins, A-ODN(280-299) and A-ODN(741-760) downregulated the levels of 
      the DOR protein, whereas the corresponding mODN were inactive. The 24-day 
      infusion of A-ODN(280-299) inhibited the rat locomotor response to [D-Ala2, 
      Glu4]deltorphin but not to DPDPE. Intracerebroventricular (i.c.v.) infusion of 
      A-ODN(741-760) reduced the locomotor responses to both delta-opioid receptor 
      agonists, whereas mODN infusion never affected agonist potencies. In conclusion, 
      these results demonstrate that 24-day continuous i.c.v. infusion of A-ODN 
      targeting the nucleotide sequence 280-299 of DOR can differentially knockdown 
      delta1 and delta2 binding sites in the rat brain.
FAU - Negri, L
AU  - Negri L
AD  - Institute of Medical Pharmacology, University La Sapienza, Rome, Italy.
FAU - Lattanzi, R
AU  - Lattanzi R
FAU - Borsodi, A
AU  - Borsodi A
FAU - Toth, G
AU  - Toth G
FAU - Salvadori, S
AU  - Salvadori S
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Antisense Nucleic Acid Drug Dev
JT  - Antisense & nucleic acid drug development
JID - 9606142
RN  - 0 (Ligands)
RN  - 0 (Oligodeoxyribonucleotides, Antisense)
RN  - 0 (RNA, Messenger)
RN  - 0 (Receptors, Opioid, delta)
RN  - 0 (Thionucleotides)
SB  - IM
MH  - Animals
MH  - Brain/*drug effects
MH  - Corpus Striatum/drug effects
MH  - Ligands
MH  - Male
MH  - Motor Activity/*drug effects
MH  - Oligodeoxyribonucleotides, Antisense/*pharmacology
MH  - RNA, Messenger/*drug effects
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Receptors, Opioid, delta/agonists/*drug effects/genetics
MH  - Thionucleotides/pharmacology
EDAT- 1999/06/04 00:00
MHDA- 1999/06/04 00:01
CRDT- 1999/06/04 00:00
PHST- 1999/06/04 00:00 [pubmed]
PHST- 1999/06/04 00:01 [medline]
PHST- 1999/06/04 00:00 [entrez]
AID - 10.1089/oli.1.1999.9.203 [doi]
PST - ppublish
SO  - Antisense Nucleic Acid Drug Dev. 1999 Apr;9(2):203-11. doi: 
      10.1089/oli.1.1999.9.203.