PMID- 2159585
OWN - NLM
STAT- MEDLINE
DCOM- 19900614
LR  - 20190824
IS  - 0169-328X (Print)
IS  - 0169-328X (Linking)
VI  - 7
IP  - 3
DP  - 1990 Apr
TI  - Ligand autoradiographic receptor screening: receptor cDNA expression in replicas 
      of transfected COS cells.
PG  - 249-59
AB  - In this paper we validate a methodology, ligand autoradiographic receptor 
      screening (LARS), for detecting expression of full length receptor cDNAs in COS 
      cells. The method involves transfection of COS cells with receptor cDNAs by 
      spheroplast fusion, production of filter replicas of the cell fragments, ligand 
      binding to the receptors expressed in the membranes, and autoradiographic 
      detection of bound ligand. A beta-adrenergic receptor cDNA cloned into a 
      eukaryotic expression vector reliably induces high levels of beta-adrenergic 
      receptor expression in 2-12% of COS cell colonies transfected with this plasmid 
      after experimental conditions are optimized. Receptor expression is monitored by 
      autoradiographic detection of 125iodocyanopindolol binding to COS cell fragments 
      immobilized on polyester filter replicas. Binding displays appropriate 
      pharmacological properties. The number of high-density binding spots per filter 
      depends on the fraction of the spheroplasts in the fusion mixture that contain 
      the beta-adrenergic receptor cDNA. A 100-plate LARS experiment can assess 
      receptor expression in more than 10(4) transfected colonies. Thus detection of 
      receptor-encoding sequences present in libraries in proportions as low as 1 in 
      10(4) should be possible. This technique may therefore be useful in detecting 
      expression of other receptor cDNAs for which selective radioligands are 
      available.
FAU - Rattray, M
AU  - Rattray M
AD  - Gene Neuroscience Unit, Addiction Research Center/NIDA, Baltimore, MD.
FAU - Lautar, S L
AU  - Lautar SL
FAU - Uhl, G R
AU  - Uhl GR
LA  - eng
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - Netherlands
TA  - Brain Res Mol Brain Res
JT  - Brain research. Molecular brain research
JID - 8908640
RN  - 0 (Receptors, Adrenergic, beta)
RN  - 83498-72-0 (Iodocyanopindolol)
RN  - 9007-49-2 (DNA)
RN  - BJ4HF6IU1D (Pindolol)
SB  - IM
MH  - Animals
MH  - Autoradiography/*methods
MH  - Cells, Cultured
MH  - DNA/*genetics
MH  - Escherichia coli/*genetics
MH  - Iodocyanopindolol
MH  - Pindolol/*analogs & derivatives/metabolism
MH  - Plasmids/*genetics
MH  - Receptors, Adrenergic, beta/*genetics/metabolism
MH  - Spheroplasts
MH  - Transfection
EDAT- 1990/04/01 00:00
MHDA- 1990/04/01 00:01
CRDT- 1990/04/01 00:00
PHST- 1990/04/01 00:00 [pubmed]
PHST- 1990/04/01 00:01 [medline]
PHST- 1990/04/01 00:00 [entrez]
AID - 0169-328X(90)90035-C [pii]
AID - 10.1016/0169-328x(90)90035-c [doi]
PST - ppublish
SO  - Brain Res Mol Brain Res. 1990 Apr;7(3):249-59. doi: 10.1016/0169-328x(90)90035-c.