Cardiovascular regulation in mice lacking α2-adrenergic receptor subtypes b and c

Science

Volumn 273, Issue 5276, 1996, Pages 803-805

  Link, Richard E ^a   Desai, Kavin ^a   Hein, Lutz ^b   Stevens, Mary E ^a   Chruscinski, Andrzej ^a   Bernstein, Daniel ^a   Barsh, Gregory S ^a   Kobilka, Brian K ^a  

^a STANFORD UNIVERSITY   (United States)

^b STANFORD UNIVERSITY SCHOOL OF MEDICINE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ALPHA 2 ADRENERGIC RECEPTOR STIMULATING AGENT; ALPHA 2B ADRENERGIC RECEPTOR; ALPHA 2C ADRENERGIC RECEPTOR; DEXMEDETOMIDINE; RECEPTOR SUBTYPE;

ANIMAL EXPERIMENT; ANIMAL TISSUE; ANTIHYPERTENSIVE ACTIVITY; ARTICLE; BLOOD PRESSURE; BRAIN STEM; CARDIOVASCULAR AUTOREGULATION; CONTROLLED STUDY; DRUG EFFICACY; DRUG SELECTIVITY; GENETIC ENGINEERING; HEART RATE; HEMODYNAMICS; HYPERTENSION; INTRAVENOUS DRUG ADMINISTRATION; MOUSE; NONHUMAN; PRIORITY JOURNAL; VASCULAR SMOOTH MUSCLE;

ANIMALIA;

EID: 0029739504     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.273.5276.803     Document Type: Article

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43. 6-7) and G protein coupling (cytoplasmic loop 3) and should be nonfunctional.
44. 32P-labeled DNA probes: for Adra2b, a 0.4-kb Sac I to Sac II fragment derived from sequences 5′ to the Adra2b coding sequence, and Neo, a 0.82-kb Pst I to Xba I fragment of the neo coding sequence.
45. + , or Adra2b / Adra2b mice with a modification of the protocol of Uhlen and Wikberg (20). Protein electrophoresis through 10%-polyacrylamide gels and transfer to nitrocellulose membranes were done as described (100 μg of membrane protein per lane) (21). After transfer to nitrocellulose and blocking in BLOTTO (5% nonfat dried milk protein, 0.1% Tween 20 in phosphate-buffered saline), membranes were incubated sequentially with affinity-purified primary antibody to Adra2b (D. Daunt, Stanford University, Stanford, CA; 1:500 dilution) and then with peroxidase-labeled secondary antibody to rabbit immunoglobulin G (Amersham: 1:1000 dilution). After washing in 0.1 % Tween 20 in phosphate-buffered saline, proteins were detected by enhanced chemiluminescense (ECL, Amersham).
46. + mouse. Nonspecific binding was determined by incubation with 100 μM WB4101. The data were analyzed with a nonlinear least-squares curve-fitting technique (GraphPad software; GraphPad Software, San Diego, CA).
47. R. Virtanen, J. M. Savola, V. Saano, Arch. Int. Pharmacodyn. Ther. 297, 190 (1989).
48. Supported by National Institutes of Health grant HL48638 (Project 4). R.E.L. is supported by an MSTP training grant (GM07365); G.S.B. and B.K.K. are assistant investigators of the Howard Hughes Medical Institute.