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1
-
-
9444266931
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2AR subtypes present on resistance arterioles is unknown.
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8
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9444262308
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note
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+ animais (4.3 ± 1.4 pups per litter; n = 4 litters).
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9
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9444288532
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note
-
Mice were anesthetized with inhaled methoxyflurane (1 to 2%). The left carotid artery was isolated and secured through a midline neck incision. An Intramedic PE-10 polyethylene catheter (Clay-Adams, Parsippany, NJ) was stretched to approximately one-half of its original diameter, inserted into the artery, and secured in place with 4-0 silk. The catheter was tunneled to the back of the neck and flushed with heparinized saline, and the end was sealed with glue and buried under the skin for later retrieval. The mice were allowed to recover in a standard rodent cage for at least 12 hours with food and water available ad libitum. For hemodynamic measurements, the arterial catheter was flushed with saline and connected to a Spectramed DTX Plus pressure transducer with a side port for infusing medications. The analog input was amplified with a Gould (Cleveland, OH) model 11-1202-25 preamplifier and model 13-4615-52 amplifier and digitized with a Data Translation (Marlboro, MA) DT2801 analogto-digital converter. The waveform was analyzed to derive mean blood pressure and heart rate through use of Dataflow data acquisition software (Crystal Biotech, Hopkinton, MA) on a Gateway 2000 486DX2 66 MHz microcomputer (Sioux City, SD). For agonist studies, boluses of dexmedetomidine (5 μg/kg) or phenylephrine (10 μg/kg) were administered through the arterial catheter. For blocking studies, intra-arterial atipamezole (200 μg/kg) was administered 5 min before dexmedetomidine.
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28
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9444228664
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note
-
+ animals derived from the 129Sv/J × C57BL/6J background.
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-
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-
31
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9444256789
-
-
note
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+ mice mean blood pressure increased by 25 ± 8% (n = 2) after intravascular administration of phenylephrine.
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B. Meister, A. Dagerlund, A. P. Nicholas, T. Hokfelt, J. Pharmacol. Exp. Ther. 268, 1605 (1994); M. Scheinin et al., Brain Res. Mol. Brain Res. 21, 133 (1994); D. E. Handy, C. S. Flordellis, N. N. Bogdanova, M. R. Bresnahan, H. Gavras, Hypertension 21, 861 (1993); A. P. Nicholas, V. Pieribone. T. Hokfelt, J. Comp. Neurol. 328, 575 (1993); M. Perala et al., Mol. Brain Res. 16, 57 (1992); L. Sun, S. Umemura, M. Chun, W. A. Pettinger, Eur. J. Pharmacol. 226, 367 (1992); D. Zeng and K. R. Lynch, Mol. Brain Res. 10, 219 (1991); W. Lorenz et al., Mol. Pharmacol. 38, 599 (1990).
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43
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9444276404
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note
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6-7) and G protein coupling (cytoplasmic loop 3) and should be nonfunctional.
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44
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9444295153
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note
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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.
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45
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9444233746
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note
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+ , 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).
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9444252135
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+ 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).
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48
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9444284781
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note
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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.
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