title: Response to prejunctional adenosine receptors is dependent on stimulus frequency.
abstract: OBJECTIVE
  To characterize the adenosine receptor modulation of norepinephrine (NE) release from sympathetic neurons in the isolated rabbit iris/ciliary body.
  
  
  METHODS
  Iris/ciliary bodies were isolated from New Zealand White rabbits and incubated in the presence of 3H-NE. Norepinephrine release was elicited by field stimulation with varied frequencies from 5 to 30 Hz. The effects of adenosinergic and alpha 2 adrenergic compounds on NE release were determined and compared.
  
  
  RESULTS
  At a stimulation frequency of 5 Hz, the addition of the adenosine A1 agonist CHA did not significantly alter 3H-NE release. However, in the presence of the alpha 2 adrenergic antagonist yohimbine, the addition of CHA produced a dose-related reduction in 3H-NE release. The EC50 for this reduction was 14 nM. At a stimulus frequency of 20 Hz, the addition of CHA alone (10(-6) M) produced a significant reduction in 3H-NE release of 41%. The EC50s for the adenosine A1 agonists CHA- and R-PIA-induced suppression of 3H-NE release at 20 Hz were 32 and 24 nM, respectively. The adenosine A2 agonist CV-1808 did not alter 3H-NE release at stimulation frequencies of 5 or 20 Hz. Pretreatment of tissues with the adenosine A1 antagonist CPT or pertussis toxin reversed the suppression of 3H-NE release induced by CHA. Comparison of the inhibitory responses of CHA to the alpha 2 adrenergic agonist UK-14,304 at stimulus frequencies of 5 to 30 Hz demonstrated that this adenosine A1 agonist was effective only in suppressing NE release at frequencies of 20 Hz or greater. In contrast, the alpha 2 adrenergic agonist UK-14,304 was most effective in reducing NE release at 5 Hz.
  
  
  CONCLUSIONS
  These results provide evidence that adenosine agonists inhibit 3H-NE release in the iris/ciliary body via prejunctional adenosine A1 receptors linked to Gi/o-protein. However, the expression of this response was dependent on the frequency of neuronal stimulation. Hence, prejunctional adenosine A1 receptors may act to selectively limit high-frequency neurotransmission.
authors.affiliation: Department of Ophthalmology and Visual Sciences, Texas Tech University Health Sciences Center, Lubbock 79430, USA.
authors.forename: C E
authors.initials: CE
authors.lastname: Crosson
authors.forename: T
authors.initials: T
authors.lastname: Gray
mesh_terms: D000241:Adenosine / Q000819:agonists / Q000037:antagonists & inhibitors; D000316:Adrenergic alpha-Agonists / Q000494:pharmacology; D000818:Animals; D002924:Ciliary Body / Q000294:innervation*; D004558:Electric Stimulation / Q000379:methods; D066298:In Vitro Techniques; D007498:Iris / Q000294:innervation*; D009474:Neurons / Q000378:metabolism*; D009638:Norepinephrine / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011817:Rabbits; D018047:Receptors, Purinergic P1 / Q000502:physiology*; D013564:Sympathetic Nervous System / Q000166:cytology / Q000378:metabolism*
chemical_list: D000316:Adrenergic alpha-Agonists; D018047:Receptors, Purinergic P1; D000241:Adenosine; D009638:Norepinephrine