Nucleotide-induced restoration of conjunctival chloride and fluid secretion in adenovirus type 5-infected pigmented rabbit eyes.
We evaluated the role of extracellular UTP and other nucleotides in the regulation of chloride (JCl) and fluid secretion (JCl) across the pigmented rabbit conjunctiva. Jv was determined in freshly excised conjunctival tissues mounted between two buffer reservoirs maintained in an enclosed environment at 37 degrees C. Short circuit current (Isc) and 36Cl flux were measured using modified Ussing-type chambers. Fluid flux measurements were made with a pair of capacitance probes. After observing the baseline for 15 to 30 min, fluid flux was measured in the presence of mucosally applied nucleotides (10 microM) for a period of 30 min. Mucosal application of 10 microM each of UTP, UDP, ATP, ADP, AMP, adenosine, and ATP-gamma-S transiently stimulated fluid secretion across the conjunctiva to a significant extent for 10 to 15 min. Other nucleotides did not show any significant effect. The stimulation of fluid secretion correlated well with the stimulation in Isc (r2 = 0.85). UTP (0.1-1000 microM) led to a maximal increase in fluid secretion by 11.72 +/- 0.48 microl/(h x cm2) with an EC50 value of 10.39 +/- 1.08 microM. ATP (0.1-1000 microM) caused a maximal increase in fluid secretion by 11.89 +/- 0.88 microl/(h x cm2) with an EC50 value of 17.23 +/- 2.63 microM. Adenovirus type 5 (Ad5) infection significantly decreased both net 36Cl secretion across the conjunctiva by approximately 56% and the rate of fluid secretion by approximately 56%. UTP (10 microM), but not 1 mM 8-bromo-cAMP, was able to elicit a normal stimulatory response in the Ad5-infected tissues. In conclusion, mucosal application of purinergic nucleotides may be therapeutically important in restoring ion and fluid secretion in the diseased conjunctiva.
Kulkarni
Ashutosh A
AA
Department of Pharmaceutical Sciences, School of Pharmacy, 1985 Zonal Ave., University of Southern California, Los Angeles, CA 90089-9121, USA.
Trousdale
Melvin D
MD
Stevenson
Douglas
D
Gukasyan
Hovhannes J
HJ
Shiue
Michael H I
MH
Kim
Kwang-Jin
KJ
Read
Russell W
RW
Lee
Vincent H L
VH
D000241:Adenosine / Q000494:pharmacology*; D000256:Adenoviridae; D000257:Adenoviridae Infections / Q000473:pathology; D000818:Animals; D001826:Body Fluids / Q000378:metabolism; D002712:Chlorides / Q000378:metabolism; D003228:Conjunctiva / Q000187:drug effects* / Q000378:metabolism; D004195:Disease Models, Animal; D015817:Eye Infections / Q000473:pathology; D010858:Pigmentation; D058913:Purinergic Agonists; D011817:Rabbits; D014529:Uridine / Q000494:pharmacology*; D014544:Uridine Triphosphate / Q000494:pharmacology
D002712:Chlorides; D058913:Purinergic Agonists; D000241:Adenosine; D014544:Uridine Triphosphate; D014529:Uridine