Single-cell contraction assay for human ciliary muscle cells. Effect of carbachol.
OBJECTIVE
The authors developed an assay to observe the contraction of a single human ciliary muscle cell.


METHODS
Cultured human ciliary muscle cells were partially detached from the culture dish by incubation with a nonenzymatic dissociation buffer and treated with carbachol or pilocarpine. Contraction was quantified by measuring the cross-sectional surface areas of the cells.


RESULTS
Carbachol decreased the cell surface area in a time-dependent manner. Contraction was observed within 1 min after the addition of carbachol and completed in less than 15 min. The effect of carbachol was dose dependent. For example, at 10 min after treatment with 10 mumol/l carbachol, the relative surface areas of cells decreased to 47% +/- 4% (mean +/- standard error of the mean, n = 7, with surface area at 0 min defined as 100%). The relative surface areas were 74% +/- 4% (n = 7) after 1 mumol/l and 100% +/- 9% (n = 7) after 0.1 mumol/l carbachol treatment. This contractile effect was antagonized by pretreatment with atropine, a specific muscarinic antagonist.


CONCLUSIONS
A simple method was established to study the functional changes of human ciliary muscle cells.
Pang
I H
IH
Glaucoma Research, Alcon Laboratories, Fort Worth, TX 76134.
Shade
D L
DL
Tamm
E
E
DeSantis
L
L
D001681:Biological Assay / Q000379:methods*; D002217:Carbachol / Q000494:pharmacology*; D002478:Cells, Cultured; D002924:Ciliary Body / Q000166:cytology / Q000502:physiology*; D006801:Humans; D009119:Muscle Contraction / Q000187:drug effects; D009132:Muscles / Q000166:cytology / Q000502:physiology*; D010862:Pilocarpine / Q000494:pharmacology
D010862:Pilocarpine; D002217:Carbachol