Effects of histamine on ciliary beat frequency of ciliated cells from guinea pigs nasal mucosa

Abstract

We aimed to investigate the effect of histamine on ciliary beat frequency (CBF) through combining high-speed digital microscopy and patch-clamp technology. Ciliated cells were obtained from septum and turbinate of 90-120-day-old healthy male guinea pigs. Tight seal was formed by applying negative pressure on the glass electrode after the drawing and pushing progress. Then, we enrolled high-speed digital microscopy to measure CBF before and after treatment with histamine of different concentrations ranging from 10(-6) to 10(-1) mol/L in Hank's solution and D-Hank's solution as well as after administrating adenosine triphosphate. One-way ANOVA, Student's t test or Kruskal-Wallis test was used for statistical comparisons. Glass electrode fix up ciliated cell is available at tip diameter of 2-5 μm and negative pressure of 10-20 cmH2O column. The baseline CBF in Hank's solution was higher than in D-Hank's solution. Treatment with 10(-6)-l0(-3) mol/L histamine of concentrations can stimulate a rise of CBF. Nevertheless, CBF in all groups decreased to baseline CBF within 20 min. Generally, 10(-2) mol/L histamine can stimulate a rise of CBF; meanwhile, the high concentration of histamine killed 50% ciliated cell. Histamine at 10(-1) mol/L killed all ciliated cells. Ciliary beating activity decreased in Ca(2+)-free solution. Moreover, adenosine triphosphate could increase CBF effectively after the stimulation effect of histamine. We construct an effective technology integrating patch-clamp technique with CBF measurements on ciliated cells. Extracellular histamine stimulation could increase CBF effectively.