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Isolation of Salivary Epithelial Cells from Human Salivary Glands for In Vitro Growth as Salispheres or Monolayers
Published on: July 15, 2019
Mycoplasma orale infection affects K+ and Cl- currents in the HSG salivary gland cell line
K T Izutsu1, S Fatherazi, C M Belton
1Department of Oral Biology, School of Dentistry, University of Washington, Seattle 98195, USA.
Abstract:
The relations between K+ channel and Cl- channel currents and mycoplasma infection status were studied longitudinally in HSG cells, a human submandibular gland cell line. The K+ channel currents were disrupted by the occurrence of mycoplasma infection: muscarinic activation of K+ channels and K+ channel expression as estimated by ionomycin- or hypotonically induced K+ current responses were all decreased. Similar decreases in ionomycin- and hypotonically induced responses were observed for Cl- channels, but only the latter decrease was statistically significant. Also, Cl- currents could be elicited more frequently than K+ currents (63% of cases versus 0%) in infected cells when tested by exposure to hypotonic media, indicating that mycoplasma infection affects K+ channels relatively more than Cl- channels. These changes occurred in the originally infected cells, were ameliorated when the infection was cleared with sparfloxacin, and recurred when the cells were reinfected. Such changes would be expected to result in hyposecretion of salivary fluid if they occurred in vivo.
Insights
Mycoplasma infection disrupts potassium (K+) and chloride (Cl-) channel currents in human salivary gland cells. These ion channel changes, particularly in K+ channels, may lead to reduced salivary fluid secretion in vivo.
Area of Science:
- Cell Biology
- Ion Channel Physiology
- Microbiology
Background:
- Mycoplasma infections can affect cellular functions.
- Ion channels play critical roles in cell physiology, including secretion.
Purpose of the Study:
- To investigate the impact of Mycoplasma infection on potassium (K+) and chloride (Cl-) channel currents in human submandibular gland (HSG) cells.
- To determine if Mycoplasma infection differentially affects K+ and Cl- channels.
Main Methods:
- Longitudinal study of HSG cells with controlled Mycoplasma infection and clearance.
- Electrophysiological measurements of K+ and Cl- channel currents.
- Assessment of channel activity using muscarinic activation, ionomycin, and hypotonic challenges.
Main Results:
- Mycoplasma infection significantly decreased muscarinic activation and ionomycin-induced K+ channel currents.
- Decreases in ionomycin- and hypotonic-induced Cl- currents were observed, with hypotonic-induced decrease being statistically significant.
- K+ channels were affected more severely than Cl- channels by Mycoplasma infection.
- Changes were reversible upon antibiotic treatment (sparfloxacin) and recurred upon reinfection.
Conclusions:
- Mycoplasma infection disrupts ion channel function in HSG cells, with a notable impact on K+ channels.
- These ion channel alterations are reversible, suggesting a direct effect of the infection.
- The observed changes in ion channel activity could lead to hyposecretion of salivary fluid in vivo.
