Making human nasal cilia beat in the cold: a real time assay for cell signalling.
Human nasal epithelium must adapt to cold climates, and yet, in vitro, human ciliary beat frequency (CBF) is zero at 4 degrees C. Similarly, hibernating mammals do not die of pneumonia despite a core body temperature as low as 6 degrees C, implying that cilia continue to beat. Here, we show that protein kinase C (PKC) and Ca(2+)/calmodulin-dependent kinase II (CaMKII) regulate the profile of human nasal CBF in response to rising temperature from 4 degrees C. Onset of ciliary beat was at 10 degrees C in Medium 199, 7 degrees C in the presence of the PKC activator phorbol 12-myristate 13-acetate (PMA), the calcium ionophore ionomycin, or the CAMKII blocker myristoylated autocamtide-2 related inhibitory peptide (MACI), and 6 degrees C for the myristoylated peptide PKC inhibitor EGF-R Fragment 651-658 (MyrPKCI). During cell warming to 32 degrees C, the thermal profile was sigmoid in all solutions except those containing MACI+PMA. Surprisingly, cilia continued to beat despite 4 degrees C and were significantly more responsive to rising temperature with either MACI+PMA, or MACI+MyrPKCI. Our data suggest that CaMKII and PKC regulate the thermal slope and profile of CBF in vitro, and that when these protein kinases are manipulated, cilia can continue to beat despite hypothermia. These findings may relate to adaptive responses to cold climates.
Mwimbi
Xowi K M S
XK
Tayside Institute of Child Health (Centre for Research into Human Development), Ninewells Hospital and Medical School, Dundee, DD1 9SY, UK.
Muimo
Richmond
R
Green
M W
MW
Mehta
Anil
A
D000222:Adaptation, Physiological; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000502:physiology; D002923:Cilia / Q000502:physiology; D003080:Cold Temperature; D006801:Humans; D007527:Isoenzymes / Q000502:physiology; D009079:Mucociliary Clearance / Q000502:physiology*; D009297:Nasal Mucosa / Q000201:enzymology / Q000502:physiology*; D011493:Protein Kinase C / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology*
D007527:Isoenzymes; D011493:Protein Kinase C; D017871:Calcium-Calmodulin-Dependent Protein Kinases