Figure 8. Cilia and cytosolic Ca 2 are... | Rockefeller University Press

Figure 8.

Cilia and cytosolic Ca2+are regulated by a common mechanism. (A) TIRF microscopy recording of Smo-GCaMP5G-mCh fluorescence from a β-cell within an intact mouse islet during an increase in the glucose concentration from 3 to 11 mM. Notice the appearance of regular oscillations in both cilium and soma. Kymograph above shows GCaMP5G/mCh ratio changes in a small part of the soma and along the cilium. (B) Color coded spectrogram of ciliary (top) and somatic (bottom) frequency components with superimposed traces (black) of the whole compartments Ca2+ activity recorded from one cell during 11 mM glucose exposure. Note the prevalence of a slow component with a period of about 100 s in both cilium and soma. (C) GCaMP5G fluorescence change from a mouse islet cell stimulated with 11 mM glucose. The cilium has been segmented and the fluorescence change within the segments is shown to the right. Notice that there is a gradual change of the oscillatory pattern as the segments becomes more distal to the cell body. (D) Scatter plots showing a positive correlation between the relative Ca2+ concentration changes in the cilia and soma (left) and between the duration of the cytosolic Ca2+ increase and the change in cilia Ca2+ concentration (right). (E) TIRF microscopy recording of GCaMP5G fluorescence change in the cilium (black) and soma (red) of islets cells following exposure to three 3-min KCl depolarizations (arrows). Data are means ± SEM for 8 cells from 1 islet and representative of 15 islets. (F–H) Color coded power spectra of ciliary (top) and somatic (bottom) frequency components with superimposed traces (black) of the whole compartments Ca2+ activity recorded from individual cells exposed to different treatments (see horizontal lines above plots). (I) TIRF microscopy recording of GCaMP5G fluorescence from 12 cilia from 1 mouse islet (means ± SEM) following addition of 5 mM α-KIC. Shown to the right are recordings of cilia and cytosolic Ca2+ from a matched cilia–soma pair within the islet. Representative of eight islets.

Cilia and cytosolic Ca ²⁺ are regulated by a common mechanism. (A) TIRF microscopy recording of Smo-GCaMP5G-mCh fluorescence from a β-cell within an intact mouse islet during an increase in the glucose concentration from 3 to 11 mM. Notice the appearance of regular oscillations in both cilium and soma. Kymograph above shows GCaMP5G/mCh ratio changes in a small part of the soma and along the cilium. (B) Color coded spectrogram of ciliary (top) and somatic (bottom) frequency components with superimposed traces (black) of the whole compartments Ca²⁺ activity recorded from one cell during 11 mM glucose exposure. Note the prevalence of a slow component with a period of about 100 s in both cilium and soma. (C) GCaMP5G fluorescence change from a mouse islet cell stimulated with 11 mM glucose. The cilium has been segmented and the fluorescence change within the segments is shown to the right. Notice that there is a gradual change of the oscillatory pattern as the segments becomes more distal to the cell body. (D) Scatter plots showing a positive correlation between the relative Ca²⁺ concentration changes in the cilia and soma (left) and between the duration of the cytosolic Ca²⁺ increase and the change in cilia Ca²⁺ concentration (right). (E) TIRF microscopy recording of GCaMP5G fluorescence change in the cilium (black) and soma (red) of islets cells following exposure to three 3-min KCl depolarizations (arrows). Data are means ± SEM for 8 cells from 1 islet and representative of 15 islets. (F–H) Color coded power spectra of ciliary (top) and somatic (bottom) frequency components with superimposed traces (black) of the whole compartments Ca²⁺ activity recorded from individual cells exposed to different treatments (see horizontal lines above plots). (I) TIRF microscopy recording of GCaMP5G fluorescence from 12 cilia from 1 mouse islet (means ± SEM) following addition of 5 mM α-KIC. Shown to the right are recordings of cilia and cytosolic Ca²⁺ from a matched cilia–soma pair within the islet. Representative of eight islets.