Antisense oligodeoxynucleotide inhibition of a swelling-activated cation channel in osteoblast-like osteosarcoma cells.
By patch-clamp analysis, we have shown that chronic, intermittent mechanical strain (CMS) increases the activity of stretch-activated cation channels of osteoblast-like UMR-106.01 cells. CMS also produces a swelling-activated whole-cell conductance (Gm) regulated by varying strain levels. We questioned whether the swelling-activated conductance was produced by stretch-activated cation channel activity. We have identified a gene involved in the increase in conductance by using antisense oligodeoxynucleotides (ODN) derived from the alpha 1-subunit genes of calcium channels found in UMR-106.01 cells (alpha1S, alpha1C, and alpha1D). We demonstrate that alpha 1C antisense ODNs abolish the increase in Gm in response to hypotonic swelling following CMS. Antisense ODNs to alpha1S and alpha1D, sense ODNs to alpha1C, and sham permeabilization had no effect on the conductance increase. In addition, during cell-attached patch-clamp studies, antisense ODNs to alpha1c completely blocked the swelling-activated and stretch-activated nonselective cation channel response to strain. Antisense ODNs to alpha1S treatment produced no effect on either swelling-activated or stretch-activated cation channel activity. There were differences in the stretch-activated and swelling-activated cation channel activity, but whether they represent different channels could not be determined from our data. Our data indicate that the alpha1C gene product is involved in the Gm and the activation of the swelling-activated cation channels induced by CMS. The possibility that swelling-activated cation channel genes are members of the calcium channel superfamily exists, but if alpha1c is not the swelling-activated cation channel itself, then its expression is required for induction of swelling-activated cation channel activity by CMS.
Duncan
R L
RL
Renal Division, Jewish Hospital, Washington University Medical Center St. Louis, Missouri 63110, USA.
Kizer
N
N
Barry
E L
EL
Friedman
P A
PA
Hruska
K A
KA
D000818:Animals; D001483:Base Sequence; D015220:Calcium Channels / Q000502:physiology*; D048429:Cell Size; D004553:Electric Conductivity; D015640:Ion Channel Gating; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000737:chemistry; D010006:Osteoblasts / Q000502:physiology*; D012516:Osteosarcoma; D018408:Patch-Clamp Techniques; D010507:Periodicity; D051381:Rats; D013314:Stress, Mechanical; D014407:Tumor Cells, Cultured; D014882:Water-Electrolyte Balance
D015220:Calcium Channels; D016376:Oligonucleotides, Antisense