Article Figures & Tables
Figures
- Fig. 1.
Scratching monolayers stimulates cAMP synthesis in MDCK cells. Confluent monolayers of MDCK cells were maintained in 1.8 mM Ca2+ Ringer's solution containing 100 µM IBMX, a phosphodiesterase inhibitor. Cells were then wounded by scratching monolayers with a 27G needle 20 times, or treated with 100 µM ATP or 100 µM forskolin. The amount of cAMP was quantified by immunoassay and normalized to the amount of total protein. The number of experiments is indicated in parentheses. *P=0.0006.
- Fig. 2.
Cell membrane disruption stimulates cAMP production in neighboring cells. (A) ‘w’ in the fluorescence image of MDCK cells expressing Green Upward cADDis indicates a wounded cell. Cells adjacent to the wounded cell were labeled with numbers in order of their proximity to the wounded cell. A cell was wounded at time zero with a glass needle in 1.8 mM Ca2+ Ringer's solution, and the time course of changes in fluorescence intensity of cADDis was plotted for neighboring cells (1–3). The image shown in this figure was acquired 90 s after cell membrane disruption. See also Movie 1. (B) Cells were wounded at time zero with a glass needle in the absence or presence of 20 U/ml apyrase, and changes in fluorescence intensity in neighboring cells were compared. The number of observed cells is indicated in parentheses. P=0.0007 (a–a′); P=0.0427 (b–b′); P=0.0197 (c–c′).
- Fig. 3.
ATP and forskolin stimulate cAMP synthesis in MDCK cells. Cells expressing Green Upward cADDis were treated with either 100 µM ATP or 100 µM forskolin at the time indicated by arrows, and the changes in fluorescence intensity of cADDis were recorded. The arrowhead indicates the transient decrease in fluorescence intensity. The number of observed cells is indicated in parentheses.
- Fig. 4.
Cell membrane disruption induces Ca2+ mobilization in neighboring MDCK cells. (A) Cells loaded with Calcium Green-1 AM were wounded at time zero with a glass needle in the presence or absence of extracellular Ca2+, and changes in fluorescence intensity in the cytoplasmic region were compared. Cells were numbered as per Fig. 2A. The number of observed cells is indicated in parentheses. See also Movie 2. (B) To compare the initial phase of increase in [Ca2+]i, data from cell #1 in A were expanded. *P=0.0333; NS, not significant.
- Fig. 5.
Cell membrane disruption potentiates membrane resealing in neighboring cells in a PKA- and PKC-dependent manner. Cells loaded with Calcein Red-Orange AM were initially wounded with a glass needle, and changes in fluorescence intensity of Calcein Red-Orange were monitored. Neighboring cells were wounded 5 min later and changes in fluorescence intensity were recorded. The resealing rate was defined as the reciprocal of resealing time in seconds. For cells that failed to reseal, the rate was defined as zero. The number of observed cells is indicated in parentheses. *P=0.002.
- Fig. 6.
Potentiation of membrane resealing induced by ATP is suppressed by PKA and PKC inhibitors in MDCK cells. Cells were pretreated for 10 min with either kinase inhibitors or DMSO. Then, cells were wounded with a glass needle after the addition of 100 µM ATP in the presence or absence of inhibitors, respectively, and the resulting resealing rates were analyzed. As a control, cells treated with 100 µM AMP were wounded with a glass needle. Resealing rates were analyzed 5–20 min after the addition of nucleotides. The number of observed cells is indicated in parentheses. *P=0.0007; †P<0.0001.
Article Navigation
Related articles
Cited by...
More in this TOC section
Similar articles
Other journals from The Company of Biologists
Cover article - Apolipoprotein L9 interacts with LC3/GABARAP and is a microtubule-associated protein with a widespread subcellular distribution
Arvind A. Thekkinghat, Kamlesh K. Yadav and Pundi N. Rangarajan demonstrate the versatile properties of apolipoprotein L9, a lipid-binding protein, and its influence on a variety of activities taking place inside an animal cell.
James J. Dowling and his colleagues conducted the first large-scale screening efforts for NEBLUN-related nemaline myopathy to address the urgent need for effective treatment for patients affected by the disease.
Have you seen our interviews with the early-career first authors of our papers? Recently, we caught up with first authors Arvind Thekkinghat, Shannon Taylor and Daisuke Takao.
Biology Open has strong credentials and publishing with us is easy and fast. BiO aims to provide rapid publication for scientifically sound observations and valid conclusions in developmental, cell, experimental and translational biology. Submit your paper here – you’ll be in good company!
Recent developmental biology highlights in BiO – Anna Nele Herdina and her colleagues define the spectrum of phenotypic abnormalities linked with Col4a2 disruption, and demonstrate the opportunities the Deciphering the Mechanisms of Developmental Disorders (DMDD) program offers for exploring rare human diseases.
Transfer to Biology Open
Did you know: if your submission to one of our sister journals, Development, Journal of Cell Science, Journal of Experimental Biology or Disease Models & Mechanisms, is unsuccessful, you can transfer your paper and any reviews directly to Biology Open. The majority of papers transferred with reviews are accepted for publication. Read here to find out more.
BiO partners with Publons.
Biology Open is pleased to announce a new partnership with Publons. This allows reviewers to easily track and verify every review by choosing to add the review to their Publons profile when completing the review submission form. Publons makes it simple for reviewers to showcase their peer review contributions in a format that can be included in job and funding applications (without breaking reviewer anonymity). Read the official announcement here!
preLights - preLights reaches 500 posts!
The success of preLights is down to the hard work of our preLighters. Head over to the preLights website to find out more about our most active early-career researchers.