Open Access Highly Accessed Research article

Regulation of aggregate size and pattern by adenosine and caffeine in cellular slime molds

Pundrik Jaiswal1, Thierry Soldati2, Sascha Thewes3 and Ramamurthy Baskar1*

Author Affiliations

1 Department of Biotechnology, Indian Institute of Technology-Madras, Chennai-600036, India

2 Département de Biochimie, Faculté des Sciences, Université de Genève, Sciences II, 30 quai Ernest Ansermet, CH-1211 Genève-4, Switzerland

3 Institute for Biology-Microbiology; Department of Biology, Chemistry, Pharmacy; Freie Universität Berlin; D-14195 Berlin, Germany

For all author emails, please log on.

BMC Developmental Biology 2012, 12:5  doi:10.1186/1471-213X-12-5

Published: 23 January 2012

Additional files

Additional file 1:

Figure S1: The morphology of the slug (Polysphondylium pallidum) in the presence of adenosine and caffeine. Adenosine favours large slug formation whereas caffeine induces emergence of many slugs from the individual aggregate. Morphology of slugs: a) Control; adenosine at (b) 0.5 mM (c) 3.5 mM and caffeine at (d) 1 mM (e) 2.5 mM (f) 3.5 mM. Scale bar = 200 μm.

Format: PDF Size: 99KB Download file

This file can be viewed with: Adobe Acrobat Reader

Open Data

Additional file 2:

Figure S2: The effect of adenosine deaminase on aggregation. A) Aggregation pattern: Adenosine deaminase induced formation of several small sized aggregates. We treated PN500 cells with 50 unit/ml of adenosine deaminase for 2 hours and subsequently plated on non-nutrient agar surface (absence of adenosine deaminase). Aggregation was observed under the light microscope. B) Time of aggregation: Adenosine deaminase delayed the aggregation for the 4 hours. C) Number of aggregates were formed with adenosine deaminase were more than control (absence of it) (Student's t-test, *p < 0.001).

Format: PDF Size: 40KB Download file

This file can be viewed with: Adobe Acrobat Reader

Open Data

Additional file 3:

Effect of caffeine and adenosine on streaming and aggregation pattern. Aggregation pattern and expression analysis of early gene diffrentiation are mentioned.

Format: DOC Size: 35KB Download file

This file can be viewed with: Microsoft Word Viewer

Open Data

Additional file 4:

Figure S3: Aggregation pattern and streaming of AX2 cells in starvation buffer in the presence of caffeine and adenosine. Both adenosine and caffeine inhibited spiral wave formation in aggregates. The arrow sign in 2nd lane of Figure 2D indicates arrangement of the cells at aggregation centres. The aggregates were developed in 90 mm petri dish submerged with Sorensen buffer containing either 3 mM adenosine or 3 mM caffeine.

Format: PDF Size: 326KB Download file

This file can be viewed with: Adobe Acrobat Reader

Open Data

Additional file 5:

Figure S4: Cytosolic glucose levels of developing cells in the presence or absence of 5 mM glucose. The glucose level was estimated as described in materials and methods. The values represent mean ± standard deviation (n = 6), *P < 0.001 (Student's t-test).

Format: PDF Size: 8KB Download file

This file can be viewed with: Adobe Acrobat Reader

Open Data

Additional file 6:

Figure S5: A) The Effect of 2 mM and 5 mM caffeine on the aggregates and fruiting bodies of AX2 cells. B) Effect of 2 mM and 5 mM caffeine on the aggregates and fruiting bodies of smlA mutant cells. Scale bar = 200 μm.

Format: PDF Size: 430KB Download file

This file can be viewed with: Adobe Acrobat Reader

Open Data

Additional file 7:

Figure S6: A) Effect of 2 mM and 5 mM adenosine of the aggregates and fruiting bodies of AX2 cells. B) Effect of 2 mM and 5 mM adenosine of the aggregates and fruiting bodies of ctnA mutant cells. Scale bar = 200 μm.

Format: PDF Size: 450KB Download file

This file can be viewed with: Adobe Acrobat Reader

Open Data

Additional file 8:

Figure S7: Caffeine shown early gene differentiation in cells pulsed with cAMP. A) AX2 cells were incubated at a density of 1 × 107 cells/ml in the Sorensen buffer containing either adenosine or caffeine and were pulsed with 30 nM cAMP for 5 hours for every six minutes intervals. The cAMP treated cells were developed on non nutrient agar plate in the presence or absence of either adenosine or caffeine. B) We checked the expression of acaA and cAR1 genes of pulsed AX2 cells in the presence or absence of either caffeine or adenosine by performing qRT-PCR. The values represent mean ± standard deviation from three independent experiments. C) Cell adhesion protein (Cad-1 and CsaA) and the early gene induction marker protein (CsaA) expressions: To check expression levels, we performed Western blot of these proteins in AX2 cells pulsed with 30 nM cAMP in the presence or absence of adenosine or caffeine.

Format: PDF Size: 488KB Download file

This file can be viewed with: Adobe Acrobat Reader

Open Data

Additional file 9:

Description of Quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). Method and primer sequences used in this study are mentioned [60].

Format: DOC Size: 30KB Download file

This file can be viewed with: Microsoft Word Viewer

Open Data