Abstract
Human placental extracts are known to help wound healing. Rapid migration of neutrophils to the wound site is a prerequisite to the wound healing process. Gel filtration analysis of heat-treated placental extract gave the initial cue to the small nature of the migration promoting factor of the extract. HPLC analysis of the extract revealed glutamate to be the predominant free amino acid. Our studies show that glutamate at an optimum concentration of 8 μM induced phenotypic neutrophil chemotaxis, as seen in the time lapse- and transwell assays. Glutamate was also found to induce chemokinesis of the neutrophil, though the stimulation of chemotaxis was more pronounced. The glutamate induced chemotaxis was accompanied by polarization of the actin cytoskeleton, and by polymerization of F-actin. These data indicate that glutamate has a strong chemotactic functionality in the neutrophil, which could be of interest both therapeutically and in further investigation of the molecular basis of chemotaxis.
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Abbreviations
- F-actin:
-
Filamentous actin
- GPCR:
-
G-protein coupling receptor
- HBSS:
-
Hanks balanced salt solution
- OPA:
-
O-phthalaldehyde
- MCF:
-
Mean cellular fluorescence
- Plx:
-
Human placental extract
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Acknowledgments
The authors are highly obliged to Prof. Anjan Das Gupta, Department of Biochemistry, Calcutta University and Dr. Sandipan Ganguly, NICED(National Institute Of Cholera And Enteric Diseases), Calcutta for the time lapse video microscopy and confocal microscopy, respectively. The authors gratefully acknowledge Prof. Gopal Kundu, NCCS (National Centre For Cell Science), Pune, India for rendering the lab facilities available for some of the initial experiments. We also thank Dr. Alok K. Sil for the positive suggestions during the work. Special thanks to Mr. Sanjoy Dudhoria for the HPLC of free amino acid. We are extremely thankful to Dr. S. Nath and Dr. G. P. Srivasatava, Albert David, Ltd., Calcutta, for the outstanding cooperation received during the project. We also most gratefully acknowledge Dr. Amitabha Majumdar, (Weill Medical College of Cornell University, New York, USA) for the Alexa Phalloidin. Finally, the authors owe much to the members of the Chattopadhay laboratory for the kind cooperation during the work and the manuscript preparation. This work was financially supported by Albert David Research Unit, Calcutta.
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An erratum to this article can be found at http://dx.doi.org/10.1007/s00726-009-0456-z
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Gupta, R., Chattopadhyay, D. Glutamate is the chemotaxis-inducing factor in placental extracts. Amino Acids 37, 271–277 (2009). https://doi.org/10.1007/s00726-008-0143-5
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DOI: https://doi.org/10.1007/s00726-008-0143-5