Cryopreserved Cytoplasts From Human Polymorphonuclear Leukocytes (Cytokineplasts) Are Chemotactic at Speeds Comparable to Those of Fresh Intact Cells

Malawista, Stephen E.; Boisfleury-Chevance, A de

doi:10.1002/jlb.50.3.313

Cited by 10 publications

(4 citation statements)

References 7 publications

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“…A microscopic field containing an erythrocyte, or adjacent ones, and PMN was chosen. Erythrocytes were destroyed by a ruby laser microbeam (wavelength, 694.3 min; duration of flash, 0.5 msec) focused backward through the optics of the microscope to a diameter of Ϸ5 m, creating a chemotactic gradient that lasts for 20 to 45 min, and the PMN were observed and recorded in time-lapse video microscopy (ϫ16 real time) (5).…”

Section: Methodsmentioning

confidence: 99%

Random locomotion and chemotaxis of human blood polymorphonuclear leukocytes (PMN) in the presence of EDTA: PMN in close quarters require neither leukocyte integrins nor external divalent cations

Malawista

Chevance

1997

Proc. Natl. Acad. Sci. U.S.A.

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Divalent cations are thought essential for motile function of leukocytes in general, and for the function of critical adhesion molecules in particular. In the current study, under direct microscopic observation with concomitant time-lapse video recording, we examined the effects of 10 mM EDTA on locomotion of human blood polymorphonuclear leukocytes (PMN). In very thin slide preparations, EDTA did not impair either random locomotion or chemotaxis; motile behavior appeared to benefit from the close approximation of slide and coverslip (''chimneying''). In preparations twice as thick, PMN in EDTA first exhibited active deformability with little or no displacement, then rounded up and became motionless. However, on creation of a chemotactic gradient, the same cells were able to orient and make their way to the target, often, however, losing momentarily their purchase on the substrate. In either of these preparations without EDTA, specific antibodies to ␤2 integrins did not prevent random locomotion or chemotaxis, even when we added antibodies to ␤1 and ␣v␤3 integrins and to integrin-associated protein, and none of these antibodies added anything to the effects of EDTA. In the more turbulent environment of even more media, effects of anti-␤2 integrins became evident: PMN still could locomote but adhered to substrate largely by their uropods and by uropod-associated filaments. We relate these findings to the reported independence from integrins of PMN in certain experimental and disease states. Moreover, we suggest that PMN locomotion in close quarters is not only integrin-independent, but independent of external divalent cations as well.Divalent cations are thought to be essential for motile function of leukocytes in general, and specifically for the function of adhesion molecules critical to a number of their motile activities (1, 2). In examining effects of concentrations of EDTA in the anticoagulant range (Ն1.8 mM) for other reasons, we noted that polymorphonuclear leukocytes (PMN) in warmed sealed slide preparations were not impaired in either random locomotion or chemotaxis. In the current study we have pushed the concentration of EDTA to a point (10 mM) where [Ca 2ϩ ] i or [Mg 2ϩ ] i in the medium would be ϽnM, with similar results.In the slide preparations, a chemotactic gradient is created by the targeted destruction of one or a few erythrocytes resulting from a brief laser flash (3). We examine the behavior of PMN under direct microscopic observation with concomitant time-lapse video recording. Hence, we can follow directly and continuously the orientation and trajectory of PMN before, during, and after establishment of a chemotactic gradient (4).In a sense, the results with EDTA make additional work on specific adhesion molecules redundant, as the function of all of them is thought to be divalent-cation-dependent. However, an examination of relevant leukocyte integrins through the use of mAbs was of interest so that their effects in this system, or lack of effects, might be placed in context wit...

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Section: Methodsmentioning

confidence: 99%

Random locomotion and chemotaxis of human blood polymorphonuclear leukocytes (PMN) in the presence of EDTA: PMN in close quarters require neither leukocyte integrins nor external divalent cations

Malawista

Chevance

1997

Proc. Natl. Acad. Sci. U.S.A.

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“…Recent experiments have demonstrated active migration of platelets (Kraemer et al, 2010;Kraemer et al, 2011). Cell fragments from leukocytes are able to respond to chemotactic signals in the same direction as the mother leukocytes and can migrate to inflammatory lesions in vivo (Malawista and de, 1991;Malawista et al, 2006). Glioma cells in vitro spontaneously shed anucleate fragments which are independently motile.…”

Section: Migration Of Cell Fragmentsmentioning

confidence: 99%

cAMP and cGMP Play an Essential Role in Galvanotaxis of Cell Fragments

Zhu

Sun

Miu

et al. 2015

Journal Cellular Physiology

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Cell fragments devoid of the nucleus and major organelles are found in physiology and pathology, for example platelets derived from megakaryocytes, and cell fragments from white blood cells and glioma cells. Platelets exhibit active chemotaxis. Fragments from white blood cells display chemotaxis, phagocytosis, and bactericidal functions. Signaling mechanisms underlying migration of cell fragments are poorly understood. Here we used fish keratocyte fragments and demonstrated striking differences in signal transduction in migration of cell fragments and parental cells in a weak electric field. cAMP or cGMP agonists completely abolished directional migration of fragments, but had no effect on parental cells. The inhibition effects were prevented by pre-incubating with cAMP and cGMP antagonists. Blocking cAMP and cGMP downstream signaling by inhibition of PKA and PKG also recovered fragment galvanotaxis. Both perturbations confirmed that the inhibitory effect was mediated by cAMP or cGMP signaling. Inhibition of cathode signaling with PI3K inhibitor LY294002 also prevented the effects of cAMP or cGMP agonists. Our results suggest that cAMP and cGMP are essential for galvanotaxis of cell fragments, in contrast to the signaling mechanisms in parental cells.

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“…Keratocyte fragments [9, 10] are the simplest motile units. Cell fragments from leukocytes are able to respond to chemotactic signals [11], but whether cell fragments are galvanotactic was unknown. We found that keratocyte fragments are the smallest motile electric field-sensing unit: they migrate to the anode, in the opposite direction of whole cells.…”

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Keratocyte Fragments and Cells Utilize Competing Pathways to Move in Opposite Directions in an Electric Field

et al. 2013

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Summary Sensing of an electric field (EF) by cells—galvanotaxis—is important in wound healing [1], development [2], cell division, nerve growth, and angiogenesis [3]. Different cell types migrate in opposite directions in EFs [4], and the same cell can switch the directionality depending on conditions [5]. A tug-of-war mechanism between multiple signaling pathways [6] can direct Dictyostelium cells to either cathode or anode. Mechanics of motility is simplest in fish keratocytes, so we turned to keratocytes to investigate their migration in EFs. Keratocytes sense electric fields and migrate to the cathode [7, 8]. Keratocyte fragments [9, 10] are the simplest motile units. Cell fragments from leukocytes are able to respond to chemotactic signals [11], but whether cell fragments are galvanotactic was unknown. We found that keratocyte fragments are the smallest motile electric field-sensing unit: they migrate to the anode, in the opposite direction of whole cells. Myosin II was essential for the direction sensing of fragments but not for parental cells, while PI3 kinase was essential for the direction sensing of whole cells but not for fragments. Thus, two signal transduction pathways, one depending on PI3K, another on myosin, compete to orient motile cells in the electric field. Galvanotaxis is not due to EF force and does not depend on cell or fragment size. We propose a “compass” model according to which protrusive and contractile actomyosin networks self-polarize to the front and rear of the motile cell, respectively, and the electric signal orients both networks toward cathode with different strengths.

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Cryopreserved Cytoplasts From Human Polymorphonuclear Leukocytes (Cytokineplasts) Are Chemotactic at Speeds Comparable to Those of Fresh Intact Cells

Cited by 10 publications

References 7 publications

Random locomotion and chemotaxis of human blood polymorphonuclear leukocytes (PMN) in the presence of EDTA: PMN in close quarters require neither leukocyte integrins nor external divalent cations

Random locomotion and chemotaxis of human blood polymorphonuclear leukocytes (PMN) in the presence of EDTA: PMN in close quarters require neither leukocyte integrins nor external divalent cations

cAMP and cGMP Play an Essential Role in Galvanotaxis of Cell Fragments

Keratocyte Fragments and Cells Utilize Competing Pathways to Move in Opposite Directions in an Electric Field

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