Time-lapse Video Recording Research Articles

Evidence for long-range chemoreceptive tracking of food odour in deep-sea scavengers by scanning sonar data

There is much speculation about chemoreception being involved in food-finding strategies of deep-sea scavengers in the literature (Dahl, 1979; Meador, 1981; Busdosh et al., 1982; Sainte-Marie, 1992). Most of these ideas have emerged from analysing time-lapse photographs and video recordings of bait deployments in the deep sea (Thurston, 1979; Lampitt et al., 1983; Hargrave, 1985; Priede et al., 1990). However, optical instruments have considerable restrictions in spatial coverage, thus all past efforts in determining any directionality in the appearance of scavengers have been limited from 0.9 to 4 m 2 only (Smith, 1985; Wilson and Smith, 1984). In this work, we present data obtained by using a scanning sonar system (SSS) which allows detection of single objects larger than 2 cm at a maximum distance of 50 m in a horizontal plane. Together with the SSS, a baited time-lapse camera attached to a free falling lander was used in the Arctic deep sea at two locations in the Fram Strait at about 2500 m water depths. We would like to point out that this combination of optical and acoustical measurements allowed, for the first time, the long-range detection of approaching scavenging amphipods in the deep sea. Eurythenes gryllus (Lichtenstein, 1822), a cosmopolitan deep-sea scavenging amphipod, was recorded to attend our bait experiments with a maximum of 618 individuals with first arrival 12 min after deployment, which is one of the fastest arrivals ever observed. We found a significant temporal correlation between integrated backscatter energy (IBE), based on measurements of the SSS and amphipod individuals counted on photographs.

Establishment of Feeder-Independent Cloned Caprine Trophoblast Cell Line Which Expresses Placental Lactogen and Interferon Tau

A feeder-independent cloned trophoblast cell line, HTS-1, was established from a mature placenta of Shiba goat (Capra hircus). During the growth phase, single HTS-1 cells exhibited ruffled membranes or lamellipodia often accompanied by elongated cell shape, indicating highly motile nature of the cells. At or near confluence, HTS-1 cells formed monolayers with few sign of cellular overlapping. Binucleate cells were found at a high frequency especially in the peripheral regions of monolayers. In small colonies and the monolayers, majority of HTS-1 cells assumed polygonally shaped cobble-stone like morphology characteristic to epithelial cells, although considerable variations in cellular morphology were observed despite of repeated cloning. Time-lapse video recordings of HTS-1 cells during culture revealed that not only the small colonies but also the monolayers near or at confluence were remarkably motile, often causing extreme elongation of the cells within them. The extremely plastic nature of HTS-1 cells in vitro is likely to be the reflection of the extraordinary capacity of caprine trophoblast cells to be stretched to extreme thiness in vivo as shown by electron microscopy. HTS-1 cells cultured on matrigel are highly invasive, and express MT1-MMP which, in the mouse, has been known to be expressed at the invasive edge of trophoblast both in vitro and in vivo. HTS-1 cells express placental lactogen (PL) and interferon-τ (IFNτ), as confirmed by immunocytochemistry, Western blotting and RT-PCR analysis. Both PL and IFNτ expression in the cells appeared to be down-regulated by cell-cell contact. In the medium conditioned by HTS-1 cells, the presence of secretory form of PL and IFNτ was confirmed by Western blotting. The HTS-1 cell line will serve as a useful in vitro model for the analysis of the molecular and/or cellular mechanisms underlying synepitheliochorial placentation in bovidae animals.

Semaphorin 3E/Collapsin-5 Inhibits Growing Retinal Axons

During development, the formation of neural networks is reflected by the oriented extension of neurites. Using retinal ganglion cells (RGCs) as a model, we identified the yet uncharacterized chick semaphorin Sema3E/collapsin-5 as a repulsive cue for outgrowing axons. Sema3E/collapsin-5 was highly regulated during retinal histogenesis, with peak expression during the period of intraretinal axon growth. Polymerase chain reaction analysis demonstrated Sema3E/collapsin-5 mRNA in retina layers, from which RGC axons are excluded. Neither isolated RGCs nor purified retinal Müller glia cells synthesized Sema3E/collapsin-5. Sema3E/collapsin-5 receptor sites were visualized by alkaline phosphatase fusion proteins in the axon-rich optic fiber layer. Time-lapse video recording of chick in vitro cultures revealed a growth cone collapsing activity of recombinant Sema3E/collapsin-5. This effect was specific for RGCs, since dorsal root ganglia (DRG) neurons of the peripheral nervous system were not affected. Comparison with Sema3A/collapsin-1 displayed a reciprocal specificity, because Sema3A/collapsin-1 hampered exclusively DRG but not RGC growth cones. The collapsing effect was mediated by low cGMP levels, but not cAMP, as revealed by a set of agonists. In summary, the data suggest a possible role of chick Sema3E/collapsin-5 in restricting growth of retinal ganglion cell axons to the optic fiber layer.

Spermatophore transfer in the hermit crab Clibanarius vittatus (Crustacea, Anomura, Diogenidae).

Although mating has been described in several hermit crab species, the mechanics of spermatophore transfer have not previously been demonstrated. Evidence from pleopod and gonopore morphology, video observations, and inseminated females indicates that in Clibanarius vittatus the male applies a spermatophoric mass directly onto the female via the gonopores rather than with modified pleopods 1-2 (gonopods) and/or genital papillae as in many other decapods. The single second pleopod of males of C. vittatus has a simple endopod with no apparent modifications for sperm transfer. There are no genital papillae extending from the male gonopores. The globular spermatophores are aligned in rows surrounded by a seminal secretion in the male ducts (vasa deferentia that terminate in ejaculatory ducts opening to the exterior via the gonopores). During copulation, described from time-lapse video recordings, the ventral surface of the last thoracic segment of the male, bearing the gonopores, was apposed to the ventral cephalothorax of the female. A massive amount of seminal secretion containing spermatophore ribbons, termed here the spermatophoric mass and described for the first time in a hermit crab species, was observed covering the sternites and coxae of pereopods 1-5 of a recently copulated female. It is suggested that during copulation the male emits the contents of the ejaculatory ducts directly onto the female without the aid of gonopods or genital papillae. Although spermatophore transfer is simple in C. vittatus, the presence of modified anterior pleopods or elongate genital papillae (sexual tubes) in other paguroidean species suggests the possibility of a more complex insemination process in these other hermit crabs.

Behavioural Responses to Castration in Lambs

AbstractWe examined the effects of two different methods of castration on the behaviour of lambs of two age groups. Rubber ring castration and combined rubber ring and Burdizzo clamp castration were compared in groups of one-week-old lambs and four-to six-week-old lambs. Behaviour was assessed by examining time-lapse video recordings made continuously over four days. Postures and behavioural states were instantaneously sampled and the mean daily frequencies of these behaviours calculated. Behaviours were assigned to different groups: standing still, moving, eating, lying, playing and abnormal postures. The times spent performing each group of behaviours on the days following castration (days 2, 3 and 4) were compared to those on the day preceding castration (day 1). Castration resulted in significant reductions in the times spent performing play behaviour in one-week-old lambs and lying behaviour in four-to six-week-old lambs. There was also a significant increase in abnormal postures following castration in four- to six-week-old lambs. There were no detectable differences between castration methods. The results show that lambs exhibit changes in behaviour over the three-day observation period that are suggestive of the presence of prolonged acute pain. These results have important implications regarding the sheep welfare recommendations of the Farm Animal Welfare Council, which propose to extend the maximum legal age for rubber ring castration from one week to six weeks of age.

Relationship between postural behaviour and gestation stall dimensions in relation to sow size

The extent to which the size of the gestation stall, relative to the size of the sow, affects the normal basic postural behavioural needs of sows like standing, sitting and lying was assessed using 25 sows randomly selected from four farms. The postural behaviour of the animals was recorded using a time-lapse video recorder. The relationships of duration of postures, time taken for various postural changes and frequency of postural changes with stall measurements in relation to sow measurements were analysed. Negative correlations (P<0.01) were found between stall length relative to sow length and the duration of time for which the sows were standing. The relationships between the duration of postures and stall width relative to animal breadth were not different (P>0.05). The time taken to change from a standing to lying posture was negatively correlated (P<0.05) with stall length relative to animal length. Similar correlations (P<0.05) were noted between stall width relative animal breadth and the duration of postural change from standing to sitting and from sitting to standing. Stall width relative to animal breadth was negatively related to the frequency of postural change from standing to sitting (P<0.05). The results suggested that the freedom of movement of pregnant sows in stalls could be improved by a little increase in the space allowance within the stall.

New insights to the functional role of the T cell-antigen presenting cell immunological synapse.

Three innovative and complementary morphological approaches were employed to study the T cell/antigen presenting cell (APC) interaction: (i) high resolution three-dimensional confocal microscopy of the T cell-APC contact site; (ii) time lapse video recording in living T cells of [Ca2+]I and changes in distribution of various GFP fusion proteins with TCR/CD3-zeta complex associated- and other signaling components; (iii) measurement of lateral TCR mobility and that of recruited signaling components using techniques based on fluorescence recovery after photo-bleaching. These approaches were combined with biochemical and functional experiments to investigate two principal issues: (A) Recruitment and the three-dimensional arrangement of receptors and signaling components at the contact site between human CD4+ T lymphocytes and APCs, (B) Structure of the immunological synapse formed at the contact site between cytotoxic T lymphocytes (CTLs) and target cells. We discuss evidence indicating that TCR engagement and triggering can occur in the absence of large-scale molecular segregation into the T cell-APC contact site. Taken together our results indicate that although not required for TCR engagement and triggering, formation of the IS is important to reinforce TCR-mediated signal transduction and achieve full T cell activation.

Time-lapse Video Opens Our Eyes to Cow Behaviour and Comfort

Rather than chase bugs with drugs, dairy producers are adopting housing and management practices to reduce environmental risks and improve cattle health. In this presentation, 24-hour time-lapse video recordings and colour photographs show several examples of ways producers are using their knowledge of cow behaviour and ergonomics to improve dairy cattle health and performance.

Roles for 147 embryonic lethal genes on C.elegans chromosome I identified by RNA interference and video microscopy.

Early embryonic development involves complex events such as the regulation of cell division and the establishment of embryonic polarity. To identify genes involved in these events, we collected four-dimensional time-lapse video recordings of the first three cell divisions and analysed terminal phenotypes after RNA interference of 147 embryonic lethal genes previously identified in a systematic screen of Caenorhabditis elegans chromosome I. Over half gave defects in early processes such as meiosis, the assembly or position of the first mitotic spindle, cytokinesis, and proper nuclear positioning. For some phenotypic classes, the majority of genes are involved in a shared biochemical process. In addition, we identified loss-of-function phenotypes for genes of unknown function, but for which homologues exist in other organisms, shedding light on the function of these uncharacterized genes. When applied to the whole genome, this approach should identify the vast majority of genes required for early cell processes, paving the way for a greatly improved understanding of these processes and their regulation at the molecular level.

Intraguild predation by aphidophagous predators on parasitised aphids: the use of multiple video cameras

AbstractA video technique that allows simultaneous behavioural observations of several experimental replicates under field and laboratory conditions is described. The technique was used to analyse predation risk of parasitised aphids in a sugar beet field. The images of 16 black and white video cameras were recorded by a video multiplexer in combination with a time‐lapse video recorder. Each camera was weather protected and equipped with a single infrared diode to allow observations during night times. Single leaves carrying aphid mummies only or mummies and unparasitised aphids were monitored. All colonies were exposed to predation and parasitation by the community of natural enemies in the field. Colonies with mummies and unparasitised aphids were visited significantly more often by predators than those without additional aphids. Predators also stayed significantly longer in patches with unparasitised aphids. Although an equal proportion of aphid mummies were destroyed in both treatments, the video analysis showed differences in predator species spectrum between treatments. In patches with aphids, coccinellid and hemipteran predators preyed on mummies, while in patches with only mummies, chrysopids accounted for most of the damage. The decrease in parasitoid survival could be attributed to the increasing number of predator visits in aphid patches and to a lesser extent to the decreasing number of unparasitised aphids (alternative prey). Parasitoid survival in colonies without alternative prey was correlated with the number of predator visits and the time predators spent on a leaf.Continuous video observations gave additional behavioural information for the interpretation of field data. Other prospective research fields for the use of the multi video camera technique are outlined and general advantages and disadvantages are discussed.

Developmental kinetics of in vitro produced bovine embryos: the effect of sex, glucose and exposure to time-lapse environment.

In this study, a simple time-lapse video recording system was used to compare developmental kinetics of female and male bovine embryos produced in vitro. Following embryo sex determination, the timing of each cleavage up to the 4-cell stage was compared between the sexes from the videotapes after culture in the presence and absence of glucose. In the second experiment, the consequences of exposure to a time-lapse video recording (TL) environment were studied by culturing embryos further until day 7 in an incubator, followed by collection and sex determination of morulae and blastocysts. In the absence of glucose, female embryos cleaved earlier than male ones. In the presence of glucose, however, male embryos cleaved earlier than female ones. There was no difference in the number of morulae/blastocysts in the absence of glucose, but in the presence of glucose more male than female embryos reached the morula and blastocyst stage. Exposure to the TL environment itself also had a sex-related effect, being more detrimental to male than female embryos. The difference in the number of functional X chromosomes between the sexes during early preimplantation development could explain these findings. In females, an increased capacity for oxygen radical detoxification through the pentose phosphate pathway could result in a reduced cleavage rate. Furthermore, glucose may influence the expression of enzymes located on the X chromosome. According to these results, a simple time-lapse video recording system is suitable for investigating embryo developmental kinetics and perhaps for the selection of embryos with the greatest developmental potential.

Tissue-specific neuro-glia interactions determine neurite differentiation in ganglion cells

Guided formation and extension of axons versus dendrites is considered crucial for structuring the nervous system. In the chick visual system, retinal ganglion cells (RGCs) extend their axons into the tectum opticum, but not into glial somata containing retina layers. We addressed the question whether the different glia of retina and tectum opticum differentially affect axon growth. Glial cells were purified from retina and tectum opticum by complement-mediated cytolysis of non-glial cells. RGCs were purified by enzymatic delayering from flat mounted retina. RGCs were seeded onto retinal versus tectal glia monolayers. Subsequent neuritic differentiation was analysed by immunofluorescence microscopy and scanning electron microscopy. Qualitative and quantitative evaluation revealed that retinal glia somata inhibited axons. Time-lapse video recording indicated that axonal inhibition was based on the collapse of lamellipodia- and filopodia-rich growth cones of axons. In contrast to retinal glia, tectal glia supported axonal extension. Notably, retinal glia were not inhibitory for neurons in general, because in control experiments axon extension of dorsal root ganglia was not hampered. Therefore, the axon inhibition by retinal glia was neuron type-specific. In summary, the data demonstrate that homotopic (retinal) glia somata inhibit axonal outgrowth of RGCs, whereas heterotopic (tectal) glia of the synaptic target area support RGC axon extension. The data underscore the pivotal role of glia in structuring the developing nervous system.

Migration and metalloproteinases determine the invasive potential of mouse melanoma cells, but not melanin and telomerase

The biological characteristic cell locomotion and invasion, melanin content, metalloproteinases and telomerase activity were studied in a parental mouse melanoma cell line B16 and two descendents B16BL6 and B16F10. The invasive potential of melanoma cells was assayed in a transwell cell culture chamber. Melanin content was determined by the absorbance value at 470 nm per 10 6 cells. Tumor cells migration within the 3-D collagen matrix was microscopically recorded with a time-lapse video recorder and analyzed by computer-assisted cell tracking. Gelatin zymography was adopted to assay the metalloproteinases secretion. A polymerase chain reaction-based telomeric repeat amplification protocol (TRAP) was used for measuring telomerase activity. The results demonstrated that B16BL6 and B16F10 cells were highly invasive compared to B16 cells, but the melanin content of B16F10 was very low. B16F10 and B16BL6 were hypermotile and secreted much more metalloproteinases than B16. No differences were observed in telomerase activity among the three melanoma cell lines. Invasion of mouse melanoma was closely correlated to tumor cell migration and secretion of metalloproteinases. Melanin content and telomerase activity were phenotypically not related to invasiveness in these three mouse melanoma cell lines.

疲労 ＳＳ４００炭素鋼のモードＩＩ，モードＩＩＩ疲労き裂の進展挙動

Fatigue characteristics are of fundamental importance when choosing a structural materials, especially when deciding upon the materials for reactor structure where long-term safety is primary importance. In this field, recently, the propagation behavior of mode II and mode III fatigue cracks has attracted special interest. The mode II and mode III fatigue testings involve greater difficulties compared with the mode I testing and the testing procedure has not been established. SS400 carbon steel was fatigue tested by a torsion fatigue testing machine. In the torsion test, fatigue cracks can propagate in all modes. To identify their behavior, surfaces of the specimen were observed by CCD camera and the behavior of surface cracks were recorded in a time-lapse video recorder which can record up to for 40 days. To reveal the morphology of the mode II and mode III crack propagation, some of test interrupted specimens were sliced and observed by a two dimension measuring microscope.To investigate the crack morphology three dimension crack demonstrate computer program was developed. In the torsion test of plain surface specimen the mode II crack initiates first, but it transits to mode I in the early stage. However, according to the 3D view a large mode III crack propagates into inside in this stage. Persistent slip band like patterns were observed on the surface and the cross section of the fatigue cracked specimen and all mode III cracks propagate in the band. Some of them grew across the center of the specimen. The da/dN-ΔK diagram of the surface mode II fatigue crack propagation was compared with that of mode I by CT type specimen of the same material. mode II propagation rate agreed with mode I propagation rate where the stress intensity factor range was converted as ΔKI=2ΔKII. It means that to design the structure by mode I crack propagation data alone may be acceptable under above conversion. To establish this relationship and to elucidate the behavior of mode III fatigue cracking, more investigations are required.

The scavenging behaviour of ferrets (Mustela furo), feral cats (Felis domesticus), possums (Trichosurus vulpecula), hedgehogs (Erinaceus europaeus) and harrier hawks (Circus approximans) on pastoral farmland in New Zealand: Implications for bovine tuberculosis transmission

Aims: To identify species that scavenge carcasses in pastoral habitats in New Zealand; to determine whether there were interspecific or intraspecific differences in scavenging behaviour and; to document any interspecific or intraspecific interactions occurring at carcasses. Methods: Scavenging by ferrets (Mustela furo), feral cats (Felis domesticus), possums (Trichosurus vulpecula), hedgehogs (Erinaceus europaeus) and harrier hawks (Circus approximans) was studied from autumn to midwinter on pastoral farmland near Palmerston (45S, 170E), Otago, New Zealand. Time-lapse video recorders and camera lens mounted with infra-red light illumination were used to monitor carcasses of 10 ferrets, 12 possums, 2 hedgehogs and 7 rabbits (Oryctolagus cuniculus) until they were totally scavenged. Results: Ferrets scavenged 5/8 ferret carcasses, 8/9 possum carcasses and 6/7 rabbit carcasses encountered. Feral cats scavenged 3/8 ferret carcasses, 5/7 rabbit carcasses, and 3/8 possum carcasses encountered. Possums scavenged 1/2 ferret carcasses and 3/4 rabbit carcasses encountered. The proportion of encounters resulting in feeding on ferret carcasses differed between ferrets (45.7%) and possums (6.3%), and between possums and cats (29.7%). Similarly, for possum carcasses, differences were found between ferrets (76.6%) and possums (0%), ferrets and cats (60.6%) and possums and cats. No interspecific differences were found in the proportion of encounters that resulted in feeding on rabbit carcasses between ferrets (85.2%), possums (75%) and cats (73.1%). In 8/12 instances of ferrets coming into contact with other ferrets whilst feeding, ferrets fed together at the carcass. On 1 occasion, 4 ferrets were recorded feeding together. In 7/8 instances where cats and ferrets came into contact over carrion, ferrets maintained possession or displaced the cat from the carcass. Conclusions: Communal carrion feeding by ferrets may facilitate intraspecific and interspecific transmission of bovine tuberculosis (caused by Mycobacterium bovis) by the consumption of contaminated carrion, fighting, or close-contact activities. Cannibalism may be one mechanism by which tuberculosis is transmitted within ferret populations. Our results also suggest that possums may acquire infection from carrion, despite being mainly herbivorous.

Opposing views on tensegrity as a structural framework for understanding cell mechanics.

Donald E. Ingber: Important new theories in science often ignite heated debates. If they do not, they are probably of little significance. Thus a strong argument in support of the importance of the tensegrity model of cell and tissue architecture first proposed almost 20 years ago ([23][1], [24][2

Transmission of Mycobacterium bovis from experimentally infected ferrets to non-infected ferrets (Mustela furo)

Aims: To demonstrate the transmission of Mycobacterium bovis infection from experimentally infected ferrets (Mustela furo) to non-infected ferrets in a laboratory setting, using three different isotypes of M. bovis, and to observe ferret behaviour that might be implicated in disease transmission. Methods: Three female ferrets, each experimentally infected with a unique strain of M. bovis, were housed together with six female and two male non-infected ferrets in an isolation facility. Transmission of infection was monitored clinically, serologically (using an ELISA test), bacteriologically, histologically, and by isotype analysis of M. bovis isolates using spoligotyping to determine whether or not transmission of each strain occurred. Ferret behaviour was observed using a time-lapse video recorder. Results: Transmission of M. bovis infection was confirmed in two male and four female ferrets. Isotype analysis showed that of the experimentally infected females, one did not infect any other ferret, another transmitted M. bovis to one ferret before it died prematurely 49 days post-infection, and the third, which was cannibalised, appears to have transmitted M. bovisto both males and three females. However, two of these latter three females had died before the event of cannibalism took place. One female was infected with two strains. Several behavioural interactions were observed that could have resulted in M. bovis transmission, including den sharing, sniffing of orifices and faeces, cannibalism and aggressive breeding behaviour. Conclusions: Horizontal transmission of M. bovis infection was demonstrated in ferrets under experimental housing conditions. Routes of transmission may involve cannibalism and factors such as den sharing, playing, fighting, mating, and sniffing of faeces.

Transition of the blastomere cell cycle from cell size-independent to size-dependent control at the midblastula stage in Xenopus laevis.

Dissociated animal cap blastomeres of Xenopus laevis blastulae were cultured at a low Ca level (1 microM) from 9th to 18th cell cycle at 22 +/- 1 degrees C and observed by a time-lapse video recorder. Blastomeres cleaved unequally to increase variability in cell size as cell cycles progressed, but synchronously at a constant cell cycle time of about 30 min up to the 12th cleavage in diploid cells, and up to the 13th cleavage in haploid cells, regardless of their cell sizes. Thereafter, blastomeres cleaved asynchronously at varying cell cycle times in proportion to the inverse square of their radii. The transition from the cell size-independent to -dependent cell cycles occurred at the critical cell radius, 37.5 microm for the diploid and 27.9 microm for the haploid. While the protein synthesis inhibitor, cycloheximide (CHX) lengthened cell cycle times two- to six-fold, epidermal growth factor (EGF) had no significant effect on the cell cycle. CHX-treated blastomeres synchronously cleaved at a constant cell cycle time of 60 min up to the 12th cleavage. Thereafter, cell cycle times became variable in proportion to the inverse square of radii in the presence of CHX at 0.10-0.14 microg/ml, but to the inverse cube of radii at 0.18 microg/ml. The critical cell size of CHX-treated blastomeres for the transition from cell size-independent to -dependent cell cycles remained the same as that of untreated blastomeres. Frequency distributions of cell cycle times of synchronous cell cycles were monomodal with the peak at 30 min, except for CHX-treated blastomeres with the peak at 60 min. In contrast, frequency distributions of asynchronous cell cycles were polymodal with peaks at multiples of a unit time of 30-35 min. To explain these results, we propose that blastomere cytoplasm has 30-min cycles that repeatedly produce mitosis promoting factor (MPF) in a quantity proportional to the cell surface area. MPF is neutralized when it titrates a nuclear inhibitor present in a quantity proportional to the genome size, and sequestered in the nucleus. When the total amount of MPF produced exceeds the threshold required to titrate all of the inhibitor, mitosis is initiated.

Individual cell motility studied by time-lapse video recording: influence of experimental conditions.

Eukaryotic cell motility plays a key role during development, wound healing, and tumour invasion. Computer-assisted image analysis now makes it a realistic task to quantify individual cell motility of a large number of cells. However, the influence of culture conditions before and during measurements has not been investigated systematically. We have evaluated intraassay and interassay variations in determinations of cellular speed of fibroblastoid L929 cells and investigated the effects of a series of physical and biological parameters on the motile behavior of this cell line. Cellular morphology and organization of filamentous actin were assessed by means of phase-contrast and confocal laser scanning microscopy and compared to the corresponding motility data. Cell dissociation procedure, seeding density, time of cultivation, and substrate concentration were shown to affect cellular speed significantly. pH and temperature of the medium most profoundly influenced cell motility and morphology. Thus, the mean cell speed was 40% lower at pH 7.25 than at pH 7.6; at 29 degrees C, it was approximately four times lower than at 39 degrees C. Of the parameters evaluated, cell motility was most strongly affected by changes in pH and temperature. In general, changes in cell speed were accompanied by alterations in cell morphology and organization of filamentous actin, although no consistent phenotypic characteristics could be demonstrated for cells exhibiting high cell speed.

Random locomotion and chemotaxis of human blood polymorphonuclear leukocytes from a patient with leukocyte adhesion deficiency-1: normal displacement in close quarters via chimneying.

The beta2 integrins are known to be important in the motile function of leukocytes in general and in the adhesive response to inflammatory stimuli in particular. In the current study, under direct microscopic observation with concomitant time-lapse video recording, we examined the locomotion of human blood PMN from a patient with Leukocyte Adhesion Deficiency-1 (LAD), a disorder in which beta2 integrins on the cell surface are markedly deficient in number or function. In thin slide preparations such that the leukocytes were somewhat compressed between slide and cover slip, PMNLAD exhibited normal random locomotion and chemotaxis, apparently by using the opposing surfaces to generate the force for locomotion (chimneying). In thicker preparations, an adherence deficit was evident, but chemotaxis still occurred, even by PMNLAD anticoagulated in EDTA. Consistent with the paucity of beta2 integrins on the surface of the PMNLAD was their failure to aggregate in the presence of antibodies to beta2 integrins, even when they had been brought together by chemotaxis. We relate these findings to the reported independence from integrins of PMN in the lung vasculature in LAD, as well as in certain experimental conditions.