Evaluation of the effectiveness of enrichment for a group of captive squirrel monkeys (Saimiri sciureus) using time-lapse recording and instantaneous scan sampling

One aim of providing enrichment to captive animals is to promote the expression of behavioural patterns similar to their wild conspecifics. We evaluated the effectiveness of four types of simple feeding enrichment, using surveillance cameras to record the behaviour of 11 captive squirrel monkeys housed in a single enclosure at Alma Park Zoo in Brisbane, Australia. The enrichment involved differences in presentation (whole/chopped) and distribution (localised/scattered) of fruit and vegetables that were part of the normal diet of these animals. Distinguishing between individual squirrel monkeys was not possible from the videos, so Instantaneous Scan Sampling was used to record the numbers of animals performing particular behaviours every 15 minutes over the 24 hour period as well as every 5 minutes for the hour following provision of enrichment. This provided an estimation of the percentage of time spent by the group in various activities. As a result of the enrichment, the activity budget of the group more closely approximated that of wild squirrel monkeys. However on a number of occasions where the enrichment required the squirrel monkeys to work to obtain their food (whole fruit and vegetables), a number of individuals became aggressive towards the zookeepers. This result highlights the variation in responses of individual animals towards enrichment and indicates that in enclosures with large numbers of animals, the response of each individual should be evaluated in addition to the overall benefit of the enrichment for the group. Furthermore, this variation also suggests that it may be beneficial to provide the animals with choices of enrichment as opposed to providing single forms of enrichment that may only be effective for a proportion of the animals in the enclosure, and may even result in undesirable responses from some individuals.

Cannibalism in juvenile blue-swimmer crabs Portunus pelagicus (Linnaeus, 1766): effects of body size, moult stage and refuge availability

Factors influencing the rate of cannibalism in juvenile blue-swimmer crabs Portunus pelagicus were investigated under controlled conditions using time-lapse video recordings. This study was undertaken to improve blue-swimmer crab culture and experimentally addressed (1) prey vulnerability (2) cannibal-victim interactions, and (3) activity patterns of juveniles in varying degrees of refuge. Crabs used in the study were aged 15 weeks and sorted into two size classes; small (less than or equal to 60 mm carapace width (CW)) and large (greater than or equal to65 mm CW) of a similar sex ratio. Vulnerability and thus survival was influenced by body size variation, moult stage and refuge availability. Crabs with carapace width less than or equal to 60 mm were more vulnerable than larger individuals, as indicated by significant differences in survival rates. As predicted, juveniles in transition stages associated with ecdysis were especially vulnerable. Premoult (redliner) crabs appeared to be in a high state of agitation as evidenced by the frequency of agonistic encounters and this may be a contributing factor to the high mortality observed at this critical premoult stag. increases in refuge density increased survival of juveniles proportionally, indicating that the quantity of shelter is important for reducing cannibalism in this species. Cannibal-victim interactions were frequently asymmetrical in terms of size and moult stage. Cannibals were significantly heavier than victims, and were predominantly at intermoult stage. Sexual biases among cannibals and victims were not found in this study. Activity patterns of juveniles were influenced by the experimental conditions. Crabs provided with high refuge showed reduced aggressive activity and increased time spent resting, but unchanged locomotion or feeding activity. Regular grading as well as the presence of suitable shelter for newly moulted crabs is recommended for improving culture of P. pelagicus. Research into inducing synchronous moulting may also yield promising results. (C) 2004 Elsevier B.V. All rights reserved.

The tyrosine phosphatase DEP-1 induces cytoskeletal rearrangements, aberrant cell-substratum interactions and a reduction in cell proliferation

The receptor protein tyrosine phosphatase density-enhanced phosphatase-1 (DEP-1) has been implicated in aberrant cancer cell growth and immune cell function, however, its function within cells has yet to be properly elucidated. To investigate the cellular function of DEP-1, stable cell lines inducibly expressing DEP-1 were generated. Induction of DEP-1 expression was found to decrease PDGF-stimulated tyrosine phosphorylation of a number of cellular proteins including the PDGF receptor, and to inhibit growth factor-stimulated phosphorylation of components of the MAPK pathway, indicating that DEP-1 antagonised PDGF receptor signalling. This was supported by data showing that DEP-1 expression resulted in a reduction in cell proliferation. DEP-1-expressing cells had fewer actin-containing microfilament bundles, reduced vinculin and paxillin-containing adhesion plaques, and were defective in interactions with fibronectin. Defective cell-substratum adhesion correlated with lack of activation of FAK in DEP-1-expressing cells. Time-lapse interference reflection microscopy of live cells revealed that although small focal contacts at the leading edge were generated in DEP-1-expressing cells, they failed to mature into stable focal adhesions, as found in control cells. Further motility analysis revealed that DEP-1-expressing cells retained limited random motility, but showed no chemotaxis towards a gradient of PDGF. In addition, cell-cell contacts were disrupted, with a change in the localisation of cadherin from discrete areas of cell-cell contact to large areas of membrane interaction, and there was a parallel redistribution of beta-catenin. These results demonstrate that DEP-1 is a negative regulator of cell proliferation, cell-substratum contacts, motility and chemotaxis in fibroblasts.

Live cell imaging of heavy-ion-induced radiation responses by beamline microscopy

To study the dynamics of protein recruitment to DNA lesions, ion beams can be used to generate extremely localized DNA damage within restricted regions of the nuclei. This inhomogeneous spatial distribution of lesions can be visualized indirectly and rapidly in the form of radiation-induced foci using immunocytochemical detection or GFP-tagged DNA repair proteins. To analyze faster protein translocations and a possible contribution of radiation-induced chromatin movement in DNA damage recognition in live cells, we developed a remote-controlled system to obtain high-resolution fluorescence images of living cells during ion irradiation with a frame rate of the order of seconds. Using scratch replication labeling, only minor chromatin movement at sites of ion traversal was observed within the first few minutes of impact. Furthermore, time-lapse images of the GFP-coupled DNA repair protein aprataxin revealed accumulations within seconds at sites of ion hits, indicating a very fast recruitment to damaged sites. Repositioning of the irradiated cells after fixation allowed the comparison of live cell observation with immunocytochemical staining and retrospective etching of ion tracks. These results demonstrate that heavy-ion radiation-induced changes in sub-nuclear structures can be used to determine the kinetics of early protein recruitment in living cells and that the changes are not dependent on large-scale chromatin movement at short times postirradiation. © 2005 by Radiation Research Society.

Visualisation of macropinosome maturation by the recruitment of sorting nexins

We report that phosphoinositol-binding sorting nexin 5 ( SNX5) associates with newly formed macropinosomes induced by EGF stimulation. We used the recruitment of GFP-SNX5 to macropinosomes to track their maturation. Initially, GFP-SNX5 is sequestered to discrete subdomains of the macropinosome; these subdomains are subsequently incorporated into highly dynamic, often branched, tubular structures. Time-lapse videomicroscopy revealed the highly dynamic extension of SNX5-labelled tubules and their departure from the macropinosome body to follow predefined paths towards the perinuclear region of the cell, before fusing with early endosomal acceptor membranes. The extension and departure of these tubular structures occurs rapidly over 5-10 minutes and is dependent upon intact microtubules. As the tubular structures depart from the macropinosome there is a reduction in the surface area and an increase in tension of the limiting membrane of the macropinosome. In addition to the recruitment of SNX5 to the macropinosome, Rab5, SNX1 and EEA1 are also recruited by newly formed macropinosomes, followed by the accumulation of Rab7. SNX5 forms heterodimers with SNX1 and this interaction is required for endosome association of SNX5. We propose that the departure of SNX5-positive tubules represents a rapid mechanism of recycling components from macropinosomes thereby promoting their maturation into Rab7-positive structures. Collectively these findings provide a detailed real-time characterisation of the maturation process of the macropinocytic endosome.