The aim of this study was to describe and analyze the regulation and spatio-temporaldynamics of melanocyte migration in vitro and its coupling to celldivision and interaction with the matrix. The melanocyte lineage is particularlyinteresting because it is involved in both embryonic development andoncogenesis/metastasis (melanoma). Biological experiments were performed on two melanocytecell lines established from wild-type and β-catenin-transgenic mice(bcat*). The multi-functional β-catenin molecule is known to be able toregulate the transcription of various genes involved in cell proliferation and migration,particularly in the melanocyte lineage. We also investigated fibronectin, anextra-cellular matrix protein that binds integrins, thereby providing adhesion points forcells and encouraging migration. As the migration of individual cells were followed,automated methods were required for processing the large amount of data generated by thetime-lapse video-microscopy. A model-based approach for automated cell tracking wasevaluated on a sample by comparison with manual tracking. This method was found reliablein studying overall cell behaviour. Its application to all the observed sequences providedinsight into the factors affecting melanocyte migration in vitro:melanocytes of mutated form of β-catenin showed higher division rates andno contact inhibition of growth was induced by the resulting increase in cell density.However, cell density limited the amplitude of cell displacements, although their motilitywas less affected. The high fibronectin concentration bound to substratum promoted cellmigration and motility, the effect being more intense for wild-type cells than for cellswith β-catenin over-expression. During the division process, cellmigration speed increased rapidly then decreased slowly. Analyses of such data is expectedto lead both to biological answers and to a framework for a better modeling processes inthe future.

Giardia lamblia is a flagellated protozoan of great medical and biological importance. It is the causative agent of giardiasis, one of the most prevalent diarrheal disease both in developed and third-world countries. Morphological studies have shown that G. lamblia does not present structures such as peroxisomes, mitochondria, and a well-elaborated Golgi complex. In this review, special emphasis is given to the contribution made by various microscopic techniques to a better knowledge of the biology of the protozoan. The application of video microscopy, immunofluorescence confocal laser scanning microscopy, and several techniques associated with transmission electron microscopy (thin section, enzyme cytochemistry, freeze-fracture, deep-etching, fracture-flip) to the study of the cell surface, peripheral vesicles, endoplasmic reticulum–Golgi complex system, and of the encystation vesicles found in trophozoites and during the process of trophozoite-cyst transformation are discussed.

Xenopus laevis eggs are surrounded by an extracellular matrix consisting of a vitelline envelope, and three jelly layers, J1, J2, and J3 (from egg surface outward). The jelly layers vary in thickness (about 150, 15 and 200 μm for J1, J2 and J3 respectively) but all are translucent allowing observation of sperm penetration. Video microscopy demonstrated that sperm are able to penetrate and traverse J3 at velocities approaching 30 μm/s. Sperm swim through jelly in a corkscrew-like manner with their rotational and forward velocities being tightly coupled at about 30°/μm forward travel. They are propelled by whip-like power strokes involving hairpin bends in the flagellum that are generated every 180° of rotation and which are propagated from base to tip. The overall trajectories of individual sperm are quite variable. Many sperm head directly for J2 but some do not, these swimming circumferentially, or even away from the egg surface. Most sperm (over 97%) that enter the jelly do not get to the egg surface but are stopped at a variety of positions within J3 or at the outer surface of J2. Efficient sperm penetration and passage through the jelly layers requires a low electrolyte concentration in the surrounding medium, and is inhibited by the lectin wheat germ agglutin (WGA) in a dose-dependent manner. WGA does not block sperm penetration of J3 but does block further progression towards the egg surface. This observation suggests that sperm motility within the jelly is dependent on the carbohydrate moieties of the large glycoconjugates present, and that their alteration by WGA binding accounts for the inability of sperm to reach the egg surface and fertilise the egg.