Golgi clusters and vesicles mediate mitotic inheritance independently of the endoplasmic reticulum

We have examined the fate of Golgi membranes during mitotic inheritance in animal cells using four-dimensional fluorescence microscopy, serial section reconstruction of electron micrographs, and peroxidase cytochemistry to track the fate of a Golgi enzyme fused to horseradish peroxidase. All three approaches show that partitioning of Golgi membranes is mediated by Golgi clusters that persist throughout mitosis, together with shed vesicles that are often found associated with spindle microtubules. We have been unable to find evidence that Golgi membranes fuse during the later phases of mitosis with the endoplasmic reticulum (ER) as a strategy for Golgi partitioning (Zaal, K.J., C.L. Smith, R.S. Polishchuk, N. Altan, N.B. Cole, J. Ellenberg, K. Hirschberg, J.F. Presley, T.H. Roberts, E. Siggia, et al. 1999. Cell. 99:589-601) and suggest that these results, in part, are the consequence of slow or abortive folding of GFP-Golgi chimeras in the ER. Furthermore, we show that accurate partitioning is accomplished early in mitosis, by a process of cytoplasmic redistribution of Golgi fragments and vesicles yielding a balance of Golgi membranes on either side of the metaphase plate before cell division.     

Reflective properties of different eyespot types in dinoflagellates

The reflective properties of different types of dinoflagellate eyespots were investigated using confocal laser scanning microscopy in the epireflection contrast mode. Although the eyespots studied differed with respect to localization (cytosol or plastid) and organization of the globule layer(s), all types effectively absorbed and reflected blue-green laser light (principal lines of 488/514 nm). The relative orientation of the eyespot surface towards the light source strongly influenced the reflective properties. Maximal reflection occurred when the eyespot surface was approximately perpendicular to the light source and rapidly decreased at increasing angles of light incidence. Horizontal and vertical optical sectioning of live and fixed cells resolved differences in the reflection patterns. Focusing of reflected light on the basal portion of the longitudinal flagellum was observed for the cytosolic eyespot of Glenodinium sp. and the triple membrane-bounded eyespot of Peridinium foliaceum, presumably a vestige of a host plastid. This flagellum is thought to be mainly involved in mediating orientational movement responses. In contrast, the reflection patterns obtained from the eyespot of Woloszynskia pascheri, which represents the third and most commonly observed dinoflagellate eyespot type within a plastid, point to only minor focusing. Reflection signals could be followed a considerable distance into the sulcus in all cases, indicating that in dinoflagellate eyespots, irrespective of the presumed receptor location (plasma membrane overlying the eyespot and/or the basal part of the longitudinal flagellum), back reflection of non-absorbed light can enhance the excitation probability of the photoreceptor(s). Such a combined reflection/absorption screen allows maximal contrast modulation and will, in conjunction with the specialized geometry of the dinoflagellate eyespots, increase the directionality of these eyespot aparatuses considerably.     

Calcium release from the Golgi apparatus and the endoplasmic reticulum in HeLa cells stably expressing targeted aequorin to these compartments

Extracellular agonists mobilize Ca2+ from SERCA-comprising intracellular Ca2+ stores located in both the Golgi apparatus and the endoplasmic reticulum. Ca2+ release from both these compartments was studied in HeLa cells stably expressing the luminescent Ca2+ indicator aequorin specifically targeted to these compartments. Changes in lumenal [Ca2+] as detected by the aequorin measurements were correlated with parallel changes in total Ca2+ content of the stores. The latencies and initial rates of Ca2+ release from the Golgi apparatus and the endoplasmic reticulum were quite similar. However, maximal Ca2+ release measured with Golgi-targeted aequorin terminated faster than that from the endoplasmic reticulum. The rate and extent of Ca2+ depletion from both compartments correlated well with the peak amplitude of the cytosolic [Ca2+] rise. Time-course experiments further revealed that the peak of the cytosolic Ca2+ response occurred before the lumenal [Ca2+] reached its lowest level. We conclude that both the Golgi apparatus and the endoplasmic reticulum contribute to the rise in cytosolic [Ca2+] upon agonist stimulation, but the kinetics of the Ca2+ release are different.     

Dosimetric analysis of the carousel setup for the exposure of rats at 1.62 GHz

The so-called carousel setup has been widely utilized for testing the hypotheses of adverse health effects on the central nervous system (CNS) due to mobile phone exposures in the frequency bands 800-900 MHz. The objectives of this article were to analyze the suitability of the setup for the upper mobile frequency range, i.e., 1.4-2 GHz, and to conduct a detailed experimental and numerical dosimetry for the setup at the IRIDIUM frequency band of 1.62 GHz. The setup consists of a plastic base on which ten rats, restrained in radially positioned tubes, are exposed to the electromagnetic field emanating from a sleeved dipole antenna at the center. Latest generation miniaturized dosimetric E field and temperature probes were used to measure the specific absorption rate (SAR) inside the brain of three rat cadavers of the Lewis strain and two rat cadavers of the Fisher 344 strain. A numerical analysis was conducted on the basis of three numerical rat phantoms with voxel sizes between 1.5 and 0.125 mm3 that are based on high resolution MRI scans of a 300 g male Wistar rat and a 370 g male Sprague-Dawley rat. The average of the assessed SAR values in the brain was 2.8 mW/g per W antenna input power for adult rats with masses between 220 and 350 g and 5.3 mW/g per W antenna input power for a juvenile rat with a mass of 95 g. The strong increase of the SAR in the brain with decreasing animal size was verified by simulations of the absorption in numerical phantoms scaled to sizes between 100 and 500 g with three different scaling methods. The study also demonstrated that current rat phantom models do not provide sufficient spatial resolution to perform absolute SAR assessment for the brain tissue. The variation of the SAR(brain)(av) due to changes in position was assessed to be in the range from +15% to -30%. A study on the dependence of the performance of the carousel setup on the frequency revealed that efficiency, defined as SAR(brain)(av) per W antenna input power, and the ratio between SAR(brain)(av) and SAR(body)(av) are optimal in the mobile communications frequency range, i.e., 0.8-3 GHz.     

Morphofunctional study of the bill and hyoid apparatus of Momotus momota (Aves, Coraciiformes, Momotidae): implications for omnivorous feeding adaptation in motmots

The present study contrasts available biological data and results of morphofunctional analyses of the bill and hyoid apparatus in motmots. It shows that these omnivorous birds, which take relatively large food items, possess osteomuscular peculiarities that enable them to process these items as a whole in order to soften or cut them, and make them suited for easy ingestion. For that, they use the crenate edges of their rhamphotheca. Their jaws work as a highly mobile saw-like system. Their mutual movements, enhanced by the fact that particular dispositions of the hyoid apparatus rise the tongue and the supported items high up into buccal cavity, facilitate an effective clamping of items that can be moved along the jaws and be quite appropriately processed.     

Stridulation variability and morphology: an examination in dung beetles of the genus<Emphasis Type="Italic"> Trypocopris</Emphasis> (Coleoptera,Geotrupidae)

Distress signals produced by dung beetles of the genus Trypocopris (Coleoptera, Geotrupidae) were analysed to test whether interspecific and intraspecific acoustic variability are species- and subspecies-specific and to ascertain to what extent bioacoustic parameters depend upon the morphology of the stridulatory organs (pars stridens). Bioacoustic analyses showed that the three species were clearly differentiated, despite the fact that disturbance stridulations presented the same stereotyped spectrographic pattern. Within each species, most of the subspecies and populations considered were also bioacoustically distinguishable. Subspecies and populations within each species were differentiated with regard to body size and stridulatory organ, and the length of the pars stridens was positively correlated with the width of the coxa, in turn positively correlated with body size. A few spectrographic measures were significantly constrained by the morphology of the stridulatory apparatus; in particular the duration of sound emission was positively correlated with the length of the apparatus and the sub-pulse rate was negatively related to the distance between two consecutive crests. For T. pyrenaeus, with the largest number of populations sampled, there was no significant correlation between morphological and spectrographic distances, but there were significant positive correlations between morphological and geographical distances and between morphological and genetic distances. It is hypothesized that genetic differentiation might directly affect variability of the stridulatory apparata which would also be indirectly influenced and constrained by external morphological traits (like the width of the coxa and body size). Stridulatory organs, in turn, would affect the ways a few stridulatory traits (especially the temporal ones) change in time and space.     

Widespread polyphyly among Alopiinae snail genera: when phylogeny mirrors biogeography more closely than morphology

Consider a group of species that is evenly divided by an easily identifiable complex morphological character. Most biologists would assume that this character should provide better phylogenetic information than, say, the spatial distribution of these species over a fairly continuous 500-km radius area. Paradoxically, this is not the case among terrestrial snail genera in the clausiliid subfamily Alopiinae. Phylogenetic analysis using the nuclear markers ITS1/ITS2 and mitochondrial markers COI/12S reveals widespread homoplasy in the clausilial apparatus (a complex aperture-closing mechanism), and concomitant extensive polyphyly among Carinigera, Isabellaria, and Sericata. In contrast, phylogenetic relationships as revealed by molecular data are closely congruent with biogeography at a relatively small scale. A combination of extremely low vagility and extremely high morphological convergence has conspired to produce this unexpected result. Implications as to the function of the clausilial apparatus are discussed.     

Association of the calpain/calpastatin network with subcellular organelles

The calcium-activated cysteine protease calpain is intimately involved in modulating cell adhesion and migration. The two ubiquitous isoforms of this protease, calpain I and II, are considered to be cytosolic proteins that can translocate to both focal complexes/adhesions or the plasma membrane. Using confocal microscopy and isopycnic density centrifugation, the results demonstrate that calpain I and II, the 30kDa regulatory subunit, and calpastatin associate with the endoplasmic reticulum and Golgi apparatus. Confocal microscopy reveals that calpain II colocalizes with the subcellular proteins calnexin and Rab6 in cells bound to laminin. To further verify this association, cell lysates prepared from laminin stimulated and unstimulated cells were subjected to isopycnic density centrifugation. The results reveal an increased association of calpain I, II, calpastatin, and the 30kDa regulatory subunit with the endoplasmic reticulum and Golgi apparatus as evidenced by their position in the gradient relative to calnexin, Rab6, caveolin, and beta1 integrin after laminin stimulation. This correlates with the accumulation of inducible calpain activity at the endoplasmic reticulum-Golgi apparatus interface. Further experiments established that calpain II colocalizes with phosphatidylinositol 4,5-bisphosphate. Finally, calpain II associates with membrane lipid rafts. These results provide new insights into how the calpain/calpastatin network is spatially and temporally regulated in cells binding to the extracellular matrix.     

Protein kinase D: a family affair

The protein kinase D family of enzymes consists of three isoforms: PKD1/PKCmu PKD2 and PKD3/PKCnu. They all share a similar architecture with regulatory sub-domains that play specific roles in the activation, translocation and function of the enzymes. The PKD enzymes have recently been implicated in very diverse cellular functions, including Golgi organization and plasma membrane directed transport, metastasis, immune responses, apoptosis and cell proliferation.     

Transport of protein toxins into cells: pathways used by ricin,cholera toxin and Shiga toxin

Ricin, cholera, and Shiga toxin belong to a family of protein toxins that enter the cytosol to exert their action. Since all three toxins are routed from the cell surface through the Golgi apparatus and to the endoplasmic reticulum (ER) before translocation to the cytosol, the toxins are used to study different endocytic pathways as well as the retrograde transport to the Golgi and the ER. The toxins can also be used as vectors to carry other proteins into the cells. Studies with protein toxins reveal that there are more pathways along the plasma membrane to ER route than originally believed.