scholarly journals Calcium transport into the cells of the sea urchin larva in relation to spicule formation

2016 ◽  
Vol 113 (45) ◽  
pp. 12637-12642 ◽  
Author(s):  
Netta Vidavsky ◽  
Sefi Addadi ◽  
Andreas Schertel ◽  
David Ben-Ezra ◽  
Muki Shpigel ◽  
...  

We investigated the manner in which the sea urchin larva takes up calcium from its body cavity into the primary mesenchymal cells (PMCs) that are responsible for spicule formation. We used the membrane-impermeable fluorescent dye calcein and alexa-dextran, with or without a calcium channel inhibitor, and imaged the larvae in vivo with selective-plane illumination microscopy. Both fluorescent molecules are taken up from the body cavity into the PMCs and ectoderm cells, where the two labels are predominantly colocalized in particles, whereas the calcium-binding calcein label is mainly excluded from the endoderm and is concentrated in the spicules. The presence of vesicles and vacuoles inside the PMCs that have openings through the plasma membrane directly to the body cavity was documented using high-resolution cryo-focused ion beam-SEM serial imaging. Some of the vesicles and vacuoles are interconnected to form large networks. We suggest that these vacuolar networks are involved in direct sea water uptake. We conclude that the calcium pathway from the body cavity into cells involves nonspecific endocytosis of sea water with its calcium.

Author(s):  
H.J. Ryu ◽  
A.B. Shah ◽  
Y. Wang ◽  
W.-H. Chuang ◽  
T. Tong

Abstract When failure analysis is performed on a circuit composed of FinFETs, the degree of defect isolation, in some cases, requires isolation to the fin level inside the problematic FinFET for complete understanding of root cause. This work shows successful application of electron beam alteration of current flow combined with nanoprobing for precise isolation of a defect down to fin level. To understand the mechanism of the leakage, transmission electron microscopy (TEM) slice was made along the leaky drain contact (perpendicular to fin direction) by focused ion beam thinning and lift-out. TEM image shows contact and fin. Stacking fault was found in the body of the silicon fin highlighted by the technique described in this paper.


1997 ◽  
Vol 200 (22) ◽  
pp. 2881-2892 ◽  
Author(s):  
P Leong ◽  
D Manahan

Early stages of animal development have high mass-specific rates of metabolism. The biochemical processes that establish metabolic rate and how these processes change during development are not understood. In this study, changes in Na+/K+-ATPase activity (the sodium pump) and rate of oxygen consumption were measured during embryonic and early larval development for two species of sea urchin, Strongylocentrotus purpuratus and Lytechinus pictus. Total (in vitro) Na+/K+-ATPase activity increased during development and could potentially account for up to 77 % of larval oxygen consumption in Strongylocentrotus purpuratus (pluteus stage) and 80 % in Lytechinus pictus (prism stage). The critical issue was addressed of what percentage of total enzyme activity is physiologically active in living embryos and larvae and thus what percentage of metabolism is established by the activity of the sodium pump during development. Early developmental stages of sea urchins are ideal for understanding the in vivo metabolic importance of Na+/K+-ATPase because of their small size and high permeability to radioactive tracers (86Rb+) added to sea water. A comparison of total and in vivo Na+/K+-ATPase activities revealed that approximately half of the total activity was utilized in vivo. The remainder represented a functionally active reserve that was subject to regulation, as verified by stimulation of in vivo Na+/K+-ATPase activity in the presence of the ionophore monensin. In the presence of monensin, in vivo Na+/K+-ATPase activities in embryos of S. purpuratus increased to 94 % of the maximum enzyme activity measured in vitro. Stimulation of in vivo Na+/K+-ATPase activity was also observed in the presence of dissolved alanine, presumably due to the requirement to remove the additional intracellular Na+ that was cotransported with alanine from sea water. The metabolic cost of maintaining the ionic balance was found to be high, with this process alone accounting for 40 % of the metabolic rate of sea urchin larvae (based on the measured fraction of total Na+/K+-ATPase that is physiologically active in larvae of S. purpuratus). Ontogenetic changes in pump activity and environmentally induced regulation of reserve Na+/K+-ATPase activity are important factors that determine a major proportion of the metabolic costs of sea urchin development.


PLoS ONE ◽  
2013 ◽  
Vol 8 (2) ◽  
pp. e57405 ◽  
Author(s):  
Bohumil Maco ◽  
Anthony Holtmaat ◽  
Marco Cantoni ◽  
Anna Kreshuk ◽  
Christoph N. Straehle ◽  
...  

1946 ◽  
Vol 23 (1) ◽  
pp. 47-70 ◽  
Author(s):  
J. D. SMYTH

A technique has been elaborated that enabled the plerocercoid larvae of Schistocephalus solidus to be removed from the body cavity of Gasterosteus aculeatus without bacterial contamination. Larvae were cultured in plugged test-tubes under completely aseptic conditions in a variety of balanced salines, glucose salines and nutrient peptone broth. The most successful results were obtained with peptone broth at room temperatures (16-19° C) in which plerocercoids remained active and showed normal behaviour for periods up to 300 days. In ¾ strength Locke's solution, which was found by experiment to be approximately isotonic with Schistocephalus (δ = -0.44 ± 0.02° C), the mean period of normal behaviour was 114 days. In the remaining saline and saline-glucose media, the mean viability and period of normal behaviour was considerably less. In the plerocercoid, histological examination revealed that the genitalia are in an immature condition. During cultivation at room temperatures, the genitalia remained in this undifferentiated condition and showed no signs of undergoing spermatogenesis, oogenesis or vitellogenesis. Plerocercoids were induced to develop into sexually mature adults by raising the temperature of cultivation in peptone broth to 40° C. (i.e. the body temperature of the final host in the natural life cycle). Oviposition took place after 48-60 hr. at this temperature, and histological examination revealed that spermatogenesis, oogenesis, vitellogenesis and shell formation had taken place in a normal manner. The viability of artificially matured Schistocephalus was 4-6 days in vitro--a period equivalent to the viability of the adult in vivo. The eversion of the cirris was observed in each proglottid after 40 hr. cultivation at 40° C. During the sexual process the cirris everted and invaginated at the rate of about once per second. Cross-fertilization between segments of the same worm or with segments of another worm was not observed. Except for one specimen in ¾ strength Locke's solution which underwent spermatogenesis and partial vitellogenesis, larvae cultured in salines or glucose salines at 40° C. died within 1-3 days without further development. Attempts to hatch out the eggs produced by the cultivation of larvae in peptone broth at 40° C. proved unsuccessful. Histological examination revealed that spermatozoa had not been taken into the vagina. It was concluded that the eggs were not fertilized owing to the failure of normal copulation to take place.


Author(s):  
Steven B. Herschbein ◽  
Kyle M. Winter ◽  
Carmelo F. Scrudato ◽  
Brian L. Yates ◽  
Edward S. Hermann ◽  
...  

Abstract Focused Ion Beam (FIB) chip circuit editing is a well-established highly specialized laboratory technique for making direct changes to the functionality of integrated circuits. A precisely tuned and placed ion beam in conjunction with process gases selectively uncovers internal circuitry, create functional changes in devices or the copper wiring pattern, and reseals the chip surface. When executed within reasonable limits, the revised circuit logic functions essentially the same as if the changes were instead made to the photomasks used to fabricate the chip. The results of the intended revision, however, can be obtained weeks or months earlier than by a full fabrication run. Evaluating proposed changes through FIB modification rather than proceeding immediately to mask changes has become an integral part of the process for bringing advanced designs to market at many companies. The end product of the FIB process is the very essence of handcrafted prototyping. The efficacy of the FIB technique faces new challenges with every generation of fabrication process node advancement. Ever shrinking geometries and new material sets have always been a given as transistor size decreases and overall packing density increases. The biggest fundamental change in recent years was the introduction of the FinFET as a replacement for the venerable planar transistor. Point to point wiring change methodology has generally followed process scaling, but transistor deletions or modifications with the change to Fins require a somewhat different approach and much more careful control due to the drastic change in height and shape. We also had to take into consideration the importance of the 4th terminal, the body-tie, that is often lost in backside editing. Some designs and FET technology can function acceptably well when individual devices are no longer connected to the bulk substrate or well, while others can suffer from profound shifts in performance. All this presents a challenge given that the primary beam technology improvements of the fully configured chip edit FIB has only evolved incrementally during the same time period. The gallium column system appears to be reaching its maximum potential. Further, as gallium is a p-type metal dopant, there are limitations to its use in close proximity to certain active semiconductor devices. Amorphous material formation and other damage mechanisms that extend beyond what can be seen visually when endpointing must also be taken into account [1]. Device switching performance and even transmission line characteristics of nearby wiring levels can be impacted by material structural changes from implantation cascades. Last year our lab participated in a design validation exercise in which we were asked to modify the drive of a multi-finger FinFET device structure to reduce its switching speed impact on a circuit. The original sized device pulled the next node in the chain too fast, resulting in a timing upset. Deleting whole structures and bridging over/around them is commonly done, but modifications to the physical size of an FET device is a rare request and generally not attempted. It requires a level of precision in beam control and post-edit treatment that can be difficult to execute cleanly. Once again during a complex edit task we considered the use of an alternate ion beam species such as neon, or reducing the beam energy (low kV) on the gallium tool. Unfortunately, we don’t yet have easy access to a versatile viable replacement column technology grafted to a fully configured edit station. And while there should be significantly reduced implant damage and transistor functional change when a gallium column FIB is operated at lower accelerating potential [2], the further loss of visual acuity due to the reduced secondary emission, especially when combined with ultra-low beam currents, made fast and accurate navigation near impossible. We instead chose the somewhat unconventional approach of using an ultra-low voltage electron beam to do much of the navigation and surface marking prior to making the final edits with the gallium ion beam in a dual-beam FIB tool. Once we had resolved how to accurately navigate to the transistors in question and expose half of the structure without disturbing the body-tie, we were able to execute the required cut to trim away 50% of the structure and reduce the effective drive. Several of the FIB modified units functioned per the design parameters of a smaller sized device, giving confidence to proceed with the revised mask set. To our surprise, the gallium beam performed commendably well in this most difficult task. While we still believe that an inert beam of similar characteristics would be preferable, this work indicates that gallium columns are still viable at the 14 nm FinFET node for even the most rigorous of editing requirements. It also showed that careful application of e-beam imaging on the exposed underside of FinFET devices could be performed without degrading or destroying them.


PLoS ONE ◽  
2020 ◽  
Vol 15 (2) ◽  
pp. e0228893 ◽  
Author(s):  
Luigi Inguglia ◽  
Marco Chiaramonte ◽  
Vincenzo Arizza ◽  
Lilla Turiák ◽  
Károly Vékey ◽  
...  

2020 ◽  
Vol 382 (3) ◽  
pp. 609-625
Author(s):  
Takayuki Miyaki ◽  
Yuto Kawasaki ◽  
Akira Matsumoto ◽  
Soichiro Kakuta ◽  
Tatsuo Sakai ◽  
...  

AbstractThe excretory system produces urine by ultrafiltration via a filtration epithelium. Podocytes are widely found as filtration epithelial cells in eucoelomates. In some animal taxa, including insects and crustaceans, nephrocytes serve to separate toxic substances from the body fluid, in addition to podocytes. Drosophila nephrocytes have been recently utilized as a model system to study podocyte function and disease. However, functionality and cellular architecture are strikingly different between Drosophila nephrocytes and eucoelomate podocytes, and the phylogenetic relationship between these cells remains enigmatic. In this study, using focused-ion beam-scanning electron microscopy (FIB-SEM) tomography, we revealed three-dimensional architecture of decapod nephrocytes with unprecedented accuracy—they filled an enormous gap, which can be called “missing link,” in the evolutionary diversity of podocytes and nephrocytes. Thus, we concluded that nephrocytes are part of the spectrum of filtration epithelial diversity in animal phylogeny.


2005 ◽  
Vol 872 ◽  
Author(s):  
John M. Maloney ◽  
Sara A. Lipka ◽  
Samuel P. Baldwin

AbstractLow pressure chemical vapor deposition (LPCVD) and plasma enhanced chemical vapor deposition (PECVD) silicon oxide and silicon nitride films were implanted subcutaneously in a rat model to study in vivo behavior of the films. Silicon chips coated with the films of interest were implanted for up to one year, and film thickness was evaluated by spectrophotometry and sectioning. Dissolution rates were estimated to be 0.33 nm/day for LPCVD silicon nitride, 2.0 nm/day for PECVD silicon nitride, and 3.5 nm/day for PECVD silicon oxide. A similar PECVD silicon oxide dissolution rate was observed on a silicon oxide / silicon nitride / silicon oxide stack that was sectioned by focused ion beam etching. These results provide a biostability reference for designing implantable microfabricated devices that feature exposed ceramic films.


2000 ◽  
Vol 113 (22) ◽  
pp. 3947-3958 ◽  
Author(s):  
J.H. Cho ◽  
Y.S. Oh ◽  
K.W. Park ◽  
J. Yu ◽  
K.Y. Choi ◽  
...  

Calsequestrin is the major calcium-binding protein of cardiac and skeletal muscles whose function is to sequester Ca(2+)in the lumen of the sarcoplasmic reticulum (SR). Here we describe the identification and functional characterization of a C. elegans calsequestrin gene (csq-1). CSQ-1 shows moderate similarity (50% similarity, 30% identity) to rabbit skeletal calsequestrin. Unlike mammals, which have two different genes encoding cardiac and fast-twitch skeletal muscle isoforms, csq-1 is the only calsequestrin gene in the C. elegans genome. We show that csq-1 is highly expressed in the body-wall muscles, beginning in mid-embryogenesis and maintained through the adult stage. In body-wall muscle cells, CSQ-1 is localized to sarcoplasmic membranes surrounding sarcomeric structures, in the regions where ryanodine receptors (UNC-68) are located. Mutation in UNC-68 affects CSQ-1 localization, suggesting that the two possibly interact in vivo. Genetic analyses of chromosomal deficiency mutants deleting csq-1 show that CSQ-1 is not essential for initiation of embryonic muscle formation and contraction. Furthermore, double-stranded RNA injection resulted in animals completely lacking CSQ-1 in body-wall muscles with no observable defects in locomotion. These findings suggest that although CSQ-1 is one of the major calcium-binding proteins in the body-wall muscles of C. elegans, it is not essential for body-wall muscle formation and contraction.


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