Correction: A bioenergetic mechanism for amoeboid-like cell motility profiles tested in a microfluidic electrotaxis assay.

Correction for 'A bioenergetic mechanism for amoeboid-like cell motility profiles tested in a microfluidic electrotaxis assay' by Hagit Peretz-Soroka et al., Integr. Biol., 2017, DOI: .

A bioenergetic mechanism for amoeboid-like cell motility profiles tested in a microfluidic electrotaxis assay.

The amoeboid-like cell motility is known to be driven by the acidic enzymatic hydrolysis of ATP in the actin-myosin system. However, the electro-mechano-chemical coupling, whereby the free energy of ATP hydrolysis is transformed into the power of electrically polarized cell movement, is poorly understood. Previous experimental studies showed that actin filaments motion, cytoplasmic streaming, and muscle contraction can be reconstituted under actin-activated ATP hydrolysis by soluble non-filamentous myosin fragments. Thus, biological motility was demonstrated in the absence of a continuous protein network. These results lead to an integrative conceptual model for cell motility, which advocates an active role played by intracellular proton currents and cytoplasmic streaming (iPC-CS). In this model, we propose that protons and fluid currents develop intracellular electric polarization and pressure gradients, which generate an electro-hydrodynamic mode of amoeboid motion. Such energetic proton currents and active streaming are considered to be mainly driven by stereospecific ATP hydrolysis through myosin heads along oriented actin filaments. Key predictions of this model are supported by microscopy visualization and in-depth sub-population analysis of purified human neutrophils using a microfluidic electrotaxis assay. Three distinct phases in cell motility profiles, morphology, and cytoplasmic streaming in response to physiological ranges of chemoattractant stimulation and electric field application are revealed. Our results support an intrinsic electric dipole formation linked to different patterns of cytoplasmic streaming, which can be explained by the iPC-CS model. Collectively, this alternative biophysical mechanism of cell motility provides new insights into bioenergetics with relevance to potential new biomedical applications.

The morphology of the male reproductive system, spermatogenesis and the spermatozoon of Daphnia magna (Crustacea: Branchiopoda).

This study analyses the histological and cellular morphology of the testis and sperm development in the male Daphnia magna Straus 1820. Due to the rarity of males and predominately parthenogenetic lifecycle of Daphnia, there has been limited detailed information on males in contrast to the well-studied female. Using light and electron microscopy approaches, we describe the morphology of the testis during the progression from an immature to mature testis. The testis has an encasing muscular mesh sheath outside the basal lamina, beneath which is a thin somatic epithelial cell layer. Internal to the epithelium are the spermatogonial stem cells and subsequent syncytial clusters of the germ cells as they progress through spermatogenesis; spermatozoa occupy the entire testis in sexually mature D. magna. We describe the structure of developing and mature spermatozoa; mature spermatozoa are non-flagellated, ovoid in shape with plasmalemma filapodia and are encased in an extracellular capsule.

Not of African Descent: Dental Modification among Indigenous Caribbean People from Canímar Abajo, Cuba.

Dental modifications in the Caribbean are considered to be an African practice introduced to the Caribbean archipelago by the influx of enslaved Africans during colonial times. Skeletal remains which exhibited dental modifications are by default considered to be Africans, African descendants, or post-contact indigenous people influenced by an African practice. Individual E-105 from the site of Canímar Abajo (Cuba), with a direct 14C AMS date of 990-800 cal BC, provides the first unequivocal evidence of dental modifications in the Antilles prior to contact with Europeans in AD 1492. Central incisors showing evidence of significant crown reduction (loss of crown volume regardless of its etiology) were examined macroscopically and with a scanning electron microscope (SEM) to determine if the observed alterations were due to deliberate modification or other (unintentional) factors considered: postmortem breakage, violent accidental breakage, non-dietary use of teeth, and wear caused by habitual or repeated actions. The pattern of crown reduction is consistent with deliberate dental modification of the type commonly encountered among African and African descendent communities in post-contact Caribbean archaeological assemblages. Six additional individuals show similar pattern of crown reduction of maxillary incisors with no analogous wear in corresponding mandibular dentition.

Microscopic identification of novel cell types in the integument of larval lake sturgeon, Acipenser fulvescens.

Osmoregulation, respiration, nutrient/mineral transport, and defense mechanisms are all evident in the integument of fish. The role of the integument in these physiological processes is particularly important during early life history in larval fishes, as functional systems such as the gills and gastrointestinal tract are not fully developed. Using a variety of microscopy techniques, we describe the morphology of keratinocytes, mitochondria rich cells, ciliated cells and mucous cells of the skin, yolk sac, and gills. The cytology we observed was similar to previous studies describing the integument of larval fish, however, we have also identified two novel cell types on the integument of larval Lake Sturgeon, Acipenser fulvescens, between 9 and 34 days post fertilization. Our detailed analysis included a multifaceted microscopy approach using scanning electron, transmission electron, and light microscopy to elucidate the histology of the tissue and cellular morphology in addition to quantification and distribution of these novel cell types. The first cell type had a characteristic ampullary shape with a central cavity and a pore opening at the surface. The second, located on the free surface of the epidermis, had an uneven plasma membrane surface. Based on the abundance of secretory vesicles, organelles necessary for protein synthesis, and the lack of neural connection in both cell types, we propose these cells to be involved in the release of semiochemicals that may act as a pheromone, alarm substance, or chemical defense mechanism.

Identification and comparison of the 3 early stages of resynthesis for the lichen Cladonia rangiferina.

A lichen is an association between a biotrophic fungal partner and a green algal and (or) cyanobacterial partner, which may be considered a "controlled" parasitic interaction. While controlled parasitism implies benefit to both interacting partners, a parasitism that is not controlled implies that one partner benefits to the detriment of the other partner. The objective of this study was to compare morphological development of the interaction between Cladonia rangiferina with its compatible algal partner (Asterochloris glomerata/irregularis) and incompatible algae (Coccomyxa peltigerae and Chloroidium ellipsoideum) at 3 early resynthesis stages. The fungus was co-inoculated with each alga separately and the stages of development were compared using quantitative measures. The first 3 stages of development of the lichen thallus were identified in the compatible interaction as the "pre-contact" stage (1 day post co-inoculation (PCI)), "contact" stage (8 days PCI), and "growth together" stage (21 days PCI). Compatible interactions showed significantly shorter internode length, significantly more new lateral hyphal branches, significantly greater appressorial frequency, and no reduction in cell diameter of the algal cells, compared with incompatible interactions. At 21 days PCI, a parasitic interaction was observed between Cladonia rangiferina and Chloroidium ellipsoideum. These findings support the importance of recognition between compatible partners for successful lichenization. This study also revealed a strategy that may explain the success of this species in northern habitats. Identification of the resynthesis stages of Cladonia rangiferina is required before expression of the proteins involved in recognition and defense can be understood.

UVB radiation affects growth, reproduction and tissue structure of Daphnia magna across several temperatures.

We examined the effects of daily exposure to UVB on growth, reproduction and histological characteristics of Daphnia magna over two generations at 20, 22, 25 and 30°C. Animals were exposed to 16 h of UVA and photosynthetically active radiation daily. Treated animals received 6 h of UVB during the light phase. Parental (P) generation growth and reproduction was impaired by exposure to UVB at all temperatures, with the poorest production at 30°C. First brood size decreased with UVB exposure; it was lowest at 30°C. Although F1 length at birth increased with P generation age, F1 produced by UVB-exposed mothers were smaller at all temperatures. The F1 generation was followed at 20 and 25°C; at both temperatures UVB exposure reduced F1 growth and reproduction. F1 growth and F2 production were lowest when both P and F1 generations were exposed to UVB. UVB exposure damaged ovarian and gut tissue at both 25 and 30°C; the consequences of this exposure were more severe at 30°C. The observed tissue damage may relate directly to the UVB-induced impairment of growth and reproduction. This study provides new insights into the effects of UVB on an important component of the pelagic zooplankton.

Evolution and development of polarized germ cell cysts: new insights from a polychaete worm, Ophryotrocha labronica.

Polarized oogenic cysts are clonal syncytia of germ cells in which some of the sister cells (cystocytes) differentiate not as oocytes, but instead as nurse cells: polyploid cells that support oocyte development. The intricate machinery required to establish and maintain divergent cell fates within a syncytium, and the importance of associated oocyte patterning for subsequent embryonic development, have made polarized cysts valuable subjects of study in developmental and cell biology. Nurse cell/oocyte specification is best understood in insects, particularly Drosophila melanogaster. However, polarized cysts have evolved independently in several other animal phyla. We describe the differentiation of female cystocytes in an annelid worm, the polychaete Ophryotrocha labronica. These worms are remarkable for their elegantly simple cysts, which comprise a single oocyte and nurse cell, making them an appealing complement to insects as subjects of study. To elucidate the process of cystocyte differentiation in O. labronica, we have constructed digital 3D models from electron micrographs of serially sectioned ovarian tissue. These models show that 2-cell cysts arise by fragmentation of larger "parental" cysts, rather than as independent units. The parental cysts vary in size and organization, are produced by asynchronous, indeterminate mitotic divisions of progenitor cystoblasts, and lack fusome-like organizing organelles. All of these characteristics represent key cytological differences from "typical" cyst development in insects like D. melanogaster. In light of such differences and the plasticity of female cyst structure among other animals, we suggest that it is time to reassess common views on the conservation of oogenic cysts and the importance of cysts in animal oogenesis generally.

Development of polarized female germline cysts in the polychaete, Ophryotrocha labronica.

Polarized oogenic cysts are clonal syncytia in which cellular compartments (cystocytes) differentiate as either oocytes or nurse cells-supportive germ cells that transport material to the oocyte through cytoplasmic bridges. Although polarized cysts have evolved independently in several animal phyla, most research on this phenomenon has focused on insects, particularly Drosophila melanogaster. To provide a foundation for comparative studies of cyst polarization, we have examined the coelomic development of polarized cysts in an annelid worm, Ophryotrocha labronica, using a variety of microscopic techniques. This species has elegantly simple cysts, each consisting of two interconnected cells (one oocyte and one nurse cell) free of somatic follicular tissue. These cysts arise from segmental ovaries, but are released to the coelom early in oogenesis, where they carry out most of their development. Although the oocyte remains transcriptionally active through oogenesis, nurse cells in this species are robust biosynthetic factories, with highly polyploid nuclei, enormous nucleoli, numerous mitochondria, and abundant rough endoplasmic reticulum. Ultimately, the nurse cell dumps its cytoplasm into oocyte, detaches, and undergoes programmed cell death. Although in insects, and possibly some polychaetes, the overall polarity of the cyst is linked to the specification of future embryonic axes in the oocyte, in Ophryotrocha this appears not to be the case. Our findings provide a detailed analysis of cyst development, addressing earlier misconceptions in the literature and adding new insights. This foundation shows the potential of Ophryotrocha labronica as a model system for further experimental analysis of nurse cell-oocyte differentiation.

Dexrazoxane (ICRF-187) protects cardiac myocytes against doxorubicin by preventing damage to mitochondria.

The clinically approved antioxidant cardioprotective agent dexrazoxane (ICRF-187) was examined for its ability to protect neonatal rat cardiac myocytes from doxorubicin-induced damage. Doxorubicin is thought to induce oxidative stress on the heart muscle, both through reductive activation to its semiquinone form, and by the production of hydroxyl radicals mediated by its complex with iron. Hydrolyzed dexrazoxane metabolites prevent site-specific iron-based oxygen radical damage by displacing iron from doxorubicin and chelating free and loosely bound iron. The mitochondrial stain MitoTracker Green FM and doxorubicin were shown by epifluorescence microscopy to accumulate in the myocyte mitochondria. An epifluorescence microscopic image analysis method to measure mitochondrial damage was developed using the mitochondrial membrane potential sensing ratiometric dye JC-1. This method was used to show that dexrazoxane protected against doxorubicin-induced depolarization of the myocyte mitochondrial membrane. Dexrazoxane also attenuated doxorubicin-induced oxidation of intracellular dichlorofluorescin. Annexin V-FITC/propidium iodide staining of myocytes was used to demonstrate that, depending on the concentration, doxorubicin caused both apoptotic and necrotic damage. These results suggest that doxorubicin may be cardiotoxic by damaging the mitochondria and dexrazoxane may be protective by preventing iron-based oxidative damage.

Embryonic development of the hemipteran insect Rhodnius prolixus.

The embryonic development of the hemipteran insect Rhodnius prolixus was studied by use of contemporary light and electron microscopy. Embryos were staged according to days postoviposition. Eggs laid on day one complete blastoderm formation and anatrepsis, the first phase of blastokinesis, by day 5. The embryo develops in a cephalocaudal orientation which is 180° to the anteroposterior axis of the egg. Subsequent development, prior to the second phase of blastokinesis (katatrepsis), leads to segmentation of the germ band, evagination of appendages, and histogenesis of germ layers. Concomitantly with these events, the amnion undergoes dramatic change. By day 7 the embryo begins a 180° revolution while migrating to the ventral surface of the yolk. This restores its polarity with respect to that of the egg and facilitates hatching. The serosa contracts, pulling the amnion and embryo anteriorly. Eventually the serosa is internalized at a point dorsal to the head and the lateral walls of the embryo grow up and surround the yolk. Development continues until day 15 when the embryo hatches as a first instar larva.

Spermathecal ultrastructure of the insect Rhodnius prolixus stal.

The paired spermathecae of Rhodnius are simple tubular out-pocketings of the common oviduct. Each consists of a short muscular proximal duct and the distal glandular region with a blind tapering end. The spermathecal wall has a cuticular intima, slender columnar epithelial cells and ensheathing longitudinal striated muscle, connective tissue, tracheoles, and nerves. Glandular epithelial cells possess an elaborate apical secretion-filled tubular inpocketing with an extensively folded plasma membrane. Laterally, cells interact by desmosomes, septate desmosomes, and extensive interdigitations. The cytoplasm is rich in longitudinally oriented microtubules associating with membrane densities along the invagination, lateral, and basal plasmalemmae. Apical concentration of mitochondria suggests their role in secretion or ion transport. The possible role of the spermathecae in maintaining the stored luminal sperm and its role in transmitting the mating stimulus is considered in light of the epithelial ultrastructure. The ultrastructure of the spermathecae of Rhodnius differs significantly from that of other insects.

Histological and ultrastructural specialization of the digestive tract of the intestinal air breather hoplosternum thoracatum (teleost).

The digestive tract of Hoplosternum thoracatum consists of an esophagus, gastric area, anterior digestive intestine with elaborate folds, digestive intestine with decreasing folds and thin, smooth-surfaced respiratory intestine. The upper tract has a mucoid columnar lining which is gently folded, whereas the gastric area has numerous pits opening into the tubular secretory glands. Striated muscle comprises the anterior muscularis but is replaced by inner circular and outer longitudinal smooth muscle layers in the gastric region. The digestive intestinal mucosa is elaborately folded, consisting of columnar cells with prominent brush borders. Mucosa, submucosa, circular and longitudinal muscularis and serosa layers are present throughout the tract. Goblet cells occur in both the digestive and respiratory intestine. Major changes that appear in the respiratory intestine are a drastic reduction in mucosa epithelial thickness and the penetration of an elaborate capillary bed into the epithelium. The other basic layers are not significantly reduced in thickness. The air-blood barrier consists of the thin epithelium, basement lamina and very thin capillary endothelium. Regional cellular composition and ultrastructural features are correlated with respective digestive and respiratory functions.