Structural Repair and Functional Recovery Following Cerebral Ganglion Removal in the Pulmonate Snail Melampus

SYNOPSIS. Regeneration of the nervous system of Melampus followingcerebral ganglion removal proceeds through tract, bud, and ganglionstages Each stage can represent a terminal condition in someanimals Early events of regeneration appear to include rolesfor chemotactic and growth-promoting agents and axonal guidanceby preferential adhesion to a connective tissue sheath Thislatter proposed mechanism accounts for the observed sequencein which the neural elements unite in the tract stage and forthe pattern of failures that result when the sheath is disruptedIn the tract stage of regeneration, communication through thesite of the missing ganglion is restored within the centralnervous system, and between neurons of non-excised ganglia andthe denervated periphery Some behavioral recovery results Thebud stage of regeneration is characterized by neuropil developmentand associated swelling at the site of confluence of the tractsSerotonin immunohistochemistry of bud stage preparations andretrograde dye transport via bud nerves show tracts and numeroussynaptic vancosities, but the neuron somata that are labeledare located in other ganglia Ultrastructural examination oflate bud/early ganglion stage tissue reveals the presence ofsmall undifferentiated cells By six to seven months postoperative,some snails have clearly reached a ganglion stage of regenerationcharacterized by the appearance of differentiated neurons withinthe bud The origin of these new neurons is currently under investigation.     

Schedule of Spermatogenesis in the Pulmonate Snail Helix aspersa, with Special Reference to Histone Transition

The schedule of spermatogenesis is determined from the times necessary for cells labeled with tritium thymidine during premeiotic DNA synthesis to pass through the successive spermatogenic stages. A transition from a typically somatic histone rich in lysine, to a histone rich in arginine is shown to occur during spermatid stages. A later shift to a protamine is observed in the maturing sperm. These changes are characterized by the use of in situ staining methods. The transition to an arginine-rich histone is accompanied by incorporation of tritium-labeled arginine, hence reflects synthesis of new protein. Comparison of the timing of arginine and thymidine incorporation, and independent measurements of DNA, show that in contrast to the case of premitotic chromosome duplication, the histone synthesis in the spermatid is unaccompanied by DNA synthesis. During the initial histone change, fine filaments are formed within the nucleus, which aggregate to form lamellae. This fine structure is lost during maturation of the sperm.     

Formation and Fate of Extranuclear Chromatin Bodies in Tetrahymena

The fate of extranuclear chromatin bodies (ECBs) formed by exclusion of macronuclear material at the time of karyokinesis was followed quantitatively in Tetrahymena pyriformis strain GL-I. In a logarithmic growth phase culture, 51% of the dividing cells produced one (43%) or more (8%) ECBs. Most of these gradually disappear before the next cell division, but ? 13% are retained and carried into subsequent cell cycles. The random distribution of ECBs into anterior or posterior daughter cells, their staining and morphological characteristics, and their rapid loss in cells in starvation medium, all indicate that ECBs play no more of a role in cellular activity than that of an internally produced food vacuole.     

Osmotic Balance in a Marine Pulmonate,Amphibola crenata

The pulmonate mud‐snail Amphibola crenata is an osmoconformer in 25%‐125% sea water, its haemolymph being slightly hyperosmotic and hyperionic to the medium. It maintains its haemolymph markedly hyperosmotic in media below 25% sea water and in freshwater, in which it survives for a week or longer. On exposure to dilute or concentrated media, water which enters or leaves the tissues preferentially enters or leaves the cells. Regulation of cell volume towards initial values is accomplished within 5 days. Measurements of changes in cell volume, and of distribution of tissue water between the intracellular and extracellular compartments, in snails moved from 100% sea water into dilute or concentrated media, and in snails moved back to 100% sea water, indicate that mechanisms of cell volume regulation are minimal in ca 60–65% sea water. Changes in the concentration of tissue free amino acids are in accord with current theories of mechanisms of isosmotic intracellular regulation. The results are discussed in the light of the taxonomic, evolutionary and ecological status of Amphibola as a shore‐living, primitive, marine representative of a predominantly terrestrial and freshwater group of gastropod molluscs.     

Spermiogenesis in a Pycnogonid Achelia echinata Hodge

Spermiogenesis and structure of the mature sperm are described in Achelia echinata Hodge. The sperm differ from previously described types and perhaps represent an unspecialised type.     

Spermiogenesis in a Scutariellid (Platyhelminthes)

Spermiogenesis and spermatozoa were studied by transmission and scanning electron microscopy in Troglocaridicolasp., a scutariellid epizoic on a cavernicolous freshwater shrimp. Spermiogenesis involves elongation of the spermatid in which the nucleus elongates, but remains close to the common cytoplasmic mass. Flagella first grow in opposite direction and at a right angle to the cytoplasmic shaft, and centrioles show associate structures. Later, the two centrioles rotate and the flagella emerge parallel, but still perpendicular to the shaft. An apical process elongates at the extremity of the spermatid shaft. The spermatozoon shows active flagellar beating and undulations of the sperm body. The spermatozoon comprises an anterior ‘corkscrew’ region, the flagellar insertion region, a cytoplasmic region and a posterior nuclear region. The corkscrew contains an electron dense structure, not membrane-bound, originating from the apical process of the spermatid. The flagella show the 9+‘1’ pattern, usual in Platyhelminthes. The cytoplasmic and nuclear regions show a cortical row of about 50 twisted longitudinal microtubules surrounding a row of electron dense, and not membrane-bound, 25-nm granules. These granules are original structures and seem to be known only in a few Platyhelminthes species in which a non-flagellar movement of the spermatozoon occurs. Thus, it is hypothesised that the 25-nm granules play a role in cellular motility. Sperm ultrastructure in Troglocaridicolashows major differences to that in the temnocephalids. It is therefore concluded that the phylogenetic position of the scutariellids within the Temnocephalidea should be reinvestigated.     

Spermiogenesis in the anthozoan Aiptasia pallida

Fine structural aspects of spermiogenesis in the anthozoan Aiptasia pallida have been examined. Nuclear condensation is similar to that reported for other anthozoans. The middle piece is formed by fusion of a few small mitochondria and is doughnut-shaped. A well-developed centriolar complex is attached to the nuclear envelope by fine filamentous material and is surrounded by the mitochondrial collar. Electron-dense vesicles, which correspond to 'proacrosomal vesicles', are scattered in the peripheral cytoplasm at the nuclear-mitochondrial junction. Cytochemical procedures indicate that these vesicles contain carbohydrates and basic proteins, suggesting they may be acrosomal-like.     

Formation of the CenH3-Deficient Holocentromere in Lepidoptera Avoids Active Chromatin

1. Download : Download high-res image (148KB)
2. Download : Download full-size image

     

Evidence for Intrinsic Differences in the Formation of Chromatin Domains in Drosophila Melanogaster

The results of an investigation into intrinsic differences in the formation of two different heterochromatic domains are presented. The study utilized two different position effect variegation mutants in Drosophila melanogaster for investigating the process of compacting different stretches of DNA into heterochromatin. Each stretch of DNA encodes for a gene that affects different aspects of bristle morphology. The expression of each gene is prevented when it is compacted into heterochromatin thus the genes serve as effective reporter systems to monitor the spread of heterochromatin. Both variegating mutants are scored in the same cell such that environmental and genetic background differences are unambiguously eliminated. Any differences observed in the repression of the two genes must therefore be the result of intrinsic differences in the heterochromatic compaction process for the two stretches of DNA. Studies of the effects different enhancers of variegation have upon the compaction of the two genes indicate each compaction event occurs independently of the other, and that different components are involved in the two processes. These results are discussed with regard to spreading heterochromatin and the role this process may play in regulating gene expression.     

中华蟾蜍精子的发生和形成

中华蟾蜍精子的发生可以分为精原细胞期、初级精母细胞期、次级精母细胞期、精子细胞期和精子期5个时期,精子的形成过程包括核固缩、细胞质丢失、鞭毛形成等阶段.曲细精管中可以分出4种形态的精原细胞,其中有3种为增殖型精原细胞,另1种形态的精原细胞通过分裂形成1个精原干细胞和1个增殖型精原细胞,前者分裂以维持精原细胞的数量,后者发育分裂成精母细胞.Mallory三色法染色能清晰的区分精原干细胞和增殖型精原细胞.毕特氏器是异性生殖腺的残余,与精巢以被膜为界.性成熟个体的毕特氏器之卵母细胞圆形或椭圆形,多处于Ⅲ时相;滤泡细胞扁平或矮立方状;卵母细胞间富含微血管、间质细胞、嗜苦味酸细胞等.     

Amiodarone Induces Overexpression of Similar to Versican b to Repress the EGFR/Gsk3b/Snail Signaling Axis during Cardiac Valve Formation of Zebrafish Embryos

Although Amiodarone, a class III antiarrhythmic drug, inhibits zebrafish cardiac valve formation, the detailed molecular pathway is still unclear. Here, we proved that Amiodarone acts as an upstream regulator, stimulating similar to versican b (s-vcanb) overexpression at zebrafish embryonic heart and promoting cdh-5 overexpression by inhibiting snail1b at atrioventricular canal (AVC), thus blocking invagination of endocardial cells and, as a result, preventing the formation of cardiac valves. A closer investigation showed that an intricate set of signaling events ultimately caused the up-regulation of cdh5. In particular, we investigated the role of EGFR signaling and the activity of Gsk3b. It was found that knockdown of EGFR signaling resulted in phenotypes similar to those of Amiodarone-treated embryos. Since the reduced phosphorylation of EGFR was rescued by knockdown of s-vcanb, it was concluded that the inhibition of EGFR activity by Amiodarone is s-vcanb-dependent. Moreover, the activity of Gsk3b, a downstream effector of EGFR, was greatly increased in both Amiodarone-treated embryos and EGFR-inhibited embryos. Therefore, it was concluded that reduced EGFR signaling induced by Amiodarone treatment results in the inhibition of Snail functions through increased Gsk3b activity, which, in turn, reduces snail1b expression, leading to the up-regulation the cdh5 at the AVC, finally resulting in defective formation of valves. This signaling cascade implicates the EGFR/Gsk3b/Snail axis as the molecular basis for the inhibition of cardiac valve formation by Amiodarone.     

Learning the Formation Mechanism of Domain-Level Chromatin States with Epigenomics Data

Epigenetic modifications can extend over long genomic regions to form domain-level chromatin states that play critical roles in gene regulation. The molecular mechanism for the establishment and maintenance of these states is not fully understood and remains challenging to study with existing experimental techniques. Here, we took a data-driven approach and parameterized an information-theoretic model to infer the formation mechanism of domain-level chromatin states from genome-wide epigenetic modification profiles. This model reproduces statistical correlations among histone modifications and identifies well-known states. Importantly, it predicts drastically different mechanisms and kinetic pathways for the formation of euchromatin and heterochromatin. In particular, long, strong enhancer and promoter states grow gradually from short but stable regulatory elements via a multistep process. On the other hand, the formation of heterochromatin states is highly cooperative, and no intermediate states are found along the transition path. This cooperativity can arise from a chromatin looping-mediated spreading of histone methylation mark and supports collapsed, globular three-dimensional conformations rather than regular fibril structures for heterochromatin. We further validated these predictions using changes of epigenetic profiles along cell differentiation. Our study demonstrates that information-theoretic models can go beyond statistical analysis to derive insightful kinetic information that is otherwise difficult to access.     

Snail1a and Snail1b cooperate in the anterior migration of the axial mesendoderm in the zebrafish embryo

The Snail genes are implicated in processes that involve cell movement, both during embryonic development and tumour progression. In teleosts, the vertebrate Snail1 gene is represented by two distinct genes, snail1a and snail1b (previously snail1 and snail2). These genes are expressed in complementary mesodermal domains and their combined expression matches that of their mammalian counterpart. By analysing their loss and gain of function, we found that the most-anterior axial mesendodermal cells, the precursors of the polster, move in a cohesive manner directed by the activity of snail1a- and snail1b-expressing cells surrounding these precursors. The cell-autonomous function of Snail1 proteins regulates cell motility and influences the behaviour of Snail-negative neighbouring cells. Snail1a is required by the prechordal plate for it to reach its normal position, whereas Snail1b controls the acquisition of its normal shape. These non-redundant functions of Snail1a and Snail1b in controlling axial mesendoderm migration comply with the duplication-degeneration-complementation model, and indicate that Snail genes not only act as inducers of epithelial-to-mesenchymal transition, but also as more general regulators of cell adhesion and movement.     

Eye Structure and Vision in the Freshwater Pulmonate Mollusc Planorbarius corneus

The structure of the mollusc Planorbarius corneus eye was studied using light and electron microscopy. The eye consists of the cornea, eye lens of non-spherical shape, and the vitreous body tightly bound to it, as well as of a monolayer non-inverted retina composed of photoreceptor and supporting (pigmented) cells. Its inner surface has two invaginations. The apices of several hundreds of photoreceptors directed to the vitreous body have groups of microvilli with a parallel orientation to each other. The eye optic system places the image into the base of the retinal microvillar layer. Its angle resolution provided by density of distribution of photoreceptor cells and the value of the index F = 1.71 is to be about 2.06° with a correction for the light spreading between microvilli of neighbor cells. Under conditions of a V-shaped labyrinth, the P. corneus molluscs show positive phototaxis by moving to the light source. The angular acuity of vision was assessed from differences in the choice by the molluscs of direction of movement to the pattern of vertical black bands with different periods of alternation. The differential threshold obtained is within an interval of 1.43–1.91°, which is close to the calculated value of angular resolution of the retina.     

Spermiogenesis in the Australian cockatiel Nymphicus hollandicus

Information on the ultrastructure of parrot spermatids and spermatozoa is limited to only four species with no comprehensive study of spermiogenesis conducted within the order Psittaciformes. The present study was undertaken to describe the development of the cockatiel spermatid using electron microscopy. Four phases of spermatid maturation were documented on the basis of nuclear morphology, development of the acrosome, perforatorium, and axial filament. These phases included 1) round nuclei, 2) irregular nuclei, 3) elongated nuclei with granular chromatin, and 4) elongated nuclei with homogenous chromatin. While development of the cockatiel spermatid was comparable to that of other domestic avian species, we have noted the hollow nature of some chromatin granules, an abnormal formation of the axoneme, the absence of the fibrous sheath around the axoneme of the principal piece, and the absence of an annulus. J. Morphol. , 2012. © 2012 Wiley Periodicals, Inc.     

大鼠老化过程小脑神经元染色质核小体线性和空间排布特征分析

阐明真核细胞染色质核小体线性和空间排布特征及其机制是染色质结构和功能研究的核心内容．近年来随着染色质分子生物学研究的深入,人们发现染色质核小体不仅作为真核基因组三维结构的基本结构单元,而且在细胞核内线性和空间排布（ｌｉｎ－ｅａｒａｎｄｓｐａｃｉａｌａ...     

K<Superscript>+</Superscript> Channels in Cardiomyocytes of the Pulmonate Snail <Emphasis Type="Italic">Helix</Emphasis>

We used the patch-clamp technique to identify and characterize the electrophysiological, biophysical, and pharmacological properties of K⁺ channels in enzymatically dissociated ventricular cells of the land pulmonate snail Helix. The family of outward K⁺ currents started to activate at –30 mV and the activation was faster at more depolarized potentials (time constants: at 0 mV 17.4 ± 1.2 ms vs. 2.5 ± 0.1 ms at + 60 mV). The current waveforms were similar to those of the A-type family of voltage-dependent K⁺ currents encoded by Kv4.2 in mammals. Inactivation of the current was relatively fast, i.e., 50.2 ± 1.8% of current was inactivated within 250 ms at + 40 mV. The recovery of K⁺ channels from inactivation was relatively slow with a mean time constant of 1.7 ± 0.2 s. Closer examination of steady-state inactivation kinetics revealed that the voltage dependency of inactivation was U-shaped, exhibiting less inactivation at more depolarized membrane potentials. On the basis of this phenomenon, we suggest that a channel encoded by Kv2.1 similar to that in mammals does exist in land pulmonates of the Helix genus. Outward currents were sensitive to 4-aminopyridine and tetraethylammonium chloride. The last compound was most effective, with an IC₅₀ of 336 ± 142 µmol l^–1. Thus, using distinct pharmacological and biophysical tools we identified different types of voltage-gated K⁺ channels. Present address for S.A.K.: Brigham and Womens Hospital, Cardiovascular Division, Harvard Medical School, 75 Francis St., Thorn 1216, Boston, MA 02115.