Ultrastructure of the ciliary pits in the Geocentrophora group (Platyhelminthes,Lecithoepitheliata)

The ultrastructure of the paired lateral ciliary pits in several endemic species of Geocentrophora from Lake Baikal and in one cosmopolitan species, G. baltica, has been compared and the possible functional significance is discussed. The pit is composed of two distinctive parts; the bottom of the pit is an extensive sensitive area, filled with uni-and biciliary sensory receptors with reduced rootlets and numerous neurotubules. The walls of the pit are formed by several large dark cells, characterized by a dark cytoplasm with numerous mitochondria, a large nucleus, intracellular canaliculi, basal infoldings of the cell membrane, glycogen granules and a varying number of cilia. A protruding, densely ciliated ridge occurs along the anterior wall of the pit. The cilia have a strengthened rootlet system and seem to provide a strong water current into the pit. Dark cell processes penetrate the basement membrane of the pit and come into the vicinity of large cells with a cytoplasm similar to that of the dark cells of the pit. These large cells in their turn come close to the terminal parts of the protonephridial canals, containing a weir. Smaller protonephridial capillaries without a weir seem to open directly into the pit lumen. The morphological data obtained suggest that the ciliary pit in not only a sensory structure, but plays a part in osmoregulation and ion exchange as well.     

Cytoplasmic Ca2+ inhibits the ryanodine receptor from cardiac muscle

Ca²⁺-dependent inhibition of native and isolated ryanodine receptor (RyR) calcium release channels from sheep heart and rabbit skeletal muscle was investigated using the lipid bilayer technique. We found that cytoplasmic Ca²⁺ inhibited cardiac RyRs with an average K _m = 15 mm, skeletal RyRs with K _m = 0.7 mm and with Hill coefficients of 2 in both isoforms. This is consistent with measurements of Ca²⁺ release from the sarcoplasmic reticulum (SR) in skinned fibers and with [³H]-ryanodine binding to SR vesicles, but is contrary to previous bilayer studies which were unable to demonstrate Ca²⁺-inhibition in cardiac RyRs (Chu, Fill, Stefani &; Entman (1993) J. Membrane Biol. 135, 49–59). Ryanodine prevented Ca²⁺ from inhibiting either cardiac or skeletal RyRs. Ca²⁺-inhibition in cardiac RyRs appeared to be the most fragile characteristic of channel function, being irreversibly disrupted by 500 mm Cs⁺, but not by 500 mm K⁺, in the cis bath or by solublization with the detergent CHAPS. These treatments had no effect on channel regulation by AMP-PNP, caffeine, ryanodine, ruthenium red, or Ca²⁺-activation. Ca²⁺-inhibition in skeletal RyRs was retained in the presence of 500 mm Cs⁺. Our results provide an explanation for previous findings in which cardiac RyRs in bilayers with 250 mm Cs⁺ in the solutions fail to demonstrate Ca²⁺-inhibition, while Ca²⁺-inhibition of Ca²⁺ release is observed in vesicle studies where K⁺ is the major cation. A comparison of open and closed probability distributions from individual RyRs suggested that the same gating mechanism mediates Ca²⁺-inhibition in skeletal RyRs and cardiac RyRs, with different Ca²⁺ affinities for inhibition. We conclude that differences in the Ca²⁺-inhibition in cardiac and skeletal channels depends on their Ca²⁺ binding properties.     

Theristus (Penzancia) anoxybioticus n. sp. (Nematoda: Xyalidae) from Sublittoral Methane Seepages in the Northern Kattegat,Denmark

The free-living marine nematode Theristus (Penzancia) anoxybioticus n. sp. is described from specimens collected in muddy sediment at 10-12 m water depth in the northern Kattegat, Denmark, where the benthic environment is influenced by methane seepages. Mean body length of the male is 1,121 μm and of the female 1,159 μm. Theristus (Penzancia) anoxybioticus n. sp. has one crown of 10 cephalic setae and a clavate tail tip without setae. The three caudal gland cells are prominent. The intestinal lumen is hexaradiate in cross section and the lining is devoid of microvilli. Reproductive adults have so far only been found in the uppermost centimeter of sediment, and their presence is restricted to April and May. Juveniles are found in deep anoxic sediment layers during other months of the year.     

THE FLAGELLAR APPARATUS OF CHRYSOCHROMULINA SP. (PRYMNESIOPHYCEAE)1

The flagellar apparatus of an undescribed species of Chrysochromulina Lackey that bears “eyelash” scales is reconstructed. The transitional region consists of two transitional plates each with an axosome, with no stellate pattern between them. Fine osmiophilic rings lie between the flagellar membrane and the outer doublets in the transitional region. The two jagella and the haptonema are inserted in a subapical depression that is lined ventrally by a spine-like projection formed by one of the parietal chloroplasts. The angles of insertion are similar to those of some other Chrysochromulina species in that both the haptonema and the right basal body lie at an extreme angle to the left basal body. The connectives of the apparatus consist of a striated distal band with a dorsal extension to the R1 and a ventral extension overlying the R2, a striated distal accessory band, an auxiliary connective from the right basal body to the adjacent ventral chloroplast, a well-developed intermediate band, two striated proximal bands, and a striated proximal accessory band. Of the microtubular roots in this Chrysochromulina species, three are associated with the left side of the cell (an R1 of 8+3; a small crystalline compound root, the R1C, associated with the R1; an R2 of three micro-tubules), and two are associated with the right basal body (an R3 of 2/2 microtubules with which the single-stranded R4 converges to form a 2/2+1 and then a 2/3 tiered arrangement). Comparisons are drawn with other species in the genus and related genera, particularly Prymne-sium.     

Activin A and transforming growth factor-β stimulate heart formation in axolotls but do not rescue cardiac lethal mutants

In the Mexican axolotl (salamander), Ambystoma mexicanum, a recessive cardiac lethal mutation causes an incomplete differentiation of the myocardium. Mutant hearts lack organized sarcomeric myofibrils and do not contract throughout their lengths. We have previously shown that RNA purified from normal anterior endoderm or from juvenile heart tissue is able to rescue mutant embryonic hearts in an in vitro organ culture system. Under these conditions as many as 55% of formerly quiescent mutant hearts initiate regular contractions within 48 hours. After earlier reports that transforming growth factor-1 and, to a lesser extent, platelet-derived growth factor-BB could substitute for anterior endoderm as a promoter of cardiac mesodermal differentiation in normal axolotl embryos, we decided to examine the effect of growth factors in the cardiac mutant axolotl system. In one type of experiment, stage 35 mutant hearts were incubated in activin A, transforming growth factors-1 or 2, platelet-derived growth factor, or epidermal growth factor, but no rescue of mutant hearts was achieved. Considering the possibility that growth factors would only be effective at earlier stages of development, we tested transforming growth factors-1 and 5, and activin A on normal and mutant precardiac mesoderm explanted in the absence of endoderm at neurula stage 14. We found that, although these growth factors stimulated heart tube formation in both normal and mutant mesodermal explants, only normal explants contained contractile myocardial tissue. We hypothesize that transforming growth factor- superfamily peptides initiate a cascade of responses in mesoderm that result in both changes in cell shape (the basis for heart morphogenesis) and terminal myocardial cytodifferentiation. The cardiac lethal mutation appears to be deficient only in the latter process.This work was supported by NIH grants HL-32184 and HL-37702 and a grant-in-aid from the American Heart Association to L.F.L.F.J. Mangiacapra and M.E. Fransen contributed equally to this work     

Compensatory responses and development of the nodose ganglion following ablation of placodal precursors in the embryonic chick (Gallus domesticus)

The nodose ganglion is the distal cranial ganglion of the vagus nerve which provides sensory innervation to the heart and other viscera. In this study, removal of the neuronal precursors which normally populate the right nodose ganglion was accomplished by ablating the right nodese placode in stage 9 chick embryos. Subsequent histological evaluation showed that in 54% of lesioned embryos surviving to day 6, the right ganglion was absent. Most embryos surviving to day 12, however, had identifiable right ganglia. In day 12 embryos, the right ganglion which developed was abnormal, with ganglion volume and ganglion cell diameter reduced by 50% and 20%, respectively, compared to control ganglia. To investigate the source of the neuron population in the regenerated ganglion, we combined nodose placode ablation with bilateral replacement of chick with quail cardiac neural crest (from mid-otic placode to somite 3). These cells normally provide only non-neuronal cells to the nodose ganglion, but produce neurons in other regions. At day 9, quail-derived neurons were identified in the right nodose ganglia of these chimeras, indicating that cardiac neural crest cells can generate neurons in the ganglion when placode-derived neurons are absent or reduced in number. On the other hand, we found that sympathetic neural crest (from somites 10 to 20) does not support ganglion development, suggesting that only neural crest cells normally present in the ganglion participate in reconstituting its neuronal population. Our previous work has shown that right nodose placode ablation produces abnormal cardiac function, which mimics a life-threatening human heart condition known as long QT syndrome. The present results suggest that the presence of neural crest-derived neurons in the developing right nodose ganglion may contribute to the functional abnormality in long QT syndrome.This work was supported by grant PO1 HL 36059     

Exceptional preservation in Lower Devonian coalified fossils from the Welsh Borderland: a new genus based on reniform sporangia lacking thickened borders

Reniform sporangia, comprising two equal valves and containing retusoid spores, recovered from Lower Old Red Sandstone strata of Devonian age ( micrornatus-newportensis Spore Biozone: lower Gedinnian lower Lochkovian) on North Brown Clee Hill in the Welsh Borderland are placed in Resilitheca salopensis gen. et sp. nov. Conventional compression fossils from Targrove, Ludlow of fertile axes showing isotomous branching with limited overtopping are considered conspecific because the terminal reniform sporangia contain the same spores. Spore ultrastructure is described using scanning and transmission electron microscopy. Sections show faint traces of lamellae. Particles associated with spores and sporangium wall are compared with the globules of pteridophytes and Ubisch bodies of angiosperms, and related to the development of the sporangium. The new plants are compared with Cooksonia caledonica Edwards known only from impressions, and with Renalia Gensel showing far more pronounced pseudomonopodial branching.     

Relations between development and regeneration of tadpole spinal cord

Summary 1. The developing spinal cords of bullfrogs and transected cords of stage IV tadpoles were subjected to two-dimensional gel electrophoresis and histological analysis. During development, the level of actin,-tubulin or-tubulin in the 7–10th spinal segments increased with time and reached a maximum around stage XIII followed by a decrease, as shown from quantitative assay on protein spots of 2-dimensional gels of cord homogenates. In contrast, the level of 68 kD neurofilament subunit (NF68) was low in tadpoles but high in frog.2. Following a complete transection made at the level of the 8th spinal segment, the cord tissue of the lesion zone degenerated; regeneration from each cut end then occurred, which lengthened for approximate 0.35 mm by 28 days after transection. The content of actin,-tubulin and-tubulin in the cord within 1–2 mm of the transection site was elevated to 124–192% of control values 7–28 days post-transection, whereas NF68 declined to near non-detectable extent.3. The regeneration of each cord stump included outgrowth of neuroepithelial cells and nerve fibers, reconstituting a newly regenerated cord segment. Ultrastructural examination revealed that features of the regrowth of fibers and guidance of neuroepithelial cells to the axonal growth resembled that seen in the developing cord. Thus the biochemical and morphological data support that the regeneration of the nervous system recaptulates its developmental events, providing evidence for molecular mechanisms underlying central axonal regeneration.     

Anatomy and ultrastructure of the sporophyte of Takakia ceratophylla (Bryophyta)

In this study, morphogenesis and structure of the sporophyte of Takakia ceratophylla are characterized beginning with the late embryo and culminating in the fully dehisced capsule. Information is presented on the development, ultrastructure, and anatomy of the three organographic regions of the sporophyte, namely capsule, seta, and foot. Diagnostic features that identify Takakia as a moss include the gradual elongation of seta, persistence of an apical calyptra, expansion of the capsule after cessation of seta elongation, existence of a columella, monoplastidic meiosis, spore ultrastructure (including a perine layer deposited late in spore wall (development), and the structure of the foot. Commonalities with the capsule of the Andreaeopsida include sporogenous tissue that overarchs a central columella, absence of stomata, and lack of a peristome and operculum. Peculiarities of the genus are seen in the internal structure of the capsule, the disintegration of the columella with spore maturation, and the dehiscence of the capsule along a single, spiralled, longitudinal suture line. Passive spore dispersal through longitudinal splitting of the capsule occurs in andreaeopsid mosses, liverworts, hornworts, and seedless vascular plants. The precise mechanism of dehiscence along a spiralled suture is unparalleled in extant archegoniates but finds counterparts in ancestral land plants such as the pteridophyte Tortilicaulis.