Aminergic innervation of the blood vessels of Octopus vulgaris

Evidence is presented from fluorescence histochemistry studies that blood vessels in the viscera of octopus vulgaris are innervated by nerve fibres containing catecholamines. This, with other evidence, suggests that cephalopods, like vertebrates, may be capable of regulating their peripheral vasculature by central neural control.     

Measurement of gas exchange rates in plant tissue culture vessels

The aerial microenvironment in culture vessels has a significant effect on the growth and development of plantlets in vitro. Since the gas exchange between outside air and inner air can influence the microenvironment of culture vessel, it is necessary to measure the air exchange rate for various vessels. In this study, water vapor was used as the tracer gas, and the change of absolute humidity inside the vessel was calculated continuously by the measured values of a relative humidity sensing element. The outside environment was maintained at constant humidity level by a saturated salt solution. The RH data were transformed into absolute humidity and the specific humidity ratio. The air exchange rates of several tissue culture vessels were then calculated. The exchange rate was between 0.0145 h^–1 to 0.0376 h^–1. This technique provides an inexpensive, rapid and simple way to determine the air exchange rate of a culture vessel within a short period. The effects of the air current velocities on the exchange rates of vessels were also determined.     

Ontogeny of tracheal dimensions and gas exchange capacities in the grasshopper, Schistocerca americana

How does body size affect the structure and gas exchange capacities of insect tracheae? Do insects become more oxygen-limited as they grow? We addressed these questions by measuring the dimensions of two transverse tracheae within the abdomen of American locusts of different ages, and evaluating the potential for diffusion or convection to provide adequate gas exchange. The grasshopper abdomen has longitudinal tracheae that run along the midgut, heart, nerve cord, and lateral body wall. Transverse tracheae run from each spiracle to the longitudinal tracheae. Dorsal air sacs attach near each spiracle. In both transverse tracheae studied, diffusive capacities increased more slowly than metabolic rates with age, and calculated oxygen gradients necessary to supply oxygen by diffusion increased exponentially with age. However, surgical studies demonstrated that transport of gas through these transverse tracheae occurred by convection, at least in adults. Convective capacities paralleled metabolic rates with age, and the calculated pressure gradients required to sustain oxygen consumption rates by convection were independent of age. Thus, in growing grasshoppers, tracheal capacities matched tissue oxygen needs. Our morphological and physiological data together suggest that use of convection allows older grasshoppers to overcome potential limitations on size imposed by diffusion through tracheal systems.     

The effects of tracheid dimensions on variations in maximum density of Picea glehnii and relationships to climatic factors

An investigation was made of the effects of tracheid dimensions on variations in the maximum density of Picea glehnii Mast., which were associated with climatic changes. Radial cell diameter and the thickness of the tangential cell walls of the last-formed cells in 90 annual rings of nine trees with different annual ring widths were analyzed by image analysis. Correlations between maximum density and tracheid dimensions indicated that changes in maximum density were due mainly to changes in cell wall thickness of the last-formed cells in annual rings and were not due to changes in radial cell diameter. The effects of climatic factors on tracheid dimensions were examined by application of dendroclimatological techniques. A chronology of cell wall thickness that represented common signals among trees was established. Simple correlation and response function analyses of the chronology revealed that cell wall thickness was influenced positively by summer temperature and negatively by precipitation in August, and these responses were similar to those of maximum density. The study demonstrated that variations in maximum density were due to variations in the cell wall thickness of the last-formed cells, which varied depending on the weather in summer. Received: 8 February 1999 / Accepted: 7 October 1999     

The Morphology and Adhesion Mechanism of Octopus vulgaris Suckers

The octopus sucker represents a fascinating natural system performing adhesion on different terrains and substrates. Octopuses use suckers to anchor the body to the substrate or to grasp, investigate and manipulate objects, just to mention a few of their functions. Our study focuses on the morphology and adhesion mechanism of suckers in Octopus vulgaris. We use three different techniques (MRI, ultrasonography, and histology) and a 3D reconstruction approach to contribute knowledge on both morphology and functionality of the sucker structure in O. vulgaris. The results of our investigation are two-fold. First, we observe some morphological differences with respect to the octopus species previously studied (i.e., Octopus joubini, Octopus maya, Octopus bimaculoides/bimaculatus and Eledone cirrosa). In particular, in O. vulgaris the acetabular chamber, that is a hollow spherical cavity in other octopuses, shows an ellipsoidal cavity which roof has an important protuberance with surface roughness. Second, based on our findings, we propose a hypothesis on the sucker adhesion mechanism in O. vulgaris. We hypothesize that the process of continuous adhesion is achieved by sealing the orifice between acetabulum and infundibulum portions via the acetabular protuberance. We suggest this to take place while the infundibular part achieves a completely flat shape; and, by sustaining adhesion through preservation of sucker configuration. In vivo ultrasonographic recordings support our proposed adhesion model by showing the sucker in action. Such an underlying physical mechanism offers innovative potential cues for developing bioinspired artificial adhesion systems. Furthermore, we think that it could possibly represent a useful approach in order to investigate any potential difference in the ecology and in the performance of adhesion by different species.     

Octopus photoreceptor membranes. Surface charge density and pK of the Schiff base of the pigments.

The chromophore of octopus rhodopsin is 11-cis retinal, linked via a protonated Schiff base to the protein backbone. Its stable photoproduct, metarhodopsin, has all-trans retinal as its chromphore. The Schiff base of acid metarhodopsin (lambda max = 510 nm) is protonated, whereas that of alkaline metarhodopsin (lambda max = 376 nm) is unprotonated. Metarhodopsin in photoreceptor membranes was titrated and the apparent pK of the Schiff base was measured at different ionic strengths. From these salt-dependent pKs the surface charge density of the octopus photoreceptor membranes and the intrinsic Schiff base pK of metarhodopsin were obtained. The surface charge density is sigma = -1.6 +/- 0.1 electronic charges per 1,000 A2. Comparison of the measured surface charge density with values from octopus rhodopsin model structures suggests that the measured value is for the extracellular surface and so the Schiff base in metarhodopsin is freely accessible to protons from the extracellular side of the membrane. The intrinsic Schiff base pK of metarhodopsin is 8.44 +/- 0.12, whereas that of rhodopsin is found to be 10.65 +/- 0.10 in 4.0 M KCl. These pK values are significantly higher than the pK value around 7.0 for a retinal Schiff base in a polar solvent; we suggest that a plausible mechanism to increase the pK of the retinal pigments is the preorganization of their chromophore-binding sites. The preorganized site stabilizes the protonated Schiff base with respect to the unprotonated one. The difference in the pK for the octopus rhodopsin compared with metarhodopsin is attributed to the relative freedom of the latter's chromophore-binding site to rearrange itself after deprotonation of the Schiff base.     

The effects on shells of drilling by Octopus

The holes drilled by Octopus vulgaris are compared with structures concerned with drilling– the radula, the muscular salivary papilla, and the terminal process of the posterior salivary gland duct, and with the teeth which they all bear. Comparison of the shell surface after drilling and after fracture has shown that some chemical dissolucion of the shell occurs during drilling. Rasp marks found on the surface of drill holes have been shown to be due to the structural formation of the gastropod shell.     

Octopus Senescence: The Beginning of the End

Senescence is a normal stage of an octopus's life cycle that often occurs before death. Some of the following symptoms typify it: lack of feeding, retraction of skin around the eyes, uncoordinated movement, increased undirected activity, and white unhealing lesions on the body. There is inter- and intraspecific variability. Senescence is not a disease or a result of disease, although diseases can also be a symptom of it. Both males and females go through a senescent stage before dying-the males after mating, the females while brooding eggs and after the eggs hatch. There are many aspects of octopus senescence that have not yet been studied. This study discusses the ecological implications of senescence.     

The fine structure of the retinal plexus in Octopus vulgaris

Summary The plexiform layer of the octopus retina was investigated by means of electron microscopy. The axonic processes of visual cells contain closely packed microtubules, 300 Å in diameter, running parallel to one another along the long axis of the processes. Visual cells also send out a large number of thin axon collaterals. Each of them forms presynaptic knobs with numerous clear vesicles along their course. These are assumed to be concerned with the reciprocal communication between visual cells. Nerve endings with dense-cored vesicles form synaptic contacts with visual cells. The visual cells show some spherical protrusions into the perivascular spaces.Octopuses, captured off the coast of Onagawa (Miyagi-ken, Japan) in autumn of 1963 and 1964, were offered for this research through the kindness of Dr. Kyoji Tasaki, Assistant Professor of Physiology in Tohoku University School of Medicine.I wish to thank Professor Toshiyuki Yamamoto for his encouragement and suggestions throughout all stages of this work, and also Mr. Masae Kato for his technical assistance in drawing.     

Quantitative analysis of the density and pattern of adrenergic innervation of blood vessels

Summary Automated quantitative image analysis (QIAF) was used to measure and compare the adrenergic nerve plexuses of 4 blood vessels from the guinea pig, demonstrated by glyoxylic acid fluorescence (GAF). The results showed considerable quantitative variation of plexus density, size of bundles, and numbers of varicosities. A range of alternative procedural and anatomical sources of variability were investigated and assessed. The carotid artery was found to have a dense plexus with more nerves than that of the mesenteric artery; the mesenteric vein and abdominal aorta had sparse plexuses. The carotid artery plexus, despite the density of its nerves, possessed only half the number of varicosities of the mesenteric artery plexus. This sparse varicosity population was shown to have a similar density to the varicosities demonstrated by QIAF in the scattered nerves of the mesenteric vein and abdominal aorta. QIAF confirmed visual estimates of adrenergic plexus density, and was able to demonstrate less obvious differences of nerve density and size, and varicosity populations, between the different plexuses studied. The method is applicable to stretch preparations and transverse sections of many adrenergically innervated tissues.     

Immunohistochemical Study of Leucine-Enkephalin and Delta Opioid Receptor in Mantles and Feet of the Octopus Octopus ocellatus Gray

The study was supposed to determine whether leucine-enkephalin (leu-Enk) or delta opioid receptor was presented in mantles and feet of the Octopus Octopus ocellatus. The results showed that they were found in the mantles, wrists, interbrachial membrane, and funnels of the Octopus Octopus ocellatus. The immunoreactivities of leu-Enk and delta opioid receptor were observed in the epithelial cells of dorsal mantle, wrist and interbrachial membrane. Strong one was presented in both the connective tissue which was close to the epithelium of dorsal mantle, wrist and interbrachial membrane and a small amount of connective tissue cells and nerve fibers, while less positive in other parts. A little of immunoreactive material was also observed in the ventral mantle and funnel, for example, weak one was in the epithelial tissue, the adjacent cells and nerve fibers. The different densities of leu-Enk and delta opioid receptor in mantles and feet of the Octopus O. ocellatus may be related to the different functions.     

Ontogeny of tracheal dimensions and gas exchange capacities in the grasshopper, Schistocerca americana

How does body size affect the structure and gas exchange capacities of insect tracheae? Do insects become more oxygen-limited as they grow? We addressed these questions by measuring the dimensions of two transverse tracheae within the abdomen of American locusts of different ages, and evaluating the potential for diffusion or convection to provide adequate gas exchange. The grasshopper abdomen has longitudinal tracheae that run along the midgut, heart, nerve cord, and lateral body wall. Transverse tracheae run from each spiracle to the longitudinal tracheae. Dorsal air sacs attach near each spiracle. In both transverse tracheae studied, diffusive capacities increased more slowly than metabolic rates with age, and calculated oxygen gradients necessary to supply oxygen by diffusion increased exponentially with age. However, surgical studies demonstrated that transport of gas through these transverse tracheae occurred by convection, at least in adults. Convective capacities paralleled metabolic rates with age, and the calculated pressure gradients required to sustain oxygen consumption rates by convection were independent of age. Thus, in growing grasshoppers, tracheal capacities matched tissue oxygen needs. Our morphological and physiological data together suggest that use of convection allows older grasshoppers to overcome potential limitations on size imposed by diffusion through tracheal systems.     

Chemical exchange in two dimensions in the 1H NMR assignment of cytochrome c.

The important role played by chemical exchange in solving the proton assignment problem for oxidized and reduced horse cytochrome c is described. Some novel approaches for establishing oxidation-reduction exchange correlations in combinations of several two-dimensional spectra were used. Unambiguous chemical exchange correlations were established for 55 NH-C alpha H resonances and all the aromatic and side chain methyl resonances. Consistent although not fully unambiguous main chain proton correlations were observed for 47 of the remaining 49 residues. The many exchange correlations found serve to multiply cross-connect the two extensive, individually self-consistent networks of assignments found for the oxidized and reduced forms, and thus help to confirm both sets of assignments.     

Beak and radula growth in Octopus vulgaris

The dry weight and the crest length of the upper and lower beak, the length of the radula ribbon, the average width of the base of the six proximal and distal rachidian teeth as well as the total number of these teeth have all been related to the live body weight of octopuses between 1.1 and 4440 g. From any one of these parameters it is possible to estimate the size and approximate age of the animal.     

Influence of long-term experimental orthostatic body position on innervation density in extremity vessels

The aim of this study was to quantitate the density of nerve terminals as well as their synaptic vesicle population in the adventitia of saphenous (SV and SA) and brachial veins and arteries (BV and BA) obtained from rats maintained in a horizontal control or a tilted position. Adult animals were kept individually in tube-like cages in a 45 degrees head-up position. After 2 wk, both tilted and control animals were anesthetized, and the whole body was perfused with fixative solution at physiological pressure. Vessels segments were then excised for electron microscopy and immunohistochemistry. The nerve terminal density (NTD) of SA was 8.20 +/- 1.46 nerve terminals/100 microm(2) cross section of adventitia and that of SV was 4.53 +/- 0.61 nerve terminals/100 microm(2) cross section of adventitia in control rats. Tilting caused a significant increase in NTD of both SA (70%) and SV (52%). The synaptic microvesicle density (SyVD) was larger in SA than SV in control rats (30.48 +/- 4.41 vs. 13.38 +/- 2.61 synaptic vesicles/10 terminal sections), but tilting resulted in more pronounced changes in SyVD of SV (95%) than SA (54%). No significant changes in NTD and SyVD of BA were found after tilt (-3.6% relative to 4.99 +/- 0.33 compared with 0.4% relative to 24.89 +/- 3.7, respectively). Whereas NTD of BV exhibited a tendency to increase (3.73 +/- 0.86 vs. 2.31 +/- 0.29 nerve terminals/100 microm(2) cross section of adventitia), SyVD did not change significantly (18.96 +/- 2.74 vs. 22.85 +/- 3.17 synaptic vesicles/10 terminal sections). A large number of nerve terminals of all vessels were tyrosine hydroxylase immunoreactive (containing norepinephrine). These findings support the hypothesis that long-term gravitational load causes adaptive morphological and functional remodeling of sympathetic innervation in blood vessels of the extremities.     

Influence of long-term head-down body position on innervation density in extremity blood vessels

The aim of the present study was to quantitate and compare the density of nerve terminals (NTD), as well as of their synaptic vesicle population (SyVD) in saphenous and brachial vein and artery, obtained from rats maintained in the horizontal or head-down tilted (HDT) position for two weeks. The same technique was applied as that for the head-up tilt study.     

Effects of visual conditions and prey density on feeding kinetics of paralarvae of Octopus vulgaris from a laboratory spawning

Consumption rate and feeding success of newly hatched paralarvaeof the cephalopod Octopus vulgaris preying on Artemia larvaewere investigated in relation to visual conditions and preydensity. Each paralarva was tested individually using a small-scaleexperimental setup; consumption over one day was measured at20°C. A factorial experiment was designed to investigatethe effects on the consumption rate of two predictor variables:illumination/background (three levels: 7.5 Wm^–2 whitelight + white background, 7.5 Wm^–2 white light +blue background, darkness) and prey density (four levels: 2.35,4.70, 9.40 and 14.10 Artemia metanauplii ml^–1). Consumptionrate varied significantly between different conditions of illuminationand prey density. Light enhanced consumption rate, but differentbackgrounds yielded similar rates. The maximal consumption rateunder illumination was close to 16 Artemia paralarva^–1day^–1, and it was around 5 Artemia paralarva^–1 day^–1for assays in darkness. The predatory efficiency, measured asthe proportion of prey consumed, was significantly affectedby prey density, pointing to a type III functional response.The number of nonfeeding paralarvae was significantly higherin darkness and at low prey density. (Received 14 February 2006; accepted 23 December 2006)