Effects of Organic Solvents on the Coupling Between ATP Hydrolysis and Sliding of Microtubules in Axonemes from Chlamydomonas Flagella

ABSTRACT. The effects of organic solvents on the ATPase activity and the sliding disintegration of axonemes from Chlamydomonas were investigated. The axonemal ATPase was markedly activated by methanol accompanying with marked inhibition of the sliding disintegration of axonemes. On the contrary, glycerol inhibited the ATPase activity without serious inhibition of the sliding disintegration. As far as the axonemes are not irreversibly denatured by extremely high concentration of solvents, the effects of solvents both on the ATPase and the ability of sliding are reversible. Therefore, the inhibition of sliding accompanied by the activation of ATPase is probably due to an inability to couple the hydrolysis of ATP to sliding between dynein and microtubule in the presence of methanol. The axonemal ATPase was less sensitive to vanadate inhibition after exposure to methanol. This indicates that methanol makes the dyneinADP.Pi complex unstable and increases product release. On the other hand, glycerol and ethylene glycol seem to stabilize the force generation responsible for the sliding through stabilizing the dynein.ADP.Pi complex.     

TRANSPORT OF CATIONS FROM COTYLEDON TO SEEDLING OF THE EMBRYONIC PLANTS OF VIGNA SESQUIPEDALIS

Decrease of dry material, water, potassium and divalent cationsin the cotyledon (catabolic organ and accumulation of thesesubstances in the seedling (anabolic organ) were traced duringthe germination stage of Vigna sesquipedalis, cultured at 15°,25° and 30° in the dark on the sand with the supplyof tap water. One half of the divalent cations finally accumulatedin the seedling came from the root, whereas most of the potassiumfrom the cotyledon. Practically all the calcium in the cotyledonremained untransported, while 90% of K stored in the cotyledonwas transported into seedling. As for divalent cations otherthan Ca, 50–60% of the total divalent cations was transportable.The concentration of K, transportable dry material and transportabledivalent cations decreased linearly with the culture age. Fromthe temperature dependency of the rate of this decrease, itwas inferred that the transport of cations from cotyledon toseedling involved at least two component processes having activationenergies of approximately 5 and 16 Kcal, respectively. The rateof transport at 30° was found to be 10–20 times greaterthan that calculated from the simple diffusion model in water.Fractionation of cell inclusions in the cotyledon by the useof a centrifuge (1,500 and 16,000 rpm) suggested that the amountsof ions in the precipitable fractions, being bound to certaincell components, precisely corresponded to the untransportablefraction. ¹Main part of this work was reported at the Annual Meeting ofthe Botanical Society of Japan held at Fukuoka on Oct. 25. 1958. (Received October 16, 1961; )     

A new method in growth-electrophysiology: Pressurized intra-organ perfusion

Abstract A new experimental system was devised for the simultaneous measurement of elongation rate and the activity of the spatially separate electrogenic ion pumps of a hypocotyl segment excised from a seedling of Vigna unguiculata L. Walp. under enforced intra-organ perfusion by artificial solutions. The pathway of the perfusion medium was apoplastic space, including xylem vessels as main routes. The elongation rate of the segment was highly dependent on the perfusion pressure applied. It was possible to increase the growth rate under pressurized perfusion by 10-30 times as much as that without perfusion. Elongation rate was also dependent on respiration under perfusion, being retarded reversibly by anoxia a few minutes after the activities of the electrogenic ion pumps were stopped. Perfusion pressure had a little influence on the membrane potential (V_px) below a breakdown level (c. 130 kPa). Perfusion of mannitol or sorbitol solution of appropriate concentration reduced the elongation rate reversibly.     

Spontaneous Membrane Potential Changes Associated With the Zooid and Vacuolar Contractions In Vorticella Convallaria

SYNOPSIS Interacellular membrane potential and spontaneous changes associated with motile responses in the zooid of Vorticella convallaria Linnaeus were recorded by conventional electrophysiological technics. an all-or-none large transient depolarization (large pulse) occurs in association with a spontaneous contraction of the zooid. A small transient potential change (small pulse) was observed in association with periodic contraction of the contractile vacuole.     

Morphological variation in relation to flower use in bumblebees

To understand resource partitioning in a bumblebee community, we analyzed various morphological characters. A total of 1269 individuals of six bumblebee species, Bombus ardens, B. hypocrita, B. diversus, B. ignitus, B. honshuensis and B. beaticola, were examined and principal component analysis showed that the bumblebee species were clearly differentiated. Glossa, prementa and head lengths were positively correlated with the second component, and a longer proboscis was associated with a narrower body, which may help bees to intrude into and access deep‐lying nectar sources. Bombus diversus, with a long proboscis and narrow body, preferred flowers with a long corolla tube, whereas B. hypocrita and B. ignitus, which have short proboscises and wide bodies, visited flowers with short corollas or dish‐shaped flowers. Two pairs of consubgeneric species that have similar morphological characteristics, B. ardens and B. beaticola, and B. hypocrita and B. ignitus, divided flower resources by habitat selection and seasonal partitioning. For resource partitioning among bumblebee species, not only morphology but also other factors, such as habitat and seasonal preference, flower use, foraging behavior, and interspecific interactions, are responsible.     

Antimelanoma Activity of Chloroquine,an Antimalarial Agent With High Affinity for Melanin

The antimalarial agent chloroquine is known for high affinity for melanin. This 4-aminoquinoline derivative was examined for anti-melanoma activity and uptake into melanoma cells. Chloroquine inhibited growth of cultured melanoma cells; the effect was much greater to a moderately pigmented cell line HMV-II than to a nonpigmented HMV-I. Treatment with chloroquine at a dose of 62 mg/kg i.p. for 12 days prolonged by 71% the life span of mice bearing B16 melanoma, while 24-day treatment at 31 mg/kg resulted in a 81% increase in life span. HMV-II cells showed a two-fold increase in up-take of chloroquine as compared with HMV-I cells. Chloroquine, 24 hr after administration to mice implanted s.c. with B16 melanoma, was selectively accumulated in the pigmented tissues, melanoma and eyes. Other nonpigmented tissues such as the liver, lung, and kidney showed rapid uptake (within 1 hr) and release. These results suggest that chloroquine is toxic to pigmented melanoma cells, the process being partly mediated by binding to melanin     

Effects of channelisation on stream habitat in relation to a spate and flow refugia for macroinvertebrates in northern Japan

1. The effects of channelisation on macroinvertebrates were examined in relation to a spate and flow refugia. Habitat components that can function as flow refugia were identified in a small, low‐gradient stream in northern Hokkaido, Japan. 2. Macroinvertebrates and their habitat characteristics (depth, current velocity and substratum) were sampled and measured in natural and channelised sections on three occasions: before, during and immediately after a spate. For macroinvertebrate sampling and habitat measurements, five (riffle, glide, pool, backwater and inundated habitats) and three (channelised‐mid, channelised‐edge and inundated habitats) habitat types were classified in the natural and channelised section, respectively. 3. The rate of velocity increase with discharge was compared among habitat types to determine which habitat types were less affected by increased discharge. The rate was the highest in riffles followed by glides and channelised‐mids. Backwaters maintained low current velocity even at high flow. In addition, current velocity in both natural and channelised inundated habitats was low relative to other habitat types during the spate. 4. Through the spate, total density of macroinvertebrates in channelised‐mids and taxon richness in both channelised‐mids and edges decreased. In the natural section, however, such a significant decrease was not found except for taxon richness in pools. This indicated that the spate had a greater impact on assemblages in the channelised section. Riffle assemblages exhibited a rapid recovery immediately after the spate, suggesting the existence of flow refugia in the natural section. Among the habitat types we examined, backwaters and inundated habitats appeared to have acted as flow refugia, because these habitats accumulated macroinvertebrates during the spate. 5. The lower persistence of the macroinvertebrate assemblage in the channelised section was attributable to the lower availability of flow refugia such as backwaters and inundated habitats. Our results emphasised the importance of considering flow fluctuations and refugia in assessing the effects of channelisation. In addition, the lateral heterogeneity of stream channels should be considered in stream restoration and management.     

Distribution of electric potential and ion transport in the hypocotyl of Vigna sesquipedalis I. Distribution of overall ion concentration and the role of hydrogen ion in generation of potential difference

Overall concentration of free inorganic ions distributes inthe hypocotyl of a bean seedling {Vigna sesquipedalis) at aconstant level (H⁺) or decreases monotonously from the cotyledonarynode towards the base (K⁺, Na⁺, Ca⁺⁺ and Mg⁺⁺, phosphate, NO₃^–).According to our theory, this is inconsistent with the distributionof electric potential having a definite minimum in the elongatingregion. The discrepancy can not be explained by regional variancein radial potential difference or histological differentiationin passive ionic permeability of the cell membrane. Short circuitcurrent observed through a hypocotyl segment corresponded toa net flux in ions of 10–24 pEq/cm².sec. It is questionable,however, whether this is due to active ion transport, whichcan be the source of electric potential difference, or is apassive flow due to histological heterogeneity in ion concentration. In order to investigate the latter possibility, pH of sap exudingfrom stumps made at various intervals along the hypocotyl axiswas measured, since H⁺ is the ion electro-osmotically most effective.pH Values of acropetal exudates distributed along the axis closelycorresponding to the distribution of electric potential. Thissuggests that potential distribution is determined by a passiveflow of H⁺ through a specific channel in the vascular system.The fact that H⁺ production and the uptake of ions and waterare most active at the elongating zone of hypocotyl is discussedfrom a physiological point of view. (Received December 3, 1969; )