Processing and Secretion of Lysosomal Acid α-Glucosidase In Tetrahymena Wild Type and Secretion-Deficient Mutant Cells

ABSTRACT. The proteolytic processing and secretion of a lysosomal enzyme, acid α-glucosidase, was studied by pulse-chase labeling with [³⁵S]methionine in Tetrahymena thermophila CU-399 cells treated with ammonium chloride. This cell secreted a large amount of acid α-glucosidase into the cultured medium during starvation. the secretion was found to be repressed by addition of ammonium chloride (NH₄Cl). Acid α-glucosidase was produced as a precursor form (108 kDa) and then processed to a mature polypeptide (105 kDa) within 60 min. This mature enzyme was secreted into the media within 2-3 h after chase, whereas the precursor form was not secreted by either control cells or NH₄Cl-treated cells. NH₄Cl did not affect the processing of the precursor acid α-glucosidase. Processing profile of this enzyme was apparently indistinguishable from that of the mutant MS-1 defective in lysosomal enzyme secretion. Furthermore, the purified extracellular (CU-399) and intracellular (MS-1) acid a-glucosidases were the same in molecular mass (105 kDa) and enzymatic properties. They contained no mannose 6-phosphate residues in N-linked oligosaccharides. These results suggested that unlike mammalian cells, Tetrahymena acid α-glucosidase may be transferred to lysosomes by a mannose 6-phosphate receptor-independent mechanism, and also that low pH was not essential for the proteolytic processing of precursor polypeptide.     

PHYSIOLOGICAL ACTIVITIES OF HELMINTHOSPOROL AND HELMINTHOSPORIC ACID: I. EFFECTS ON GROWTH OF INTACT PLANTS

Helminthosporol (H-ol) and helminthosporic acid (H-acid) wereeffective in promoting elongation of leaf sheaths of rice, Japanesebarnyard grass and dwarf maize (d-2 and d-5) and of hypocotylsof taisai (Brassica chinensis), but inactive in leaf sheathsof oat and wheat, hypocotyls of sesame and morning glory (Pharbitisnil) and epicotyls of Pharbitis and dwarf and tall peas. Onthe elongation of the leaf sheath of maize d-1, H-ol was promotivebut the activity of H-acid was doubtful. On hypocotyls of lettuceand daikon (Raphanus sativus), only H-acid was active. Multiplicationrate and size of fronds of Lemna perpusila were not affectedby either of the substances. Compared with gibberellic acid for the effect on the shoot growth,H-ol and H-acid were weak in activity and narrower in the scopeof plants that responded. H-ol and H-acid characteristicallypromoted the elongation of the primary root. Comparative effectivenessof H-ol and H-acid varied with plant species or parts examined. ¹ This study was supported in part by grant-in aid of the Ministryof Education (No. 0417). The results reported here were presentedat the Annual Meeting of the Botanical Society of Japan at Kanazawain 1964 (S).     

Respiratory nervous activity in the isolated nerve cord of the larval dragonfly, and location of the respiratory oscillator

ABSTRACT. Rhythmic respiratory nerve activity was recorded in the dragonfly larvae, Anax parthenope Julius Brauer (Anisoptera). Alternating expiratory and inspiratory bursts of spikes occurred in abdominal nerve cords isolated from all peripheral connections. These bursts are similar to the activity recorded in semi-intact preparations, suggesting that the respiratory rhythm can be generated without peripheral sensory feedback. Expiratory bursts started and ended at the same time in different segments of semi-intact preparations. When connectives were severed, the nerve cord separated from the last abdominal ganglion did not normally show rhythmic bursts; the last ganglion alone and the nerve cord connected to the last ganglion exhibited the rhythmic bursts. However, in a few cases the nerve cord separated from the last ganglion exhibited the rhythm. The results suggest that the last ganglion contains the main oscillator, but that other weak oscillators occur elsewhere.     

A comparative study on insemination dependency of circadian activity pattern in mosquitoes

Abstract. Two forms of mosquitoes in the Culex pipiens complex, C.p.pallens and C.p.molestus , were compared for the insemination-dependency of entrained female circadian activity in LD 16:8h. When virgin, Pallens is nocturnal and molestus is, in addition, diurnal. Insemination and injection of extracts of the male accessory gland affects virgin activity in that nocturnality is strengthened in both mosquitoes, and the diurnal activity of molestus is depressed. This was so whether the treatments were intra- or interspecific. These findings are compared with those already published for other mosquitoes. A coherent view is proposed that the mosquito, nocturnal/crepuscular, nocturnal and diurnal, or diurnal, has potentially a similar entrained waveform which is affected by insemination in a common way, and by lighting conditions in species specific ways.     

Sensory Responses of Reinnervated Muscle Spindles

EXTENSIVE morphological, histochemical and electrophysiological studies have been made on skeletal muscles at varying stages of motor reinnervation. But in spite of the physiological significance of muscle receptors in controlling skeletal muscle activity, there have been no detailed studies of their reinnervation. We have therefore investigated the functional and structural characteristics of muscle spindles associated with their sensory reinnervation.     

Extraorgan freezing in wintering flower buds of Cornus officinalis Sieb. et Zucc.

Abstract. Extraorgan freezing as a mechanism for increasing cold hardiness was shown using flower buds of Cornus officinalis Sieb. et Zucc. Differential thermal analysis (DTA) revealed that florets in flower buds of C. officinalis owed their cold hardiness to deep supercooling and also that slower cooling rates increased the supercooling ability of florets. During slow stepwise cooling (5°C h⁻¹), the water content of florets decreased and that of scales (involucral bracts) increased, which resulted in accumulation of ice within the scales. This was more extensive in early winter and early spring buds than mid-winter ones. Flower buds with silicone oil in the space between florets and scales also showed a similar decrease in water content of florets and an increase in that of scales. This indicated that water migration from the florets to the scales probably took place by way of the peduncles and the receptacle, possibly through their vascular traces, and not directly from the surface of the florets to the ice sink in the form of vapour. Possible mechanisms of extraorgan freezing are postulated along with this finding.