The phosphoinositide PI(3,5)P2 mediates activation of mammalian but not plant TPC proteins: functional expression of endolysosomal channels in yeast and plant cells

Two-pore channel proteins (TPC) encode intracellular ion channels in both animals and plants. In mammalian cells, the two isoforms (TPC1 and TPC2) localize to the endo-lysosomal compartment, whereas the plant TPC1 protein is targeted to the membrane surrounding the large lytic vacuole. Although it is well established that plant TPC1 channels activate in a voltage- and calcium-dependent manner in vitro, there is still debate on their activation under physiological conditions. Likewise, the mode of animal TPC activation is heavily disputed between two camps favoring as activator either nicotinic acid adenine dinucleotide phosphate (NAADP) or the phosphoinositide PI(3,5)P₂. Here, we investigated TPC current responses to either of these second messengers by whole-vacuole patch-clamp experiments on isolated vacuoles of Arabidopsis thaliana. After expression in mesophyll protoplasts from Arabidopsis tpc1 knock-out plants, we detected the Arabidopsis TPC1-EGFP and human TPC2-EGFP fusion proteins at the membrane of the large central vacuole. Bath (cytosolic) application of either NAADP or PI(3,5)P₂ did not affect the voltage- and calcium-dependent characteristics of AtTPC1-EGFP. By contrast, PI(3,5)P₂ elicited large sodium currents in hTPC2-EGFP-containing vacuoles, while NAADP had no such effect. Analogous results were obtained when PI(3,5)P₂ was applied to hTPC2 expressed in baker’s yeast giant vacuoles. Our results underscore the fundamental differences in the mode of current activation and ion selectivity between animal and plant TPC proteins and corroborate the PI(3,5)P₂-mediated activation and Na⁺ selectivity of mammalian TPC2.     

Monoaminergic innervation of the pulmonary vessels in various laboratory animals (rat,rabbit, cat)

Zusammenfassung Mit Hilfe der histochemischen Fluoreszenzmethode nachFalck ³ wurde die monoaminergische Innervation der Lungengefässe bei der Ratte, beim Kaninchen und bei der Katze untersucht. Während die Pulmonalarterie der Ratte keine Nervenendigungen aufweist, wurden solche an der Oberfläche der Media festgestellt. Bronchial- und Pulmonalarterien der Katze besitzen die monoaminergische Innervation nur an der Oberfläche der Media. Im Unterschied zur Ratte besitzt das Kaninchen eine typisch muskuläre Pulmonalarterie und reiche monoaminergische Innervation an der Oberfläche der Media.     

Long-term administration of a substance P antagonist, (D-Pro2, D-Trp7,9)-SP,abolishes the response to ocular trauma

Summary The long-term effect of ocular administration of a substance P (SP) antagonist, (D-Pro², D-Trp^7,9)-SP, was studied in the rabbit. After 2–3 months of topical administration of the antagonist twice daily, a mild trauma was applied in the form of infrared irradiation of the iris. The control eye responded with disruption of the blood-aqueous barrier, the eye treated with the antagonist did not. Two days after termination of treatment, the response to ocular injury was still reduced. Another 2 days later, ocular injury evoked a normal response, which shows that the protection was reversible. No adverse reaction to the SP antagonist was noted.     

Sexual dimorphism in the lipid fraction of the defensive secretion ofGastrophysa viridula (Coleoptera: Chrysomelidae)

The lipid fraction of the defensive secretion of adultGastrophysa viridula was analyzed by gas-liquid chromatography and gas chromatography-mass spectrometry. Three long chain acetates and two methyl-branched hydrocarbons were identified and their amounts in the secretion of single field-collected and laboratory-reared beetles determined. The composition of the secretion was affected by the sex, reproductive status, and age of the beetle. The ratio of (Z)-11-eicosenyl acetate to (Z)-13-docosenyl acetate ranged from 0.30 to 0.78 in male beetles and from 1.05 to 2.06 in female beetles. The biological significance of this sexual dimorphism is discussed.     

Insect photoperiodism: various ways of regulating univoltinism in lacewings (Planipennia: Chrysopidae)

Summary Reproduction inTjederina gracilis adults depends on short-day followed by long-day conditions. InNineta pallida, development of overwintering first-instar larvae is retarded by short days; later on, that of second-and third-instar by long days.Nineta flava andChrysopa perla prepupae automatically enter diapause, but inN. flava, photoperiod regulates diapause eermination, and long days may retard egg laying     

On the multiple sex chromosome mechanism in a lygaeid,Oxycarenus hyalinipennis (Costa)

Résumé Oxycarenus hyalinipennis (Costa) possède vraisemblablement des chromosomes sexuels multiples du typeX ₁ X ₂ Y. Les métaphases diploïdes renferment, soit 17, soit 19 éléments. Les 2 éléments supplémentaires des cinèses à 19, bien que comparables auxm-chromosomes des autres Lygéides, diffèrent de ces derniers en certains points de leur comportement qui évoque celui des «surnuméraires» trouvés chez d'autres insectes. La méiose est du type habituel chez les Hétéroptères.     

Chromosomal translocations in 2 Italian populations ofReticulitermes lucifugus (Rossi) (Insecta,Isoptera: Rhinotermitidae)

Summary The chromosomes of 2 Italian populations ofReticulitermes lucifugus (Rossi) were studied in both spermatogonial and oogonial divisions. The diploid chromosome complement is 2=42. All the first meiotic metaphases in Sardinian males show a complex translocation arranged in chains of 4 bivalents. The Apulian males show either 21 bivalents or 19 bivalents and a chain of 2.The authors express their thanks to Prof. A. Springhetti for his kind help in supplying the materials and for criticism in preparation of this paper, and to Prof. G. Colombo for valuable advice and criticism.     

Is the variation in the susceptibility of various species to atherosclerosis due to inborn differences in prostacyclin (PGI2) formation?

Summary Species exhibiting a higher susceptibility to the development of atherosclerosis have a reduced prostacyclin (PGI₂)-generation in the arterial wall, which differs in various parts of the vascular system. As the difference in PGI₂-formation in various diseases is a generalized vascular effect, the changes can be detected in all vessels. This is a very important point for diagnostic purposes in humans.This study was supported by a grant of the «Fonds 600 Jahre Wiener Universität der Kammer der gewerblichen Wirtschaft für Wien».     

Evidence of urinary glomerular basement membrane (GBM) antigens excretion in 3 rodents (rat, mouse, and guinea-pig) (author's transl)]

Immunodiffusion technique, with rabbits antibodies against rat glomerular basement membrane (GBM), permits to evidence the presence of GBM antigens into normal urines of 3 rodents (rat, mouse, and guinea-pig). These results confirm the earlier works in human and rabbit urines and show the antigenic communauty existing between the 3 rodents GBM, since we can evidence antigens of 3 different mammals with the same antibody. The origin and the nature of this patterns and the signification of this presence into mammalian urines are discussed.     

The probable significance of the differential occurrence of protein in various castes of the termiteOdontotermes assmuthi (Isoptera: Termitidae)

Résumé L'évaluation de la somme de protéine contenue dans les représentants des diverses castes du termiteOdontotermes assmuthi donne les résultats suivants: 1. Les formes reproductrices potentielles, mâles et femelles, ont des quantités significatives (47,9 et 37%) que requirèrent leur activité reproductrice. 2. Les rois issus de mâles ailés n'atteignant leur pourcent maximum de protéine qu'à l'âge adulte. 3. Les reines en augmentent leur quantité (à 65,7%) en proportion de leur intense production d'ufs. 4. Les ouvriers et les soldats soumis à des tâches ardues contiennent beaucoup de protéine (33,5 et 44,5%).

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Acknowledgment. The authors are indebted to ProfessorC. J. George (C.S.I.R.) for his valuable guidance and thanks to ProfessorM. Appaswamy Rao, for the facilities provided.