Homologous recombination-mediated repair of DNA double-strand breaks operates in mammalian mitochondria

Mitochondrial DNA is frequently exposed to oxidative damage, as compared to nuclear DNA. Previously, we have shown that while microhomology-mediated end joining can account for DNA deletions in mitochondria, classical nonhomologous DNA end joining, the predominant double-strand break (DSB) repair pathway in nucleus, is undetectable. In the present study, we investigated the presence of homologous recombination (HR) in mitochondria to maintain its genomic integrity. Biochemical studies revealed that HR-mediated repair of DSBs is more efficient in the mitochondria of testes as compared to that of brain, kidney and spleen. Interestingly, a significant increase in the efficiency of HR was observed when a DSB was introduced. Analyses of the clones suggest that most of the recombinants were generated through reciprocal exchange, while ~ 30% of recombinants were due to gene conversion in testicular extracts. Colocalization and immunoblotting studies showed the presence of RAD51 and MRN complex proteins in the mitochondria and immunodepletion of MRE11, RAD51 or NIBRIN suppressed the HR-mediated repair. Thus, our results reveal importance of homologous recombination in the maintenance of mitochondrial genome stability.     

Dynamics of proteins in Golgi membranes: comparisons between mammalian and plant cells highlighted by photobleaching techniques

In less than a decade the green fluorescent protein (GFP) has become one of the most popular tools for cell biologists for the study of dynamic processes in vivo. GFP has revolutionised the scientific approach for the study of vital organelles, such as the Golgi apparatus. As Golgi proteins can be tagged with GFP, in most cases without altering their targeting and function, it is a great substitute to conventional dyes used in the past to highlight this compartment. In this review, we cover the application of GFP and its spectral derivatives in the study of Golgi dynamics in mammalian and plant cells. In particular, we focus on the technique of selective photobleaching known as fluorescence recovery after photobleaching, which has successfully shed light on essential differences in the biology of the Golgi apparatus in mammalian and plant cells.     

Induction of neovascularization in vivo by glycerol

Glycerol, injected into a site between the femoral vessels of the rat, induced neovascularization, both from the preexisting microcirculation and from the side of the femoral vein facing the artery-vein interstitium where the glycerol was administered. The use of glycerol together with a known angiogenic substance (PGE₂) did not modify the neocapillary density (NCD) obtained with glycerol alone. In contrast, the lower level of NCD achieved with an acylglycerol (triacetylglycerol) was increased when the latter was associated with PGE₂. Values reached were similar to, but never higher than, those for glycerol alone, or combined with PGE₂. The results suggest that glycerol and some substances containing glycerol, amongst which 1-butyrylglycerol has been previously considered¹, may stimulate angiogenesis by a direct or indirect mechanism of action.     

Induction of the formation of a coli-bacterio-phage and colicin by hydroperoxide

Zusammenfassung Die Arbeit gibt die Resultate der Induktion der Lysogenie und der Colicinogenie im selben StammeE. coli (L II-18) mit Wasserstoffperoxid an. Der Phagentiter wurde 35mal, der Colicintiter 7mal erhöht. Die optimale Konzentration des H₂O₂ für Phagenbildung warM/500, die für ColicinbildungM/2000. Im Vergleich mit der Induktion desselben Stammes mit UV-Licht ist die Wirkung des H₂O₂ verhältnismässig schwach.     

Induction of morphological changes in bacteria by optochin

Zusammenfassung Es wird die Wirkung des Optochins auf die Zellmorphologie desBacterium anitratum beschrieben und aus Versuchsergebnissen geschlossen, dass Optochin zur Gruppe chemischer Substanzen gehört, die zu hochgradigen morphologischen Veränderungen der Bakterienzelle führen.     

Uptake by mammalian cells of nucleic acids combined with a basic protein

Résumé Les acides nucléiques se combinent avec l'albumine méthylée et forment des complexes qui ne sont pas hydrolysés par les nucléases, mais dissociés par des ions inorganiques. Ces nucléoprotéines pénètrent dans les cellules en quantité beaucoup plus élevée que les acides nucléiques correspondants, et contrairement à ceux-ci, elles ne sont pas catabolisées. La combinaison avec l'albumine méthylée rend inactif l'acide ribonucléique viral: les ions inorganiques, et non les cellules vivantes, sont capables de dissocier ce complexe et de libérer l'ARN infectieux.     

Induction of uncoiled chromosomes by vibration

Summary Chromatin condensation during metaphase can be removed by simple vibration of metaphase cells prior to fixation. Uncoiled chromosome arms consist of long threads with dense regions at irregular distances each from the other.Acknowledgment. This work is supported by the Hellenic Anticancer Institute.     

Induction of delayed hypersensitivity by carrageenan

Zusammenfassung Nachweis einer Sensibilisierung des Meerschweinchens durch intradermale Injektion von Carrageenan, was zu «passiv übertragbaren» Spätreaktionen führte.

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The authors wish to thank to MissI. Kawana, MissA. Saito and MissA. Chida for their technical assistance and Prof.Y. Horiuchi for his advice.     

Inhibition of mammalian acetylcholinesterase by phenylmethanesulfonyl fluoride

Resumen El fenilmentano sulfonil fluoro (FMSF) 4×10^–5 M inhibe a la acetilcolinesterasa (E. C. 3.1.1.7) de cerebro de rata en más de 50%, y 9×10^–4 M más de 90%. La constante de reacción inhibidor enzima es de 4.7×10² (1 mol^–1 min^–1). La colinesterasa (E. C. 3.1.1.8) de cerebro de rata es insensible a FMSF.

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Supported by NIMH Grant No. 1 RO3 MH 19125, by Comisión Nacional de Investigación Cientiffica Proyecto No. 80, and by Comisión de Investigación Cientiffica Universidad de Chile, Proyecto No. 95.     

Induction of immunological memory in mice by RNA extract

Summary RNA extract isolated from spleens of mice immunized with lipopolysaccharide fromE. coli induced an immunological memory in normal mice. Application of small amounts of corresponding antigen provoked a specific secondary immune response in RNA primed mice.     

Induction of developmental anomalies in mice by maternal stress

Summary The short-time restraint of pregnant mice on day 8 of gestation led to a significant increase of the anomaly rate in fetuses. This effect may be due to stress factors of endocrine origin.The technical assistance of Mrs E. Frei is gratefully acknowledged. Supported by the Swiss National Foundation for Scientific Research.     

Induction of developmental anomalies in mice by maternal stress

The short-time restraint of pregnant mice on day 8 of gestation led to a significant increase of the anomaly rate in fetuses. This effect may be due to stress factors of endocrine origin.     

Induction in the chick by quail Hensen's node

Résumé Des greffons du nud de Hensen prélevés à des embryons de caille aux stades 3+ à 4 furent introduits sous l'ectoblaste d'embryons de poulet de mêmes stades cultivês selon la mêthode deNew ¹⁰. Les greffons se sont différenciés en divers tissus et en même temps ont provoqéu une différentiation neurale dans l'ectoblaste des hôtes. Les cellules des greffons de la partie postérieure de la ligne primitive 5e sont dispersés dans lemésoblaste extraembryomnaire des hôtes et n'ont exercé aucune action sur l'ectoblaste.

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Work supported bya grant toD. J. McCallion from the National Research Council of Canada.

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Supported by the Canadian International Development Agency, Ottawa.     

Induction of epidermal cell proliferation by a tumour promoter in vitamin B6-deficient mice

Summary Topical application of the tumour promoter 12-0-tetradecanoyl-phorbol-13-acetate to skin caused a marked enhancement of mitotic acitivity both in mice maintained on a complete diet or on a vitamin B₆-deficient diet.Acknowledgment. The author thanks Mr P. Daenke for excellent technical assistance.This investigation was supported by the Australian Research Grants Committee and the Australian Tobacco Research Foundation.