Isolation of a cAMP receptor protein from yeast mitochondria (Mr 45000) and comparison with mitochondrial RNA polymerase (Mr 45000)

We have isolated a cAMP-binding protein from highly purified yeast mitochondria by affinity chromatography. It is a lipophilic protein of molecular mass 45 000 Da, which is tightly membrane-bound and localized on the outer surface of the inner membrane. It can be solubilized in active form under mild conditions. The cAMP receptor resembles mitochondrial RNA polymerase prepared as described by Levens et al. [(1981) J. Biol. Chem. 256, 1474] in a surprisingly large number of properties including molecular mass. Comparison of the two proteins revealed that the polypeptide previously considered as RNA polymerase is, in fact, a mitochondrial cAMP receptor protein.     

Oriented axon outgrowth from avian embryonic retinae in culture

The neural retina of avian embryos was spread on a membrane filter and cut in any desired orientation. Strips cut across the retina of 4- to 7-day chick or 3- to 6-day quail embryos were explanted onto collagen gels. Vigorous neurite outgrowth was seen for about 3 days, by which time many neurites were 3 mm long. Horseradish peroxidase (HRP) labeling showed that the cells producing the neurites were large and formed a layer near the inner limiting membrane, indicating that the neurites in vitro were axons of retinal ganglion cells. The size of the neurite population and the regions from which neurites emerged vaired with the donor age, while most neurites sprouted from the side of the explant formerly closest to the optic fissure. This pattern closely resembled that of axon growth in the normal retina, as revealed by SEM, silver staining, and HRP labeling. Mitotic inhibitors (Ara-C and FUdR) did not alter the neurite outgrowth. Pretreatment of retinae with trypsin or collagenase did not disorganize axons at the time of explantation, but tended to equalize neurite emergence on each side of the retinal strips. We suggest that microenvironmental factors, especially the enzyme-labile inner limiting membrane, are important for axon guidance in the retina.     

Elektronenmikroskopische Untersuchungen an Lysosomen im Blastem der Extremitätenknospe der Ratte

Zusammenfassung Die Extremitätenknospen und die Schwanzknospe von Rattenembryonen enthalten ein mesenchymales Zellretikulum, in dessen Achse sich die Zellen polygonal abrunden und zu einem Blastem zusammenlegen. In den umgebenden mesenchymalen Zellen kommen keine Lysosomen vor. Dagegen enthalten die Zellen des zentralen Blastems Lysosomen. Die primären Lysosomen sind cytoplasmatische Granula mit einem Durchmesser von 0,1–0,2 und einem elektronendichten granulären Inhalt. Auffällig ist das Vorkommen von multivesikulären Einschlußkörpern in der Nähe der Golgizone. Wegen ihrer Funktion können sie auch zu den primären Lysosomen gerechnet werden. Die häufigsten Lysosomen sind runde Cytolysosomen (Durchmesser von 0,5–2 ), die von einer Membran umgeben sind und polymorphen Inhalt besitzen. Dicht gepackte Granula, die wie zusammengeflossene Ribosomen aussehen, kommen vor. Außerdem liegen andere Zellorganellen (Mitochondrien, Membranstrukturen) in den sekundären Lysosomen. Schließlich kommen auch Myelinfiguren, allein oder mit anderen Strukturen kombiniert, in ihnen vor. Die verschiedenen Stadien dieser Lysosomen sind der Ausdruck für eine partielle Autodigestion von Zellorganellen im Blastem. Eine Autodigestion von Ribosomen könnte eine Bedeutung haben. Nach dieser Autodigestion der alten Information und der Ribosomen beginnt nämlich eine Zelldifferenzierung zu Vorknorpel mit neuer Information und neuen Ribosomen für die jetzt neu auftretende Zwischensubstanz. Die beschriebenen Lysosomen zeigen eine positive elektronenmikroskopisch nachweisbare Reaktion auf saure Phosphatase.

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Summary The limb buds and the tail bud of rat embryos contain a mesenchymal cellular reticulum. In its axis the cells become polygonal and group to a blastema. There are no lysosomes in the cells of the surrounding mesenchyma. The cells of the central blastema, however, possess lysosomes. The primary lysosomes are cytoplasmic granules with a diameter of 0,1–0,2 . Their granular contents are electron dense. There is a striking occurrence of multivesiculated bodies near the Golgi zone. For their function they may be counted to the primary lysosomes. The most frequent lysosomes are the cytolysosomes. These are of circular shape with a diameter of 0,5–2 and are enclosed by a membrane, their contents are polymorphic. Tightly stacked granules looking like conglomerated ribosomes can be found. Apart from these other cell organelles like mitochondria and membrane structures lie in the secondary lysosomes. Finally, there are also myelin figures alone or combined with other structures in the lysosomes. The different stages of these lysosomes are the result of a partial autodigestion of cell organelles within the blastema. The autodigestion of ribosomes may be important. After this autodigestion of the old template and ribosomes a new phase of cyto-differentiation to precartilage with new template and ribosomes begins. The lysosomes show a positive reaction for acid phosphatase.

     

Behavioural specialization in pre-reproductive colonies of the allodapine beeExoneura bicolor (Hymenoptera: Anthophoridae)

Summary Exoneura bicolor is a univoltine allodapine bee common in montane forests of southern Australia, where it exhibits a semisocial/quasisocial colony organization. Within-nest behaviour in postemergence autumn nests ofExoneura bicolor was recorded with the aim of studying behavioural specialization in pre-reproductive colonies. Ten complete colonies were transferred to purpose-built observation nests shortly before brood eclosion in late summer. Behaviour within observation nests was recorded for periods of up to 44 days after establishment, covering a period when colonies are preparing for overwintering. Dispersal of females and brood rearing do not occur at this time, although some females may become inseminated. Analyses of data using multivariate techniques indicated four distinguishable behavioural castes, designated here as Guards, Nest Absenters, Nest Modifiers and Non-recruits. This represents a higher degree of behavioural specialization than recorded to date for other allodapines. Behaviours performed by Guards and Nest Absenters are likely to involve considerable risks, but benefit the colony as a whole, so that some nestmates in prereproductive colonies exhibit altruism that frequently aids adult siblings or cousins. The males in our study were fed by females via trophallaxis and two of the males participated in nest maintenance tasks. Our results suggest that autumn colonies ofE. bicolor form well-integrated behavioural units even though brood rearing does not commence until the following spring.     

Modification of sodium inactivation in myelinated nerve by Anemonia toxin II and iodate. Analysis of current fluctuations and current relaxations

(1) Na+ currents and Na+ current fluctuations were measured in single myelinated nerve fibres of Rana esculenta under voltage-clamp conditions. The process of Na+ inactivation was modified by external treatment with 7 microM Anemonia Toxin II or by internal application of 20 or 40 mM IO3(-). (2) At depolarization of 24 and 32 mV the spectral density of Na+ current fluctuations could be described as the sum of two contributions, Sh(f) and Sm(f), representing the spectrum from fluctuations of the inactivation (h) and activation (m) gates, respectively. At higher depolarizations of 40 and 48 mV the low frequency (h) fluctuations could be better fitted by the sum, Sh1(f)+Sh2(f), of two separate Lorentzian functions. (3) The Na+ current and the variance of Na+ current fluctuations between 150 and 450 ms after depolarization are increased by one order of magnitude after application of Anemonia Toxin II or IO3(-). (4) The kinetics of Na+ current inactivation were described as A1 x exp(-t/tau h1) + A2 x exp(-t/tau h2) + B. The constant, tau h1, of fast Na+ inactivation was the same in normal and modified nerve fibres. The slow inactivation time constant, tau h2, increased with increasing depolarizations in modified fibres but decreased under control conditions. In all cases tau h2 showed a similar voltage dependence as the time constant found by fitting the low frequency fluctuations of Na+ current with one Lorentzian function, Sh(f). (5) It is concluded that Anemonia Toxin II and IO3(-) modify a fraction of Na+ channels in an all-or-none manner. A lower limit of the number of modified Na+ channels is estimated from the Na+ current and the variance Na+ current fluctuations. 7 microM external Anemonia Toxin II modifies more than 17% and 20 or 40 mM internal IO3(-) more than 8% of all Na+ channels. The inactivation gates in modified channels experience an electric field different from that in normal fibres.