Wave frequency recognition and absolute pitch for water waves in the clawed frog,Xenopus laevis

Summary The ability of adultXenopus laevis to identify water wave frequencies was demonstrated by go/no-go conditioning. The acuity of frequency recognition is of absolute-pitch quality.Abbreviations S+ stimulus with response reward - S- stimulus with response punishment - u discrimination index     

The effect of centrifugal accelerations on the polarity of statocytes and on the graviperception of cress roots

The structural polarity of statocytes of Lepidium sativum L. is converted to a physical stratification by a root-tip-directed centrifugal acceleration. Sedimentation of amyloplasts and nucleus to the centrifugal (distal) cell pole and the lateral displacement of the distal endoplasmic reticulum (ER) complex occur after centrifugation for 20 min at an acceleration of 50 g. With higher doses (20 min, 100-2,000 g), smaller organelles become increasingly displaced. From the centrifugal to the centripetal cell pole, the following stratification is observed: 1) amyloplasts with mitochondria; 2) nucleus with mitochondria and a few dictyosomes, as well as laterally located ER; 3) dictyosomes with a few mitochondria; 4) vacuoles; and 5) lipid droplets. Within the first 7.5 min, after the roots have been returned to 1 g, the original arrangement of the amyloplasts sedimented on the underlying ER complex is reestablished in 66% of the statocytes. When roots previously centrifuged in an apical direction are exposed in a horizontal position to 1 g, the latent period of the graviresponse is increased by 7.5 min relative to the non-centrifuged controls. The kinetics of the response are identical to the controls. Roots centrifuged first in an apical direction and then for 2 h in a lateral direction (1,000 g) have statocytes with a physical stratification perpendicular to the root axis. A gravitropic curvature does not take place during the lateral centrifugation. These results support the hypothesis that the distal ER complex is necessary and sufficient for graviperception.     

Degradation of aniline and monochloroanilines by Rhodococcus sp. An 117 and a pseudomonad: a comparative study

Two newly isolated aniline-degrading bacterial strains were characterized with regard to their enzyme systems responsible for aniline catabolism. One of them identified as a Rhodococcus sp. metabolized aniline exclusively via the beta-ketoadipate pathway by means of inducible enzymes. The aniline-degrading enzyme system of the second isolate, presumably a pseudomonad, was shown to consist of an inducible aniline-converting enzyme and constitutive meta-pathway enzymes. Both isolates failed to metabolize monochlorinated anilines in the absence of additional carbon sources. To explain this the ring-cleaving enzymes of both isolates were examined for their substrate specificities. Furthermore, the effect of 4-chlorocatechol on the enzymes catalyzing aniline conversion and catechol oxygenation was investigated.     

Versuche zur Frage der Erhaltungszüchtung beim Kulturchampignon

Zusammenfassung 1. Die Arbeit befaßt sich mit Fragen der Erhaltungszüchtung beim Kulturchampignon. Von den drei Möglichkeiten der Vermehrung, Mycelteilung, Gewebekultur und Aussaat, wird die Mycelteilung untersucht.2. Es wurden zu diesem Zweck Vielsporkulturen (viele Kerntypen) und Einsporkulturen (nur 2 Kerntypen) miteinander verglichen.3. Eine Ein- und eine Vielsporkultur wurden bei Kultur auf Agar-Nährboden fortlaufend durch Mycelteilung vermehrt. Während die Einsporkultur auch nach 40maliger Teilung keine Mycelveränderung zeigte, traten bei der Vielsporkultur nach 13maliger Vermehrung Degenerationserscheinungen in Form von langsam wachsendem belagartig anliegendem Mycel auf. Später bildete sich teilweise wieder schneller wachsendes Mycel.In Ertragsprüfungen zeigte die Vielsporkultur in der höchsten geprüften Vermehrungsstufe (27. V.) einen starken Ertragsabfall. Bei der Einsporkultur trat kein Ertragsrückgang ein.4. Beide Stämme wurden auch durch Überimpfungen von Körnern vermehrt. Nach achtmaliger Vermehrung war kein Ertragsabfall nachzuweisen. Der in einem zweiten Versuch mit der Einsporkultur allein festgestellte Ertragsrückgang nach 15 Vermehrungen konnte nicht statistisch gesichert werden.5. Von einer Schale der 22. Vermehrung der Vielsporkultur, die belagartige langsamwachsende und fädige schnellwachsende Sektoren enthielt, wurden von beiden Mycelarten Stücke abgeimpft. Es war dadurch möglich, die verschiedenen Wuchstypen voneinander zu trennen. Jedoch bildeten sich manchmal wieder fädige Sektoren im belagartigen Typ und umgekehrt. Insgesamt wurde sechsmal hintereinander Mycel der verschiedenen Typen abgeimpft (6 Teststufen).6. Die Trennung der verschiedenen Wuchstypen wird mit Kernentmischung erklärt. Die Frage bleibt offen, ob Kerne vom Typ des langsam wachsenden Mycels schon von der Aussaat an in der Vielsporkultur waren oder erst später durch Mutation oder Modifikation entstanden. In diesem Falle hätten sich die Degenerationserscheinungen genausogut in der Einsporkultur wie in der Vielsporkultur zeigen können.7. Daß die Entmischung unvollkommen war, liegt vermutlich an den großen Mycelstücken, die abgeimpft wurden. In Fortsetzung der Versuche wird mit zerkleinertem Mycel gearbeitet. Dann werden einzelne Zellen, die unter dem Phasenkontrastmikroskop als kernarm bestimmt wurden, übergeimpft.8. Vier Viel- und vier Einsporkulturen wurden in Wachstumstesten bei Auslese auf schnell- und lang-samwachsendes Mycel miteinander verglichen. Es wurden von jedem Stamm aus zehn Kulturschalen die am wenigsten durchsponnene Schale (l-Gruppe) und die am weitesten durchsponnene Schale (s-Gruppe) ausgelesen und vermehrt. Von der l-Gruppe wurde fünfmal hintereinander aus der am wenigsten durchsponnenen Schale, von der s-Gruppe genausooft aus der am weitesten durchsponnenen Schale Mycel auf zehn neue Schalen abgeimpft.Die Differenz zwischen dem durchschnittlichen Myceldurchmesser der l- und s-Gruppe war unterschiedlich und vergrößerte sich nicht mit der Zahl der Überimpfungen. Die l-Gruppen der Vielsporkulturen wuchsen immer langsamer als die s-Gruppen, während es bei den Einsporkulturen auch umgekehrte Fälle gab. Auch war die Differenz zwischen der l-und s-Gruppe im Mycelwachstum bei den Vielsporkulturen doppelt so häufig statistisch gesichert als bei den Einsporkulturen.Für die Erhaltungszüchtung kann daraus gefolgert werden, daß sich Vielsporkulturen bei Vermehrung durch Teilung leichter verändern können als Einsporkulturen.9. Mycelkulturen auf drei verschiedenen Nährböden (Biomalz-Agar, Weizen-Agar und Kompost-Agar) zeigten je nach Nährboden unterschiedliches Wachstum. Auf Biomalz-Agar wuchsen die Kulturen am langsamsten. Auf Kompost-Agar, dem nährstoff-reichsten der drei Nährböden, waren die Unterschiede im Aussehen des Mycels am deutlichsten ausgeprägt.Es wird über einige Arbeiten anderer Autoren berichtet, in denen ein großer Einfluß des Nährbodens auf das Mycel festgestellt wurde.Die Ursachen dieses Einflusses werden diskutiert.Für die Erhaltungszüchtung wird gefolgert, daß die Stammkulturen auf einem möglichst nährstoffreichen Substrat, z. B. Kompost, gehalten werden sollten.10. In Untersuchungen des schlecht wachsenden belagartigen Mycels auf Krankheitsbefall konnten weder Bakterien noch Schadpilze nachgewiesen werden. Die Virusteste (Fusionsversuche, Wärmeschock) fielen unterschiedlich aus. Das Material wird noch mit Hilfe der Elektronenmikroskopie genauer untersucht werden. Über das Ergebnis wird später berichtet.

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Experiments on maintenance of strains of the cultivated mushroomI. Propagation by mycelium transfer

Summary 1. The paper deals with problems of maintaining strains of the cultivated mushroom. There are three possibilities for propagation: Transfer of mycelium, tissue culture and multispore culture. Of these the transfer of mycelium was investigated.2. Multispore cultures (many types of nuclei) and monospore cultures (only two types of nuclei) were compared.3. Mono- and multispore cultures were continuously propagated on agar media by transfers of mycelium. After 40 transfers the monospore culture showed no change in the mycelium. In the multispore culture degenerative symptoms in the form of slowly growing, matted mycelium appeared after 13 transfers, though the faster growing mycelium reappeared later in some cases. After the last (27th) transfer tested, the multispore culture showed a strong decrease in yield; none was found in the monospore culture.4. Both strains were also propagated by grain transfer. After eight transfers there was no decrease in yield, and the one noticed after 15 transfers in a repeat experiment with the monospore culture proved to be statistically not significant.5. From a petri dish containing, after the 22nd transfer of the multispore culture, matted, slow growing and stringy, fast growing mycelia, pieces of both kinds were taken. Thus it was possible to separate the two types of growth. However, sometimes stringy sectors reappeared in the matted type, and vice versa. Mycelia of the different types were transferred six consecutive times.6. The separation of the various types is explained by a separation of types of nuclei. The question whether nuclei of the type of the slow growing mycelium were already present in the multispore culture before starting the culture or originated later by mutation or modification remains open.In case of a modification or mutation the degeneration symptoms could have occurred in the monospore culture as well as in the multispore culture.7. The reason for an incomplete separation could be the transfer of large pieces of mycelium. In further experiments disintegrated mycelium will be used. Then single cells selected under phase contrast microscope as poor in nuclei will be transferred.8. Four multi- and four monospore cultures were compared for growth rates by selection for fast and for slow growing mycelium. For each strain the least overgrown (l-group) and the most overgrown (s-group) were chosen from ten culture plates and subcultured by five successive transfers to ten fresh plates.The difference in average diameter of mycelium from the l-group and s-group varied and did not increase with the number of transfers. The mycelium of the l-groups of multispore cultures always grew slower than that of the s-groups; in monospore cultures the opposite also occurred. The difference between the l- and s-group in the multispore culture was twice as often significant as in the monospore culture. From this one can conclude that, in propagation with frequent transfer, multispore cultures change more easily than do monospore cultures.9. Mycelium cultures on three different media (biomalt agar, wheat agar, compost agar) showed different growth rates. The slowest growth was found on biomalt agar. On compost agar, richest in nutrients, the difference in appearance of the mycelium was most obvious. Papers by other authors who found a great influence of the medium on the mycelium and the reasons for this influence are discussed. For the maintenance of strains original cultures should be kept on a substrate rich in nutrients, i.e. compost.10. Tests for bacteria and fungi on slow growing, matted mycelium were negative. Tests for virus (fusion tests, heat shock) showed different results. Results of electron microscopic studies on this material will be reported later.

     

Binding of the J 1 Adhesion Molecules to Extracellular Matrix Constituents

The J1 glycoproteins can be obtained in multiple forms in the soluble fraction of developing and adult mouse brain tissue. They are recovered as two forms of apparent molecular weights of 160,000 and 180,000 (J1-160) from adult mouse brain and as forms of apparent molecular weights of 200,000 and 220,000 (J1-220) from developing brain. J1-160 and J1-220 share common epitopes but are considered as separate entities, with J1-220 being immunochemically closely related if not identical to tenascin. Based on the observation that J1 immunoreactivity appears on basement membrane and interstitial collagens after denervation of the neuromuscular junction in adult rodents, we became interested in investigating the binding properties of J1 glycoproteins to extracellular matrix constituents in vitro. Both J1-160 and J1-220 bound to collagens type I-VI and IX but not to laminin, fibronectin, bovine serum albumin, or gelatin under hypotonic buffer conditions. Under isotonic buffer conditions, J1-220 bound to all collagen types, whereas J1-160 bound only to collagen types V and VI with values that could be examined by Scatchard analysis. Binding of J1-220 to collagens displayed two binding constants (KD) between 1.5 and 4.4 X 10(-9) and 1.8 and 5.5 X 10(-8) M, respectively, under hypotonic buffer conditions and a single KD of 2.1-8.0 X 10(-8) M under isotonic buffer conditions. Binding of J1-160 to collagens had an apparent KD of 1.9-8.0 X 10(-9) M under hypotonic buffer conditions. Under isotonic buffer conditions, binding constants of J1-160 to collagen types V and VI were approximately 2 X 10(-8) M. Binding of J1-220 to collagen type I could be inhibited by J1-220, J1-160, and collagen type VI but not by fibronectin or gelatin. Conversely, binding of J1-160 was inhibited by J1-220, J1-160, and collagen type VI (in order of decreasing efficacy of competition). J1-160 and J1-220 were retained on a heparin-agarose column and eluted in a salt gradient at approximately 0.5 M NaCl. The formation of the J1-heparin complexes was inhibited 100-fold more efficiently by heparin than by chondroitin sulfate. These experiments show that J1 glycoproteins resemble in many respects the extracellular matrix constituents fibronectin, laminin, vitronectin, and von Willebrand factor.     

Single chloride channels in endosomal vesicle preparations from rat kidney cortex

Summary Endocytotic vesicles from rat kidney cortex, isolated by differential centrifugation and enriched on a Percoll gradient, contain both an electrogenic H⁺ translocation system and a conductive chloride pathway. Using the dehydration/rehydration method, we fused vesicles of enriched endosomal vesicle preparations and thereby made them accessible to the patch-clamp technique. In the fused vesicles, we observed Cl^– channels with a single-channel conductance of 73±2 pS in symmetrical 140mm KCl solution (n=25). The current-voltage relationship was linear in the range of –60 to +80 mV, but channel kinetic properties dependended on the clamp potential. At positive potentials, two sublevels of conductance were discernible and the mean open time of the channel was 10–15 msec. At negative voltages, only one substate could be resolved and the mean open time decreased to 2–6 msec. Clamp voltages more negative than –50 mV caused reversible channel inactivation. The channel was selective for anions over cations. Ion substitution experiments revealed an anion permeability sequence of Cl^–=Br^–=I^–>SO ₄ ^2– F^–. Gluconate, methanesulfonate and cyclamate were impermeable. The anion channel blockers 4,4-diisothiocyanatostilbene-2,2-disulfonic acid (DIDS, 1.0mm) and 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB, 0.1mm) totally inhibited channel activity. Comparisons with data obtained from radiolabeled Cl^–-flux measurements and studies on the H⁺ pump activity in endocytotic vesicle suspensions suggest that the channel described here is involved in maintenance of electroneutrality during ATP-driven H⁺ uptake into the endosomes.     

Analysis of extracellular calcium and volume changes in the compound eye of the honeybee drone,Apis mellifera

Summary Superfused slices of drone retina were used for a quantitative analysis of light-induced changes in extracellular Ca²⁺ concentration ([Ca²⁺]_o) and extracellular space (ECS) volume. 20-ms light flashes elicited biphasic changes in [Ca²⁺]_o. For a saturating flash a brief, initial decrease was followed by a transient increase of 120±34 M. Long, dim steps of light (5 min) produced either a decrease or an increase in [Ca²⁺]_o depending strongly on the previous illumination. Brighter continuous lights caused the [Ca²⁺]_o to increase transiently by 1.4 mM to a peak from which it decayed to a plateau, up to 0.6 mM above the dark concentration.Light flashes (20 ms) caused a shrinkage in ECS volume not exceeding 4%. Thus, changes in [Ca²⁺]_o were almost completely due to Ca²⁺ fluxes between the ECS and adjacent cells. Continuous lights caused a shrinkage in ECS volume rarely exceeding 16%–20%. Thus, less than 15% of the measured Ca²⁺ changes could be attributed to shrinkage of the ECS. These data confirm that the ECS functions as a source and a sink for Ca²⁺ mobilized by light. For comparison, we also made a few measurements of changes in [Ca²⁺]_o in the retina ofCalliphora.Abbreviations [Ca ²⁺]_i intracellular free Ca²⁺ concentration - [Ca ²⁺]_o extracellular free Ca²⁺ concentration - ECS extracellular space - ER endoplasmic reticulum - TMA ⁺ tetramethylammonium ion     

Hypervariability of intronic simple (gt)n(ga)m repeats in HLA-DRB genes

We have investigated the extent of DNA variability in intronic simple (gt)_n(ga)_m repeat sequences and correlated this to sequence polymorphisms in the flanking exon 2 of HLA-DRB genes. The polymerase chain reaction (PCR) was used to amplify a DNA fragment containing exon 2 and the repeat region of intron 2. The PCR products were separated on sequencing gels in order to demonstrate length hypervariability of the (gt)_n(ga)_m repeats. In a parallel experiment, the PCR products were cloned and sequenced (each exon 2 plus adjacent simple repeats) to characterize the simple repeats in relation to the HLA-DRB sequences. In a panel of 25 DRB1, DRB4, and DRB5 alleles new sequences were not detected. Restriction fragment length polymorphism (RFLP) subtyping of serologically defined haplotypes corresponds to translated DNA sequences in 85% of the cases, the exceptions involving unusual DR/DQ combinations. Many identical DRB1 alleles can be distinguished on the basis of their adjacent simple repeats. We found group-specific organization of the repeats: the DRw52 supergroup repeats differ from those of DRB1*0101, DRB4*0101, and DRB5*0101 alleles and from those of pseudogenes. Finally, we amplified baboon DNA and found a DRB allele with extensive similarity to DRB1 sequences of the DRw52 supergroup. The simple repeat of the baboon gene, however, resembles that of human pseudogenes. In addition to further subtyping, the parallel study of polymorphic protein and hypervariable DNA alleles may allow conclusions to be drawn on the relationships between the DRB genes and perhaps also on the theory of trans-species evolution.The nucleotide sequence data reported in this paper have been submitted to the GenBank nucleotide sequence database and have been assigned the accession number M 34258.