A mammary gland EST showing linkage disequilibrium to a milk production QTL on bovine Chromosome 14

As part of a genome scan, ESTs derived from mammary gland tissue of a lactating cow were used as candidate genes for quantitative trait loci (QTL), affecting milk production traits. Resource families were genotyped with 247 microsatellite markers and 4 polymorphic ESTs. It was shown by linkage analysis that one of these ESTs, KIEL_E8, mapped to the centromeric region of bovine Chromosome (Chr) 14. Regression analysis revealed the presence of a QTL, with significant effect on milk production, in this chromosome region, and analysis of variance showed no significant interaction of marker genotype and family. The estimated significant differences between homozygous marker genotypes were 140 kg milk, −5.02 kg fat yield, and 2.58 kg protein yield for the first 100 days of lactation. Thus, there was strong evidence for a complete or nearly complete linkage disequilibrium between KIEL_E8 and the QTL. To identify the biological function of KIEL_E8, we extended the sequence for 869 bp by 5′-RACE. A 560-bp fragment of this shows a 90.9% similarity to a gene encoding a cysteine- and histidine-rich cytoplasmic protein in mouse. Although such a protein may have a regulatory function for lactation and a linkage disequilibrium between the EST marker and the QTL has been observed, it remains to be elucidated whether they are identical or not. Nevertheless, KIEL_E8 will be an efficient marker to perform marker-assisted selection in the Holstein-Friesian population. Received 20 October 2000 / Accepted: 11 April 2001     

Nitrogenase activity in composting horse bedding and leaves

To date little or no nitrogen fixation has been found in composting materials. However, some materials have high C:N ratios and thus might be expected to support nitrogen fixation. We examined windrows of composting horse bedding and leaves for nitrogenase activity via acetylene reduction and used carbon monoxide controls to differentiate ethylene evolved by nitrogenase from that evolved by endogenous processes. In both piles temperatures were substantially elevated and partial pressures of oxygen greatly reduced thus indicating vigorous microbial activity. In the horse bedding, there were only low rates of ethylene evolution and little of this was due to nitrogen fixation. In contrast, much higher rates of ethylene evolution were measured in the leaf pile and 94% of this was due to nitrogenase activity. We estimate that the leaf pile fixed approximately 0.062 mg N/g leaves during a 5-week period.     

Professor Dr. Geoffrey Burnstock (1929–2020)

Purinergic Signalling -     

Arguments against the significance of the Fenton reaction contributing to signal pathways under in vivo conditions

One of the common explanations for oxidative stress in the physiological milieu is based on the Fenton reaction, i.e. the assumption that radical chain reactions are initiated by metal-catalyzed electron transfer to hydrogen peroxide yielding hydroxyl radicals. On the other hand — especially in the context of so-called “iron switches” — it is postulated that cellular signaling pathways originate from the interaction of reduced iron with hydrogen peroxide.

Using fluorescence detection and EPR for identification of radical intermediates, we determined the rate of iron complexation by physiological buffer together with the reaction rate of concomitant hydroxylations of aromatic compounds under aerobic and anaerobic conditions. With the obtained overall reaction rate of 1,700 M^-1s^-1 for the buffer-dependent reactions and the known rates for Fenton reactions, we derive estimates for the relative reaction probabilities of both processes.

As a consequence we suggest that under in vivo conditions initiation of chain reactions by hydroxyl radicals generated by the Fenton reaction is of minor importance and hence metal-dependent oxidative stress must be rather independent of the so-called “peroxide tone”. Furthermore, it is proposed that — in the low (subtoxic) concentration range — hydroxylated compounds derived from reactions of “non-free” (crypto) OH radicals are better candidates for iron-dependent sensing of redox-states and for explaining the origin of cellular signals than the generation of “free” hydroxyl radicals.     

Characterization of new G protein-coupled adenine receptors in mouse and hamster

The nucleobase adenine has previously been reported to activate G protein-coupled receptors in rat and mouse. Adenine receptors (AdeR) thus constitute a new family of purine receptors, for which the designation “P0-receptors” has been suggested. We now describe the cloning and characterization of two new members of the AdeR family from mouse (MrgA10, termed mAde1R) and hamster (cAdeR). Both receptors were expressed in Sf9 insect cells, and radioligand binding studies were performed using [³H]adenine. Specific binding of the radioligand was detected in transfected, but not in untransfected cells, and K _D values of 286 nM (mAde1R, B _max 1.18 pmol/mg protein) and 301 nM (cAdeR, B _max 17.7 pmol/mg protein), respectively, were determined. A series of adenine derivatives was investigated in competition binding assays. Minor structural modifications generally led to a reduction or loss of affinity, with one exception: 2-fluoroadenine was at least as potent as adenine itself at the cAdeR. Structure–activity relationships at all AdeR orthologs and subtypes investigated so far were similar, but not identical. For functional analyses, the cAdeR was homologously expressed in Chinese hamster ovary (CHO) cells, while the mAde1R was heterologously expressed in 1321N1 astrocytoma cells. Like the previously described AdeRs from rat (rAdeR) and mouse (mAde2R), the mAde1R (EC₅₀ 9.77 nM) and the cAdeR (EC₅₀ 51.6 nM) were coupled to inhibition of adenylate cyclase. In addition, the cAdeR from hamster expressed in CHO cells produced an increase in intracellular calcium concentrations (EC₅₀ 6.24 nM) and was found to be additionally coupled to G_q proteins.     

Buchbesprechung (Book Review)

Die Bedeutung insektenpathogener Viren als “Biologische Pflanzenschutzmittel” im System des Integrierten Pflanzenschutzes wird erläutert. Es wird dabei insbesondere auf die “Lückenindikationen” in Kulturpflanzenarten mit geringem Anbauumfang hingewiesen. Zu ihnen werden u.a. Gemüse, Zierpflanzen, Sonderkulturen wie Hopfen, Tabak und Wein, aber auch der Obstbau, gerechnet; daneben gehören aber auch Heil‐und Gewürzpflanzen sowie Rohstoffe für Diät‐und Säuglingsnahrungsmittel dazu. Schließlich wird der Forstschutz als ein wichtiges Refugium für die Anwendung biologischer Pflanzenschutzmittel im allgemeinen und damit auch für insektenpathogene Viren angesehen.

Einen hohen Stellenwert besitzen Unter‐Glas‐Kulturen, da hier biologische Bekämpfungsverfahren schon in einem erheblichen Umfang zur Anwendung kommen.

Die nachfolgenden Beispiele sollen stellvertretend für einen erfolgreichen Einsatz insektenpathogener Viren stehen:

• Bekämpfung von Spodoptera exigua in Chrysanthemenbeständen unter Glas in den Niederlanden mit dem autochthonen Virus und von Mamestra brassicae mit dem spezifischen Kernpolyeder‐Virus in Gewächshauskulturen von Rosen und Paprika (Wirkungsgrad 80 bis 100%) sowie an Kohl im Freiland (Wirkungsgrad 68, 9 bis 100%) in Deutschland.

• Das Granulose‐Virus der Wintersaateule (Agrotis segetum) ergab bei Anwendung gegen den Schädling an Astern Mortalitätswerte zwischen 90, 5 und 94, 1%.

In allen Versuchen erwiesen sich die Viren den als Standard mitgeführten chemischen Insektiziden als gleichwertig.

Am Zusammenbruch lokaler Gradationen der Kiefernbuschhornblattwespe Diprion similis in verschiedenen deutschen Bundesländern (Sachsen‐Anhalt, Sachsen) war ein spezifisches Kernpolyeder‐Virus wesentlich mitbeteiligt.     

Parental care and UV coloration in blue tits: opposite correlations in males and females between provisioning rate and mate’s coloration

Parental investment and sexually‐selected signals can be intimately related, either because the signals indicate the amount of investment that an individual is prepared to make, and hence its value as a mate (the ‘good parent process’), or because individuals are selected to vary their own investment in relation to their mate’s signals (‘differential allocation’ or ‘reproductive compensation’). Correlations between parental investment and the sexually selected signals of both an individual and its mate are therefore of central interest in sexual selection. Blue tits Cyanistes caeruleus are an ideal study species to investigate such correlations because they provide substantial amounts of biparental care and possess sexually‐selected structural UV coloration that seems to signal attractiveness in both sexes. We investigated whether feeding rates of male and female blue tits were correlated with either their own or their mate’s UV coloration, and whether any such correlation was affected by the sex ratio of the brood. We also investigated whether any such correlations were reflected in offspring phenotype. Feeding rates were not correlated with either sex of parent’s own UV coloration. However, they were correlated with the mate’s UV coloration, but in opposite directions in males and females: females had higher feeding rates when mated to bright UV males, implying differential allocation, while males had lower feeding rates when mated to bright UV females, implying reproductive compensation. These relationships were unaffected by the sex ratio of the brood. In addition, fledgling tarsus length, but not mass, was related to male UV coloration, and to female UV coloration in interaction with male age. These results suggest that both male and female attractiveness influence parental investment of the mate, and that this in turn affects offspring phenotype. We found no evidence for differential sex allocation.