A re-investigation of the ribonuclease sensitivity of a DNA demethylation reaction in chicken embryo and G8 mouse myoblasts.

Recently published results (Nucleic Acids Res. 26, 5573-5580, 1998) suggest that the ribonuclease sensitivity of the DNA demethylation reaction may be an experimental artifact due to the possible tight binding of the nucleases to the methylated DNA substrate. Using an improved protocol we show for two different systems that demethylation of hemimethylated DNA is indeed sensitive to micrococcal nuclease, requires RNA and is not an experimental artifact. The purified 5-MeC-DNA glycosylase from chicken embryos and G8 mouse myoblasts was first incubated for 5 min at 37 degrees C with micrococcal nuclease in the presence of Ca2+ in the absence of the DNA substrate. Upon blocking the nuclease activity by the addition of 25 mM EGTA, the DNA demethylation reaction was initiated by adding the labeled hemimethylated DNA substrate to the reaction mixture. Under these conditions the DNA demethylation reaction was abolished. In parallel controls, where the purified 5-MeC-DNA glycosylase was pre-incubated at 37 degrees C with the nuclease, Ca2+ and EGTA or with the nuclease and EGTA, RNA was not degraded and no inhibition of the demethylation reaction was obtained. As has already been shown for chicken embryos, the loss of 5-MeC-DNA glycosylase activity from G8 myoblasts following nuclease treatment can also be restored by the addition of synthetic RNA complementary to the methylated strand of the substrate DNA. No reactivation of 5-MeC-DNA glycosylase is obtained by complementation with a random RNA sequence, the RNA sequence complementary to the non-methylated strand or DNA, thus ruling out a non-specific competition of the RNA for the binding of the nuclease to the labeled DNA substrate.     

Slow actions of hyperpolarization on sodium channels in the membrane of myelinated nerve.

The mean sodium current, I, and the variance of sodium current fluctuations, var, were measured in myelinated nerve during a depolarization to V = 40 mV applied from the resting potential (VH = 0) or from a hyperpolarizing holding potential VH = -28 mV. From I and var the relative variations in the number N and the conductance gamma of sodium channels following changes of the holding potential were calculated. Hyperpolarizing the membrane from VH = 0 to -28 mV increased N by a factor of 3.7, whereas gamma decreased by a factor of 0.53. These actions of holding potential on sodium channels develop slowly since 500 ms prepulses to 0 or -28 mV do not alter the values of N and gamma.     

Latitudinal variation in blue tit and great tit nest characteristics indicates environmental adjustment

Aim The laying of eggs and the building of a nest structure to accommodate them are two of the defining characteristics of members of the class Aves. Nest structures vary considerably across avian taxa and for many species the structure of the completed nest can have important consequences both for parents and their offspring. While nest characteristics are expected to vary adaptively in response to environmental conditions, large‐scale spatial variation in nest characteristics has been largely overlooked. Here, we examine the effects of latitudinal variation in spring temperatures on nest characteristics, including insulatory properties, and reproductive success of blue tits, Cyanistes caeruleus, and great tits, Parus major. Location Great Britain. Methods Nests and reproductive data were collected from seven study sites, spread over 5° of latitude. The nest insulatory properties were then determined before the nests were separated into nest base material and cup lining material. Results As spring temperatures increased with decreasing latitude, the mass of the nest base material did not vary in either species, while the mass of the cup lining material and nest insulatory properties decreased in both species. This suggests that in response to increasing temperatures the breeding female reduces the mass of the cup lining material, thereby maintaining an appropriate microclimate for incubating and brooding. The mean first egg date of both species advanced with decreasing latitude and increasing spring temperatures, although clutch size and brood size at hatching and fledging did not vary. Main conclusions This is the first study to demonstrate that the nest‐construction behaviour of birds varies in response to large‐scale spatial variation in ambient temperatures. Therefore, nest composition reliably indicates environmental conditions and we suggest that studies of nest structure may be sentinels for the early signs of rapid climate change.     

Why Movement Is Captured by Music,but Less by Speech: Role of Temporal Regularity

Music has a pervasive tendency to rhythmically engage our body. In contrast, synchronization with speech is rare. Music’s superiority over speech in driving movement probably results from isochrony of musical beats, as opposed to irregular speech stresses. Moreover, the presence of regular patterns of embedded periodicities (i.e., meter) may be critical in making music particularly conducive to movement. We investigated these possibilities by asking participants to synchronize with isochronous auditory stimuli (target), while music and speech distractors were presented at one of various phase relationships with respect to the target. In Exp. 1, familiar musical excerpts and fragments of children poetry were used as distractors. The stimuli were manipulated in terms of beat/stress isochrony and average pitch to achieve maximum comparability. In Exp. 2, the distractors were well-known songs performed with lyrics, on a reiterated syllable, and spoken lyrics, all having the same meter. Music perturbed synchronization with the target stimuli more than speech fragments. However, music superiority over speech disappeared when distractors shared isochrony and the same meter. Music’s peculiar and regular temporal structure is likely to be the main factor fostering tight coupling between sound and movement.