Thermal adaptation in the honeybee (Apis mellifera) via changes to the structure of malate dehydrogenase.

In honeybees there are three alleles of cytosolic malate dehydrogenase gene: F, M and S. Allele frequencies are correlated with environmental temperature, suggesting that the alleles have temperature-dependent fitness benefits. We determined the enzyme activity of each allele across a range of temperatures in vitro The F and S alleles have higher activity and are less sensitive to high temperatures than the M allele, which loses activity after incubation at temperatures found in the thorax of foraging bees in hot climates. Next, we predicted the protein structure of each allele and used molecular dynamics simulations to investigate their molecular flexibility. The M allozyme is more flexible than the S and F allozymes at 50°C, suggesting a plausible explanation for its loss of activity at high temperatures, and has the greatest structural flexibility at 15°C, suggesting that it can retain some enzyme activity at cooler temperatures. MM bees recovered from 2 h of cold narcosis significantly better than all other genotypes. Combined, these results explain clinal variation in malate dehydrogenase allele frequencies in the honeybee at the molecular level.

Effects of cold narcosis on memory acquisition, consolidation and retrieval in honeybees (Apis mellifera).

In learning and memory studies on honeybees (Apis mellifera), cold-induced narcosis has been widely used to temporarily immobilize honeybees. In this study, we investigated the effects of cold narcosis on the associative memories in honeybees by using the proboscis extension response (PER) paradigm. Severe impairments in memory acquisition was found when cold narcosis was performed 30 min, instead of 1 h before training. Locomotor activities were reduced when honeybees were tested 15 min, instead of 30 min after cold narcosis. These results indicate that cold narcosis impairs locomotor activities, as well as memory acquisition in a time-dependent manner, but by comparison no such effects on memory retrieval have yet been observed.[0].