Aedes aegypti juvenile hormone acid methyl transferase, the ultimate enzyme in the biosynthetic pathway of juvenile hormone III, exhibits substrate control.

We report on the cloning, sequencing, characterization, 3D modeling and docking of Aedes aegypti juvenile hormone acid methyl transferase (AeaJHAMT), the enzyme that converts juvenile hormone acid (JHA) into juvenile hormone (JH). Purified recombinant AeaJHAMT was extensively characterized for enzymatic activity and the Michaelis Menten kinetic parameters Km, Vmax, k(cat) (turn over number) and k(cat)/Km (catalytic efficiency) using JHA and its analogues as substrates. AeaJHAMT methylates JHA III 5-fold faster than farnesoic acid (FA). Significant differences in lower methyl transferase (MT) activities towards the cis/trans/trans, cis/trans/cis and the trans/cis/cis isomers of JHA I (1.32, 4.71 and 156-fold, respectively) indicate that substrate chirality is important for proper alignment at the catalytic cavity and for efficient methyl transfer by S-adenosyl methionine (SAM). Our 3D model shows a potential binding site below the main catalytic cavity for JHA analogues causing conformational change and steric hindrance in the transfer of the methyl group to JHA III. These, in silico, observations were corroborated by, in vitro, studies showing that several JHA analogues are potent inhibitors of AeaJHAMT. In vitro, and in vivo studies using [(3)H-methyl]SAM show that the enzyme is present and active throughout the adult life stage of A. aegypti. Tissue specific expressions of the JHAMT gene of A. aegypti (jmtA) transcript during the life cycle of A. aegypti show that AeaJHAMT is a constitutive enzyme and jmtA transcript is expressed in the corpora allata (CA), and the ovary before and after the blood meal. These results indicate that JH III can be synthesized from JHA III by the mosquito ovary, suggesting that ovarian JH III may play an important physiological role in ovarian development and reproduction. Incubating AeaJHAMT with highly pure synthetic substrates indicates that JHA III is the enzyme's preferred substrate, suggesting that AeaJHAMT is the ultimate enzyme in the biosynthetic pathway of JH III.

Recovery from retinal lesions: molecular plasticity mechanisms in visual cortex far beyond the deprived zone.

In cats with central retinal lesions, deprivation of the lesion projection zone (LPZ) in primary visual cortex (area 17) induces remapping of the cortical topography. Recovery of visually driven cortical activity in the LPZ involves distinct changes in protein expression. Recent observations, about molecular activity changes throughout area 17, challenge the view that its remote nondeprived parts would not be involved in this recovery process. We here investigated the dynamics of the protein expression pattern of remote nondeprived area 17 triggered by central retinal lesions to explore to what extent far peripheral area 17 would contribute to the topographic map reorganization inside the visual cortex. Using functional proteomics, we identified 40 proteins specifically differentially expressed between far peripheral area 17 of control and experimental animals 14 days to 8 months postlesion. Our results demonstrate that far peripheral area 17 is implicated in the functional adaptation to the visual deprivation, involving a meshwork of interacting proteins, operating in diverse pathways. In particular, endocytosis/exocytosis processes appeared to be essential via their intimate correlation with long-term potentiation and neurite outgrowth mechanisms.

Sublethal effects of crop protection on honey bee pollination: foraging behaviour and flower visits.

Crop protection strategies essential for pest and disease control can pose risk to pollinators. Fruits cannot be grown commercially without the use of crop protection agents, either from organic or chemical origin. The use of products with toxic effects is banned during flowering, and precise pre-flowering intervals have to be respected in Good Agricultural Practice. Bee pollination is essential for fruit crops to guarantee maximal fruit quality (shape and size) and quantity (yield weight). Fruit growers in Belgium depend mostly on non-commercial beekeepers to provide sufficient colonies for adequate pollination. Under optimal circumstances, beekeepers and fruit growers have mutual benefits from this cooperation as both honey and fruit yield increase. In those European countries with a monitoring scheme, acute bee poisoning incidents have decreased considerably and hardly cause problems at present. In recent years, some concerns arose around sublethal effects (i.e., behavioural changes) of chemical crop protection on bees, especially with regard to increased winter mortality. Even though short-term effects can indeed be induced in individually exposed bees, studies that exposed complete colonies did not reveal any long-term consequences at colony level. However, from the fruit growers' viewpoint, potential short-term effects on foraging behaviour are relevant as they can bear on pollination efficacy.