Development of the protozoan pathogen Trypanosoma brucei involves regulated changes in parasite structure, biochemistry, and the cell cycle. The transition of slender blood forms into stumpy bloodforms includes cell cycle arrest and a decrease in protein synthesis. The next stage in the development cycle, the procyclic form, shows increased protein synthesis and proliferates. To address the mechanism of the cyclical changes in protein synthesis, we examined two parameters: polyadenylation of mRNA and ribosome loading. We developed a method for analytical polyribosome analysis in T. brucei which provided excellent results with regard to reproducibility, yield of mRNA densely loaded with ribosomes, and separation of mRNA associated with different numbers of polyribosomes. Use of this technique allowed us to determine that the polysome profiles of the different developmental stages are distinctly different, with higher ribosome loading in the proliferating stages. The lengths of the poly(A) tails on the total population of RNA from the different developmental stages showed no significant variation. These data indicate that changes in polysome loading of mRNAs accompany development, and that they do not reflect bulk changes in polyadenylation. We speculate that developmental changes in translation reflect reduced translational initiation.

The Etruscan shrew, Suncus etruscus, is one of the smallest mammals. Etruscan shrews can recognize prey shape with amazing speed and accuracy, based on whisker-mediated tactile cues. Because of its small size, quantitative analysis of the Etruscan shrew cortex is more tractable than in other animals. To quantitatively assess the anatomy of the Etruscan shrew's brain, we sectioned brains and applied Nissl staining and NeuN (neuronal nuclei) antibody staining. On the basis of these stains, we estimated the number of neurons of 10 cortical hemispheres by using Stereoinvestigator and Neurolucida (MBF Bioscience) software. On average, the neuron number per hemisphere was found to be ~1 million. We also measured cortical surface area and found an average of 11.1 mm² (n = 7) and an average volume of 5.3 mm³ (n = 10) per hemisphere. We identified 13 cortical regions by cytoarchitectonic boundaries in coronal, sagittal, and tangential sections processed for Nissl substance, myelin, cytochrome oxidase, ionic zinc, neurofilaments, and vesicular glutamate transporter 2 (VGluT2). The Etruscan shrew is a highly tactile animal with a large somatosensory cortex, which contains a barrel field, but the barrels are much less clearly defined than in rodents. The anatomically derived cortical partitioning scheme roughly corresponds to physiologically derived maps of neocortical sensory areas.