Description, measurement, and automatic classification of the Plasmodium berghei oocyst morphology during early differentiation in vitro

Abstract

After colonization of the mosquito midgut by the malaria parasite, Plasmodium differentiates from an invasive, motile ookinete to a multiplicative, sessile oocyst. Despite their importance in establishing the infection and increasing its population, relatively little is known about the early morphological transformation associated with these changes in function. Oocyst differentiation begins with the formation of a spherical protrusion near the center of the crescent-shaped ookinete. As this protuberance grows, it engulfs the content of the two distal ends, thus rounding the cell. In this work, scrutinized observations of the overall changes in shape, coupled with the migration of the malaria pigment granules and the nucleus into the protuberance, revealed that the movement of the cell content happens in an anteroposterior manner. The resulting data, formalized as morphometric measurements, led to the identification of 5 transitional stages and to the development of a computer training algorithm that automatically classifies them. Since cell differentiation has been associated with redox fluctuations, the classification algorithm was tested with parasites stained with a glutathione-specific fluorescent probe. This revealed changes in the glutathione content during differentiation that are suggestive of a redox modulation during transformation. • Plasmodium ookinetes differentiate into reproductive oocysts after mosquito invasion. • Visual scrutiny of the transformation led to the recognition of 5 transitional stages. • Transformation and organelle migration happens in an anteroposterior manner. • A machine learning algorithm was developed to automatically classify these stages. • The discretization into stages allowed the determination of GSH oscillations.