Ultrastructural analysis of apoptosis and autophagy in the midgut epithelium of Piscicola geometra (Annelida, Hirudinida) after blood feeding.

M M Rost-Roszkowska,M Kszuk-Jendrysik,W Rupik,E Swadźba,I Poprawa,P Świątek

doi:10.1007/s00709-015-0774-9

Abstract

Cell death in the endodermal region of the digestive tract of the blood-feeding leech Piscicola geometra was analyzed using light and transmission electron microscopes and the fluorescence method. Sexually mature specimens of P. geometra were bred under laboratory conditions and fed on Danio rerio. After copulation, the specimens laid cocoons. The material for our studies were non-feeding juveniles collected just after hatching, non-feeding adult specimens, and leeches that had been fed with fish blood (D. rerio) only once ad libitum. The fed leeches were prepared for our studies during feeding and after 1, 3, 7, and 14 days (not sexually mature specimens) and some weeks after feeding (the sexually mature). Autophagy in all regions of the endodermal part of the digestive system, including the esophagus, the crop, the posterior crop caecum (PCC), and the intestine was observed in the adult non-feeding and feeding specimens. In fed specimens, autophagy occurred at very high levels—in 80 to 90 % of epithelial cells in all four regions. In contrast, in adult specimens that did not feed, this process occurred at much lower levels—about 10 % (esophagus and intestine) and about 30 % (crop and PCC) of the midgut epithelial cells. Apoptosis occurred in the feeding adult specimens but only in the crop and PCC. However, it was absent in the non-feeding adult specimens and the specimens that were collected during feeding. Moreover, neither autophagy nor apoptosis were observed in the juvenile, non-feeding specimens. The appearance of autophagy and apoptosis was connected with feeding on toxic blood. We concluded that autophagy played the role of a survival factor and was involved in the protection of the epithelium against the products of blood digestion. Quantitative analysis was prepared to determine the number of autophagic and apoptotic cells.

Highlights

During digestion in blood-feeding animals, the blood generates many molecules which may be toxic or even lethal for the organism (Dunkov et al 2002; Taketani 2005)
The aims of this study were (1) to study which kind(s) of cell death occur(s) in nonfeeding juveniles, non-feeding adults, and feeding adult specimens; (2) to analyze the relationship between cell death and the time after feeding; (3) to describe cell death at the ultrastructural level; and (4) to state whether cell death participates in the neutralization of toxic substances which originate from blood
The material for our studies Non-feeding adult specimens was non-feeding juveniles collected just after hatching, non-feeding adult speci- Apoptosis mens, adult specimens dur- Adult feeding specimens ing feeding, and Autophagy leeches that had been fed with fish blood (D. rerio) only once Apoptosis ad libitum

Summary

Introduction

During digestion in blood-feeding animals, the blood generates many molecules which may be toxic or even lethal for the organism (Dunkov et al 2002; Taketani 2005). The perimicrovillar or the peritrophic membranes, which separate the midgut epithelium from the midgut lumen, protect the organism against the entrance of pathogens or toxic substances (Terra 2001; Albuquerque-Cunha et al 2004; Terra and Ferreira 2012). They can be discharged from the midgut epithelium into its lumen As a consequence, such cells will not affect the entire epithelium and the organism will be protected against the effects of stress factors (e.g., toxic substances) (Franzetti et al 2012; RostRoszkowska et al 2012b; Wilczek et al 2014).

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