The analysis of cell composition in the hemolymph of three Planorbidae species (Gastropoda: Pulmonata)

Abstract

РЕЗЮМЕ: Изучен клеточный состав гемолимфы трёх видов пульмонат семейства Planorbidae: Biomphalaria glabrata, Planorbarius corneus и Planorbis planorbis. По данным морфологического, морфометрического и цитофлуориметрического анализов гемоциты этих моллюсков представлены популяцией гиалиноцитов и гетерогенной фракцией, состоящей из двух популяций гранулоцитов-крупных и мелких клеток. С использованием специфического лизосомного красителя LysoTracker и красителя нуклеиновых кислот SYTO62 в составе каждой популяции определены четыре субпопуляции клеток. Между клетками, относящимися к разным морфологическим популяциям, обнаружены достоверные различия в содержании лизосом и накоплении нуклеинового красителя. Гиалиноциты содержат меньшее количество лизосом, чем гранулоциты. При этом крупные гранулоциты характеризуются наибольшей концентрацией лизосом и метаболической активностью, а мелкие сильнее варьируют по степени гранулярности и более гетерогенны по составу гранул.

Changes in the cell composition of the hemolymph in the snail Planorbarius corneus after infection with the trematode Plagiorchis sp.

“…Using these dyes, we distinguished two main subpopulations of hyalinocytes, differing in the metabolic activity and the functional state (Figure 9). In a previous study, we also identified subpopulations of granulocytes based on morphological features (Prokhorova, Serebryakova, Tokmakova, Kudryavtsev, et al, 2018). However, no functional cell groups among granulocytes were found in this study.…”

“…Flow cytofluorometry is now actively used for studying molluscan hemolymph in addition to morphological analysis (Accorsi et al, 2013; Ataev et al, 2016; Cueto et al, 2015; Martins‐Souza et al, 2009; Ottaviani & Montagnani, 1989; Portet et al, 2019; Prokhorova, Serebryakova, Tokmakova, & Ataev, 2018; Prokhorova, Serebryakova, Tokmakova, Kudryavtsev, et al, 2018; Rolton et al, 2020; Tokmakova et al, 2020). However, most studies register only forward scatter and side scatter values, which reflect cell size and granularity.…”

“…In this study, we applied a previously developed method for analyzing the population composition of hemocytes in pulmonates, which is based on the estimation of the amount of nucleic acids and lysosomes in cells (Prokhorova, Serebryakova, Tokmakova, & Ataev, 2018; Prokhorova, Serebryakova, Tokmakova, Kudryavtsev, et al, 2018). The nucleic acid and lysosome content may indicate functional and metabolic properties of hemocytes of different types.…”

Hemolymph of molluscan origin: from biochemistry to modern biomaterials science

“…The smallest molluscan hyalinocytes reported in the literature were found in tropical freshwater snail Bulinus africanus (Krauss, 1848), 2.6 μm in diameter [114], and the largest in air-breathing freshwater snail Bulinus truncates (Audouin, 1827), 28 μm in diameter [115]. In contrast, granulocytes have numbers of cytoplasmic granules and exhibit effective phagocytosing activity [106,116]. According to the most scientific reports, this type constitutes the dominant hemocyte population [9].…”

“…Over the years, the classification of molluscan hemocytes has been carried out by various methods, for example: light and electron microscopy [38,117,[126][127][128], differential centrifugation [129], flow cytometry [111,112,116,130], as well as lectin and antibody binding [88,131]. Also various kinds of staining, including Giemsa [108,112,113,127,132], hematoxylin-eosin [37,38,112], calcein AM [36,37], neutral red [112], Methylene Blue-eosin (Diff-Quick or Hemacolor) [113,133] and acridine orange staining [134], have been used for hemocyte classification and identification.…”