Microscopic analysis of spermatogenesis and mature spermatozoa in the amphibian leech Batracobdella algira (Annelida, Clitellata, Hirudinida).

Abstract

Spermatogenesis and spermatozoa ultrastructure of the amphibian leech Batracobdella algira Moquin-Tandon, 1846 (Hirudinida: Glossiphoniidae) have been investigated by means of electron and fluorescent microscopy. In B. algira, there are seven pairs of testisacs (testes) that are located latero-ventrally throughout the body. Each testis contains numerous cysts with developing germ cells. The germ cells in a given cyst are in the same developmental stage (i.e., there are spermatogonial, spermatocytic, and spermatid cysts); however, there is no developmental synchrony between the cysts, and therefore, all of the developmental stages occur simultaneously in the same testis. In the cysts, each germ cell is connected to acentral cytoplasmic mass, the cytophore, by one intercellular bridge. The spermatozoa of the studied species conform to the general organization plan that is known for Hirudinida: they are filiform cells that are formed in sequence by an elongated and twisted acrosome that consists of an anterior and posterior acrosome, a fully condensed and helicoid nucleus, a midpiece composed of a single and twisted mitochondrion that is characteristically surrounded by an electron-dense sheath, and a flagellum with the conventional 9 × 2 + 2 axonemal pattern. Using a comprehensive approach, we compared our findings with the ultrastructural data that had been obtained from the spermatozoa of the other hirudinids that have been studied to date. Only minor differences in the length and shape of the studied organelles were found which seems to be connected with the different ways of insemination, specific properties of female reproductive tracts, and physiology of fertilization. Additionally, we studied the organization of the microtubular cytoskeleton in male germline cysts at consecutive stages of spermatogenesis using fluorescent and electron microscopy. By comparing the present data with those from Oligochaeta, Branchiobdellida, and Acanthobdellida, we found that only the presence of an anterior acrosome characterizes the true leeches and that, at present, should be regarded as an autapomorphic character of Hirudinida. Our results showed that the arrangement of the microtubules changed dynamically during spermatogenesis.