In fish leeches (Piscicolidae) indirect (hypodermic) insemination has evolved, thus the spermatophores are released in the specialised region of the body wall known as a copulatory area or a copulatory region. The way in which the spermatozoa reach the ovaries is not fully understood. In piscicolids beneath the copulatory area there is a specialized connective tissue (vector tissue), which is thought to guide the spermatozoa toward the ovaries. To date the structure of the vector tissue has not been observed in copulating specimens, which have spermatophores implanted in their coplulatory area. Here we present the first ultrastructural observation of massive sperm transfer from the spermatophore throughout the vector tissue to the ovaries. Our results show that the sperm transfer is both massive and rapid. The migrating spermatozoa form huge aggregations which push aside the vector tissue cells, in such a way that between these cells voluminous gaps are formed. Unexpectedly to our previous suggestions, the ultrastructural pictures show that the long cytoplasmic processes of granular cells, which constitute the main mass of the vector tissue, are not engaged in sperm transport. We suggest that the sperm is pumped with a high pressure from the spermatophore into the vector tissue, and as a result the vector tissue cells are pushed aside and spermatozoa can freely pass between them.
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