Mieke Boone

Ontogeny of the complex sperm in the macrostomid flatworm Macrostomum lignano (Macrostomorpha, Rhabditophora)

Spermiogenesis in Macrostomum lignano (Macrostomorpha, Rhabditophora) is described using light‐ and electron microscopy of the successive stages in sperm development. Ovoid spermatids develop to highly complex, elongated sperm possessing an undulating distal (anterior) process (or “feeler”), bristles, and a proximal (posterior) brush. In particular, we present a detailed account of the morphology and ontogeny of the bristles, describing for the first time the formation of a highly specialized bristle complex consisting of several parts. This complex is ultimately reduced when sperm are mature. The implications of the development of this bristle complex on both sperm maturation and the evolution and function of the bristles are discussed. The assumed homology between bristles and flagellae questioned. J. Morphol., 2009.

Spermatogenesis and the structure of the testes in Nemertodermatida

The ultrastructure of the testes in two representatives of the enigmatic taxon Nemertodermatida was studied using transmission electron microscopy. Nemertoderma westbladi has paired testes, which are delineated by lining cells. Within each testis, different follicles, each surrounded by a membrane-like structure, are found. Flagellophora apelti has genuinely follicular testes, consisting of several follicles, each containing a certain stage of spermatogenesis. As the gametes are not enclosed by a structure that can be called a true gonad, the structure of the testes differs from most bilaterian animals, but resembles the organization of gametogenic areas of ctenophores. Each stage of spermatogenesis in F. apelti is described, enabling the inference of the origin of the structures seen in mature spermatozoa. The overall structure of the mature spermatozoa is similar in all nemertodermatids and unique within the Metazoa: an elongated head containing the nucleus; a middle piece containing an axoneme, mitochondrial derivatives and in F. apelti granular derivatives; and a flagellar tail.

Use of freeze-cracking in ontogenetic research in Macrostomum lignano (Macrostomida, Rhabditophora)

A method for studying whole mount flatworm embryos based on freeze-cracking of the eggs is described. This method allows successful immunohistological and immunocytological studies of whole mount embryos. It does not require the use of sharpened needles or a microinjection system to puncture the eggshell. Moreover, this method is more practical and less time-consuming than classical puncturing and much cheaper than the use of a microinjection system. The advantages of this method are illustrated by results of several immunolocalisation experiments in the macrostomid flatworm Macrostomum lignano. The optimal procedure and crucial steps for this method are discussed.