Cytogenetic and molecular studies in a lungfish, Protopterus annectens (Osteichthyes, Dipnoi)

A neontological approach to the problem of the origin of tetrapods consists in the examination of the available cytological and molecular data on the genome of these vertebrates. Dipnoans are a group of osteichthyian fishes, the evolutionary relationships of which with tetrapods have been disputed since their discovery. In the past, they were variously considered as being related to actinistians, tetrapods, and lower actinopterygians, though nowadays they are considered a monophyletic group, the sister group of crossopterygians. Dipnoans first appeared in the geologic record in the Early Devonian with 50 extinct genera, surviving up to date, with only three genera: Lepidosiren, Neoceratodus and Protopterus, including only six recognized species. Nothing is known of the genome of the early tetrapods, except that they and the Choanoichthyes exhibited a remarkable interspecific variability of the karyotype and of DNA content. These characteristics are often found in dipnoans and in the extant lissamphibians. Very little is known about the evolutionary karyology in the four Protopterus species and in the dipnoan clade in general. In this paper, we karyotyped ten male and female specimens of P. annectens (2n=34) from Nigeria. Moreover, we localized heterochromatin and nucleolar organizer regions by using base-specific fluorochromes and detected the human telomeric (TTAGGG)(n) sequences on all the telomeric sites of P. annectens chromosomes. DNA was also extracted and digested with seven restriction enzymes, which revealed the probable presence of almost three different families of satellite DNA. Nuclear DNA content was identified from blood samples by flow cytometry. New genomic and karyological data were compared and discussed with those on closer genera and taxa available in literature.