Lepidosirenidae is a clade of freshwater lungfishes that comprise the South American Lepidosiren paradoxa and four African species of the genus Protopterus. These two genera have been geographically separated since the Early Cretaceous break-up of Gondwana, but they share similar biology and skeletal morphology. The lepidosirenid species traditionally were distinguished by a combination of features such as head-to-body ratios, the number of pairs of vertebral ribs, and the presence or absence of external gills, but there are no published discrete skeletal characteristics and no published comparative studies including all extant species. I used High Resolution X-Ray Computed Tomography (CT), X-Ray photography, and alcohol-preserved, cleared-and-stained, and dry skeletal specimens from museum collections to describe the skeletal morphology of all species of lepidosirenid lungfishes in a comparative context. I digitally disarticulated the bones in each CT scan to compile a comprehensive comparative atlas of the cranial and pectoral elements of all extant lungfish. I discovered that the anocleithrum in Lepidosiren paradoxa, which was previously thought to be lacking, is actually present. I also identified skeletal differences between species in the frontoparietal, parasphenoid, supraorbital, and suboperculum. I incorporated those characters into the first morphological phylogenetic analysis to determine the interrelationships of the lepidosirenids. I also used previously published molecular sequence data from the ribosomal RNA gene 16s to run combined morphological and molecular phylogenetic analyses. To generate phylogenetic hypotheses using different types of data and different methods of determining phylogeny, I employed the maximum parsimony, maximum likelihood, and Bayesian inference methods. Lepidosirenidae is monophyletic in almost all analyses, Protopterus is monophyletic in each analysis, and Protopterus annectens and Protopterus aethiopicus are sister taxa in every analysis. The phylogenetic positions of Protopterus dolloi and Protopterus amphibius are incongruent in many of the analyses, which indicates that further examination of the skeletal variation and addition of molecular sequences of different genes is needed. Based on the comparative morphological atlas and the phylogenetic analyses, questions of lepidosirenid biogeography, morphological variation within lungfish, and better identification of lungfish fossils can now be investigated in a more rigorous context. / text

Submitted by Cássio da Cruz Nogueira (cassionogueirakk@gmail.com) on 2017-01-26T13:59:31Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_AnaliseMorfologicaMolecular.pdf: 1631378 bytes, checksum: 5757c5c8705000ab31e5fe11f2a6a2d0 (MD5) / Approved for entry into archive by Edisangela Bastos (edisangela@ufpa.br) on 2017-01-27T13:33:58Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_AnaliseMorfologicaMolecular.pdf: 1631378 bytes, checksum: 5757c5c8705000ab31e5fe11f2a6a2d0 (MD5) / Made available in DSpace on 2017-01-27T13:33:58Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_AnaliseMorfologicaMolecular.pdf: 1631378 bytes, checksum: 5757c5c8705000ab31e5fe11f2a6a2d0 (MD5) Previous issue date: 2015-12-18 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A espécie Lepidosiren paradoxa pertence à ordem Dipnoi, juntamente com mais dois gêneros, sendo considerados os peixes pulmonados verdadeiros. Os machos adultos de L. paradoxa diferenciam-se das fêmeas através da presença de filamentos nas nadadeiras pélvicas. Estes filamentos assemelham-se àqueles encontrados em brânquias de peixes e de salamandras neotênicas. Estes filamentos desenvolvem-se e tornam-se vascularizados durante o período de reprodução. Neste trabalho, propomos testar a hipótese de que os filamentos pélvicos de L. paradoxa compartilham características morfológicas e moleculares com filamentos brânquiais. Para tanto, realizamos análise morfológica e molecular dos filamentos das nadadeiras pélvicas entre as estações de estiagem e chuvosa. A análise morfológica ocorreu através de coloração de hematoxilina e eosina (HE) e microscopia eletrônica de varredura (MEV). Por fim, foi feita a quantificação de expressão gênica de marcadores enriquecidos em brânquias de peixes, através de PCR em Tempo Real (RT-PCR), utilizando a nadadeira peitoral como referência. O comprimento dos filamentos da estação chuvosa e de estiagem apresentaram os valores de média e desvio padrão de, 4,31 mm±0,186 e 1,63 mm±0,104, respectivamente. Nas imagens de MEV foram observadas algumas células com microvilosidades e/ou microciliadas e algumas células menores. Nas análises de HE os filamentos apresentaram uniformidade no seu epitélio formada com 4 camadas de células, sendo preenchido por tecido conjuntivo e por fim tornam-se mais vascularizados na estação chuvosa. Na analise molecular de RT-PCR os marcadores selecionados não apresentaram variação quando comparados com a nadadeira peitoral e entre as estações. Em conclusão, apesar de existirem semelhanças morfológicas entre filamentos pélvicos de L. paradoxa e filamentos brânquiais de peixes e anfíbios, os dados moleculares aqui obtidos não suportam a hipótese de que estes filamentos realizem trocas gasosas. / The Lepidosiren paradoxa species belongs to Dipnoi order, along with two genre and are considered true lungfishes. Adult males of L. paradoxa differ from females by the presence of filaments in the pelvic fin. These filaments resemble those found in gills of fish and neotenic salamanders. These filaments are grow and become vascularized during the reproduction period. In this work we tested the hypothesis that the pelvic filament of L. paradoxa share morphological and molecular characteristics with gill filaments. Thus, we performed morphological and molecular analyses of the filaments of the pelvic fins between the dry and rainy seasons. Morphological analysis was perfomed through hematoxylin and eosin (HE) and scanning electron microscopy (SEM). Finally, we quantified gene expression of gill markers by RT-PCR using pectoral fin as reference. The mean length and standard deviation of the filaments during rainy and dry season were 4,31mm of ± 0.186 and 1,63mm ± 0.104, respectively. In the SEM images, we observed with a few cell microvilli and/or microplicae and some smaller cells. In the HE analyzes we found that the filaments have uniform epithelium formed with four layers of cells being filled by connective tissue and finally become more vascularized the rainy season. Molecular analysis through RT-PCR did not show any change when compared to the pectoral fin and between the stations. In conclusion, despite morphological and molecular similarities between gill filaments and pelvic fin filaments, our findings do not support a role for L. paradoxa pelvic filaments in gas exchange.