Reversal of diabetic-induced myopathy by swimming exercise in pregnant rats:a translational intervention study

Gestational diabetes mellitus (GDM) plus rectus abdominis muscle (RAM) myopathy predicts long-term urinary incontinence (UI). Atrophic and stiff RAM are characteristics of diabetes-induced myopathy (DiM) in pregnant rats. This study aimed to determine whether swimming exercise (SE) has a therapeutic effect in mild hyperglycemic pregnant rats model. We hypothesized that SE training might help to reverse RAM DiM. Mild hyperglycemic pregnant rats model was obtained by a unique subcutaneous injection of 100 mg/kg streptozotocin (diabetic group) or citrate buffer (non-diabetic group) on the first day of life in Wistar female newborns. At 90 days of life, the rats are mated and randomly allocated to remain sedentary or subjected to a SE protocol. The SE protocol started at gestational day 0 and consisted of 60 min/day for 6 days/week in a period of 20 days in a swim tunnel. On day 21, rats were sacrificed, and RAM was collected and studied by picrosirius red, immunohistochemistry, and transmission electron microscopy. The SE protocol increased the fiber area and diameter, and the slow-twitch and fast-twitch fiber area and diameter in the diabetic exercised group, a finding was also seen in control sedentary animals. There was a decreased type I collagen but not type III collagen area and showed a similar type I/type III ratio compared with the control sedentary group. In conclusion, SE during pregnancy reversed the RAM DiM in pregnant rats. These findings may be a potential protocol to consider in patients with RAM damage caused by GDM

Estudo filogenetico na superfamilia Loricarioidea (Teleostei: Siluriformes) com base na ultraestrutura dos espermatozoides

Orientadores: Irani Quagio-Grassiotto, Claudio de OliveiraDissertação (mestrado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: O resumo poderá ser visualizado no texto completo da tese digitalAbstract: The abstract is available with the full electronic digital documentMestradoBiologia CelularMestre em Biologia Celula

Phylogenetic analysis of superfamily Loricarioidea (Teleostei: Siluriformes) based on spermiogenesis and spermatozoa ultrastructure

Orientador: Irani Quagio-Grassiotto, Claudio de OliveiraTese (doutorado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: A ordem Siluriformes compreende um grupo diverso e amplamente distribuído dentre os Ostariophysi, apresentando 36 famílias com aproximadamente 480 gêneros e mais de 3.000 espécies. Entre os grupos de Siluriformes neotropicais, reconhecidamente monofiléticos, está a superfamília Loricarioidea. Os relacionamentos filogenéticos entre as famílias de Siluriformes, sugerem que Loricarioidea é grupo irmão de Amphiliidae, uma família africana de Siluriformes. A superfamília Loricarioidea encontra-se atualmente constituída por seis famílias: Nematogenyidae, Trichomycteridae, Callichthyidae, Scoloplacidae, Astroblepidae e Loricariidae. Apesar do conhecimento atual sobre os relacionamentos entre as famílias de Siluriformes vir sendo obtido com base em caracteres osteológicos, outros dados como a ultra-estrutura da espermiogênese e dos espermatozóides parecem ser potencialmente úteis na elucidação dos relacionamentos de grupos. O objetivo deste estudo foi a realização da análise filogenética da superfamília Loricarioidea, com base nas características ultra-estruturais da espermiogênese e dos espermatozóides, visando testar a real habilidade destes dados na resolução dos relacionamentos filogenéticos intra e inter-familiar, bem como na ordem Siluriformes. Foi feita a descrição das características ultra-estruturais das células germinativas masculinas em 27 representantes de diferentes famílias de Loricarioidea. A análise geral dos dados obtidos revelou que quando essa classe de caracteres ultra-estruturais reprodutivos é utilizada em grupos mais restritos como a superfamília Loricarioidea, observa-se que ela pode ser mais informativa e pode corroborar o monofiletismo de alguns grupos. Entretanto, o uso desses caracteres nas análises filogenéticas em nível de ordem não é informativo, uma vez os grupos sugeridos são muito incongruentes com a hipótese de relacionamento disponível para os Siluriformes. Além disso, alguns caracteres que poderiam representar sinapomorfias, tornam-se homoplasias quando se considera a ocorrência da mesma característica em outros grupos não relacionados. Portanto, o emprego desses caracteres ultra-estruturais reprodutivos em análises filogenéticas deverá ser criteriosamente planejado, evitando-se interpretações equivocadasAbstract: The order Siluriformes comprises the most diverse and widely distributed ostariophysan groups, presenting thirty-six families with approximately 480 genera and over 3.000 species. Among the neotropical siluriform lineages likely to be monophyletic is the superfamily Loricarioidea. The relationships among catfish families suggest that Loricarioidea is sister group of Amphiliidae, an African siluriform family. The Loricarioidea is constituted by six families: Nematogenyidae, Trichomycteridae, Callichthyidae, Scoloplacidae, Astroblepidae, and Loricariidae. Although the current knowledge of the relationships among siluriform families has been acquired on the basis on osteological characters, other data such as the spermiogenesis and spermatozoal ultrastructure seem be potentially useful in the clarification of the relationships of the groups. The aim of the present study was to develop a phylogenetic analysis of the superfamily Loricarioidea, using the ultrastructural characteristics of both spermiogenesis and spermatozoa as a test to evaluate the ability of this data in resolving the phylogenetic relationships inside the families, among families and in the order Siluriformes. The description of the ultrastructural characteristics of male germinative cells in 27 specimens of different families of Loricarioidea was presented. The general analysis of the data obtained revealed that when this class of reproductive ultrastructural characters is employed in a more restrict group, as the superfamily Loricarioidea, it is really informative and can strongly support the monophyly of some groups. However, the phylogenetics analysis using these characters is not informative at order level as the suggested groups are very incongruent with the available hypotheses for Siluriformes. Moreover, some characters that could represent synapomorphies, change to homoplasies considering the occurrence of the same characters in other unrelated groups. Then, the use these reproductive ultrastructural characters in phylogenetics analysis should be carefully planed to avoid erroneous conclusionsDoutoradoBiologia CelularDoutor em Biologia Celular e Estrutura

Occurrence of biflagellate spermatozoa in Cetopsidae, Aspredinidae, and Nematogenyidae (Teleostei : Ostariophysi : Siluriformes)

In the present study spermiogenesis was investigated in Cetopsis coecutiens (Cetopsidae), and Bunocephalus amazonicus (Aspredinidae), while spermatozoa ultrastructure was investigated in C. coecutiens, B. amazonicus, and Nematogenys inermis (Nematogenyidae). Aspredinidae and Cetopsidae share a spermatogenesis of the semicystic type, and a particular type of spermiogenesis process not reported in any fish group. In the three species analyzed, spermatozoa are biflagellate with flagella having the classical axoneme formulae (9 + 2). The analysis of thirteen characters showed the presence of eight characters shared by Cetopsidae and Aspredinidae, and six characters shared by Cetopsidae and Nematogenyidae, which may suggest that these three families may be more related than actually hypothesized, comprising a very primitive siluriform lineage originated after Diplomystidae

Spermiogenesis and spermatozoal ultrastructure in Trichomycteridae (Teleostei: Ostariophysi: Siluriformes)

Spadella, M.A., Oliveira, C. and Quagio-Grassiotto, I. 2009. Spermiogenesis and spermatozoal ultrastructure in Trichomycteridae (Teleostei: Ostariophysi: Siluriformes). -Acta Zoologica (Stockholm) 91: 373-389.Siluriformes comprises the most diverse and widely distributed ostariophysan group, a fish assemblage that includes about three quarters of the freshwater fish of the world. In this study, the ultrastructural characterization of spermiogenesis and spermatozoa in specimens of Copionodontinae (the sister group to all other trichomycterids), Trichomycterinae (a derived trichomycterid group), and Ituglanis (a genus not assigned to any trichomycterid subfamily) is presented. The comparative analyses of the data show that trichomycterid species share six of seven analyzed spermiogenesis characters, reinforcing the monophyly of the group. Analyses of trichomycterid sperm ultrastructure showed that the species studied share the same character states for nine of seventeen characters analyzed. Copionodon orthiocarinatus and Ituglanis amazonicus each share more ultrastructural characters with species of Trichomycterus than with one another. Regarding the families of Loricarioidea, the species of Trichomycteridae share more characters of spermatogenesis, spermiogenesis, and sperm with representatives of the families Callichthyidae, Loricariidae, and Scoloplacidae than with Nematogenyidae, its hypothesized sister group. With the exception of the family Nematogenyidae, the character similarities observed reinforce the monophyly of the superfamily Loricarioidea.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Morphology and histology of male and female reproductive systems in the inseminating species Scoloplax distolothrix (Ostariophysi : Siluriformes : Scoloplacidae)

The morphology and histology of male and female reproductive systems were examined in Scoloplax distolothrix. Internal insemination was documented in this species by the presence of sperm within the ovaries. Mature males and females have elongated genital papillae, exhibiting a tubular shape in males and a plain heart-shape with two median protuberances in females. The testes are two elongated structures that converge ventrally, under the intestine, towards the genital papilla. They are joined at the caudal end, forming an ovoid single chamber for sperm storage. Secretory regions were not observed. In the lumen of the testicular tubules, spermatozoa can be tightly packed along their lengths, but do not constitute a spermatozeugmata. The lumen of the sperm storage chamber and spermatic duct are filled with free spermatozoa without the accompanying secretions. The ovaries are bird-wing shaped, saccular structures that converge ventrally under the intestine, towards the genital papilla. They are joined at the caudal end, forming a tubular chamber possibly destined for oocyte storage. An oviduct with an irregular outline connects the chamber to the tubular region of the genital papilla. No distinct sperm storage structure was found in the ovaries. The unique male and female genital papillae suggest that these structures are associated with the reproductive mode in scoloplacids, representing evidence for insemination. The occurrence of free spermatozoa, without the accompanying secretions and not arranged in a spermatozeugmata can be associated with the presence of a tubular male genital papilla for sperm transfer to the female genital tract. This reinforces the idea that sperm packets are not necessary for all inseminating species. The male reproductive system in scoloplacids is very different from that in auchenipterids, a second catfish family with insemination, which indicates that the occurrence of insemination is not connected to the internal morphology of reproductive organs.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Male and female reproductive morphology in the inseminating genus Astroblepus (Ostariophysi: Siluriformes: Astroblepidae)

The structure of the reproductive organs of males and females of four species of Astroblepus were studied by light microscopy and TEM. In females, the gonadal and urinary orifices are separated. The urinary pore opens at the tip of a urinary papilla. In males, the testes conform to the anastomosing tubular type. Spermatogenesis is partially cystic, with most spermatid stages taking place within the lumen of the seminiferous tubules. The sperm ducts pass through seminal vesicles. The main sperm ducts fuse and join the urinary duct to form a urogenital duct that passes through to the tip of an long urogenital papilla. A circularly arranged skeletal muscle at the base of the urogenital papilla probably serves as a sphincter. The spermatozoon is an anacrosomal introsperm with an elongate, conical nucleus and an elongate midpiece. The midpiece contains numerous fused mitochondria that form rings within the cytoplasmic collar, but no vesicles were observed. The flagellum has a classic 9 + 2 axoneme with the A-tubules of the peripheral doublets being electron-dense. Two lateral fins on the posterior part of the flagellum have electron-dense termini. Nuclear chromatin is highly condensed, and the two centrioles are oriented perpendicularly to one another and are contained within a nuclear fossa. (C) 2011 Elsevier GmbH. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

The enigmatic role of cholinergic reflex in the pathogenesis of Chagas disease

This study evaluated the inflammatory process in the colons of mice infected with Trypanosoma cruzi QM2 strain, through the analysis of muscle reactivity and the measurement of butyrylcholinesterase (BuChE) in plasma. Swiss mice were infected with T. cruzi QM2 strain and after 15 (G15), 30 (G30), 60 (G60), 90 (G90), and 210 (G210) days, each group had blood collected for the measurement of butyrylcholinesterase plasma concentrations ([BuChE]), a measure which functioned as an indicator of plasmatic Ach levels. All groups, except G15, had a segment of proximal colon removed to assess muscle reactivity to acetylcholine (Ach) and noradrenaline (NA) stimulation. After reactivity tests, the tissues were then fixed and stained with hematoxylin and eosin (HE) for histological evaluation of inflammatory response. The QM2 strain did induce inflammatory process in mice colon, and demonstrated differences in muscular contraction between the G60 and G210 groups, with p < 0.05. Plasma [BuChE] increased during the acute phase of infection (p < 0.05) with subsequent heterogeneous decline in the late chronic phase. These results show that the QM2 strain has tropism to the colon of mice and causes damage characteristic of megacolon; also, Ach has an enigmatic importance in the anti-inflammatory reflex over the course of T. cruzi infection.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP