Bruno Machado Bertassoli

Diaphragm: A vital respiratory muscle in mammals

The diaphragm is a respiratory muscle that is primarily responsible for the respiratory function in normal individuals. In mammals, the diaphragm muscle has been studied from the early days of zoology, comparative and experimental anatomy, physiology, medicine, physics, and philosophy. However, even with these early advances in knowledge pertaining to the diaphragm, comprehensive morphological data on the diaphragm are still incomplete. In this review, we summarize the beginnings of the morphological description of the diaphragm, and we describe the current status of the known morphological and embryological features. In addition, we correlate how the impairment of the diaphragm muscle in Duchenne muscular dystrophy (DMD) can lead to patient deaths. DMD is the most common X-linked muscle degenerative disease and is caused by a lack of dystrophin protein. Dystrophin is an important muscle protein that links the cellular cytoskeleton with the extracellular matrix. In the absence of dystrophin, the muscle becomes susceptible to damage during muscle contraction. This review allows researchers to obtain an overview of the diaphragm, transcending the morphological data from animals described in conventional literature.

Characterization of the estrous cycle in Galea spixii(Wagler, 1831)

The Galea spixii inhabits semiarid vegetation of Caatinga in the Brazilian Northeast. They are bred in captivity for the development of researches on the biology of reproduction. Therefore, the aim of this study is characterize the estrous cycle of G. spixii, in order to provide information to a better knowledge of captive breeding of the species. The estrous cycle was monitored by vaginal exfoliative cytology in 12 adult females. After the detection of two complete cycles in each animal, the same were euthanized. Then, histological study of the vaginal epithelium, with three females in each phase of the estrous cycle was performed; five were paired with males for performing the control group for estrous cycle phases, and three other were used to monitor the formation and rupture of vaginal closure membrane. By vaginal exfoliative cytology, predominance of superficial cells in estrus, large intermediate cells in proestrus, intermediate and parabasal cells, with neutrophils, in diestrus and metestrus respectively was found. Estrus was detected by the presence of spermatozoa in the control group. By histology, greater proliferation of the vaginal epithelium in proestrus was observed. We conclude that the estrous cycle of G. spixii lasts 15.8 ± 1.4 days and that the vaginal closure membrane develops until complete occlusion of the vaginal ostium, breaking after few days. Future studies may reveal the importance of this fact for the reproductive success of this animal.

Mesenchymal stem cells: emphasis in adipose tissue

The study of stem cells has evolved rapidly in recent decades. The importance is given to the concept that these cells are potentially able to become any cell type and have the power of self-renewal throughout the life of the organism. Mesenchymal stem cells (MSCs) can be isolated from various organs of the body such as bone marrow, adipose tissue, synovium, muscle and dermis, deciduous teeth, umbilical cord, placenta, liver, spleen and thymus. After their isolation in vitro, mesenchymal stem cells have the capacity to differentiate into various mesenchymal lineages and various tissues after the use of appropriate cultures. Studies have reported that mesenchymal stem cells from adipose tissue have the potential to differentiate themselves, like the cells commonly studied bone marrow. Adipose tissue is attractive due to its easy access, rapid expansion in vitro and only one collects the large amount of tissue. This review intends to show the protocols for isolation, cell culture and means of commercial cellular differentiation most widely used with emphasis on adipose tissue.

Arquitetura comparativa dos pulmões de camundongos normais e afetados pela Distrofia Muscular de Duchenne

A Distrofia Muscular de Duchenne (DMD) e uma doenca genetica de carater recessivo que caracterizada por fraqueza muscular progressiva de cintura pelvica e escapular evoluindo para insuficiencia respiratoria e, ou cardiaca. O camundongo mdx e um modelo amplamente utilizado para estudos da DMD. Apesar do fenotipo destes animais serem mais suave, estes apresentam o principal musculo respiratorio, o diafragma com morfologia e bioquimica semelhante a DMD humana, fato este que pode comprometer a funcao respiratoria e consequentemente os pulmoes. Foi realizado um estudo anatomico descritivo do parenquima pulmonar dos pulmoes de 5 animais modelo mdx comparando estes com os pulmoes de 5 camundongos BALB/C57 (Mus musculus). Os pulmoes foram analisados macroscopicamente e atraves de microscopia de luz e eletronica de varredura. Os achados sugerem que o modelo mdx apresenta morfologia pulmonar semelhante aos camundongos BALB/C57 e que seu uso deve ser cauteloso e criterioso em ensaios clinicos que aborde este orgao.

Diaphragm morphology of guinea pig (Cavia porcellus).

The diaphragm is the main respiratory muscle. Along with other respiratory muscles, the diaphragm is responsible for the muscular contraction that generates the respiratory cycle and, as a consequence, the gaseous interchanges in the lungs. Guinea pigs (Cavia porcellus Linnaeus 1758) are largely used as experimental animals in many biology applications due to their easy management, low cost, and docile behavior. As the diaphragm exerts important effects on lung physiology and function, this study aimed at investigating the morphological characteristics of the muscle, through macroscopic, microscopic, and scanning electron microscopy to add reference data for future studies. We observed a “U”‐shaped tendineous center and its morphology was similar to other mammals. These results cooperate with the descriptive and comparative anatomy of mammals, besides can be used as control data for areas of surgery and stem cells. Microsc. Res. Tech. 76:316–320, 2013.

FGF2, FGF3 and FGF4 expression pattern during molars odontogenesis in Didelphis albiventris

Odontogenesis is guided by a complex signaling cascade in which several molecules, including FGF2-4, ensure all dental groups development and specificity. Most of the data on odontogenesis derives from rodents, which does not have all dental groups. Didelphis albiventris is an opossum with the closest dentition to humans, and the main odontogenesis stages occur when the newborns are in the pouch. In this study, D. albiventris postnatals were used to characterize the main stages of their molars development; and also to establish FGF2, FGF3 and FGF4 expression pattern. D. albiventris postnatals were processed for histological and indirect immunoperoxidase analysis of the tooth germs. Our results revealed similar dental structures between D. albiventris and mice. However, FGF2, FGF3 and FGF4 expression patterns were observed in a larger number of dental structures, suggesting broader functions for these molecules in this opossum species. The knowledge of the signaling that determinates odontogenesis in an animal model with complete dentition may contribute to the development of therapies for the replacement of lost teeth in humans. This study may also contribute to the implementation of D. albiventris as model for Developmental Biology studies.

Adrenal glands of Spix's yellow-toothed cavy (Galea spixii, Wagler, 1831): morphological and morphometric aspects

Considering the physiological importance and need of greater morphophysiological knowledge of adrenal glands, the aims of present study were compare the morphometric data between left and right adrenal of male and female; perform a histological, scanning and transmission electron microscopy study showing tissue constitution of glands; finally, in order to define the presence and correct site of the cytochrome P450c17 expression in adrenal glands, immunohistochemical study of this enzyme was performed in 18 adrenal glands (right n=9 and left n=9) of nine adult Galea spixii (four males and five females). Right adrenal was more cranially positioned than left adrenal; dimensions (weight, length and width) of right adrenal was larger than left adrenal; no differences between male and female body and adrenal measurements were found; the morphology of cells and different amounts of lipid droplets may be related to the different demands of steroid hormones production, related to each zone of the adrenal cortex; and, the cytochrome P450c17 immunolocalization in fasciculate and reticular zone may be related with synthesis of 17-hydroxy-pregnenolone, 17-hydroxy-progesterone, dehydroepiandrosterone or androstenedione.

Rat dental pulp stem cells: isolation and phenotypic characterization method aiming bone tissue bioengineering

Dental pulp stem cells (DPSC) have been showing a considerable potential for regenerative medicine. Pulps were collected from lower incisors (n=2) through direct access of the tooth pulp chamber. The isolated cells were cultured in alfa-MEM 10% FBS, in standard culture conditions. At the third passage, DPSC were characterized by flow cytometry (MHCI, CD54, CD73, CD90, CD45, CD11 and CD34); RT-PCR for Nanog gene; and their differentiation capacity in osteogenic, adipogenic and chondrogenic cell lines. Isolated cells exhibited adhesion capacity to plastic; fusiform morphology, and 80% confluence reached in approximately 3 days. These cells have also revealed positive expression for CD54, CD73 and CD90 markers; and negative expression for CD11, CD34 and CD45. Nanog expression was detected by RT-PCR, expected for a mesenchymal stem cell profile. DPSC chondrogenic differentiation was confirmed by positive staining in Alcian Blue; lipidic droplets stained with oil red confirmed their capacity to differentiate in adipogenic fate; while mineralized beads, stained with alizarin red, confirmed their differentiation in osteogenic phenotype. These results indicate the viability of the isolation and expansion of rat DPSC following this method, and osteogenic differentiation potential opens new perspectives for in vivo studies and the use of these cells in cellular therapies and tissue bioengineering, aiming bone repair.

Classificação morfofuncional dos dentes de saguis-de-tufo-branco (Callithrix jacchus, Callitrichidae), saguis-de-tufo-preto (C. penicillata) e saguis-de-cara-branca (C. geoffroyi)

A study was conducted to analyze the morphology of the teeth of white-tufted-ear-marmoset (C. jacchus) black-tufted-ear-marmoset (C. penicillata) and marmosets-white-faced (C. geoffroyi), to compare them among themselves and with other species described in the literature. Teeth of the three species were submitted to macroscopic, microscopic and ultrastructural analyzes. The results were correlated with those of other species. We concluded that: the three species have a similar dental formula, reaching a total of 32 teeth, expressed in the formula 2x: incisors 2/2; canines 1/1, pre-molars 3/3 and molars 2/2, ich are classified as diplodont, anelodont, bunodont and brachyodont.

Classificação Morfológica da Glândula Prepucial do Nasua nasua (Quati).

A proposta desta pesquisa e descrever a glândula prepucial do Nasua nasua. Utilizaram-se oito quatis provenientes do Criatόrio Cientifico Cecrimpas (Unifeob); as glândulas prepuciais foram analisadas, dissecadas e descritas macro e microscopicamente, visando estabelecer sua classificacao morfofuncional para, assim, poder compara-las com as de outras especies ja descritas na literatura. Amostras de tecido glandular foram coletadas, coradas (Hematoxilina e Eosina, Azul Toluidina, e Picrosirius) e analisadas atraves da microscopia de luz. Todos os resultados foram compilados e fotodocumentados. Na especie citada, essa glândula tem como principal funcao a demarcacao territorial e esta situada na porcao prepucial do mesmo modo que nos javalis. Em outros animais, como primatas, cervideos e roedores, essas glândulas demarcatόrias estao localizadas em diferentes regioes anatomicas, como entre os olhos, no metatarso, nas areas perianais e esternais. Em nosso resultado das medidas biometricas, obtivemos como media 3,8 ± 1,41cm de largura, 3,15 ± 0,93cm de altura e 10,26 ± 1,89cm de circunferencia, e essa glândula foi histologicamente classificada como do tipo tubuloalveolar, com secrecao holόcrina.