Immunolocalization of steroidogenic enzymes in the vaginal mucous of Galea spixii during the estrous cycle.

BackgroundThe synthesis of sex steroids is controlled by several enzymes such as17α-hydroxylase cytochrome P450 (P450c17) catalyzing androgen synthesis and aromatase cytochrome P450 (P450arom) catalyzing estrogen synthesis, both of which must complex with the redox partner NADPH-cytochrome P450 oxidoreductase (CPR) for activity. Previous studies have identified expression of steroidogenic enzymes in vaginal tissue, suggesting local sex steroid synthesis. The current studies investigate P450c17, P450aromatase and CPR expression in vaginal mucosa of Galea spixii (Spix cavy) by immuno-histochemical and western immunoblot analyses.MethodsStages of estrous cyclicity were monitored by vaginal exfoliative cytology. After euthanasia, vaginal tissues were retrieved, fixed and frozen at diestrus, proestrus, estrus and metestrus. The ovaries and testis were used as positive control tissues for immunohistochemistry.ResultsData from cytological study allowed identification of different estrous cycle phases. Immunohistochemical analysis showed different sites of expression of steroidogenic enzymes along with tissue response throughout different phases of the estrous cycle. However, further studies are needed to characterize the derived hormones synthesized by, and the enzymes activities associated with, vaginal tissues.ConclusionCurrent results not only support the expression of enzymes involved in sex steroid synthesis in the wall of the vagina, they also indicate that expression changes with the stage of the cycle, both the levels and types of cells exhibiting expression. Thus, changes in proliferation of vaginal epithelial cells and the differentiation of the mucosa may be influenced by local steroid synthesis as well as circulating androgens and estrogens

Origin and distribuition of sciatic nerves of the Galea

O preá do semiárido nordestino (Galea spixii Wagler, 1831) é um roedor pertencente à família Caviidae. Pouca literatura é encontrada sobre essa espécie em relação a sua morfologia e seu comportamento ambiental e reprodutivo. Com o objetivo de entender a morfologia geral, em foco, na inervação do membro pélvico dessa espécie, neste trabalho, foi explorado o nervo isquiático, o qual é o maior de todos os nervos do organismo. Foram utilizados 10 preás (cinco machos e cinco fêmeas) que vieram a óbito por causas naturais, oriundos do Centro de Multiplicação de Animais Silvestres da Universidade Federal Rural do Semiárido (CEMAS/UFERSA). Os animais foram fixados após o óbito em solução aquosa de formaldeído 10% e, após 48 horas de imersão nessa solução, foram dissecados para exposição do nervo isquiático. Dessa forma, os dados obtidos foram compilados em tabelas e expressos em desenhos esquemáticos e fotografias. Os pares de nervos isquiáticos originaram-se de raízes ventrais de L6L7S1 (70%) e de L7S1S2 (30%) e distribuíram-se pelos músculos glúteo profundo, bíceps femural, semitendinoso e semimembranoso.The Galea of the semi-arid northeast (Galea spixii Wagler, 1831) is a rodent belonging to the family Caviidae. Little literature is found on this species in relation to morphology, environmental and reproductive behavior. With a view to understanding the general morphology, particularly, in the innervation of the pelvic limb of this species, ponder the sciatic nerve, which is the largest of all the nerves of the body. 10 cavies were used (five males, five females) that they had died of natural causes, originating from breeders scientific, legally licensed by IBAMA, the UFRSA, Mossoró, RN. The animals were fixed after the death in an aqueous solution of formaldehyde 10% and after 48 hours of immersion in it, were dissected to expose the sciatic nerve. Thus, the data were compiled in tables and expressed in drawings and photographs. The pairs of sciatic nerves were derived from ventral roots L6L7S1 (70%) and L7S1S2 (30%) and were distributed by the deep gluteal muscles, biceps femural, semitendinosus and semimembranosus muscles

ORIGEM E DISTRIBUIÇÃO DA ARTÉRIA CELÍACA EM PREÁS (Galea spixii Wagler, 1831)

Para o estudo foram utilizados 20 animais, que estavam em freezer no Centro de Multiplicação de Animais Silvestres (CEMAS/UFERSA). Os animais foram descongelados, a cavidade abdominal aberta e a aorta torácica canulada no sentido caudal para injeção de solução de látex Neoprene 650 corado de vermelho e, em seguida, fixados em solução de formaldeído a 10%, por um período mínimo de 48 horas, para que pudessem ser dissecados, fotografados e os resultados analisados. A artéria celíaca originou-se da aorta abdominal e emitiu as artérias gástrica esquerda, lienal e esplênica. A artéria gástrica esquerda emitiu de dois a três ramos à região da curvatura menor do estômago. A artéria lienal emitiu quatro ramos ao parênquima do baço, de seis a doze ramos pancreáticos e a artéria gastroepiplóica esquerda. A artéria hepática emitiu de um a dois ramos pancreáticos e bifurcou-se em um curto tronco formado pelas artérias gástrica direita e hepática própria e ainda na artéria gastroduodenal, que originava a pancreaticoduodenal cranial e gastroepiplóica direita. O comportamento de trifurcação da artéria celíaca do preá assemelha-se ao encontrado em roedores, como o nutria, a cutia, o hamster e os ratos, o que sugere que este seja o padrão para a ordem. Palavras-chave: aorta abdominal; aparelho digestório; Galea spixii; roedor; vascularização

Collateral Arteries of the Aortic Arch in Mongolian Gerbil (Meriones unguiculatus)

Background: Gerbils (Meriones unguiculatus) are rodents belonging to the Muridae family. Recently, breeding of this species as pets has increased significantly. Animal models are being investigated to study diseases related to the human aortic arch. Despite the importance of the aortic arch in maintaining homeostasis, there is limited data available regarding its morphology in gerbils. This study was performed with the objective of describing the collateral branches of the aortic arch in this animal to establish a standard model and thus contribute to future research on cardiovascular diseases in humans.Materials, Methods & Results: This study used 20 male specimens from previous studies that were frozen and stored at the Laboratory of Veterinary Anatomy of the Federal Rural University of the Semi-Arid Region. After thawing the animals, the thoracic cavity was opened for aortic cannulation. The vascular system was washed using saline solution and Neoprene latex stained with red pigment was injected. Subsequently, the animals were fixed in 10% formaldehyde and were dissected and analyzed 72 h later. The arrangement of the collateral branches of the aortic arch in gerbils was analyzed in all animals. The brachiocephalic trunk, the left common carotid, and the left subclavian artery were observed to originate as collateral branches. The brachiocephalic trunk bifurcated into the right common carotid and the right subclavian arteries. The right and the left subclavian arteries branched into the vertebral artery, the internal thoracic artery, the superficial cervical artery, the costocervical trunk, and the axillary artery.Discussion: Several studies reported in the literature describe the collateral branches of the aortic arch in domestic and wild mammalian species. These studies examined the main arteries that originate directly from the aortic arch and their respective branches, and classified the different anatomical variants of the aortic arch in each species. Three different arrangements have been commonly described. The first type corresponds only to the brachiocephalic artery originating from the aortic arch. The right and the left common carotid arteries and the right and the left subclavian arteries originate from this brachiocephalic artery. This type has already been described in the laboratory rat, catingueiro-deer, cattle, and horses. The second type is characterized by the presence of 2 arteries - the brachiocephalic trunk and the left subclavian artery. The right and the left common carotid arteries and the right subclavian artery originate from the brachiocephalic trunk. This arrangement has been reported in most species already studied such as rodents including the paca, chinchilla, guinea pig, mocó, nutria and the preá. The third type of vascular arrangement is observed in the gerbil. In this species, 3 collateral arteries originate from the aortic arch (the brachiocephalic trunk, the left common carotid, and the left subclavian artery). The right common carotid and the right subclavian artery originate from the brachiocephalic trunk. This vascular model has been described in the manatee, in humans, mice, sauim, and the monkey-nail. Thus, we concluded that the branching pattern of the aortic arch of the gerbil was characterized by the brachiocephalic trunk, the left common carotid, and the left subclavian artery, as has been described in mice, the manatee, monkey-nail, sauim, and humans. Based on these morphological characteristics, gerbils could serve as potential experimental models to study diseases related to the human aortic arch

Morphometric Alteration of Intestinal Epithelium of Rats (Rattus norvegicus) Submitted to the Technique of Enteropexy

Background: Due to the numerous complications that enteropexy may cause in domestic animals and humans, this study aimed at investigating the microscopic level, the damage generated by this technique in mice, in order to investigate the morphometric changes caused by enteropexy technique in rats.Materials, Methods & Results: Eighteen Wistar rats were submitted to surgical technique of enteropexy. To evaluate intestinal disorders they were euthanized at 30 (Group I), 60 (group II) and 90 (group III) days after the procedure. The animals were perfused with 2.5% glutaraldehyde and 4% paraformaldehyde for the removal of intestinal fragments for analysis in conventional microscopy and scanning electron microscopy. In classical histopathology, group I presented mild lesions with necrosis of the epithelium and infiltration of mononuclear cells. After 60 days, extensive ischemic necrosis area was observed, characterized by the denudation of epithelium cells and cellular debris in the lumen surface and loss of intestinal crypts. After 90 days (group III) lesions were evident and were characterized by strong coagulative necrosis of epithelial cells and intestinal crypts. Under electron microscopy, group I showed early fusion of the adjacent intestinal walls. After 60 days there was adherence of luminal stenosis and intestinal walls. In group III loss of epithelium and substitution of necrotic tissue were identified.Discussion: In the current study it was noticed microscopically that the groups had inflammatory reactions to foreign bodies. It is known that the sutures, independent of manufacture, behave as foreign bodies that induce tissue inflammation of the recipient organism and could harm the scar repair. Among the surgical threads, the nylon type (which was chosen for the current experiment) is indicated in the approach tissues in general and bandages, including cardiovascular, ophthalmic and neurological procedures. This category wire induces minimal inflammation of the receptor tissue. In studies of the histopathological analysis of the tissue reactions produced by the wire implant or nylon thread clamp in rats, it was observed that when the local action of two implants were compared, inflammation showed qualitatively similar responses, although they had different characteristics regarding their course. Inflammation is proportional to the proliferation of fibroblasts and the presence of fibrous tissue around the suture. This quote confirms the analyzed samples, in which, in all postoperative times showed inflammatory process, although at different magnitudes. Surgical research in laboratory animals has expanded in recent decades, mainly due to better anesthetic support, the sophistication of infrastructure, material for perioperative continuous monitoring and an incessant search for species that reproduce human morbid conditions. The main focus of these studies have been improving the knowledge about the pathophysiological mechanisms of disease, undertake therapeutic trials with new drugs, studying biomarkers and evaluate new techniques to be applied prospects in man. The most common sites of occurrence of intestinal are ileocolic intussusception junction and jejunojejunal segment. The enteroplication has been used to reduce the occurrence of intussusception, promoting the adhesion of the adjacent serosa layers of intestinal segments and thereby decreasing intestinal motility, their applicability is reported in humans, dogs and cats. The enteropexy technique causes morphological changes in intestinal epithelium of rats, evidenced in conventional and electron microscopy, progress with the passing of time the procedure

Morphology and topography of the liver and pancreas of Rhea americana

As emas são aves ratitas nativas do continente sul americano, são consideradas aves primitivas do ponto de vista filogenético que constituem um grupo altamente especializado. Este estudo buscou caracterizar macro e microscopicamente o fígado e pâncreas de emas. O material foi coletado no Centro de Multiplicação de Animais Silvestres (CEMAS), na cidade de Mossoró-RN, Brasil, (Registro IBAMA n° 14.78912). Utilizaram-se 20 animais jovens com idade entre dois e seis meses independente do sexo. Em emas, o fígado se relacionava cranialmente com o ápice do coração, dorsalmente com os pulmões, esôfago e o proventrículo gástrico, caudalmente, com o ventrículo gástrico, o baço, o duodeno e parte do jejuno. Apresentava coloração vermelha escura e possuía apenas dois lobos, sendo o direito ligeiramente menor que o esquerdo. Histologicamente, era revestido por uma cápsula de tecido conjuntivo delgada e cada lóbulo hepático pôde ser identificado pela presença evidente de veias centrais, com muitos sinusoides comunicando-se com elas. O pâncreas, ventralmente, apresentava-se como uma fita fina, formado por um lobo dorsal e um lobo ventral. Longitudinalmente o pâncreas em emas localiza-se no mesentério dorsal desde o fígado até a flexura cranial do duodeno, mantendo-se preso às alças duodenais por ligamentos. Histologicamente, era composto por uma cápsula delgada de tecido conjuntivo denso, com discretos lóbulos separados por tecido conjuntivo capsular, compostos por estruturas tubuloalveolares e ductos. O fígado e pâncreas de emas apresentam padrão morfológico similar ao descrito para aves domésticas.The rhea are ratites birds native of the South American continent, which are considered primitive birds from the standpoint of the phylogenetic group that is highly specialized. This study aimed to characterize gross and microscopically the liver and rhea pancreas. The material was collected in a Wild Animals Multiplication Center, Mossoro-RN, Brazil, (Registration no 14.78912 IBAMA). It was used 20 young animals aged between two and six months independent of sex. In the rhea the liver is related to the cranial apex of the heart, dorsally with the lungs, esophagus and gastric proventriculus, caudally with the gastric ventricle, the spleen, the duodenum and part of the jejunum. It was dark red color and had only two lobes, being the right lobe slightly smaller than the left. Histologically was lined by a thin connective tissue capsule and each lobe liver could be clearly identified by the presence of central veins, with sinusoids communicating with the veins. The pancreas, ventrally, showed a thin tape formed by a dorsal lobe and a ventral lobe. Longitudinally the pancreas in rhea is located in the dorsal mesentery since the liver until the cranial duodenal flexure, remaining connected the duodenal handles by ligaments. Histologically was composed of a thin capsule of dense connective tissue, with discrete lobules separated by capsular connective tissue, composed of tubuloalveolares structures and pipelines. The liver and pancreas of rhea have morphology similar to that described for domestic poultry

Correlação entre o peso, comprimento e anexos fetais de mocós (Kerodon rupestris Wied, 1820)

Foi avaliada a existência de correlação entre diferentes anexos fetais de mocós e determinada a relação entre peso do feto e peso da placenta, relação entre o peso do feto e comprimento do cordão umbilical e a relação entre o peso do feto e comprimento do feto. Foram utilizados anexos fetais e fetos de três, cinco e seis fêmeas, respectivamente, no terço inicial, médio e final de gestação, obtidas no Centro de Multiplicação de Animais Silvestres (CEMAS-UFERSA). Os dados foram expressos em média ± desvio padrão, bem como valores mínimos e máximos, avaliados pelo programa estatístico GraphPad Prism Versão 6.0. Após análise dos pressupostos paramétricos, os dados foram submetidos aos Testes de Kruskal-Wallis e Mann-Whitney e regressão linear. A relação entre peso do feto e peso da placenta demonstrou expressão y = 33,73 + 16,38x com cálculo do coeficiente de correlação entre peso do feto e o peso da placenta elevado e positivo, evidenciando dependência entre as variáveis (R2 = 0,7251). A relação entre o peso do feto e comprimento do cordão umbilical demonstrou expressão y = 35,64 + 25,64x e a análise de correlação entre as variáveis, positiva (R2 = 0,7201) indicando elevada dependência entre as variáveis. Quanto à relação entre o peso do feto e comprimento do feto, identificou-se uma relação do tipo y = a + bx, cuja expressão definida foi y = 1,26 + 0,41x, com análise de correlação das variáveis apresentando elevada correlação considerando-se o valor de R2 = 0,7890. As variáveis analisadas demonstram uma influência direta no desenvolvimento embrionário e fetal em mocós confirmando a correlação destes com o tamanho do feto.We evaluated the correlation between different fetal membranes of rock cavies and determined the relationship between fetal weight and placental weight, the relationship between fetal weight and length of the umbilical cord and the relationship between fetal weight and length of the fetus. The fetuses and fetal membranes of three, five and six females respectively in the first third, middle and late gestation were used, obtained from Multiplication Center for Wild Animals of the Federal University Rural of Semiarid (CEMAS-UFERSA). The data were expressed as mean ± standard deviation, minimum and maximum values, evaluated by the statistical program GraphPad Prism version 6.0. After analysis of parametric assumptions, the data were subjected to Kruskal-Wallis' test and Mann-Whitney's test and linear regression. The relationship between fetal weight and placental weight showed the expression y = 33.73 + 16.38x with an estimate of the coefficient of correlation between fetal weight and placental weight high and positive, showing dependence between variables (R2 = 0.7251). The relationship between fetal weight and length of the umbilical cord showed expression y = 35.64 + 25.64x and the correlation analysis between variables, positive (R2 = 0.7201) indicating high dependence between variables. Regarding the relationship between fetal weight and length of the fetus, we identified a relationship of the type y = a + bx, whose expression was defined y = 1.26 + 0.41x, with correlation analysis of the variables showing high correlation considering the value of R2 = 0.7890. The variables analyzed demonstrate a direct influence on embryonic and fetal development in rock cavies confirming the correlation of these with the size of the fetus

Período gestacional e ciclo reprodutivo em preás (Galea spixii Wagler, 1831)

O preá é um roedor com elevado potencial biológico e zootécnico a ser explorado, sendo o conhecimento sobre os aspectos reprodutivos fundamentais para que sua criação seja satisfatória. Objetivou-se determinar a duração da gestação em preás, e a caracterização das fases do ciclo reprodutivo, verificando, se existe influência da presença do macho neste processo. Os animais foram separados em três grupos: Acompanhamento gestacional (G1), 5:1, proporção de fêmea e macho no box; Ciclo estral, 5:1, com o macho preso em gaiola (G2) e cinco fêmeas em outro box sem o macho (G3). O exame colpocitológico ocorreu diariamente, identificado o espermatozoide na lâmina como cópula. As fêmeas do G1 foram separadas assim que copulavam e contadas como dia “zero” da gestação. As fêmeas do G2 e G3 foram avaliadas ao longo de dois ciclos estrais completos, avaliados qualitativamente pelo o esfregaço vaginal. O período de gestação em preás foi de 59±2,24 dias, com um ciclo poliéstrico contínuo, com duração de 14,8±0,73 dias no G2 e 14,6±0,75 dias no G3. O proestro caracterizou-se pelo predomínio de células parabasais, intermediárias, bactérias e leucócitos; o estro, células superficiais, com predomínio das anucleadas com presença ou não de bactérias; metaestro, células parabasais e grande quantidade de células intermediárias, neutrófilos e bactérias; diestro, predomínio de células basais, parabasais e intermediárias e grande quantidade de muco vaginal, neutrófilos e bactérias. A presença do macho influenciou significativamente a duração do diestro, tornando-se mais longa, fato que pode estar atrelado a influência sobre a produção de progesterona na fêmea. Palavras-chave: Ciclo estral; estro; Galea spixii; gestação; roedor

Immunolocalization of steroidogenic enzymes in the vaginal mucous of Galea spixii during the estrous cycle

Abstract\ud \ud Background\ud The synthesis of sex steroids is controlled by several enzymes such as17α-hydroxylase cytochrome P450 (P450c17) catalyzing androgen synthesis and aromatase cytochrome P450 (P450arom) catalyzing estrogen synthesis, both of which must complex with the redox partner NADPH-cytochrome P450 oxidoreductase (CPR) for activity. Previous studies have identified expression of steroidogenic enzymes in vaginal tissue, suggesting local sex steroid synthesis. The current studies investigate P450c17, P450aromatase and CPR expression in vaginal mucosa of Galea spixii (Spix cavy) by immuno-histochemical and western immunoblot analyses.\ud \ud \ud Methods\ud Stages of estrous cyclicity were monitored by vaginal exfoliative cytology. After euthanasia, vaginal tissues were retrieved, fixed and frozen at diestrus, proestrus, estrus and metestrus. The ovaries and testis were used as positive control tissues for immunohistochemistry.\ud \ud \ud Results\ud Data from cytological study allowed identification of different estrous cycle phases. Immunohistochemical analysis showed different sites of expression of steroidogenic enzymes along with tissue response throughout different phases of the estrous cycle. However, further studies are needed to characterize the derived hormones synthesized by, and the enzymes activities associated with, vaginal tissues.\ud \ud \ud Conclusion\ud Current results not only support the expression of enzymes involved in sex steroid synthesis in the wall of the vagina, they also indicate that expression changes with the stage of the cycle, both the levels and types of cells exhibiting expression. Thus, changes in proliferation of vaginal epithelial cells and the differentiation of the mucosa may be influenced by local steroid synthesis as well as circulating androgens and estrogens.This work was supported by grants from São Paulo Research Foundation/\ud FAPESP, Sao Paulo, Brazil (Process Number: 2011/03655-2) and the National\ud Council of Scientific Researches/CNPq (Process Number: 402220/2010-2)