SUMMARY: The rabbit is considered an ideal animal model for studies that describe abnormalities in the testicles due to the similar morphogenetic mechanisms of sexual development and diseases commonly found in humans. The aim of this study was to determine the male sexual differentiation of the New Zealand rabbit (Oryctolagus cuniculus) through development. The gestational age was estimated and classified as 9, 12, 14, 16, 18, 20, 23 and 28 gestational days. The morphological and sexual determination were performed by histological analysis of the reproductive tract in the embryos and fetuses (9-28 days) as well as by immunohistochemistry- Desert hedgehog-Dhh- (testis-specific protein on Y chromosome- 16, 20, 23 days and adult rabbits). Gonads were observed from the 14th day in an undifferentiated stage and with homogeneous aspect. Sexual differentiation was observed from the 16th day with presence of cells forming gonadal cords and Dhh+ cells in the gonadal parenchyma. From the 18th gestational day testicular cords were observed, which evolved into organized seminiferous tubules. The formation of the efferent ducts and ductus deferens and epididymis was observed on the 20th and 23rd days, respectively. The differentiation of the external genitalia occurred from the 23rd days from the anogenital distance and was identified to identify the penile structures. In summary, the features of the sexual differentiation were determined by observation of the Dhh+ protein in embryos from the 16th day to adulthood, and the morphological particularities observed from the 18th gestational day, determined by differentiation of the external genitalia from the 23rd day.

RESUMEN: El conejo se considera un modelo animal ideal para estudios que describen anomalías a nivel testícular debido a que presenta mecanismos morfogenéticos similares al desa- rrollo sexual y enfermedades que se encuentran comúnmente en los seres humanos. El objetivo de este estudio fue determinar la diferenciación sexual masculina del conejo de Nueva Zelanda (Oryctolagus cuniculus) a través del desarrollo. La edad gestacional se estimó y clasificó en 9, 12, 14, 16, 18, 20, 23 y 28 días gestacionales. La determinación morfológica y sexual se realizó mediante análisis histológico del tracto reproductivo en los embriones y fetos (9 - 28 días) así como mediante inmunohistoquímica -Desert hedgehog-Dhh- (proteína testicular específica en el cromosoma Y- 16, 20, 23 días y conejos adultos). Las gónadas se observaron a partir del día 14 en un estadio indiferenciado y con aspecto homogéneo. Se observó diferenciación sexual a partir del día 16 con presencia de células formadoras de cordones gonadales y células Dhh+ en el parénquima gonadal. A partir del día 18 de gestación se observaron cordones testiculares, que evolucionaron a túbulos seminíferos organizados. La formación de los conductos eferentes, deferentes y del epidídimo se observó a los 20 y 23 días, respectivamente. La diferenciación de los genitales externos ocurrió a partir del día 23 desde la distancia anogenital y se utilizó para identificar las estructuras del pene. En conclusión, las características de la diferenciación sexual se determinaron mediante la observación de la proteína Dhh en embriones desde el día 16 hasta la edad adulta, y las particularidades morfológicas observadas a partir del día 18 de gestación, determinadas por diferenciación de los genitales externos a partir del día 23.

SUMMARY: The study reported the influence of the high and acute dose of Letrozole on the testis morphology in paca (Cuniculus paca), an aromatase inhibitor that reduces the endogenous estrogen, the essential hormone for spermatogenesis. Morphological changes were observed in seminiferous epithelium with germ cells with apoptotic characteristics and presence of vacuoles and nuclei in pycnose.

RESUMEN: El objetivo de este estudio fue analizar la influencia de una dosis alta de Letrozol en la morfología de los testículos de la paca (Cuniculus paca), un inhibidor de la aromatasa que reduce el estrógeno endógeno, la hormona esencial para la espermatogénesis. Se observaron cambios morfológicos en el epitelio seminífero con células germinales con características apoptóticas y la presencia de vacuolas y núcleos en picnosis.

Abstract Pluripotent stem cells have been studied as source of cells for regenerative medicine and acquire or genetic diseases, as an innovative therapy. Most tissues have stem cells populations, however in few quantities or impossible to be used during adult life, which lead to scientists look for new sources. Thus, this study aimed to analyze the presence of pluripotent cells in the uterus and placenta, following up non-pregnant, pregnant (begin, middle, and final), and postpartum periods in dogs. The uteri were obtained from social castration programs for population control in Pirassununga, Sao Paulo, Brazil. It was collected 20 uteri at different stages. The samples were fixed and processed for immunohistochemical analysis of NANOG, OCT4 and SOX2 expression, knowing as pluripotent stem cells makers. Our results showed positive expression for NANOG, OCT4 and SOX2 in all stages of gestation and nonpregnant uterus; however, we highlight some quantitative different between stages. OCT4 showed more expression in non-pregnant uterus than NANOG and SOX2, and its expression increased in pregnant uterus. In pregnant uterus there was more expression of NANOG than OCT4 and SOX2. Interesting, no difference was found between these markers in the other periods. In conclusion, it was possible to identify pluripotent stem cells in all periods in dog placenta and uterus, however during the early stage of pregnancy we observed more pluripotent stem cells than in all the others periods confirming the high plasticity and regeneration capacity of the uterine tissue.

SUMMARY: To contribute knowledge of an important experimental model for studies on skin embryology, a study was carried out to describe the morphological events of the skin during the intrauterine life of Cavia porcellus from the 10th to the 60th day of gestation. Embryos and fetuses were dissected, and the skin of the nasal, cranial, lumbar and anal regions was processed byoptical microscopy. At 30 days the first hairs, called lanugos, were observed in the cranial region. The morphological description showed that a few days can make a great difference in development.

RESUMEN: Con el objetivo de contribuir al conocimiento de un importante modelo experimental para estudios sobre embriología de la piel, se llevó a cabo un estudio para describir los eventos morfológicos de la piel durante la vida intrauterina de Cavia porcellus desde el día 10 hasta el día 60 de gestación. Los embriones y los fetos se disecaron y se procesó la piel de las regiones nasal, craneal, lumbar y anal, mediante microscopía óptica. A los 30 días se observaron los primeros vellos, llamados lanugos, en la región craneal. La descripción morfológica mostró que unos pocos días pueden marcar una gran diferencia en el desarrollo.

SUMMARY: The aim of this study was to know the embryonic and fetal development of the female rabbit genital system (Oryctolagus cuniculus), describing its main phases and the moment of sexual differentiation. Eleven pregnant New Zealand female rabbits were used in different gestational phases. The day of coitus was determined as day 0. For each stage a minimum of two animals was considered. The samples were obtained every two days from the ninth day post-coitus (dpc) until the 28th dpc. The gestational period was divided in two: animals with undifferentiated sex (group 1) and animals with differentiated sex (group 2). The ages of embryos and fetuses were estimated through the crown-rump method. Subsequently, embryos and fetuses were dissected, fixed and processed to be embedded in paraffin (Histosec). The histological analysis was performed on sections stained with hematoxylin and eosin. Immunohistochemical analysis to determine sexual differentiation was performed on samples from the 16th, 18th and 28th dpc. Desert Hedgehog (Dhh) and Indian Hedgehog (Ihh) primary antibodies, respectively, were used to identify cells of the male and female germinal epithelium. The immunohistochemical results showed that at the 16th dpc, female sexual differentiation was evident, since positive expression of the Ihh protein was observed. Sexual differentiation was obtained through histological analysis on the 18th dpc and through anatomical observation of the external genitalia on the 24th dpc. Knowing the characteristics of the embryonic and fetal development of the female rabbit genital system as well as the moment of sexual differentiation make it possible to establish bases for future research that address the physiology and pathology of these organs. Thus, any alteration in the chain of events of sexual determination and differentiation must search for an explanation from the knowledge of the possible normal mechanisms affected.

RESUMEN: El objetivo de esta investigación fue conocer el desarrollo embrionario y fetal del sistema genital femenino de conejo (Oryctolagus cuniculus), describiendo sus principales fases y el momento de la diferenciación sexual. Se utilizaron 11 conejos hembras gestantes neozelandesas, en diferentes fases gestacionales. El día del coito se determinó como día 0. Para cada etapa fue considerado un mínimos de dos animales. Las muestras fueron obtenidas cada dos días, a partir del noveno día post-coito (dpc) hasta el 28 dpc. El periodo gestacional fue dividido en dos: animales con sexo indiferenciado (grupo 1) y, animales con sexo diferenciado (grupo 2). Las edades de los embriones y los fetos fueron estimadas a través del método de crown-rump. Posteriormente, embriones y fetos fueron disecados, fijados y procesados para su inclusión en parafina (Histosec). El análisis histológico se realizó en secciones teñidas con Hematoxilina y Eosina. El análisis inmunohistoquímico para determinar la diferenciación sexual fue realizado en muestras de 16, 18 y 28 dpc. Para identificar células del epitelio germinativo masculino y feminino se utilizaron los anticuerpos primarios Desert Hedgehog (Dhh) e Indian Hedgehog (Ihh), respectivamente. Los resultados inmunohistoquímicos mostraron que a los 16 dpc se evidenció diferenciación sexual femenina, ya que se observó expresión positiva de la proteína Ihh. La diferenciación sexual, a través del análisis histológico fue obtenida a los 18 dpc y a través de la observación anatómica de los genitales externos a los 24 dpc. Conocer las características del desarrollo embrionario y fetal del sistema genital femenino de conejo, así como, el momento de la diferenciación sexual, permiten sentar bases para futuras investigaciones que aborden la fisiología y patología de estos órganos. Así, cualquier alteración en la cadena de eventos de la determinación y diferenciación sexual deberá buscar una explicación a partir del conocimiento de los posibles mecanismos normales afectados.

RESUMO: Buscou-se estudar o desenvolvimento do pulmão do preá ( Galea spixii ) com ênfase nas características macroscópicas, histológicas e imunohistoquimicas, bem como acompanhar a proliferação celular. Para tanto, foram utilizados 8 fetos, os quais, de acordo com o tamanho (crown-rump - CR), foram divididos em 3 grupos: Grupo I (33-35 dias), Grupo II (38-40 dias) e Grupo III (43-45 dias). Macroscopicamente, não houve diferenças quanto à morfologia do pulmão entre os grupos. Em relação à maturação pulmonar, no grupo I, observou-se o início da formação dos brônquios e bronquíolos, os quais estavam rodeados por grande quantidade de mesênquima e vasos sanguíneos de pequeno calibre, caracterizando o estágio pseudoglandular. Os indivíduos do Grupo II apresentaram maior quantidade de formações tubulares no parênquima pulmonar, assim como redução da quantidade de mesênquima, atingindo, dessa forma, a fase canalicular. No Grupo III, os pulmões apresentaram-se completamente formados, atingindo a fase alveolar. Nas análises de imunohistoquimica, os pulmões dos indivíduos dos Grupos I e II apresentaram marcação para Pcna, Oct-4 e VEGF. Por outro lado, não houve marcação no Grupo III. A anatomia do pulmão do preá se assemelha ao pulmão da cutia ( Dasyprocta sp.) quanto ao número de lobos e fissuras. No que se refere ao desenvolvimento pulmonar, o pulmão do preá apresentou maior similaridade com o pulmão de fetos humanos, visto que ambos, ao nascer, encontram-se na fase alveolar, contrário a outras espécies de roedores.

ABSTRACT: The aim of this research was to study the development of the lung in Galea spixii , by gross anatomy, histological analysis and immunohistochemical techniques. Totally, 8 fetuses were used and allocated into three groups of age: Group I (33-35 days), Group II (38-40 days), and Group III (43-45 days) with given crown-rump - CR lenghts. According to the gross morphology, there were no differences in relation to the lung morphology among groups. In relation to lung maturation, the Group I showed early formation of the bronchi and bronchioles, which were richly surrounded by mesenchyme and small blood vessels, typical features of the pseudoglandular stage. Individuals from Group II showed higher amounts of tubular formations in the lung parenchyma and reduced mesenchyme, reaching the canalicular stage. The lung from individuals of the Group III was completely formed, reaching the alveolar phase. In the immunohistochemical analysis, the lung of individuals from both Groups I and II were positive for Pcna, Oct-4, and VEGF. In contrast, there was not labbeling on samples from Group III. Thus, the G. spixii lung anatomy resembles the agouti ( Dasyprocta sp.) lung, in relation to the number of lobes and fissures. In regard to the lung development, G. spixii showed more similarity with the human fetal lung, since both are born in the stage of the alveolar phase, different to other rodent species.

Resumo: Este estudo descreve o desenvolvimento do sistema nervoso central em guinea pig do 12º dia pós-concepção (dpc) até ao nascimento. No total, 41 embriões e fetos foram analisados macroscopicamente e por microscopia de luz e eletrônica. O fechamento do tubo neural foi observado no dia 14 e o desenvolvimento da medula espinhal e diferenciação das vesículas primitivas do sistema nervoso central foram observados no dia 20. Histologicamente, o tecido cerebral indiferenciado foi observado como uma massa de tecido mesenquimal entre os dias 18 e 20 e no 25º dia o tecido no interior do canal medular apresentou maior densidade. No dia 30 o tecido cerebral apresentou-se diferenciado, período no qual a medula espinhal preenchia todo o canal vertebral e foi possível observar os estratos cerebral e cerebelar. No dia 45 as intumescências cervical e lombar foram visualizadas e os hemisférios cerebrais estavam divididos, com uma clara distinção entre substância branca e cinzenta no cérebro e cerebelo. O sulco mediano dorsal da medula espinhal e a cauda equina foram evidentes apenas no dia 50. Não houve diferenças estruturais significativas em fetos de 50 e 60 dpc e animais a termo eram todos lisencefálicos. Estudos morfológicos do sistema nervoso em guinea pig podem fornecer informações importantes para estudos clínicos em seres humanos devido ao alto grau de maturidade neurológica em relação ao seu período de gestação curto, fato que servir como excelente ferramenta em estudos neurológicos.

Abstract: This study describes the development of the central nervous system in guinea pigs from 12th day post conception (dpc) until birth. Totally, 41 embryos and fetuses were analyzed macroscopically and by means of light and electron microscopy. The neural tube closure was observed at day 14 and the development of the spinal cord and differentiation of the primitive central nervous system vesicles was on 20th dpc. Histologically, undifferentiated brain tissue was observed as a mass of mesenchymal tissue between 18th and 20th dpc, and at 25th dpc the tissue within the medullary canal had higher density. On day 30 the brain tissue was differentiated on day 30 and the spinal cord filling throughout the spinal canal, period from which it was possible to observe cerebral and cerebellar stratums. At day 45 intumescences were visualized and cerebral hemispheres were divided, with a clear division between white and gray matter in brain and cerebellum. Median sulcus of the dorsal spinal cord and the cauda equina were only evident on day 50. There were no significant structural differences in fetuses of 50 and 60 dpc, and animals at term were all lissencephalic. In conclusion, morphological studies of the nervous system in guinea pig can provide important information for clinical studies in humans, due to its high degree of neurological maturity in relation to its short gestation period, what can provide a good tool for neurological studies.

A pele representa parte importante do sistema tegumentar, pois desempenha funções como a proteção contra desidratação, lesões e infecções, além de apresentar alto poder de renovação. Histologicamente, a pele consiste de duas camadas funcionais, morfologicamente distintas, a epiderme e a derme. As glândulas mamárias, capsula ungueal e cornos também fazem parte deste sistema. Devido à escassez de dados na literatura sobre o desenvolvimento da pele e dos anexos de natureza córnea em bovinos, o objetivo deste trabalho foi descrever as características morfológicas do desenvolvimento do sistema tegumentar durante os períodos embrionário e fetal em bovinos. Os indivíduos foram divididos em três grupos: Grupo I -embriões de 20-26 dias (n=4); Grupo II -embriões 30-47 dias (n=6) e Grupo III -fetos de 74-140 dias (n=6). Durante o desenvolvimento da pele observou-se diferentes padrões morfológicos de acordo com as regiões analisadas, apresentando índice maior de diferenciação no intervalo entre 30-47 dias de gestação. O aparelho ungueal e a glândula mamária se desenvolvem mais tardiamente entre os dias 74-140 da gestação. Em resumo, estes resultados acerca do desenvolvimento do sistema tegumentar em bovinos, poderão auxiliar na interpretação e compreensão da formação dos órgãos que o constituem, assim como para a compreensão de alterações patológicas associadas à organogênese.

The skin is an important part of the integumentary system, which has important functions such as protection against dehydration, injury and infection, as well as present high potential of renewal. Histologically, the skin consists of two functional layers, that are morphologically distinct, the epidermis and dermis. The mammary glands, ungueal capsule and horn are also part of this system. Due the poor data available on literature about the development of skin and other cornea attachments in bovine, the aim of this study was to describe the morphological characteristics of the integumentary system development during the embryonic and fetal periods in bovine. Individuals were allocated in three groups: Group I -embryos from 20 to 26 days (n=4), Group II -embryos from 30 to 47 days (n=6) and Group III -fetuses from 74 to 140 days (n=6). During the development of the skin it was observed different morphological patterns according to the analyzed regions. Especially, a higher level of differentiation was observed during 30-47 days of gestation. The bovine claw and mammary gland develop later between 74 to 140 days of gestation. In summary, these results related to the development of the integumentary system in bovine may help the analysis and understanding of the development of their organs, as well as the pathological disorders associated to the organogenesis.

A avaliação da função pulmonar em cães pode ser obtida mediante exames cintilográficos, os quais incluem a injeção de radiofármacos, seguida de radiografias. Neste estudo, foram utilizados 10 cães machos hígidos da raça Rottweiler e radiograficamente normais e soronegativos para dirofilariose, com administração de doses variadas de macroagregado de albumina sérica humana marcados com tecnécio-99m ([99mTc](MAA)). Os resultados foram avaliados qualitativa e quantitativamente, considerando o índice de perfusão pulmonar e sua homogeneidade, sendo dessa maneira visualizado um aumento na homogeneidade da imagem diretamente relacionado ao número de partículas de MAA injetado. Este estudo define os critérios de normalidade e de dosagem na perfusão pulmonar em cães hígidos da raça Rottweiller estabelecidos com a técnica de cintilografia.

Pulmonary function evaluation in dogs can be gotten by scintigraphy exams, which are obtained with radiopharmacs injections, followed by radiography. In this study it was used 10 healthful male dogs of Rottweiler breed, radiographically normal and serum negative for dirofilariasis, with administration of varied doses of human serum albumin grouped marked with technetium-99m (MAA). The results were evaluated qualitatively and quantitatively, considering the index of pulmonary perfusion and its homogeneity, being found an increase on the homogeneity of image directly related with the number of MAA particles injected. This study defines the criteria of normality and the dosage of pulmonary perfusion in healthful dogs established with scintigraphy technique.

Este estudo objetivou caracterizar a presença de pneumócitos tipo II e o início da produção de lipoproteína surfactante em bovinos, correlacionando a idade gestacional com a síntese de surfactante durante o desenvolvimento fetal. Pulmões de fetos com quatro meses de idade gestacional estavam na fase canalicular de desenvolvimento, sem a presença de pneumócitos tipo II ou bandas eletroforéticas compatíveis com a presença de proteínas surfactante. No 5° mês gestacional, os pulmões dos fetos encontravam-se em fase de saculação terminal, com a presença de alvéolos por epitélio cúbico, com áreas formadas por pneumócitos I e II. Nesse período ainda não foi possível identificar proteína surfactante nos pulmões. Esses órgãos em fetos com seis meses de idade gestacional estavam em fase de saco terminal, com presença de pneumócitos tipo I e II. Nessa fase a análise para determinação protéica do surfactante de feto bovino (SDS - PAGE) demonstrou presença de bandas entre 26 e 36kDa, confirmando produção de SP - A, proteína surfactante encontrada em maior quantidade. A partir do 7° mês gestacional, a fase de saco terminal é mais evidente e complexa, com desenvolvimento de intensa vascularização. O pneumócito tipo I apresentava aspecto mais pavimentoso, e o tipo II apresentava aspecto mais globoso. Na análise SDS - PAGE do lavado bronco - alveolar, bandas de proteína surfactante com aspecto similar ao de animais recém-nascidos foram encontradas. Em recém-nascidos, pulmões na fase alveolar foram observados com pneumócitos tipo I e II característicos. O perfil das bandas do lavado bronco-alveolar dos recém-nascidos foi igual ao de animais adultos. Esses achados sugerem que um animal nascido precocemente, a partir dos sete meses de gestação, teria sua sobrevivência garantida devido a uma possível funcionalidade do sistema respiratório do feto, pois o pulmão possuiria as características necessárias para a síntese de proteínas surfactantes. Entretanto, mais estudos clínicos sobre a funcionalidade do sistema respiratório abrem novas fronteiras de experimentos sobre fisiologia respiratória em recém-nascidos bovinos.

The aim of this study was to characterize the presence of type II pneumocyte and the beginning of the surfactant protein production in bovines to the correlated gestational age. Lungs of fetuses at 4 months gestational age were found to be in the canalicular development phase, without presence of type II pneumocytes or appearance of electrophoretic bands compatible with the presence of the surfactant protein. In fetuses at 5 months of gestational age, the lungs were at the terminal sac phase, with the presence of primitive alveolus, formed by cubical epithelium and areas formed by pneumocytes I and pneumocytes II. No electrophoretic bands compatible to the surfactants proteins were identified. In contrast, lungs of fetuses at 6 months of gestational age showed, development of terminal sac phase, with the presence of type I and type II pneumocytes. In this phase, proteic determination for the SDS - PAGE analysis showed the presence of bands between 26 to 36kDa, demonstrating SP - A production, which is the surfactant protein found in highest amount. From the 7th gestational month on, the phase of terminal sac is more evident and complex, with intense sprouting vascularization. The type I pneumocyte had a more squamous aspect and type II pneumocyte were more globular. In the SDS - PAGE analysis of the bronchial - alveolar wash, surfactant protein bands had been observed with a profile similar to that of the newborn animals. In newborn animals, lungs in alveolar phase showed the development of type I and II pneumocytes. The profile of the bronchial - alveolar wash from the newborn was similar to an adult animal, presenting the same bands in triplets. These results suggest that early born fetuses, from 7 months of pregnancy could have a guarantee of surviving due to possible functionality of respiratory system because the lung presented all necessary features for surphactant protein syntheses. However, new clinical studies related to respiratory system functionality open new venues for newborn bovine's experiments respiratory physiology.