Three-dimensional digitalized and printed tongue models of the cow, dog, pig, and horse for undergraduate veterinary educationh / Modelos tridimensionales digitalizados e impresos de lengua de vaca, perro, cerdo y caballo para la educación veterinaria de pregrado

SUMMARY: This study aimed to construct three-dimensional (3D) anatomical models of the tongue of domestic mammals of veterinary interest. The tongues were obtained from the didactic collection of the Laboratory of Veterinary Macroscopic Anatomy in the Surgery Department of the School of Veterinary Medicine and Animal Science, University of Sao Paulo. Tongues from a cow, dog, horse, and pig were selected for scanning and creation of the 3D-printed models. The printer used a filamentous thermoplastic material, acrylonitrile-butadiene-styrene (ABS), which was deposited together with a support resin. In addition to the printing of models, an interactive 3D PDF was generated, creating a didactic collection for students. The anatomical characteristics and peculiarity of the tongues were easily identified in the scanned and printed images. The 3D scanning and printing offered an innovative method of visualizing different anatomical structures and, together with the existing methods, can optimize anatomy teaching in an educational context.

RESUMEN: Este estudio tuvo como objetivo construir modelos anatómicos tridimensionales (3D) de la lengua de mamíferos domésticos de interés veterinario. Las lenguas se obtuvieron de la colección didáctica del Laboratorio de Anatomía Macroscópica Veterinaria del Departamento de Cirugía de la Facultad de Medicina Veterinaria y Zootecnia de la Universidad de São Paulo. Se seleccionaron lenguas de vaca, perro, caballo y cerdo para escanear y crear los modelos impresos en 3D. La impresora utilizó un material termoplástico filamentoso, acrilonitrilo-butadieno-estireno (ABS), que se depositó junto con una resina de soporte. Además de la impresión de modelos, se generó un PDF 3D interactivo, creando una colección didáctica para los estudiantes. Las características anatómicas y la peculiaridad de las lenguas se identificaron fácilmente en las imágenes escaneadas e impresas. El escaneo e impresión 3D ofrecieron un método innovador para visualizar diferentes estructuras anatómicas y, junto con los métodos existentes, puede optimizar la enseñanza de la anatomía en un contexto educativo.

Canine Skull Digitalization and Three-Dimensional Printing as an Educational Tool for Anatomical Study.

This article aims to standardize 3D scanning and printing of dog skulls for educational use and evaluate the effectiveness of these anatomical printed models for a veterinary anatomy course. Skulls were selected for scanning and creating 3D-printed models through Fused Deposition Modeling using acrylonitrile-butadiene-styrene. After a lecture on skull anatomy, the 3D-printed and real skull models were introduced during the practical bone class to 140 students. A bone anatomy practical test was conducted after a month; it consisted in identifying previously marked anatomical structures of the skull bones. The students were divided into two groups for the exam; the first group of students took the test on the real skulls, whereas the second group of students took the test on 3D-printed skulls. The students' performance was evaluated using similar practical examination questions. At the end of the course, these students were asked to answer a brief questionnaire about their individual experiences. The results showed that the anatomical structures of the 3D-printed skulls were similar to the real skulls. There was no significant difference between the test scores of the students that did their test using the real skulls and those using 3D prints. In conclusion, it was possible to construct a dynamic and printed digital 3D collection for studies of the comparative anatomy of canine skull species from real skulls, suggesting that 3D-digitalized and-printed skulls can be used as tools in veterinary anatomy teaching.

Effect of racial crossing on the seminal parameters of rams submitted to heat stress / Efeito do cruzamento racial nos parâmetros seminais de carneiros submetidos ao estresse de calor

The aim of this study was to evaluate the effect of racial crossing on seminal parameters of eight Santa Inês and crossbred (Santa Inês x Dorper) rams submitted to heat stress, and to monitor the return of these parameters to previously reported. Before to place the insulation bags, two collects of semen through electroejaculation were performed. The insulation pouches were made with double-layer plastic, internally lined with cotton, and fixed around the spermatic funiculus and scrotum with adhesive tape and bandage remaining on the testes of the animals for seven days. The first collect was performed on the day that the pouches were taken (day 0) and thereafter, every seven days, totalizing 15 measurements. Data were submitted to analysis of variance (ANOVA). The analyzed variables were subjected to Dunnett test at 5% probability to compare the values obtained before treatment with those obtained in the following days. In this study it was found that the animals restored normal seminal parameter after the insulation effects, however, the return rate differed slightly among the studied breeds. The crossbred animals restored the seminal patterns, on average, a week before Santa Inês. It is concluded that the racial crossing influences the semen parameters of rams submitted to heat stress.

O objetivo deste estudo foi avaliar o efeito do cruzamento racial sobre parâmetros seminais de oito carneiros Santa Inês e mestiços, submetidos ao estresse térmico e monitorar o retorno desses parâmetros aos relatos anteriormente. Antes de colocar as bolsas de insulação, foram realizadas duas coletas de sêmen por meio de eletroejaculação. As bolsas de insulação foram confeccionadas com plástico de camada dupla, revestidas internamente com algodão, fixadas ao redor do funículo espermático e escroto com fita adesiva e bandagem, permanecendo nos testículos dos animais por sete dias. A primeira coleta foi realizada no dia em que as bolsas foram retiradas (dia 0) e a partir daí, a cada sete dias, totalizando 15 coletas. Os dados foram submetidos à análise de variância (ANOVA). As variáveis analisadas foram submetidas ao teste de Dunnett a 5% de probabilidade para comparar os valores obtidos antes do tratamento com aqueles obtidos nos dias seguintes. Neste estudo verificou-se que os animais restauraram os parâmetros seminais normais após os efeitos da insulação, porém, a taxa de retorno diferiu ligeiramente entre as raças estudadas. Os animais mestiços restauraram os padrões seminais, em média, uma semana antes da Santa Inês. Conclui-se que o cruzamento racial influencia os parâmetros seminais de carneiros submetidos ao estresse térmico.

Morphology of the genital organs of the female red-rumped agouti (Dasyprocta leporina, Linnaeus, 1758) during estrous cycle phases and in advanced pregnancy.

The study investigated the gross and microscopic anatomy of the genital organs of 20 agoutis at different stages of the estrous cycle and four in the final trimester of pregnancy. Specimens were euthanized and their reproductive organs were fixed in a 4% paraformaldehyde or 2.5% glutaraldehyde solution and submitted to routine histological techniques for light and scanning electron microscopy. In the ovary, during the proestrus phase, we observed developing follicles and corpus luteum (CL) in regression; during estrus, there were Graafian follicles; during metestrus, there was a hemorrhagic corpus, whereas in diestrus, there was a mature CL. The uterus was partially double because the cervix was cranially septate but caudally, the septum disappeared, forming a single ostium that opened into the vagina. Changes occurred along the estrous cycle in the uterine and vaginal epithelia, that is, an increase in the uterine epithelium height accompanied by an increase of thickness of the vaginal epithelium during the follicular phase and a decrease of thickness of both epithelia during the luteal phase. The endometrial lining was composed of a simple cuboidal epithelium to simple columnar epithelium with basal nuclei. The vaginal mucosa consisted of epithelium that varied from nonkeratinized stratified squamous (luteal phase) to keratinized stratified squamous (follicular phase). The clitoris was external to the vagina. It presented two protruding lateral keratinized spicules and a centralized urethra, with no common parts between the urinary and genital tracts. Anatomical and histological changes were observed mainly in the cervix, vagina and spicules of the clitoris during the EC.

Skin development in 14 to 60-day-old Guinea pig fetus / Desarrollo de la piel del feto de conejillo de indias de 14 a 60 días

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.

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.

Morphological tools to evaluate the digestory apparatus in rocky cavy (Kerodon rupestris).

This research describes for the first time the complete morphology of the digestive apparatus of rock cavies. Dissection, light microscopy, and scanning electron microscopy were performed. The oral cavity has: the hard palate without palatine wrinkles and the soft palate; the tongue composed by striated musculature, with presence of vallate, foliated, and fungiform papillae with taste buds and filiform papillae with mechanical function; and, 20 teeth of the hypsodonts type. Esophagus, stomach, small intestine (duodenum, jejunum, and ileum), and the large intestine (cecum, colon, and rectum) are found. The anus is present at the end of the alimentary channel. Organs of digestive tube are composed by four tunics: mucosa, submucosa, muscular, and serosa. The duodenum, jejunum and ileum have villi. Jejunum, ileum and cecum present Lieberkühn crypts. The cecum has mucous glands. Colon and rectum are folded and have goblet cells. Anus presents sebaceous glands. As associated glands it is found the liver with six lobes and gallbladder; a lobulated pancreas; and a pair of each major salivary gland (parotid, mandibular, and sublingual). Parotid glands have serous acini and mandibular and sublingual glands have mucous acini. Pancreas has adenomers. The liver has hepatocytes and portal vein, hepatic artery, and bile duct (portal triad), separated by sinusoids. It is concluded that the digestive apparatus of the rock cavy has variations in the dentition, lingual papillae, and acini of the salivary glands when compared to other rodents. Other variations refer to the well-developed cecum characteristic of herbivorous behavior.

Morphology and anatomic-surgical segmentation of the spleen of Pecari tajacu Linnaeus, 1758 / Morfologia e segmentação anátomo-cirúrgica do baço de Pecari tajacu Linnaeus, 1758

The spleen is a secondary lymphoid organ with importance in clinical surgery. Due this and the lack of data in the literature, the current paper analyzes the organ´s morphology and segmentation in collared peccaries. Twenty animals were used at the Center for the Multiplication of Wild Animals of the Universidade Federal Rural do SemiÁrido, Mossoró RN Brazil. The spleen was removed with the splenic pedicle preserved, identifying the lineal artery and vein. Fragments from four spleens were harvested to be examined under light microscopy. They were fixed in a paraformaldehyde solution 4% and buffered with sodium phosphate 0.1M, pH 7.4. Routine histological techniques were performed: the spleens were dehydrated in increasing ethanol concentrations; diaphanized in xylol; soaked in paraffin; 7µm cuts were obtained and stained by hematoxylin-eosin or Gomori trichrome technique. The intraparenchemal vascularization of sixteen spleens were analyzed by latex or vinilite acetate perfusal of the lineal artery and vein and the organ fixed, respectively, in a water solution of formaldehyde 10% or immersed in a solution of sulfuric acid 30%. The collared peccary´s spleen had a tongue-like shape. Under the microscope, the spleen featured an intermediary type, with a great amount of white pulp, a predominance of red pulp and few trabeculae. The spleen´s segments had three different regions, namely dorsal, middle and ventral, in irrigation terms with a possible surgical removal of the dorsal region. Knowledge on the angio-architecture and segmentation of the spleen will be a contribution for surgical procedures in wild species, having a great relevance when partial splenectomy is required.

O baço é um órgão linfoide secundário e com importância na clínica cirúrgica. Diante da inexistência de dados na literatura, este artigo estudou a morfologia e segmentação deste órgão em catetos. Foram utilizados 20 animais que vieram a óbito por causas naturais no Centro de Multiplicação de Animais Silvestres da Universidade Federal Rural do Semi-Árido, Mossoró-RN, Brasil. O baço foi coletado preservando-se o pedículo esplênico, pelo qual identificavam-se a artéria e a veia lienais. Fragmentos de quatro baços foram coletados para análise em microscopia de luz. Estes foram fixados em solução de paraformaldeído a 4% tamponado com fosfato de sódio 0,1M, pH 7,4 e submetidos a técnicas histológicas de rotina. Os baços foram desidratados em concentrações crescentes de etanol; diafanizados em xilol; embebidos em parafina e obtidos cortes de 7µm e corados pela técnica de hematoxilina-eosina ou tricrômio de Gomori. A vascularização intraparenquimal de vinte baços foram analisadas através da perfusão de latex ou acetato de vinilite na artéria e veia lienais e logo o órgão foi fixado, respectivamente, em solução aquosa de formaldeído a 10% ou imersos em solução de ácido sulfúrico a 30%. O baço do cateto apresentou-se como uma estrutura longa com forma similar a uma língua. Microscopicamente, o baço caracterizou-se como do tipo intermediário, possuindo quantidade considerável de polpa branca, com predominância da polpa vermelha e poucas trabéculas. A segmentação do baço em termos de irrigação apresentou três regiões distintas: dorsal, média e ventral, com a região dorsal passível de remoção cirúrgica. O conhecimento sobre a angioarquitetura e segmentação do baço de cateto contribuirá para a realização de procedimentos cirúrgicos nesta espécie silvestre e de grande importância quando se fizer necessário a esplenectomia parcial.

Sexual determination and differentiation during embryonic and fetal development of New Zealand rabbit females / Determinación y diferenciación sexual durante el desarrollo embrionario y fetal de conejos hembras de Nueva Zelanda

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.

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.

Development of urogenital system in the Spix cavy: A model for studies on sexual differentiation.

This study documented, for the first time, the morphological patterns of differentiation of male and female genital organs of Spix cavy (Galea spixii) using histological and ultrastructural analyses, with immuno-localization of steroidogenic enzymes, cytochromes P450 aromatase (P450arom) and 17α-hydroxylase/17, 20-lyase (P450c17), involved in the synthesis of estrogens and androgens respectively throughout fetal sexual development. Undifferentiated gonads of Spix cavy develop into ovaries in females after 25 days of gestation (DG), exhibiting P450arom immunoreactivity. After 25 DG, paramesonephric ducts develop and form oviducts, uterine horns and cranial portion of the vagina. The caudal portion of the vagina originates from the urogenital sinus, and a vaginal closure membrane is present at the end of gestation. Partial channeling of the urethra into the clitoris occurs after 40 DG, but complete channeling never occurs. A preputial meatus emerges near the tip of organ. In males, undifferentiated gonads develop into testes at 25 DG and develop immunoreactivity for P450c17, which is required for androgens synthesis and likely maintenance of mesonephric ducts. Mesonephric ducts develop subsequently, forming the epididymis and ductus deferens. The pelvic urethra develops after 25 DG with channeling into the penis occurring around 30 DG. This is the first morphological study describing the process of sexual differentiation during gestation in a hystricomorph rodent and one of the most comprehensive analyses conducted in any mammal. Male genital organ development follows the general pattern described in other domestic mammals, but does not include formation of the baculum as occurs in mice and rats. In females, clitoral development includes partial canalization by the urethra and development of a preputial meatus. Further studies are required to clarify the mechanisms involved in the differentiative processes described.

Influence of racial crossing on testicular structure and spermatogenesis in sheep Santa Inês and crossbred Santa Inês and Dorper.

The study aimed at evaluating the testicular structure and spermatogenesis of Santa Inês and Santa Inês and Dorper crossbred sheep. Sixteen testicles of the animals under study were used, in order to evaluate the volumetric proportion, the diameter of the tubules, the height of seminiferous epithelium, the frequency of stages of the seminiferous epithelium cycle and the spermatogenic yield. The data were submitted to Student-Newman-Keulstestat 5% significance. The tubular diameter was 173.12 ± 29.09 µm and 185.71 ± 29.7 µm, and the height of the seminiferous epithelium was 52.29 ± 9.98 µm and 56.68 ± 11.25 µm, for the crossbred and the Santa Inês, respectively. There was no difference between the testicular compartments and the frequency of the stages of the seminiferous epithelium cycle between the groups. Santa Inês sheep presented a number of 15.36 ± 4.49 spermatocytes in the pre-leptotene/leptotene stage and 27.42 ± 6.65 spermatocytes in the pachytene, whereas the crossbred presented 13.18 ± 5.19 and 23.48 ± 7.80, respectively. The crossbred showed higher meiotic yield (3.98 ± 1.28) and spermatogenesis yield (3.71 ± 1.02). It is concluded there are differences in testicular morphology and spermatogenesis between Santa Inês sheep and crossbred Dorper/Santa Inês, indicating that the crossbreeding between the Santa Inês and Dorper animals allows a gain in the reproductive potential.

Development of Adrenal Glands from the Embryonic Phase to Puberty in Spix's Yellow-toothed Cavies (Galea spixii, Wagler, 1831)

ABSTRACT Several authors have underscored the importance of establishing parameters in morphological development by gender comparison to establish clinical and pre-clinical assays through the use of experimental models. Current research compares the morphometry of right and left adrenal glands of males and females and describes differentiation of the cortex and medulla tissue during the embryonic, pre-puberty and puberty phases in Spix´s yellow-toothed cavies. Embryos aged 22 (22D), 25 (25D), 30 (30D), 40 (44D) and >50 (50D) days of gestation and neonates aged 15 (15DPN) (DPN= Days postnatal), 30 (30DPN) and 90 (90DPN) days after birth were analyzed. Comparisons included morphometric and histological analysis in all periods described. When compared the right and left adrenal glands, results show that the length and width have statistical differences (p<0.05). Statistical difference between right and left glans for weight occurred only after 30D in males and after 50D in females. When compared male and females, no statistical difference in the right and left glands was extant. In the case of tissue differences, the glomerular zone is the first to emerge after 22D, followed by the fasciculate zone after 25D and by the reticular zone during the post-natal period. Medullar tissue was spread between the cortical tissue at the onset of development, establishing itself at the center of the organ since the end of pregnancy (>50D) up to puberty. Considering tissue differentiation, there was no difference between the adrenal glands of male and female cavies or between the right and left adrenal glands.

Bioimpressão e produção de mini-órgãos com células tronco / Bioprinting with stem cells and production of mini-organs

A bioimpressão é considerada uma fonte promissora no desenvolvimento celular, e na produção de mini-órgãos, válulas, cartilagens que futuramente poderão ser utilizados na terapia para transplantes em animais e humanos. Assim, essa técnica poderá ser utilizada como uma terapia eletiva, no tratamento de injúrias e principalmente no tratamento de doenças crônico-degenerativas. Em humanos essa terapia está sendo pesquisada a fim de auxiliar a medicina no tratamento e regeneração de tecidos impressos a partir de arcabouços de células desenvolvidas a partir de células-tronco, biomateriais e impressões em 3D. O uso dessa tecnologia é também um auxiliar nas pesquisas oncológicas com o intuito de projetar e avaliar a proliferação celular de tumores, bem como a ação de novos medicamentos quimioterápicos. No entanto, a maior limitação para o uso da terapia utilizando-se a impressora de células, órgãos e tecidos em 3D é a falta de protocolos unificados com metodologias reprodutíveis e detalhadas; com o objetivo de viabilizar a utilização da impressora e a impressão de células, órgãos e tecidos em 3D. Dessa forma, esta revisão busca reunir as publicações mais atuais na área, as quais destacam os avanços no uso de bioimpressão com células-tronco, a fim de descrever as principais técnicas e os potenciais de utilização como alternativa terapêutica na medicina humana e veterinária.(AU)

The bioprinting is considered a promising source in cell development, and production of mini-organs, valves, cartilage that may eventually be used in therapy for transplantation in animals and humans. It can also be used as an elective therapy in the treatment of injuries and treatment of chronic degenerative diseases. In humans, this therapy is been studied mainly in the treatment and regeneration of tissues printed from scaffold cells developed from stem cells, biomaterials and impressions in 3D. This technology is also an aid for the study of the formation of tumors, in order to design and evaluate the cellular proliferation of the tumors and the action of new chemotherapy drugs. However, the main drawback to this therapy is the lack of standardized protocols with reproducible and detailed methodologies with the aim of enabling the use of bioprinting and printing cells, tissues and organs in 3D. Thus, this review seeks to bring together the most current publications of the bioprinting area in order to describe the technique and its potential use as a therapeutic alternative.(AU)

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

BACKGROUND: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

Follicular development and morphological changes in the vaginal epithelium during the estrous cycle of Galea spixii.

The current study aimed to determine if characteristics observed in vaginal cytology during the estrous cycle of female SYT cavies corresponded with proliferation of the vaginal epithelium, characterized by proliferating cell nuclear antigen (PCNA) immunolocalization, and with follicular development at different phases of the estrous cycle. After determining estrous cycle phases by vaginal cytology, females were euthanized at metestrus, diestrus, proestrus, and estrus. Histological study of the vaginal epithelium and ovary were then performed. Immunohistochemistry for PCNA in vaginal tissue at each cycle phase was also performed. Superficial cornified cells and early post-ovulatory follicles were found at estrus. Few nuclei below the enucleate superficial cells were immunoreactive to PCNA. At metestrus, the vaginal epithelium underwent desquamation and lost the superficial cornified cells; basal and intermediate cells appeared, and the post-ovulatory follicle formed an early corpus luteum. No PCNA immunoreactivity was observed. At diestrus, the corpus luteum was developed, and the vaginal epithelium contained basal and intermediate cells. There was PCNA immunoreactivity in the cellular nucleus in the germinative stratum of the epithelium. Because of the growth and maturation of ovarian follicles, the vaginal epithelium suffered intense proliferation at proestrus. Vaginal cytology revealed large intermediate cells and nucleated and enucleated superficial cornified cells. In the ovary, mature follicles were present. More apparent immunoreactivity of PCNA in the germinative layer was found. In summary, we inferred that vaginal exfoliative findings matched the proliferation process of the vaginal epithelium. PCNA immunolocalization occurred as well as corresponding follicular development in the ovaries.

Morphology of the adrenal glands of giant anteater (Myrmecophaga tridactyla, Linnaeus, 1758) of wild life / Morfologia das glândulas adrenais de tamanduá-bandeira (Myrmecophaga tridactyla, Linnaeus, 1758) de vida livre

The Myrmecophaga tridactyla (family Pilosa), popularly known in Brazil as the giant anteater is classified as an endangered species by the main survey agencies of biodiversity preservation around the world. The adrenal glands are important organs, which are related to homeostasis of the animal. This study aimed to describe the morphology of the adrenal glands of M. tridactyla, providing data for future studies related to the production of steroid hormones in specimens exposed to different stress factors. The adrenal glands of 14 specimens were used. The glands were measured, photodocumented and described by light microscopy. The adrenals were positioned in the cranial extremity of the kidneys (left and right) and related medially to the caudal vena cava. The right adrenal gland had an elongated shape while the left adrenal gland had pyramidal or triangular shape. The weight was 4.765±0.129g for the right adrenal and 3.975±0.213g for the left adrenal. The length was 4.50±0.14cm for the right adrenal and 4.28±0.11cm for the left adrenal. The width was 2.60±0.13cm for the right adrenal and 2.37±0.12cm for the left adrenal. The thickness was 0.45±0.11cm for the right adrenal and 0.68±0.14cm for the left adrenal. Statistical differences (p<0.05) between right and left adrenals for all values analyzed were observed. Microscopically it was observed a capsule composed by modeled dense connective tissue lining the organ. Below this capsule, from the outermost to the innermost layer, it was observed a cortical region divided into glomerular, fasciculata and reticular zones with a medullar region occupying the center of the organ. Further studies related to steroidogenesis with specimens collected at different periods of the year are necessary, aiming to observe if there are morphological or hormonal variations in the adrenal glands due to seasonal periods. This information would be of great importance because it could reflect the behavioral habits of this species.

O Myrmecophaga tridactyla (família Pilosa), conhecido popularmente no Brasil como Tamanduá- Bandeira, é uma espécie classificada como ameaçada de extinção pelos principais órgãos de levantamento e preservação da biodiversidade no mundo. As glândulas adrenais são importantes órgãos, relacionados com a homeostasia dos animais. Neste estudo, objetivou-se descrever a morfologia das glândulas adrenais de M. tridactyla, fornecendo dados para futuros estudos relacionados à produção de hormônios esteroides em espécimes expostos a diferentes fatores de stress. Esta pesquisa traz a análise das glândulas adrenais em 14 espécimes de M. tridactyla, as quais foram mensuradas, fotodocumentadas, processadas e analisadas por microscopia de luz. Encontrou-se que as glândulas adrenais pares estavam posicionadas na extremidade cranial dos rins e relacionadas medialmente com a veia cava caudal. Sendo que, a glândula adrenal direita possuía formato mais alongado que a glândula adrenal esquerda, a qual apresentou formato piramidal ou triangular. A massa da glândula adrenal direita foi de 4,765±0,129g, enquanto a esquerda pesou 3,975±0,213g. O comprimento foi de 4,50±0,14cm para a glândula direita e 4,28±0,11cm para a esquerda. A largura foi de 2,60±0,13cm para a glândula direita e 2,37±0,12cm para a esquerda. A espessura da glândula direita foi de 0,45±0,11cm e da esquerda foi 0,68±0,14. Foram observadas diferenças estatísticas (p<0,05) entre as glândulas direitas e esquerdas em todas as dimensões analisadas. Com auxílio de microscopia de luz observou-se uma cápsula de tecido conjuntivo denso modelado, revestindo o órgão. Abaixo dessa cápsula, foi possível evidenciar, da camada mais externa para a mais interna, uma região cortical, dividida em zonas glomerular, fasciculada e reticular, com uma região medular ocupando o centro do órgão. Futuros estudos relacionados à esteroidogênese adrenal com espécimes em diferentes épocas do ano poderiam demonstrar se as glândulas adrenais apresentam variações sazonais em sua conformação morfológica ou na produção de hormônios adrenais decorrentes dessas variações, sendo que, estes dados seriam de grande importância, pois poderiam refletir os hábitos comportamentais nesta espécie.

Development of the central nervous system in guinea pig (Cavia porcellus, Rodentia, Caviidae) / Desenvolvimento do sistema nervoso central em guinea pig (Cavia porcellus, Rodentia, Caviidae)

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.(AU)

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.(AU)

Lumbosacral plexus of the mongolian gerbil(Meriones unguiculatus Milne-Edwards, 1867) / Plexo lombossacral do gerbil (Meriones unguiculatus Milne-Edwards, 1867)

The Mongolian gerbil, belonging to the Muridae family, is characterized as an economic asset in the pet market due to its easy management. Current research analyzes and describes the origin of the nerves that make up the lumbosacral plexus in the species, foregrounding comparative anatomy, especially among wild rodents. The experiment, approved by the Committee of Institutional Bioethics, involved 15 animals which had been used in other studies and were donated to the Laboratory of Applied Animal Morphophysiology of the Federal Rural University of the Semi-Arid, Mossoró, Rio Grande do Norte, Brazil. The animals were fixed in a water solution of formaldehyde 10%. After 72 hours, they were dissected, the abdominal cavity was opened and eviscerated, and the psoas minor, psoas major and the quadratus lumborum muscles were removed to inspect the nerves that form the lumbosacral plexus. Results were given in percentages and grouped in tables. The lumbosacral plexus of the Mongolian gerbil frequently derived from the ventral roots of the last thorax nerves, from the seven lumbar nerves and from the four sacral nerves (Type II ­ T12-S4). The nerves of the gerbil´s lumbosacral plexus are the iliohypogastricus, ilioinguinalis, cutaneus femoris lateralis, genitofemoralis, femoralis, obturatorius, ischiadicus, gluteus cranialis, pudendus, gluteus caudalis and rectales caudales nerves. Information on the origin of the nerves of the gerbil´s lumbosacral plexus and their description is relevant for clinical and surgical studies, and for the application of techniques or anesthetic drugs.

O esquilo da Mongólia é um roedor pertencente à família Muridae, caracterizado por seu interesse econômico no mercado pet e por ser de fácil manejo. Esta pesquisa analisa e descreve a origem dos nervos que formam o plexo lombossacral da espécie. No experimento foram utilizados 15 animais, provenientes de outros estudos aprovados pelo Comitê de Bioética Institucional, que foram doados ao Laboratório de Morfofisiologia Animal Aplicada da Universidade Federal Rural do Semi-Árido, Mossoró, Rio Grande do Norte. Os animais foram fixados em solução aquosa de formaldeído a 10%. Após 72 horas, os animais foram dissecados, realizando-se a abertura da cavidade abdominal, depois, eviscerados e em seguida, os músculos psoas maior, psoas menor e quadrado lombar foram removidos para visualização dos nervos que formam o plexo lombossacral. Os resultados foram expressos em percentagens e agrupados em tabelas. O plexo lombossacral do gerbil originou-se mais frequentemente das raízes ventrais do último nervo torácico, dos sete nervos lombares e dos quatro nervos sacrais (Tipo II ­ T12-S4). Os nervos que formaram o plexo lombossacral do gerbil foram: ílio-hipogástrico, ílioinguinal, cutâneo femoral lateral, genitofemoral, femoral, obturatório, isquiático, glúteo cranial, pudendo, glúteo caudal, e nervo retal caudal. O conhecimento sobre a origem e os nervos do plexo lombossacral no gerbil mostra-se útil para estudos clínico-cirúrgicos e aplicação de técnicas ou fármacos anestésicos.

Development and morphological changes in the vaginal closure membrane throughout gestation in Galea spixii (Rodentia: Caviidae).

Present research was carried out in order to perform the monitoring of development, recognizes the type of tissue and describes histological and cellular changes of the vaginal closure membrane (VCM) throughout pregnancy in Galea spixii. The results showed that at 20 days of gestation (DG), the VCM occludes completely the external vaginal ostium. Microscopically, the VCM presented juxtaposed cells, derived from the stratum germinative of the stratified epithelium of vaginal mucosa at 20 DG and areas with cell clusters with the presence of intercellular spaces in the final stages of pregnancy (40-50 DG). At 0 DG, the stratified epithelium of vaginal mucosa presented all strata but at 20 DG presented stratified epithelium without the stratum corneum and stratum granular and showed communicant junctions by desmosomes and interdigitations in the cell membrane compound the VCM. Gradually from 40 to 50 DG the stratum germinative became barely perceptible. Many cells showed apoptotic nuclei and emerged many intercellular spacing. So, the interdigitations and desmosomes were not observed. Here, it was demonstrated for the first time that the VCM is formed after the extinction of the stratum granular and corneum of the vaginal mucosa epithelium, with the proliferation of the cells of stratum germinative and communication and junction through desmosomes and interdigitations of these cells. At the end of pregnancy, cellular apoptosis; the spread of stratum germinative; and, absence of cellular communication and junction may be responsible for the weakening of the VCM and may assist the process of rupture of this membrane.

Morphological and ultrastructural analysis of the tongue of Gracilinanus microtarsus (Wagner, 1842) / Análise morfológica e ultraestrutural da língua da Gracilinanus microtarsus (Wagner, 1842)

The G. microtarsus is a small marsupial belonging to the Didelphidae family and may be found mainly in the Atlantic Rainforest of the Brazilian Southeastern region. G. microtarsus has important ecological importance in the dispersion of seeds of several plant species. Current research characterizes morphologically six tongues of adult male and female specimens, which are fixed in 10% formaldehyde. The tongues were analyzed, measured and photographed "in situ" and "ex situ". Anatomy, light microscopy and ultrastructural analysis were performed. Macroscopically the tongue has root, a fixed body and a free apex. The tongue has fungiform papillae and high conical and acute filiform papillae across the dorsal surface. It also featured three vallate papillae located at the root and marginal fungiform papillae at the apex. Microscopically, the tongue is composed of longitudinal and transversal muscle fibers. Blood vessels, nerves, serous, mucous and mixed acini are found in non-modeled connective tissue. Ventrally, the tongue is lined by a keratinized stratified pavement epithelium. The morphology of the tongue of G. microtarsus resembles that described in carnivores and marsupials Didelphidae regarding its microscopic structure, differing only in the distribution and number of papillae.

O G. microtarsus é um pequeno marsupial pertencente à família Didelphidae, que pode ser encontrado principalmente na Mata Atlântica da região sudeste brasileira. O G. microtarsus possui importância ecológica na dispersão de sementes de diversas espécies de plantas. A pesquisa atual caracteriza morfologicamente as línguas de seis animais adultos, fixados em formol a 10%. As línguas foram analisadas, mensuradas e fotografadas. Foram realizadas análises anatômicas, microscópicas de luz e ultraestruturais. Macroscopicamente a língua possui raiz, um corpo fixo e um ápice livre. A língua possui papilas fungiformes, filiformes cônicas altas e agudas em toda a superfície dorsal. Também possui três papilas valadas localizadas na raiz e papilas fungiformes marginais no ápice da língua. Microscopicamente, a língua é composta por fibras musculares longitudinais e transversais. Os vasos sanguíneos, nervos e ácinos serosos, mucosos e mistos são encontrados em tecido conjuntivo não modelado. Ventralmente, a língua é revestida por um epitélio pavimentoso estratificado queratinizado. A morfologia da língua de G. microtarsus se assemelha ao descrito em carnívoros e marsupiais Didelphidae quanto à sua estrutura microscópica, diferindo apenas na distribuição e número de papilas.

Composition and significance of glycosaminoglycans in the uterus and placenta of mammals

Glycosaminoglycans (GAGs) are heteropolysaccharides in mammalian tissue and consist of repeated disaccharide units with mono-sulfated or non-sulfated monosaccharides. GAGs are important components of the Extracellular Matrix (ECM) with several physiological roles, in the recognition, migration, adhesion, proliferation and differentiation processes. They are also important in angiogenesis, blood homeostasis, immune reactions, follicule development and also in the development of pathologies such as infertility, tumors and metastases. It has been shown that the profile of glycosaminoglycans in the uterine and placental tissues is highly variable throughout the reproductive cycle and during pregnancy. It may be directly related to their physiological or pathological functions in the tissue. The latter has recently triggered special clinical interest. Current review collaborates for a deeper knowledge on the profile and importance of GAGs in uterine and placental tissues throughout the reproductive cycle and pregnancy. It also covers information on the involvement of these molecules in pathological processes.