RESUMEN
In viviparous teleosts, intraovarian gestation occurs intrafollicularly, as in poeciliids, or intraluminally, as in goodeids and anablepids. Furthermore, there are two different forms of embryonic nutrition: lecithotrophy and matrotrophy; depending on the species, these can be exclusive or coexist during gestation. In matrotrophic species, nutrients are transmitted from the mother to the embryo and are especially important in species with intraluminal gestation. Jenynsia lineata is a South American viviparous teleost with intraluminal gestation, characterized by eggs with scarce yolk, which is resorbed when embryos are 6 mm long, thus developing a branchial placenta. Using histological, histochemical, and immunohistochemical techniques, the present study describes the characteristics and changes of the ovarian mucosa in J. lineata during gestational and nongestational phases, and analyzes the embryonic pharyngeal epithelium in the branchial placenta. The ovaries of 30 adult female specimens were processed using histological techniques and stained with hematoxylin-eosin, Masson's trichrome, and Alcian Blue pH 2.5/periodic acid Schiff reagent. To detect cell proliferation, we used antiproliferating cell nuclear antigen antibody. In nonpregnant females, eosinophilic granular cells (EGCs) and lymphocytes were identified in the lamina propria of the tunica mucosa, and melanomacrophage centers (MMCs) and fibroblasts were identified adjacent to tissue debris in the ovarian folds'. In the cellular debris, an embryo in resorption was observed. In pregnant females, the ovarian mucosa has thin vascularization branches entering the opercular chamber of the embryos, in close contact with the forming gill processes, thereby establishing a branchial placenta. Active cell replacement was observed in these ovarian branches. The identification of fibroblasts, lymphocytes, EGCs, and MMCs adjacent to tissue debris could indicate that these cell types are involved in the embryonic resorption process. Considering the new data obtained in this study on the branchial placenta of J. lineata, we conclude that cell proliferation could be involved in the development of maternal-embryonic interaction.
Asunto(s)
Ciprinodontiformes , Ovario , Femenino , Animales , Embarazo , Placenta , Núcleo Celular , Proliferación CelularRESUMEN
The toxic effect of ferns of the genus of Pteris in bovines is caused by ptaquiloside, the main carcinogenic toxin. In this study, ten species of Pteris fern in different phenologic stages and plant conditions were collected in northwest Argentina. The phytochemical analysis showed the presence of Pt in the recent collected samples (adults and young plants) but not in the herbarium specimens. The results show a great variation of Pt concentration that depends on the phenologic stage, plant condition, and collection site. Pt was measured in 6-4326 µg/g concentration, with a mean concentration of 644 µg/g. No Pt was detected in eight species of Pteris collected from herbarium samples; such results may be a false negative. It is important to notice that analysis of herbarium samples for Pt may not be a reliable method to determine its presence. It is important to further understand the potential toxicity caused by these ferns because of their effect on animals, public health, and the environment.
Asunto(s)
Helechos , Pteris , Sesquiterpenos , Animales , Bovinos , Argentina , Indanos/toxicidadRESUMEN
The main evolutionary milestone in the oviparity-viviparity transition is placentation. The placenta is an organ with great morphological diversity among eutherians. The expression of different glycosidic residues (Gr) in the near-term placenta constitutes its glycotype. In this study, the expression of different Gr was determined by lectin histochemistry in early, midterm, and near-term placentas of the plains viscacha (Lagostomus maximus), a caviomorph rodent with the highest poliovulatory rate and embryonic resorption rate among eutherians. Besides, a matrix with the expression of each Gr in the exchange trophoblast of viscacha and other eutherians was constructed to map and infer phylogenetic and evolutionary relationships. Between early, midterm, and near-term placentas, variations in the pattern expression of Gr were observed. The glycotype of the near-term placenta is composed of a high diversity of Gr. Reconstruction of the ancestral state for each Gr present in the near-term placenta showed a diverse scenario: some sugars were common to the species of Placentalia included in this study. In the analyzed species with synepitheliochorial and epitheliochorial placentas, no differential glycosylation patterns between them were observed. In species with invasive placentas, such as the endotheliochorial placentas of Carnivora, some common Gr were detected among them, while others were species-specific. In species with hemochorial placenta, the same Gr are shared. Particularly, in the viscacha greater differences with species of the Hominidae and even Muridae families were observed. Nevertheless, greater similarities with other caviomorph rodents were detected. Placental glycotype of each species constitutes an excellent tool to achieve phylogenetic and evolutionary inferences among eutherians.
Asunto(s)
Placenta , Roedores , Embarazo , Femenino , Animales , Placenta/anatomía & histología , Filogenia , Placentación , TrofoblastosRESUMEN
The Neotropical armoured catfish Corydoras paleatus is a facultative air-breathing teleost commonly exported as ornamental fish. In this species, air breathing enables it to survive and inhabit freshwater environments with low oxygen levels. Therefore, it is important to analyse the gills from a morphological aspect and its dimensions in relation to body mass with reference to aquatic respiration. For that, the gills were analysed using a stereoscopic microscope for morphometric studies, and structural and ultrastructural studies were carried out to compare the four branchial arches. Furthermore, two immunohistochemical techniques were used to locate and identify the presence of a Na+ /K+ pump. The characterization of the potential for cell proliferation of this organ was assessed using an anti-PCNA antibody. The results show that gills of C. paleatus present some characteristics related to its diet and lifestyle, such as the limited development of gill rakers and the abundance of taste buds. In addition, other special features associated with the environment and bimodal breathing were observed: scarce and absent mucous cells (MCs) in the gill filaments and branchial lamellae, respectively, and the localization of mitochondria-rich cells (MRCs) covering the basal third of the branchial lamellae, which reduces the gill respiratory area. A peculiar finding in the gill epithelium of this armoured catfish was the presence of mononuclear cells with sarcomeres similar to myoid cells, whose functional importance should be determined in future studies. Finally, in C. paleatus, the interlamellar space of gill filaments is an important site for cell turnover and ionoregulation; the latter function is also performed by the branchial lamellae.
Asunto(s)
Bagres , Branquias , Animales , Epitelio , Mitocondrias , RespiraciónRESUMEN
Five adult Saanen goats received a single oral dose of Heterophyllaea pustulata containing 42.25 µg/kg rubiadin (anthraquinone) and 3 adult goats were untreated controls. All goats were exposed to sunlight and sequential ear skin biopsies were collected before treatment and at 32 hours, 3 days, 8 days, and 15 days after treatment. Changes at 32 hours after dosing included epidermal spongiosis, single cell death and acantholysis, an increased BAX/BCL-2 protein ratio, and dermal edema. Lesions at day 3 included epidermal and adnexal necrosis, crust formation, and acanthosis. Acanthosis, hyperkeratosis, and dermal fibrosis and neovascularization were present at day 15. The pro-apoptotic (BAX)/anti-apoptotic (BCL-2) protein ratio increased at 32 hours, whereas epidermal and dermal PCNA immunolabeling increased between days 8 and 15 after treatment. The cutaneous lesions were consistent with sunlight-induced damage, and the occurrence in treated but not control goats indicates photosensitization.
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Enfermedades de las Cabras , Trastornos por Fotosensibilidad , Animales , Enfermedades de las Cabras/inducido químicamente , Cabras , Trastornos por Fotosensibilidad/inducido químicamente , Trastornos por Fotosensibilidad/veterinaria , PielRESUMEN
Embryonic death followed by resorption is a conserved process in mammals. Among the polyovular species, Lagostomus maximus (plains viscacha) constitutes a model of early and physiological embryonic death, since out of a total of 10-12 implants, 8-10 are resorbed during early/intermediate gestation, surviving are only the most caudal implantations of each uterine horn. This regular reproductive event is unique to this species, but many characteristics of the implantations during the early gestation of L. maximus, when embryonic death processes begin are unknown. The aim of the present work was to analyze the implantation sites of this species using morphological, morphometric, histochemical, lectinhistochemical, and immunohistochemical techniques to infer the possible causes of this event. Macroscopically, the length and width of the implantation sites significantly increased in a craniocaudal direction. Histochemically, the implantation sites did not differ in the expression of glycoconjugates and glycosidic residues. Furthermore, no variations were observed in cell renewal, hormone receptor expression, and decidualization. Both the glandular and vascular areas of the implantation sites significantly increased in the craniocaudal axis. Some necrotic cells and an inflammatory response with a predominance of lymphocytes and fibrin were observed in the cranial and middle but not in the caudal implantation sites. We conclude that signs of embryonic death and resorption are already observed in the early gestation of L. maximus. Our results reaffirm the hypothesis that postulates the key potential role of uterine glands and blood vessels in the gestation of the species, with emphasis on embryonic death. This pattern of embryonic death in L. maximus makes this species an unconventional mammalian model, which adds to the peculiarities of polyovulation (200-800 oocytes/estrus) and hemochorial placentation.
Asunto(s)
Euterios , Roedores , Animales , Femenino , Hormonas , Placentación , Embarazo , ÚteroRESUMEN
During embryogenesis, teleost females do not develop Müllerian ducts, which form the oviducts in all other vertebrates. Thus, when they reach sexual maturity they do not have oviducts. In viviparous teleosts, the lack of oviducts means that the development of the embryos occurs as an intraovarian gestation, unique among vertebrates. The ovary is an unpaired hollow organ whose cavity is continuous with the caudal portion, the gonoduct, characterized by the absence of germinal cells, which opens to the exterior at the gonopore. The gonoduct attains essential function as a barrier between the germinal region of the ovary and the exterior during all reproductive stages. This study describes the functional morphology of the gonoduct in the viviparous teleost Cnesterodon decemmaculatus during non-gestation (previtellogenesis and vitellogenesis) and gestation. The ovaries were processed using histological techniques and stained with hematoxylin-eosin, and periodic acid Schiff. The gonoduct has two regions: cephalic and caudal, and is formed by three histological layers, which are, from inside to the periphery: (a) tunica mucosa; (b) tunica muscularis; and (c) tunica serosa. In the cephalic region there are mucosal folds extending into the lumen and forming a structure similar to a cervix. The histology of the gonoduct indicates essential functions, that is, (a) the control of the luminal diameter in the limit to the germinal region of the ovary by the presence of a cervix; (b) during insemination the gonoduct receives the spermatozoa, may store and transport them to the germinal region; (c) the presence of melano-macrophage centers indicates support of immunological processes, especially during gestation when these centers increase in size; (d) production of exocrine secretions; and (e) it is the birth canal, internally lined by an ciliated epithelium and surrounded by smooth musclesboth tissues supposedly supporting the birth process.
Asunto(s)
Ciprinodontiformes/anatomía & histología , Gónadas/anatomía & histología , Viviparidad de Animales no Mamíferos , Animales , Desarrollo Embrionario , Epitelio/anatomía & histología , Femenino , Masculino , Melanocitos/citología , Ovario/anatomía & histología , Reproducción/fisiología , Espermatozoides/citología , Vitelogénesis/fisiologíaRESUMEN
The uterus is an organ with great plasticity due to the morphological and physiological changes it experiences during the estrous cycle and pregnancy. In mammals, pregnancy requires diverse sex hormones, growth factors and cytokines, among others, for promoting uterine remodeling to favor implantation, placentation, and embryo/fetus survival and growth. The hystricognathi rodent Lagostomus maximus (plains viscacha) has a high rate of embryonic resorption. The cranial and middle implants are reabsorbed 25-35 days after intercourse while the caudal embryos continue with their development until two precocial offspring are born. So far, no uterine studies of non-pregnant L. maximus females were performed to determine the possible existence of variations in the organ that could be related to the differential survival of the implants. We used ultrasonography, as well as morphological, morphometric, histochemical, lectinhistochemical, and immunohistochemical methods to study differences in the uterine glands (area), vasculature (area), and musculature (thickness) along the uterine horns in non-pregnant females. Along the uterus, all these structures were in more advanced developmental condition in the caudal region as compared to more anterior positions. These regional variations could be decisive in explaining the reason why only caudal implantations come to term. In contrast, no differences in the in the luminal and glandular epithelial cells, nor in the degree of cell proliferation and apoptosis, and hormonal receptor staining were found. These parameters could be related to implantation along the uterine horns, but not to the differential survival of the implants.
Asunto(s)
Embrión de Mamíferos/anatomía & histología , Músculos/anatomía & histología , Roedores/embriología , Útero/anatomía & histología , Útero/irrigación sanguínea , Animales , Apoptosis , Proliferación Celular , Femenino , Hormonas/metabolismo , Lectinas/metabolismo , Receptores de Superficie Celular/metabolismo , Roedores/anatomía & histología , Útero/diagnóstico por imagenRESUMEN
We describe the morphology, histology, and histochemical characteristics of the uropygial gland (UG) of the monk parakeet Myiopsitta monachus. The UG has a heart-shape external appearance and adenomers extensively branched with a convoluted path, covered by a stratified epithelium formed by different cellular strata and divided into three zones (based on the epithelial height and lumen width), a cylindrical papilla with an internal structure of delicate type and two excretory pores surrounded by a feather tuft. Histochemical and lectin-histochemical techniques performed showed positivity against PAS, AB pH 2.5, AB-PAS, and some lectines, likely related to the granivorous feeding habits. Also, we describe the morphogenesis of the UG of the monk parakeet, which appears at embryological stage 34 as a pair of ectodermal invaginations. Heterochronic events in the onset development of the UG when compared with other birds could be recognized. Finally, to examine the phylogenetic occurrence of the UG within the Psittaciformes and infer its evolutionary history, we mapped its presence/absence over a molecular phylogeny. The reconstruction of the characters states at ancestral nodes revealed that the presence of the UG was the plesiomorphic feature for Psittaciformes and its loss evolved independently more than once.
Asunto(s)
Evolución Biológica , Glándulas Exocrinas/embriología , Morfogénesis , Periquitos/embriología , Animales , Embrión no Mamífero/embriología , Glándulas Exocrinas/anatomía & histología , Periquitos/anatomía & histología , FilogeniaRESUMEN
Osteoderms are present in a variety of extinct and extant vertebrates, but among mammals, the presence of osteoderms is essentially restricted to armadillos (Cingulata, Dasypodidae). Osteoderms have been proposed to exhibit a variety of functionalities in Dasypodidae, mainly protection and thermoregulation, and they have been considered as one of the synapomorphies of this group. In this study, we use high-resolution microcomputed tomography to describe the osteoderm micromorphology of several extant species of Dasypodidae in a comparative context. This study allowed the identification, 3D-reconstruction and volume quantification of different internal structures of osteoderms as well as their interrelations. This detailed characterization of the internal osteoderm morphology was compared in a phylogenetic context to assess the evolutionary trends of the species involved. This enables the identification of distinctive patterns for the most widely recognized clades, the Dasypodinae and Euphractinae with a morphological homogeneity in the microstructure of their osteoderms, in comparison with Tolypeutinae where it has not been possible to establish a common morphological pattern. The most important features for linage differentiation is the degree of compaction of the osteoderms, the number of cavities and the development of hairs. It is likely that the differential development of the various structures occurred as adaptive response to climate changes.
Asunto(s)
Armadillos/anatomía & histología , Huesos/anatomía & histología , Imagenología Tridimensional , Piel/anatomía & histología , Animales , Huesos/diagnóstico por imagen , Filogenia , Análisis de Componente Principal , Piel/diagnóstico por imagen , Microtomografía por Rayos XRESUMEN
The lectin histochemical pattern (LHP) was characterized and compared in normal and cystic ovaries of sows. Six biotinylated lectins (PNA, SBA, WGA, RCA-1, DBA and UEA-1) were used on tissue sections. In the normal ovaries, the reaction to UEA-1 and SBA was mild to moderate in mesothelial and endothelial cells. RCA-1 staining was mild to moderate in theca interna of growing follicles, corpora luteum and mesothelium. In addition, this lectin presented strong reaction in endothelial cells, granulosa cells of atretic follicles, zona pellucida of growing follicles and plasma. DBA showed strong intensity in mesothelial and endothelial cells. There was mild to moderate reactivity to WGA in granulosa cells, corpus luteum and theca interna of follicles in development, and moderate in zona pellucida, in granulosa cells of atretic follicles and mesothelium. PNA staining was mild to moderate in oocytes and in the adventitia and media of medullary arteries. Changes in the LHP of the cystic ovaries were noted; however, there were no differences in these findings between the follicular and luteinized cysts. UEA-1 reactivity in the cystic ovaries was moderately reduced in the mesothelial and endothelial cells, whereas there was mild reduction in the DBA staining in the granulosa cells. Reaction to RCA-1 and WGA in the cysts also was decreased in theca interna, zona pellucida and granulosa cells of atretic follicles. Furthermore, endothelium and theca interna in the cystic ovaries presented mild reduction of marcation to SBA, whereas there was decreased reactivity to PNA in the oocytes and adventitia and media layers of the medullary arteries. The results of the current study show that cysts modify the LHP in swine ovaries. These changes of glycoconjugates in many ovarian structures could modify diverse process and may be one of the reasons for decreased fertility in sows.
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Lectinas/metabolismo , Quistes Ováricos/veterinaria , Ovario/metabolismo , Enfermedades de los Porcinos/metabolismo , Animales , Células Endoteliales , Femenino , Células de la Granulosa , Oocitos , Quistes Ováricos/metabolismo , Porcinos , Zona PelúcidaRESUMEN
Cutaneous papillomas (CP) are one of the most common skin neoplasms in dogs. Different murine models have shown that persistent activation of the phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin (PI3K/Akt/mTOR) pathway has a central role in the development and progression of CP. The purpose of this study were to evaluate the immunohistochemical expression pattern of two key molecules involved in the PI3K/Akt/mTOR signalling pathway, pAktSer473 , and pS6Ser235/236 , on 36 canine specimens of CP using a tissue microarray. The results show that the PI3K/Akt/mTOR signalling pathway is persistently activated in CP of dogs, pointing to this pathway as a potential therapeutic target.
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Enfermedades de los Perros/metabolismo , Papiloma/veterinaria , Transducción de Señal/fisiología , Neoplasias Cutáneas/veterinaria , Serina-Treonina Quinasas TOR/metabolismo , Animales , Perros , Regulación de la Expresión Génica , Neoplasias Cutáneas/metabolismo , Serina-Treonina Quinasas TOR/genética , Análisis de Matrices TisularesRESUMEN
This study reports the histochemistry and the distribution of glycoconjugates (GCs) in the zona pellucida (ZP) of preantral, secondary, tertiary, polyovulatory and atretic follicles of ovaries from non-pregnant (NPr) and pregnant (Pr) females of Lagostomus maximus. GCs were studied using histochemical and lectin histochemical methods. The viscacha ZP was positive to all the histochemical techniques. In addition, it was observed that the intensity of staining of the ZP was constant in the different follicular stages between both female groups. The lectin histochemical study revealed that ZP was positive for certain lectins (WGA, RCA-I and CON-A) and that the labelling did not vary between the different follicular stages, but between the two groups of females. By using both histochemical techniques, it was established that the GCs present in the ZP label the complexity of the area. These results allow us to increase our knowledge on the biology of the viscacha's ovary, particularly contributing to the study of polyovulation.
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Glicoconjugados/química , Folículo Ovárico/fisiología , Ovulación/fisiología , Roedores/fisiología , Zona Pelúcida/química , Animales , Femenino , Fase Folicular/fisiología , Histocitoquímica , Lectinas/metabolismo , EmbarazoRESUMEN
Lectin binding relies on the affinity of these substances for specific terminal sugars. The method facilitates the identification of complex structures to which the terminal sugar attaches and may reveal physiological or pathological changes in cells, intracellular interactions or extracellular transport pathways. This study was carried out to investigate the effect of infection with Brucella abortus on the pattern of lectin binding in bovine fetal lungs (n=6) and bovine placentas (n=5). Fetal lungs and placenta from heifers experimentally inoculated with B. abortus, strain 2308 were examined by histological, lectin-histochemical, immunohistochemical and cultural techniques. B. abortus antigens were immunohistochemically detected in fetal lungs and placenta. An increase in the labeling with UEA-1, DBA, PNA, RCA-1 and SBA was found in the lungs and an increase in the labeling with UEA-1, ConA, PNA, DBA was found in the placentas. The present lectin histochemical study revealed a distinctive pattern of oligosaccharide distribution in the lungs and placenta of B. abortus-infected fetuses.
RESUMEN
The flagellated protist Tritrichomonas foetus is a parasite that causes bovine trichomonosis, a major sexually transmitted disease in cattle. Cell division has been described as a key player in controlling cell survival in other cells, including parasites but there is no information on the regulation of this process in T. foetus. The regulation of cytokinetic abscission, the final stage of cell division, is mediated by members of the ESCRT (endosomal sorting complex required for transport) machinery. VPS32 is a subunit within the ESCRTIII complex and here, we report that TfVPS32 is localized on cytoplasmic vesicles and a redistribution of the protein to the midbody is observed during the cellular division. In concordance with its localization, deletion of TfVPS32 C-terminal alpha helices (α5 helix and/or α4-5 helix) leads to abnormal T. foetus growth, an increase in the percentage of multinucleated parasites and cell cycle arrest at G2/M phase. Together, these results indicate a role of this protein in controlling normal cell division.
Asunto(s)
División Celular/genética , Proteínas Protozoarias/genética , Tritrichomonas foetus/fisiología , Citocinesis/genética , Proteínas Protozoarias/metabolismo , Tritrichomonas foetus/genéticaRESUMEN
The placenta is a highly vascularized organ, indispensable tothe transfer of nutrients to the growing fetuses. During gestation, there exists an expansion of the placental vascular network through active angiogenesis. The aim of this research was to study cell proliferation and apoptosis through high resolution light microscopy (HRLM) and transmission electron microscopy (TEM) ultrastructure, immunohistochemistry for Ki67and caspase-3, determination of placental vascular area,and TUNEL assay. Crossbred sows placental tissues from approximately 30±2(n=5), 40±2(n=5), 60±2 (n=5), 80±2(n=5), 90±2(n=5) and 114±2(n=5) days of gestation were used. The evaluation of cell proliferation showed the highest%Ki67 values on days 30±2 and 80±2 of pregnancy. Caspase-3 expressed the highest value on day 30±2, while the highest apoptotic indexes were found on days30±2 and 90±2. The placental vascular area was higher on day 80±2 of pregnancy. According to our results, an active vascular cell remodeling by a caspase-3 dependent apoptosis seems to be present in early pregnancy. The increase in the vascular area on day 80±2 would be the result of the intense vascular cell proliferation detected with Ki67. Further studies are needed to understand the complex processes of angiogenesis, cell proliferation and apoptosis that interact in the placenta during porcine gestation.
Asunto(s)
Apoptosis/fisiología , Proliferación Celular/fisiología , Placenta/irrigación sanguínea , Placentación/fisiología , Porcinos/fisiología , Animales , Femenino , Neovascularización Fisiológica/fisiología , Placenta/citología , Placenta/ultraestructura , EmbarazoRESUMEN
To test the hypothesis that in domestic cats, postnatal androgens induce sterility, the aims of this study were to describe the reproductive effects and the clinical safety of a postnatal administration of a long term release androgen in this species. Thirteen newborn littermate female kittens were randomly assigned to one of the following treatment groups within the first 24h of birth: testosterone enanthate 12.5mg sc (TE; n=8) or Placebo (PL; n=5). The animals were subsequently assessed for fecal sexual hormones until puberty was attained and subsequently when matings occurred. After 21 days, ovulation and gestation were diagnosed. All queens were subsequently ovario-hysterectomized. Fecal testosterone concentrations differed between the treatment groups throughout the study period (P<0.05) being greater during the first 2 postnatal weeks in those of the TE group (P<0.01). Fecal estradiol was not affected by treatment (P>0.1). While all the females were receptive during the pubertal estrus (P>0.1), two TE (2/8) compared with all (5/5) females of the PL group had ovulations (P<0.05). Only one (1/2) compared with three (3/5) of the queens of the TE and PL groups, respectively became pregnant. All kittens of the TE group had transient clitoral enlargement. Anovulatory TE-treated cats had no corpus luteum, and a significant diminution of the endometrial glands as well as of the height of the uterine epithelium. It is concluded that, in domestic cats, a single postnatal supra-physiological dose of testosterone caused a large proportion of queens to be anovulatory and there were also histological endometrial abnormalities that also occurred with this treatment that were accompanied by mild and transient side effects.
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Ovario/efectos de los fármacos , Maduración Sexual/efectos de los fármacos , Testosterona/análogos & derivados , Andrógenos , Animales , Animales Recién Nacidos , Gatos , Preparaciones de Acción Retardada , Esquema de Medicación , Heces/química , Femenino , Tamaño de los Órganos , Ovario/crecimiento & desarrollo , Embarazo , Testosterona/administración & dosificación , Testosterona/farmacologíaRESUMEN
AbstractThe Neotropical catfish Corydoras paleatus is a facultative air-breather and the caudal half of the intestine is involved in gas exchange. In South America, air-breathing fishes are found in tropical or sub-tropical freshwaters where the probability of hypoxia is high. The aim of this study was to characterize by traditional histochemical and lectinhistochemical methods the pattern of carbohydrate in the intestinal mucosa. Intestine samples were taken from 25 healthy adult specimens collected in Buenos Aires (Argentina). Samples were fixed by immersion in 10 % buffered formalin and routinely processed and embedded in paraffin wax. Subsequently, these sections were incubated in the biotinylated lectins battery. Labeled Streptavidin-Biotin (LSAB) system was used for detection, diaminobenzidine as chromogen and haematoxylin as a contrast. To locate and distinguish glycoconjugates (GCs) of the globet cells, we used the following histochemical methods: PAS; PAS*S; KOH/ PA*S; PA/Bh/KOH/PAS; KOH/PA*/Bh/PAS; Alcian Blue and Toluidine Blue at different pHs. Microscopically, the general structure of vertebrate intestine was observed and showed all the cell types characteristic of the intestinal epithelium. The cranial sector of catfish intestine is a site of digestion and absorption and its structure is similar to other fish groups. In contrast, enterocytes of the caudal portion are low cuboidal cells; and between these, globet cells and capillaries are observed, these latter may reach the mucosal lumen. Underlying the epithelium, observed a well-developed lamina propria-submucosa made of connective tissue; this layer was highly vascularized and did not exhibit glands. According to histochemistry, the diverse GCs elaborated and secreted in the intestine are associated with specific functions in relation to their physiological significance, with special reference to their role in lubrication, buffering effect and prevention of proteolytic damage to the epithelium together with other biological processes, such as osmoregulation and ion exchange. The lectinhistochemical analysis of the intestinal mucosa reveals the presence of terminal residues of glucose, mannose and galactose. In conclusion, this study has shown that GCs synthesized in the intestine of C. paleatus exhibit a high level of histochemical complexity and that the lectin binding pattern of the intestinal mucosa is characteristic of each species and the variations are related with the multiple functions performed by the mucus in the digestive tract. The information generated here may be a relevant biological tool for comparing and analyzing the possible glycosidic changes in the intestinal mucus under different conditions, such as changes in diet or different pathological stages. Rev. Biol. Trop. 64 (1): 327-340. Epub 2016 March 01.
ResumenEl pez neotropical Corydoras paleatus, de respiración aérea de tipo facultativa, utiliza el sector caudal del intestino para el intercambio gaseoso. En América del Sur, los peces con respiración aérea se encuentran en las aguas dulceacuícolas tropicales y subtropicales, donde la probabilidad de hipoxia es alta. El objetivo de este trabajo fue caracterizar mediante técnicas histoquímicas tradicionales y de lectinhistoquímica el patrón de carbohidratos de la mucosa intestinal. Para ello se utilizaron muestras de intestino de 25 ejemplares sanos adultos recolectados en la provincia de Buenos Aires (Argentina). Las muestras fueron fijadas en formol amortiguado al 10 % y se procesaron para su inclusión en parafina. Posteriormente, los cortes fueron incubados con una batería de lectinas biotiniladas. Se utilizó el sistema de marcado con estreptavidina-biotina (LSAB) para su detección, diaminobencidina como cromógeno y hematoxilina como colorante de contraste. Para localizar y diferenciar los glicoconjugados (GCs) de las células caliciformes, se utilizaron las siguientes técnicas histoquímicas: PAS, PAS*S, PAPS, KOH/PA*S, PA/Bh/KOH/PAS, KOH/PA*/Bh/PAS, Azul Alcian y Azul de Toluidina a diferentes pHs. Microscópicamente, se observa la estructura general del intestino de los vertebrados y el epitelio intestinal presenta todos los tipos celulares característicos de esta región. El sector craneal del intestino de este teleósteo, es el sitio de digestión y absorción, y posee una estructura similar a la de otros grupos de peces. En cambio, los enterocitos de la porción caudal, son células cúbicas bajas, entre ellos se observan células caliciformes y capilares sanguíneas que llegan hasta el lumen de la mucosa. Por fuera del epitelio, se observa una lámina propia-submucosa muy desarrollada compuesta por tejido conectivo, altamente vascularizada que no presenta glándulas. De acuerdo con las técnicas histoquímicas, los diversos GCs elaborados y secretados por la mucosa intestinal se encuentran asociados con funciones específicas de importancia fisiológica, como su rol en la lubricación, su efecto amortiguador y la prevención de daños proteolíticos del epitelio junto con otros procesos biológicos, tales como la osmorregulación y el intercambio iónico. El análisis lectinhistoquímico de la mucosa intestinal revela la presencia de residuos terminales de glucosa, manosa y galactosa. En conclusión, en este estudio se demuestra que los GCs sintetizados en el intestino de C. paleatus muestran un alto nivel de complejidad histoquímica y que el patrón de unión de lectina de la mucosa intestinal es característico para cada especie y las variaciones se hallan relacionadas con las múltiples funciones realizadas por el mucus en el tracto digestivo. La información brindada en este trabajo es una herramienta de relevancia biológica para comparar y analizar los posibles cambios glicosídicos del mucus intestinal bajo diferentes condiciones como los cambios en la dieta o diferentes estados patológicos.
Asunto(s)
Animales , Masculino , Femenino , Bagres/clasificación , Glicoconjugados/análisis , Intestinos/química , Histocitoquímica , Mucosa Intestinal/citología , Mucosa Intestinal/química , Intestinos/citologíaRESUMEN
The Neotropical catfish, Corydoras paleatus (Callichthyidae) is a facultative air-breathing teleost that makes use of the caudal portion of the intestine as an accessory air-breathing organ. This portion is highly modified, being well vascularized with capillaries between epithelial cells, which makes it well suited for gas exchange. Instead, the cranial portion is a digestion and absorption site, as it has a typical intestinal epithelium with columnar cells arranged in a single row, villi and less vascularized tunica mucosa. Therefore, the intestine was studied by light and electron microscopy to assess differences between the cranial, middle and caudal portions. To characterize the potential for cell proliferation of this organ, we used anti-proliferating cell nuclear antigen antibody and anti-Na(+) K(+) -ATPase monoclonal antibody to detect the presence of Na(+) /K(+) pump. In C. paleatus it was observed that cell dynamics showed a decreasing gradient of proliferation in cranio-caudal direction. Also, the intestine of this catfish is an important organ in ionoregulation: the basolateral Na(+) /K(+) pump may have an active role, transporting Na(+) out of the cell while helping to maintain the repose potential and to regulate cellular volume.
Asunto(s)
Bagres/fisiología , Mucosa Intestinal , Estructuras Animales/química , Estructuras Animales/citología , Estructuras Animales/fisiología , Estructuras Animales/ultraestructura , Animales , Femenino , Inmunohistoquímica , Mucosa Intestinal/química , Mucosa Intestinal/citología , Mucosa Intestinal/fisiología , Mucosa Intestinal/ultraestructura , Masculino , Antígeno Nuclear de Célula en Proliferación/análisis , Antígeno Nuclear de Célula en Proliferación/metabolismo , ATPasa Intercambiadora de Sodio-Potasio/análisis , ATPasa Intercambiadora de Sodio-Potasio/metabolismoRESUMEN
The Neotropical catfish Corydoras paleatus is a facultative air-breather and the caudal half of the intestine is involved in gas exchange. In South America, air-breathing fishes are found in tropical or sub-tropical freshwaters where the probability of hypoxia is high. The aim of this study was to characterize by traditional histochemical and lectinhistochemical methods the pattern of carbohydrate in the intestinal mucosa. Intestine samples were taken from 25 healthy adult specimens collected in Buenos Aires (Argentina). Samples were fixed by immersion in 10 % buffered formalin and routinely processed and embedded in paraffin wax. Subsequently, these sections were incubated in the biotinylated lectins battery. Labeled Streptavidin-Biotin (LSAB) system was used for detection, diaminobenzidine as chromogen and haematoxylin as a contrast. To locate and distinguish glycoconjugates (GCs) of the globet cells, we used the following histochemical methods: PAS; PAS*S; KOH/ PA*S; PA/Bh/KOH/PAS; KOH/PA*/Bh/PAS; Alcian Blue and Toluidine Blue at different pHs. Microscopically, the general structure of vertebrate intestine was observed and showed all the cell types characteristic of the intestinal epithelium. The cranial sector of catfish intestine is a site of digestion and absorption and its structure is similar to other fish groups. In contrast, enterocytes of the caudal portion are low cuboidal cells; and between these, globet cells and capillaries are observed, these latter may reach the mucosal lumen. Underlying the epithelium, observed a well-developed lamina propria-submucosa made of connective tissue; this layer was highly vascularized and did not exhibit glands. According to histochemistry, the diverse GCs elaborated and secreted in the intestine are associated with specific functions in relation to their physiological significance, with special reference to their role in lubrication, buffering effect and prevention of proteolytic damage to the epithelium together with other biological processes, such as osmoregulation and ion exchange. The lectinhistochemical analysis of the intestinal mucosa reveals the presence of terminal residues of glucose, mannose and galactose. In conclusion, this study has shown that GCs synthesized in the intestine of C. paleatus exhibit a high level of histochemical complexity and that the lectin binding pattern of the intestinal mucosa is characteristic of each species and the variations are related with the multiple functions performed by the mucus in the digestive tract. The information generated here may be a relevant biological tool for comparing and analyzing the possible glycosidic changes in the intestinal mucus under different conditions, such as changes in diet or different pathological stages.