Resumo
Despite the great importance of the siphons for infaunal bivalves, only a few studies have examined their tissues using histology techniques or scanning electron microscopy. In the present study, the siphons of Tellina lineata Turton, 1819 and Macoma biota Arruda & Domaneschi, 2005 were investigated. The siphon walls are composed by a series of muscle sheets of longitudinal ("L"), circular ("C") and radial ("R") fibers, with a clear pattern common to both species: there is a main median longitudinal layer (Lm), and two peripheral circular layers, one inner (Ci) and one outer (Co), near the epithelia. A median circular layer (Cm) separates an internal (Lmi) from an outer (Lmo) median longitudinal layer. Further, the Co is split by a thin outer longitudinal layer (Lo), forming Coi and Coo layers, the former being obliquely oriented. Thin radial fibers (R) delimit clear packages of Lmi and Lmo fibers. In each siphon, there are six longitudinal nerve cords, running within the Lmi layer, adjacent to the Cm. The inhalant and exhalant siphons of M. biota are very similar in structure, but the Lmo of the exhalant siphon is almost twice as thick as its Lmi, while in the inhalant siphon these layers have similar thicknesses; the Coi is very thick, especially in the exhalant siphon. The inhalant siphon of T. lineata is very similar to that of M. biota, differing only with respect to the thickness of the Coi, which in the former species is not as well developed as in the latter. The Lmo of the exhalant siphon of T. lineata is by far the most developed layer, with the Lmi represented only by uniseriate small cells; in the vicinities of the nerve cords, the Cm is split in two layers. The epithelia of both siphons of M. biota and T. lineata bear ciliated receptors, which were difficult to observe as they were frequently covered with mucus. It was possible to observe that cilia are present in both species, differing in length and in the number per receptor between the inhalant and exhalant siphons, and between the species. Detailed comparisons among the siphons of M. biota and T. lineata and other bivalve species are very difficult, because of at least two reasons. First, each investigator has used different methods to prepare and observe the siphons through histological sections; and second, different nomenclatural schemes are used to describe the musculature of the siphons, causing confusion when the same layers are compared among different species. In order to unify the nomenclature of tissue layers of the bivalve siphons, we now propose a scheme to name these layers based on topological homology.
Resumo
Despite the great importance of the siphons for infaunal bivalves, only a few studies have examined their tissues using histology techniques or scanning electron microscopy. In the present study, the siphons of Tellina lineata Turton, 1819 and Macoma biota Arruda & Domaneschi, 2005 were investigated. The siphon walls are composed by a series of muscle sheets of longitudinal ("L"), circular ("C") and radial ("R") fibers, with a clear pattern common to both species: there is a main median longitudinal layer (Lm), and two peripheral circular layers, one inner (Ci) and one outer (Co), near the epithelia. A median circular layer (Cm) separates an internal (Lmi) from an outer (Lmo) median longitudinal layer. Further, the Co is split by a thin outer longitudinal layer (Lo), forming Coi and Coo layers, the former being obliquely oriented. Thin radial fibers (R) delimit clear packages of Lmi and Lmo fibers. In each siphon, there are six longitudinal nerve cords, running within the Lmi layer, adjacent to the Cm. The inhalant and exhalant siphons of M. biota are very similar in structure, but the Lmo of the exhalant siphon is almost twice as thick as its Lmi, while in the inhalant siphon these layers have similar thicknesses; the Coi is very thick, especially in the exhalant siphon. The inhalant siphon of T. lineata is very similar to that of M. biota, differing only with respect to the thickness of the Coi, which in the former species is not as well developed as in the latter. The Lmo of the exhalant siphon of T. lineata is by far the most developed layer, with the Lmi represented only by uniseriate small cells; in the vicinities of the nerve cords, the Cm is split in two layers. The epithelia of both siphons of M. biota and T. lineata bear ciliated receptors, which were difficult to observe as they were frequently covered with mucus. It was possible to observe that cilia are present in both species, differing in length and in the number per receptor between the inhalant and exhalant siphons, and between the species. Detailed comparisons among the siphons of M. biota and T. lineata and other bivalve species are very difficult, because of at least two reasons. First, each investigator has used different methods to prepare and observe the siphons through histological sections; and second, different nomenclatural schemes are used to describe the musculature of the siphons, causing confusion when the same layers are compared among different species. In order to unify the nomenclature of tissue layers of the bivalve siphons, we now propose a scheme to name these layers based on topological homology.
Resumo
Despite the great importance of the siphons for infaunal bivalves, only a few studies have examined their tissues using histology techniques or scanning electron microscopy. In the present study, the siphons of Tellina lineata Turton, 1819 and Macoma biota Arruda & Domaneschi, 2005 were investigated. The siphon walls are composed by a series of muscle sheets of longitudinal ("L"), circular ("C") and radial ("R") fibers, with a clear pattern common to both species: there is a main median longitudinal layer (Lm), and two peripheral circular layers, one inner (Ci) and one outer (Co), near the epithelia. A median circular layer (Cm) separates an internal (Lmi) from an outer (Lmo) median longitudinal layer. Further, the Co is split by a thin outer longitudinal layer (Lo), forming Coi and Coo layers, the former being obliquely oriented. Thin radial fibers (R) delimit clear packages of Lmi and Lmo fibers. In each siphon, there are six longitudinal nerve cords, running within the Lmi layer, adjacent to the Cm. The inhalant and exhalant siphons of M. biota are very similar in structure, but the Lmo of the exhalant siphon is almost twice as thick as its Lmi, while in the inhalant siphon these layers have similar thicknesses; the Coi is very thick, especially in the exhalant siphon. The inhalant siphon of T. lineata is very similar to that of M. biota, differing only with respect to the thickness of the Coi, which in the former species is not as well developed as in the latter. The Lmo of the exhalant siphon of T. lineata is by far the most developed layer, with the Lmi represented only by uniseriate small cells; in the vicinities of the nerve cords, the Cm is split in two layers. The epithelia of both siphons of M. biota and T. lineata bear ciliated receptors, which were difficult to observe as they were frequently covered with mucus. It was possible to observe that cilia are present in both species, differing in length and in the number per receptor between the inhalant and exhalant siphons, and between the species. Detailed comparisons among the siphons of M. biota and T. lineata and other bivalve species are very difficult, because of at least two reasons. First, each investigator has used different methods to prepare and observe the siphons through histological sections; and second, different nomenclatural schemes are used to describe the musculature of the siphons, causing confusion when the same layers are compared among different species. In order to unify the nomenclature of tissue layers of the bivalve siphons, we now propose a scheme to name these layers based on topological homology.
Resumo
O objetivo deste trabalho foi verificar a presença e a localização de receptores para hormônios esteróides e gonadotróficos, através da técnica de imunohistoquímica, pelo método de peroxidase-antiperoxidase (PAP), nos diferentes tecidos que compõe o trato genital da égua e a variação de reatividade destes receptores durante o ciclo estral e no anestro fisiológico. Também se objetivou verificar se há diferença de reatividade em éguas com e sem endometrose. Foram coletadas amostras de útero, cérvice e oviduto, de 41 éguas sem raça definida e com histórico reprodutivo desconhecido, em um abatedouro. Quinze éguas se encontravam em estro, dezoito em diestro e oito éguas em anestro. Concluiu-se que a intensidade e a distribuição da coloração para os receptores de estrógeno (RE), progesterona (RP) e hormônio luteinizante (RLH) variaram de acordo com o tipo de célula e o estágio do ciclo estral. Nas amostras de endométrio observou-se imunorreatividade alta no epitélio luminal para RE e RP tanto no estro quanto no diestro; o epitélio glandular, estroma e miométrio mostraram reatividade moderada para os dois receptores durante as duas fases. Durante o anestro os resultados foram semelhantes aos encontrados durante a fase cíclica. Na avaliação da reatividade para RLH, durante o estro e diestro, o epitélio luminal mostrou reatividade de fraca a moderada, mas no diestro houve maior reatividade média. O epitélio glandular apresentou menor reatividade do que o luminal. No miométrio a coloração foi fraca durante todo o ciclo. Durante o anestro a reatividade foi fraca no epitélio luminal, ausente em quase todas as amostras no epitélio glandular e de fraca a ausente no miométrio. Neste experimento, não foi observada diferença significativa de reatividade entre os endométrios com e sem endometrose, mas as áreas afetadas mostraram coloração assíncrona para RE, RP e RLH. Na cérvice, foi observada imunorreatividade moderada a alta para RE e RP no epitélio luminal, no estroma e no músculo. A intensidade de coloração das células epiteliais e musculares variou pouco entre o estro e o diestro, mas durante o anestro houve maior reatividade no tecido muscular e no estroma. Foi observada reatividade para RLH no epitélio e camada muscular, sem variação significativa nas fases do ciclo. A intensidade de coloração foi de fraca a moderada no epitélio e fraca na camada muscular. No oviduto, observou-se imunorreatividade para RE e RP nos três tecidos, durante a fase cíclica e o anestro. No epitélio, os valores encontrados foram de moderados a altos, sem variação significativa nas três fases. A coloração das células epiteliais do oviduto foi nitidamente irregular, com o núcleo muito corado no que parecem ser células secretoras e pouco corado ou sem coloração nas células ciliadas, refletindo provavelmente as diferentes funções das células epiteliais neste órgão. No estroma a reatividade foi moderada durante a fase luteal, mostrando reatividade mais alta no estro e no anestro. A camada muscular apresentou reatividade máxima para RE no estro e no diestro. A reatividade para RLH no epitélio luminal foi de fraca a moderada durante todo o ciclo. No músculo também foi observada reatividade, porém bem mais fraca do que no epitélio. Durante o anestro somente três das oito amostras apresentaram reatividade no tecido muscular. No diestro foi observada maior reatividade do que no estro. Os resultados do presente estudo evidenciam, pela primeira vez, a presença de receptores para LH nos diferentes tecidos do trato reprodutor extragonadal da égua. Embora existam relatos da expressão e localização de RE e RP no endométrio equino, esta é a primeira vez que se utiliza a técnica de imunohistoquímica para localizar estes receptores na cérvice e no oviduto desta espécie. Foi observada variação individual bastante acentuada entre as amostras, em uma mesma fase cíclica. Provavelmente estes resultados sejam o reflexo da variação entre o dia do ciclo em que os animais se encontravam, bem como da complexidade dos mecanismos envolvidos na presença desses receptores. Os achados deste estudo indicam que tanto os hormônios gonadais quanto o LH atuam por meio de seus receptores nos diferentes tecidos do trato reprodutivo da égua, podendo servir para a elaboração de novas estratégias para melhorar a eficiência reprodutiva nesta espécie
The aim of this study was to demonstrate the presence and localization of gonadotropic and steroid hormone receptors, in different tissues of the mare genital tract and the different reactivity to these receptors during the endometrial cycle and physiologic anestrus. Another objective was to compare the reactivity to theses receptors in mares with and without endometrosis. Immunohistochemistry was performed using the peroxidase anti-peroxidase technique (PAP). Uterus, cervix and oviduct of 41 criollo mares were collected in an abattoir. There was variation in the intensity of the staining and distribution for estrogen receptors (PR), progesterone receptors (PR) and luteinizing hormone receptors (LHR) with the endometrial cycle and different tissues. The endometrial surface epithelial cells were stained strongly for ER and PR in the estrous and dioestrus; glandular epithelial cells, stromal cells and smooth muscle cells of the myometrium had moderate staining for ER and PR during these two phases and in anestrus too. The immunoreactive score for LHR in the surface epithelial cells during endometrial cycle was weak to moderate but, in general, strong staining was observed in dioestrus. More weak staining intensity was observed in the glandular epithelial cells than luminal epithelial cells. Smooth muscle cells of the myometrium showed weak staining for LHR throughout the endometrial cycle. During the anestrus, the immunoreactivity score was weak in the surface epithelial cells. In general, the glandular epithelium was not stained. Myometrium cells were weak to not staining for LHR, in this phase. In this study there was no significant difference in immunoreactive score for ER, PR and LHR in endometrium with or without endometrosis but fibrotic glands showed different expression patterns of ER, PR and LHR, could evidence for functionally glandular maldifferentiation in endometrosis. The cervical epithelial surface, stromal cells and smooth muscle cells were moderate to strongly staining for ER and PR, with little variation throughout the endometrial cycle but the immunorectivity was strongest during the anestrus in muscular and stromal cells. Surface epithelial cells of cervix were weak to moderate stained for LHR; smooth muscle cells showed weak staining for these receptors. There was no variation during cycle. In the oviduct, epithelial, stromal and muscle cells showed reactivity for RE and RP, during cycle and anestrus. Epithelial cells were moderate to strongly staining for these receptors, with evident irregularity in different types of cells. Apparently ciliated epithelial cells were stained but the intensity was much less than that observed in nonciliated epithelial cells, probably reflecting different functions of these cells. Stromal cells showed moderate staining during dioestrus and strongest reactivity in estrous and anestrus; muscle cells showed strong reactivity for ER throughout the cycle. The reactivity for LHR was weak to moderate throughout the cycle in the epithelial cells and weak in the muscle cells. During anestrus only three strains of muscle cells showed reactivity for LHR. In dioestrus the intensity was strongest. These findings evidence for the firs time the presence for LHR in extra-gonadal reproductive organs of mare. Though there were reports of ER and PR expression in equine endometrium, this is the first report of localization of these receptors in cervix and oviduct of mare using immunohistochemistry. It was found marked individual variation among the strains. These results probably were caused by the variation among the day of cycle and the complexity of mechanisms involved in the presence of these receptors. The findings of the present study allow us to infer that the ovarian steroid hormones nad LH function through their receptors in different tissues of mare reproductive tract, can help us to elaborate new strategies to improve the reproductive efficiency in this specie