RESUMEN
Hemocytes are the first line of defense of the immune system in invertebrates, but despite their important role and enormous potential for the study of gene-environment relationships, research has been impeded by a lack of consensus on their classification. Here we used flow cytometry combined with histological procedures, histochemical reactions and transmission electron microscopy to characterize the hemocytes from the oyster Crassostrea rhizophorae. Transmission electron microscopy revealed remarkable morphological characteristics, such as the presence of membranous cisternae in all mature cells, regardless of size and granulation. Some granular cells contained many cytoplasmic granules that communicated with each other through a network of channels, a feature never previously described for hemocytes. The positive reactions for esterase and acid phosphatase also indicated the presence of mature cells of all sizes and granule contents. Flow cytometry revealed a clear separation in complexity between agranular and granular populations, which could not be differentiated by size, with cells ranging from 2.5 to 25 µm. Based on this evidence we suggest that, at least in C. rhizophorae, the different subpopulations of hemocytes may in reality be different stages of one type of cell, which accumulates granules and loses complexity (with no reduction in size) as it degranulates in the event of an environmental challenge.
Asunto(s)
Hemocitos/citología , Ostreidae/citología , Animales , Citometría de Flujo , Microscopía Electrónica de TransmisiónRESUMEN
Crassostrea rhizophorae (Guilding, 1828) es una de las especies de ostras tropicales cultivadas en la costa brasilera, que presenta alto valor comercial, no existiendo información sobre las características gonadales, durante las fases reproductivas de esta especie. El objetivo de este trabajo fue analizar la morfología de los ovocitos y de los folículos gonadales de Crassostrea rhizophorae, en tres fases del ciclo gonadal. Analizando los cortes histológicos de las gónadas y considerando los ovocitos que presentaban núcleo y nucléolo evidentes, fueron determinadas tres fases del ciclo gonadal: gametogénesis inicial, crecimiento y maduración. En la fase de gametogénesis inicial, el diámetro de los folículos y de los ovocitos era de 180,29 (+ - 41,91) e 18,66 (+ - 6,85) µm, respectivamente; también identificamos gran cantidad de tejido conjuntivo interfolicular y un número mayor de ovocitos previtelogénicos. En la fase de crecimiento, el diámetro de los folículos y de los ovocitos era de 218,02 (+ - 43,19) y 25,92 (+ - 9,94) µm, respectivamente. Este fase fue caracterizada por una pequeña cantidad de tejido conjuntivo interfolicular y predominio de ovocitos vitelogénicos. En la fase de maduración, el diámetro de los folículos y de los ovocitos era de 298,16 ( + - 99,24) y 35,27 (+ - 6,2) µm, respectivamente, existiendo gran número de ovocitos maduros. De esta manera, concluimos que durante la implantación del cultivo, Crassostrea rhizophorae tolera la influencia de los factores intrínsecos y extrínsecos y no presenta alteraciones significativas en su actividad reproductiva.
Crassostrea rhizophorae (Guilding, 1828) is one of the tropical species of oysters cultivated on the Brazilian shore. Despite its high commercial value, there is no information on the gonadal characteristics during the reproductive stages of this species. The objective of this work was to analyze the morphology and morphometry of Crassostrea rhizophorae oocytes and follicles in three stages of the gonadal cycle. Were analized histological sections of gonads considering that the oocytes presented visible nuclei and nucleoli, it was observed three gonadal cycle stages: early gametogenesis, growth and maturation. In the early gametogenesis stage, follicles and oocytes presented diameters of 180.29 (± 41.91) and 18.66 (± 6.85) µm, respectively, and presence of large amount of connective tissue and previtellogenic oocyte. In the growth stage, follicles and oocytes presented diameters of 218.02 (± 43.19) and 25.92 (± 9.94) µm, respectively, this stage was characterized by a small amount of interfollicular connective tissue with vitellogenic oocyte predominate. In the maturation stage, follicles and oocytes presented diameters of 298.16 (± 99.24) and 35.27 (± 6.2) µm, respectively, and presence of large number of mature oocytes. We concluded that during culture Crassostrea rhizophorae tolerates the influence of intrinsic and extrinsic factors and does not undergo significant changes in reproductive activity.