-This study describes the histology and histochemistry of the male reproductive system in Callinectes ornatus, comparing juvenile and adult developmental stages. We also analyzed changes in the gonadosomatic (GSI) and hepatosomatic (HSI) indices, and the weights of the testis and vas deferens during the development. The results showed that all stages, beginning with the juvenile (JUV), through developing (DEV) and mature (MAT) adult males of C. ornatus produce sperm and spermatophores. During development, testicular lobes showed the same characteristics of production and release of sperm into the seminiferous duct. The vas deferens showed little histological and histochemical change in the epithelium in juvenile and adult males. The differences consisted of the larger amount of secretion in MAT males compared to JUV and DEV ones. The chemical composition of the seminal fluid was similar, but MAT males produced a more homogeneous secretion. Morphological and physiological maturation are not synchronized in C. ornatus, since JUV males produced spermatophores similar to those in DEV and MAT males. However, these JUV are not yet able to reproduce, since they still have the abdomen attached to the cephalothoracic sternum. The increase of the GSI during development was significant for MAT males, and is related to the production of sufficient volume of seminal fluid to form the sperm "plug" in the female seminal receptacle. The HSI decreased from DEV to MAT adult stages, indicating that reserves from the hepatopancreas are used to develop the reproductive system after the pubertal molt.
We described the ultrastructure and histochemistry of the reproductive system of five Callinectes species, and evaluate the seasonal variation in weight of the reproductive system and hepatopancreas by comparing annual changes of somatic indices. The somatic indices changed little throughout the year. In Callinectes, spermatogenesis occurs inside the lobular testes and, within each lobule, the cells are at the same developmental stage. Spermatogenesis and spermiogenesis follow the same development pattern in all Callinectes studied. Mature spermatozoa are released into the seminiferous ducts through the collecting ducts. Cells of the vas deferens are secretory as evidenced by rough endoplasmic reticulum, Golgi complex, and secretory vesicles that produce the seminal fluid. The anterior vas deferens shows two portions: proximal and distal. In proximal portion (AVDp), spermatozoa are clustered and embedded in an electron-dense, basophilic glycoproteinaceous secretion Type I. In the distal portion (AVDd), the spermatophore wall is formed by incorporation of a less electron-dense glycoproteinaceous secretion Type II. The secretion Type I change to an acid polysaccharide-rich matrix that separates the spermatophores from each other. The median vas deferens (MVD) stores the spermatophores and produces the granular glycoproteinaceous seminal fluid. The posterior vas deferens (PVD) has few spermatophores. Its epithelium has many mitochondria and the PVD seminal fluid changes into a liquid and homogeneous glycoprotein. Many outpocketings in the PVD and MVD help to increase the fluid production. Overall, the reproductive pattern of Callinectes is similar to other species that produce sperm plugs. The secretions of AVD, MVD, and PVD are responsible for the polymerization that forms the solid, waxy plug in the seminal receptacle. The traits identified here are common to all Portunidae species studied so far.
The ovarian development of Callinectes ornatus and Arenaeus cribrarius was described using histochemistry and ultrastructure analyses. Both species shows six stages of ovary maturation: juvenile (JUV), rudimentary (RUD), developing (DEV), intermediary (INT), mature (MAT) and spent (OV). The JUV and RUD stages showed similar histological and histochemical characteristics, in which the oocytes cytoplasm is basophilic and glycoproteic yolk nucleus is observed. The cytoplasm shows small acidophilic portions, stained for protein and reactive to PAS, which correspond to the dilated cisternae of the rough endoplasmic reticulum (RER) in ultrastructure. The exogenous phase begins at the DEV stage, with the fusion between pinocytic vesicles and vesicles of immature yolk, forming the mature yolk granules. At the INT stage, the formation of the chorion begins and the mature yolk granules increase in size and number, and the lipid and glycogen reserves are expanded, while the ER diminishes. In MAT, the oocytes are completely formed. Additionally, the cytoplasm is filled with mature yolk granules, lipids droplets and glycogen. There are no significant variations between the gonadosomatic and hepatosomatic indices, which allow us to infer the transfer of reserves from the hepatopancreas to the development of the female reproductive system in the portunids studied.
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