Assessing the ovarian development of Macrobrachium amazonicum (Heller, 1862) phenotypes by means of an integrative analysis.

Macrobrachium amazonicum is a species of economic interest with a wide distribution in the Americas and high morphological and reproductive variability. Three phenotypes can be observed in this species: i) large-size amphidromous, ii) large-size and iii) small-size hololimnetic prawns. In the present work, the morphological, histochemical and ultrastructural aspects of ovarian development in the three phenotypes were comparatively analyzed. In addition, the interaction between the ovary and the hepatopancreas was investigated in these phenotypes through the use of gonadosomatic (GSI) and hepatosomatic (HSI) indices. Despite the morphological differences and different reproductive strategies adopted by the females, the macroscopic, histochemical and ultrastructural patterns of ovarian development showed no differences between the phenotypes. The ovaries were macroscopically classified into five stages of development (I to V). In early stages (I and II), the ovaries are full of oogonia, previtellogenic oocytes and oocytes in primary or endogenous vitellogenesis. At these stages, the rough endoplasmic reticulum (RER) produces a granular electron-dense material and sends it to the Golgi apparatus, where it will be modified, compacted and transformed into immature yolk granules. From stage III, secondary or exogenous vitellogenesis begins (with no interruption of endogenous vitellogenesis), where follicular cells nourish the oocytes and extracellular material is absorbed by endocytic vesicles, which fuse with immature yolk granules (forming mature granules) or with existing mature yolk granules. In stages IV and V, secondary vitellogenesis continues and mature yolk granules progressively occupy the cytoplasm. In M. amazonicum, the patterns of increase in oocyte diameter are quite similar between phenotypes, being greater in the small-size phenotype. This is related to the formation of larger oocytes/eggs and the production of large lipid reserves for their larvae. Changes in GSI and HSI during ovarian development show strong similarity between phenotypes, supporting the results obtained by histology and ultrastructure. Females in stages III and IV mobilize hepatopancreas reserves for ovarian maturation, which justifies the higher HSI values recorded in these stages. On the other hand, females in stage V show higher GSI and lower HSI values, indicating a mobilization of resources for the end of ovarian development as the females are ready to spawn.

Is there a trade-off between sperm production and sexual weaponry in the Amazon River prawn Macrobrachium amazonicum (Heller, 1862)?

The Amazon River prawn Macrobrachium amazonicum shows populations with four well-defined morphotypes in males. Dominant males of morphotypes green claw 1 (GC1) and green claw 2 (GC2) have large bodies and chelipeds and a higher reproductive success in comparison with the submissive morphotypes - translucent claw (TC) and cinnamon claw (CC). However, recently, some populations of the species do not have dominant morphotypes. Here, we compared the patterns of spermatic production and concentration among morphotypes and populations with three different phenotypes: (i) large-size amphidromous prawns, and (ii) large-size ("i" and "ii" with dominant morphotypes) and (iii) small-size hololimnetic prawns (without morphotypes). We described the spermatogenesis and the histochemical features of vasa deferentia (VD) and evaluated the relationship between the investment in spermatic production and sexual weapons acquisition in males of different phenotypes. The spermatic production and concentration in populations with four morphotypes were similar between morphotypes. The exception was the CC morphotype in which males had the seminiferous tubules filled with spermatocytes and low spermatic concentration. The spermatogenesis, spermiogenesis, and VD structure were not different among the studied phenotypes and populations. The seminal fluid of M. amazonicum is comprised by glycoproteins and by concentric layers of secretions of types I, II (basophilic), and III (eosinophilic). We could infer that males of dominant morphotypes allocate a higher amount of energy to the development of strong sexual weapons at the expense of the energy allocated to the reproductive system during the sequential growth. Inversely, small-size males direct more energy toward the spermatic production and transference at the expense of sexual weapons. Therefore, there is a clear trade-off between the investment in the gonadal and sexual weapons development in males of M. amazonicum.

The androgenic gland in male morphotypes of the Amazon River prawn Macrobrachium amazonicum (Heller, 1862).

Sexual differentiation and primary and secondary sexual characteristics in male crustaceans are modulated by hormones produced in the androgenic gland (AG). The AG is also responsible for the determination of morphotypes in caridean shrimps, such as Macrobrachium amazonicum that shows four morphotypes: translucent claw (TC), cinnamon claw (CC), green claw 1 (GC1) and green claw 2 (GC2). Here, we verified the anatomical, histological and ultrastructural characteristics of the AG in different morphotypes of this species with both amphidromous and hololimnetic life cycles. In submissive morphotypes (TC and CC), the AGs are reduced and concentrated in the terminal expansion of the distal portion of vasa deferentia (DVD), the ejaculatory ducts (ED). In dominant morphotypes (GC1 and GC2) these glands lie along the DVD and ED. Two morphological stages (I and II) were recorded for AG cells. In submissive morphotypes stage I cells predominated in the AGs, while in dominant morphotypes stage II cells were more common. AG cells in both stages were positive for proteins, confirming the protein nature of the secreted hormone. Stage I cells have abundant rough endoplasmic reticulum (RER) with numerous parallel cisternae, whereas in stage II cells, the cisternae of RER are highly dilated. Stage II cells do not produce secretory granules, but they undergo hypertrophy and the hormone release to hemolymph probably occurs by holocrine secretion. The AGs in TC, GC1 and GC2 morphotypes increase as the animals grow and are larger in GC1 males. On the other hand, AGs decrease in the CC morphotype as the animal grows. These differences are related to the type of reproductive strategy adopted by each morphotype. In M. amazonicum, the AGs show the same morphological, histochemical and ultrastructural patterns between the different life history populations.