Reproductive cycle of Hepatus pudibundus (Herbst, 1785) (Crustacea, Decapoda, Calappidae) in Ubatuba, SP, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Testicular morphological changes during the seasonal reproductive cycle in the bullfrog Rana catesbeiana

Spermatogenesis and steroidogenesis undergo seasonal variations during the reproductive cycle in amphibians. Testicular morphological and morphometric seasonal variations as well as interstitial lipidic inclusions and intralobular glycoconjugates were evaluated during seasonal cycle of Rana catesbeiana. Testes of frogs collected during the annual seasons were weighed for calculation of GSI (Gonadosomatic index). Seminiferous lobule diameters (DSL) and volume densities of seminiferous lobules (VvSL), excretory ducts (VvED), and interstitial tissue (VvIT) were analyzed. Semithin sections were submitted to Periodic Acid-Schiff (PAS) and Alcian Blue (AB) methods for detection of glycoconjugates, while lipidic inclusions were detected by Sudan Black B. GSI showed no significant variations during the year. Since VvED and VvIT increased significantly during summer and were inversely proportional to VvSL, a compensatory effect between the testicular compartments may be related to the maintenance of GSI. During autumn/winter, larger lobular diameters were observed in comparison to spring/summer when spermiogenesis and spermiation were commonly observed. The increased VvIT and the numerous lipidic inclusions in the interstitial cells during summer suggest a relationship between spermiogenesis and steroidogenesis. Besides the structural stability variations occurring in the IT and SL, a possible paracrine interaction between ED and IT should be also involved in the IT development during summer. The presence of PAS and AB-positive globular structures were observed in the seminiferous lobules and excretory ducts. These structures containing acid glycoconjugates appear to be Sertoli cell apical portions, which are accumulated in the lumen of the seminiferous lobules mainly during spermiation. © 2004 Wiley-Liss, Inc.

Reproductive cycle, ovarian development, and vertebrate-type steroids profile in the freshwater prawn Macrobrachium rosenbergii

This study is intended to characterize the ovarian development and hemolymph vertebrate-type steroids concentration during the reproductive cycle of the freshwater prawn, Macrobrachium rosenbergii. A five-stage classification based on the external observation of the ovary's size and color as seen through the tegument was used. The results showed the existence of a direct correspondence between the ovarian stages and the gonadosomatic index, the oöcyte diameter, and the characteristics of ovarian histology. In each stage total bleeding of the prawns was conducted and the hemolymph concentrations of 17β-estradiol (E 2), testosterone (T) and 17α-hydroxyprogesterone (17-OHP; conjugated and unconjugated) were determined by solid-phase radioimmunoassay (RIA). High levels of unconjugated 17-OHP, relatively constant concentrations of unconjugated T, and null concentration of unconjugated E 2 were found throughout the five stages considered. Nonetheless, low levels of E 2 were determined in all stages, in conjugated (glucoronide) form. The highest levels were obtained in stage II and III (18.14 ± 14.52 pg/ml hemolymph) and progressively declined in the other stages (7.53 ± 6.76 pg/ml hemolymph). These results point out the possible involvement of vertebrate-type steroids in the endocrine regulation of the Macrobrachium rosenbergii's ovarian cycle. To our knowledge this is the first study concerning the vertebrate-like steroid levels throughout the ovarian cycle and is a step needed to characterize the hemolymph profile of these steroids in order to elucidate the possible role in the regulation of the reproductive crustacean cycle.

Sexual development and reproductive cycle of the Eyespot skate Atlantoraja cyclophora (Regan, 1903) (Chondrichthyes: Rajidae: Arhynchobatinae), in southeastern Brazil

Specimens o/Atlantoraja cyclophora were collected monthly from commercial fishing landings at Guarujá, São Paulo State, Brazil, from March 2005 to April 2006 at depths between 10 and 146 m. Males ranged from 13.3 to 58.5 cm TL (n = 396). Both the smallest mature male and the largest immature male were 47.0 cm long. Males' size-at-50% maturity was calculated to be 46.3 cm. Females ranged from 11.5 to 68.0 cm (n = 401). The smallest mature and the largest immature female were 51.6 and 53.0 cm long respectively. For the females, size-at-50% maturity was calculated to be 53.2 cm. In the males the hepatosomatic andgonadosomatic indices varied between 0.48 (August) and 3.54 (November) and between 0.15 (November) and 1.45 (June) respectively, with no significant variation for the fourteen-month period. In the females the hepatosomatic and gonadosomatic indices varied from 1.55 and 6.30 3.54 (both for April 2006) and from 0.08 (December) to 4.41 (October) respectively, with no significant difference among months. Egg-bearing females were found in all months with proportions varying from 0.03 (March) to 0.67 (April). Both males and females undergo an annual cycle, with slight seasonal variations in reproductive activity and a peak in the proportion of egg bearing females between April and July.

Reproductive cycle and fecundity of Potimirim glabra (Kingsley, 1954) (Caridea, Atyidae) from a littoral stream

The reproductive cycle of Potimirim glabra, especially the female size at sexual maturity, reproductive period and fecundity, was studied at two sites of one of the northern littoral streams in the state of São Paulo, Brazil. Prawns were collected monthly for one year. The minmum size at sexual maturity was estimated from the smallest ovigerous female (cephalothorax length = CL), and the reproductive period was determined monthly by examining the relative frequency of ovigerous females. Size comparisons between sampling sites was carried out using the Mann-Whitney u test. The relationship between environmental factors and reproductive aspects was verified using the Spearman's rank correlation. At Site 1, the collected ovigerous females had a CL mean size of 4.24±0.36 mm (n=481); at Site 2, ovigerous females (n=391) had a CL mean size of 5.48±0.49 mm. The reproductive pattern in Camburi is seasonal, with the highest frequency of ovigerous females in February. The frequency of ovigerous females was positively correlated to organic matter content in the substratum, rainfall and temperature. Increased percentage organic matter may indicate greater food availability whereas higher rainfall increases the velocity of the current, thereby aiding the larvae, which depend on brackish water to complete their development, to reach the estuarine region. The temperature acts on the speed of larval development. At Site 1, the mean fecundity obtained was 202±72.31 eggs (90 females; CL from 2.7 to 4.7mm PL), whereas at Site 2, it was 433.3±120.7 eggs (46 females; CL from 4.0 to 5.5mm PL). © 2010 Balaban.

Gametogenesis and reproductive cycle of Melanorivulus aff. punctatus (Boulenger, 1895) (Cyprinodontiformes, Rivulidae) in Chapada dos Guimarães, Mato Grosso, Brazil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

The reproductive cycle of the male sleep snake Sibynomorphus mikanii (Schlegel, 1837) from southeastern brazil

This study describes the male reproductive cycle of Sibynomorphus mikanii from southeastern Brazil considering macroscopic and microscopic variables. Spermatogenesis occurs during spring-summer (September-December) and spermiogenesis or maturation occurs in summer (December-February). The length and width of the kidney, the tubular diameter, and the epithelium height of the sexual segment of the kidney (SSK) are larger in summer-autumn (December-May). Histochemical reaction of the SSK [periodic acid-Schiff (PAS) and bromophenol blue (BB)] shows stronger results during summer-autumn, indicating an increase in the secretory activity of the granules. Testicular regression is observed in autumn and early winter (March-June) when a peak in the width of the ductus deferens occurs. The distal ductus deferens as well as the ampulla ductus deferentis exhibit secretory activities with positive reaction for PAS and BB. These results suggest that this secretion may nourish the spermatozoa while they are being stored in the ductus deferens. The increase in the Leydig cell nuclear diameter in association with SSK hypertrophy and the presence of sperm in the female indicate that the mating season occurs in autumn when testes begin to decrease their activity. The peak activity of Leydig cells and SSK exhibits an associated pattern with the mating season. However, spermatogenesis is dissociated of the copulation characterizing a complex reproductive cycle. At the individual level, S. mikanii males present a continuous cyclical reproductive pattern in the testes and kidneys (SSK), whereas at the populational level the reproductive pattern may be classified as seasonal semisynchronous. © 2012 Wiley Periodicals, Inc.

Female germ cell renewal during the annual reproductive cycle in Ostariophysians fish

The objective was to characterize female germ cell renewal during the annual reproductive cycle in two species of ostariophysian fish with distinct reproductive strategies: a siluriform, Pimelodus maculatus, in which oocyte development is group synchronous and the annual reproductive period is short; and a characiform, Serrasalmus maculatus, with asynchronous oocyte development and a prolonged reproductive period. These reproductive strategies result in fish determinate and indeterminate fecundity, respectively. Annual reproductive phases were determined by biometric and histologic analysis of gonads and interpreted according to new proposals for phase classification and stages of oocyte development (with special attention to germinal epithelium activity). Histologically, there were two types of oogonia in the germinal epithelium: single oogonia and those in mitotic proliferation. Oogonial proliferation and their entry into meiosis resulted in formation of cell nests (clusters of cells in the ovarian lamellae). Morphometric analysis was used to estimate germ cell renewal. Based on numbers of single oogonia in the lamellar epithelium, and nests with proliferating oogonia or early prophase oocytes throughout the annual reproductive cycle, oogonial proliferation and entrance into meiosis were more intense during the regenerating phase and developing phase, but decreased sharply (P < 0.05) during the spawning-capable phase. Oogonial proliferation gradually recovered during the regressing phase. We concluded that, independent of species or features of the reproductive cycle, germ cell renewal occurred during the regenerating phase, ensuring availability of eggs for the spawning event. © 2013 Elsevier Inc.

Annual reproductive cycle of males of the flat-faced fruit-eating bat, Artibeus planirostris (Chiroptera: Phyllostomidae)

Artibeus planirostris is an endemic species of Phyllostomid bat from the Neotropical region. Some studies have indicated that it exhibits seasonal bimodal polyestry; however, others postulate that it may be able to produce young at any time during the year. Thus, the aim of this study was to evaluate the annual variations in testicular and epididymal parameters of this species in southeast Brazil and try to understand how the reproduction of this species is regulated in this environment. Sixty mature male specimens, collected between June 2009 and May 2010, were submitted to morphometric and immunohistochemical analysis. Our study showed that A. planirostris presented a continuously active pattern of spermatogenesis throughout the year, presenting spermatozoa inside its cauda epididymis in all months, but with two pronounced peaks of spermatogenic production, one in September and other in February. We propose that the males developed these two peaks in order to produce sufficient sperm for the reproduction in a harem system and to synchronize with the female reproductive cycle, which had a bimodal polyestric pattern. Control of this variation is directly linked to the expression of the androgen receptor (AR) in Sertoli cells and to serum testosterone levels, which appear to synchronize to establish these two peaks. In the months preceding the two peaks, the testis have a higher expression of the AR, which possibly stimulates the increase in PCNA, and drives a gradual increase in the testicular parameters. Taken together the results suggest that if sperm storage happens in this species, it is of short duration. © 2013 Elsevier Inc.

Ultrastructural characteristics of the spermatogenesis during the four phases of the annual reproductive cycle of the black myotis bat, Myotis nigricans (Chiroptera: Vespertilionidae)

Myotis nigricans is an endemic species of vespertilionid bat, from the Neotropical region, that resembles temperate zone bats in their reproductive cycle; presenting an annual reproductive cycle with two periods of testicular regression, which are not linked to the apoptotic process and seems to be not directly linked to any seasonal abiotic variation. Thus, this study aimed to ultrastructurally evaluate their reproductive cycle. The process of testicular regression could be divided into four periods: active; regressing; regressed and recrudescence; with all presenting distinct characteristics. The active period was similar to that of other bats, presenting the complete occurrence of spermatogenesis, with three main types of spermatogonia (Ad, Ap, and B) and 12 steps in spermatid differentiation; however, it differed in having the outer dense fibers 1, 5, 6, and 9 larger than the others. These three types of spermatogonia undergo considerable morphologic changes from regressing to the regressed period, and in the recrudescence, they return to the basic morphology, which reactivates spermatogenesis. In conclusion, our study described the process of spermatogenesis, the ultrastructure of the spermatozoa and the distinct morphologic variations in the ultrastructure of the testicular cells of M. nigricans during the four different periods of its annual reproductive cycle. Microsc. Res. Tech., 76:1035-1049, 2013. © 2013 Wiley Periodicals, Inc.

Reproductive cycle of the Neotropical cichlid yellow peacock bass Cichla kelberi: A novel pattern of testicular development

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Dynamics of ovarian maturation during the reproductive cycle of Metynnis maculatus, a reservoir invasive fish species (Teleostei: Characiformes)

Neste estudo, avaliamos a dinâmica da maturação ovariana a desova durante o ciclo reprodutivo de Metynnis maculatus. Fêmeas adultas (n = 36) foram coletadas bimestralmente entre abril de 2010 e março de 2011. O índice gonadossomático (IGS) foi calculado e amostras de ovário e de sangue foram submetidas à avaliação morfométrica e das concentrações plasmáticas dos esteroides por ELISA, respectivamente. A espécie apresenta desenvolvimento ovariano assincrônico, com múltiplas desovas. Neste estudo revelamos que mesmo sendo de desova parcelada, os ovários do M. maculatus mostraram um padrão de desenvolvimento com predomínio de atividade vitelogênica entre abril a agosto e intensificação da desova em setembro. Em outubro houve uma diminuição nos valores médios de IGS, bem como registramos as maiores frequências de folículos pós-ovulatórios (FPOs). Observamos uma correlação positiva entre a frequência de FPOs e a concentração plasmática de 17 α-OHP. O M. maculatus tem potencial para ser usado como fonte para uso de hipófise para preparo de extrato bruto para indução hormonal, sendo o período teórico para coleta de hipófises de setembro a outubro, mas estudos específicos para esta finalidade ainda precisam ser desenvolvidos.

Extracellular matrix remodeling of the testes through the male reproductive cycle in Teleostei fish

During the fish reproductive cycle, testes undergo morphological changes related to germinal epithelium and remodeling of extracellular matrix components (ECM). ECM is degraded mainly by action of matrix metalloproteinases (MMPs). Due to the natural renewal of ECM in fish testes, we choose Pimelodus maculatus to study remodeling of ECM throughout reproductive cycle, using picrosirius (to identify type I, II, III collagen) and reticulin (type III collagen), and to immunolocalize MT1-MMP (membrane type 1-matrix metalloproteinase) and MMP-2 in testis cells. Testes were classified in four reproductive phases: regenerating, development, spawning capable and regressing. Picrosirius and reticulin demonstrated a differential distribution of total collagen fibers during the reproductive cycle. Immunohistochemistry showed MT1-MMP only in acidophilic granulocyte cells mainly inside blood vessels, in connective tissue of capsule close to the germinal compartment, and also infiltrated in interstitial connective tissue. MMP-2 was detected in fibroblast and endothelial cells of interstitial and capsule blood vessels, in epithelial cells of capsule, and in acidophilic granulocyte cells at same description for MT1-MMP. The fish testes ECM were remodeled throughout reproductive cycle in according to morphophysiological alterations. During reproductive season (spawning capable), the interstitium increased in total collagen fibers (type I, II, III). After spermiation period (regression and regenerating), the amount of collagen fibers decreased in response to action of MMPs on collagen degradation and other interstitial components (not assessed in this study). MMPs seem to be indispensable components for natural cyclic events of ECM remodeling of fish testes and for guarantee tissue homeostasis throughout reproductive cycle.

Two periods of total testicular regression are peculiar events of the annual reproductive cycle of the black Myotis bat, Myotis nigricans (Chiroptera: Vespertilionidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)