Report of 2009 Bottlenose Dolphin (Tursiops truncatus) Life History Workshop: expanding network capabilities

The Virginia Aquarium & Marine Science Center Foundation’s Stranding Response Program (VAQS) was awarded a grant in 2008 to conduct life history analysis on over 10 years of Tursiops truncatus teeth and gonad samples from stranded animals in Virginia. A major part of this collaborative grant included a workshop involving life historians from Hubbs-Sea World Research Institute (HSWRI), NOS, Texas A & M University (TAMU), and University of North Carolina Wilmington (UNCW). The workshop was held at the NOAA Center for Coastal Environmental Health & Biomolecular Research in Charleston, SC on 7-9 July 2009. The workshop convened to 1) address current practices among the groups conducting life history analysis, 2) decide on protocols to follow for the collaborative Prescott grant between VAQS and HSWRI, 3) demonstrate tissue preparation techniques and discuss shortcuts and pitfalls, 4) demonstrate data collection from prepared testes, ovaries, and teeth, and 5) discuss data analysis and prepare an outline and timeline for a future manuscript. The workshop concluded with discussions concerning the current collaborative Tursiops Life History Prescott grant award and the beginnings of a collaborative Prescott proposal with members of the Alliance of Marine Mammal Parks and Aquariums to further clarify reproductive analyses. This technical memorandum serves as a record of this workshop.

Reproductive biology of male franciscanas (Pontoporia blainvillei) (Mammalia: Cetacea) from Rio Grande do Sul, southern Brazil

The reproductive biology of male franciscanas (Pontoporia blainvillei), based on 121 individuals collected in Rio Grande do Sul State, southern Brazil, was studied. Estimates on age, length, and weight at attainment of sexual maturity are presented. Data on the reproductive seasonality and on the relationship between some testicular characteristics and age, size, and maturity status are provided. Sexual maturity was assessed by histological examination of the testes. Seasonality was determined by changes in relative and total testis weight, and in seminiferous tubule diameters. Testis weight, testicular index of maturity, and seminiferous tubule diameters were reliable indicators of sexual maturity, whereas testis length, age, length, and weight of the dolphin were not. Sexual maturity was estimated to be attained at 3.6 years (CI 95% =2.7–4.5) with the DeMaster method and 3.0 years with the logistic equation. Length and weight at attainment of sexual maturity were 128.2 cm (CI 95%=125.3–131.1 cm) and 26.4 kg (CI 95% =24.7–28.1 kg), respectively. It could not be verified that there was any seasonal change in the testis weight and in the seminiferous tubule diameters in mature males. It is suggested that at least some mature males may remain reproductively active throughout the year. The extremely low relative testis weight indicates that sperm competition does not occur in the species. On the other hand, the absence of secondary sexual characteristics, the reversed sexual size dimorphism, and the small number of scars from intrassexual combats in males reinforce the hypothesis that male combats for female reproductive access may be rare for franciscana. It is hypothesized that P. blainvillei form temporary pairs (one male copulating with only one female) during the reproductive period.

Development of artificial breeding techniques for long-whiskered catfish, Sperata aor and giant river catfish, Sperata seenghala of Bangladesh

Sperata aor and S. seenghala are the two important native catfishes of Bangladesh but commercial farming of these species is not possible due to lack of naturally collected or artificially produced seeds for stocking. Attempts were made to develop techniques for seed production by artificial breeding and nursery-rearing of fries of these catfishes. A total of 60 S. seenghala (750-1,500 g) and 10 S. aor (600-1,000 g) broods were collected from the Brahmaputra river-basin and floodplains in Mymensingh region four months prior to their breeding season. The collected brood fishes were reared in separate earthen ponds with supplementary feeds comprising of rice bran (40%), mustard oil cake (29%), fish meal (30%) and vitamin-premix (1 %). Three experiments were conducted to optimize the hormone dose. A total of nine S. seenghala females weighing from 750 to 1,500 g were given an initial and resolving dose of 12-20 and 16-24 mg PG/kg body weight, respectively. The males weighing from 650-950 g were administered a single dose of 18-26 mg PG/kg body weight at the time of the time of administering the resolving dose to the females. The females ovulated partially and the eggs were examined under a compound microscope, but most of them were found to be less ripe or damaged. Collection of milt by stripping the males was not successful. The testes were taken out and sperm were observed to be non-motile and less developed. In view of stimulating natural propagation of S. seenghala, artificial holes (nests) were constructed in the pond bottom. Each hole was 0.7 m in diameter and 0.3 m in depth. A total of 10 holes were made and then 10 pairs of S. seenghala breeders (800-1,200 g) were stocked in the pond. In mid February, 3,000 fry of S. seenghala with a mean length of 4.60 cm and weight of 0.36 g were collected by repeated netting followed by drying of the pond. The fry were then stocked in a nursery pond and fed with commercial feed (SABINCO starter-1). The average length and weight of the fingerlings were 9.01 cm and 3.95 g, respectively and the estimated survival was 60% after two months of rearing. S. aor did not respond to natural spawning. Further study is essential to develop techniques for their successful artificial and natural breeding.

Reproductive biology, artificial propagation and larval rearing of two freshwater eels, Monopterus cuchia and Mastacembelus armatus

Studies on reproductive biology and artificial propagation including larval rearing of freshwater mud eel, Monopterus cuchia and spiny eel, Mastacembelus armatus were attempted. The gonadosomatic index (GSI) of mud eel ranged from 0.41 (August) to 5.52 (June) in males and 0.53 (August) to 7.61 (June) in females. In both cases the GSI showed a peak in June. Fecundity ranged from 228 (TL - 396 mm; W - 78g) to 5510 (TL - 865 mm; W - 630 g). In case of spiny eel, the GSI varied from 0.65 (August) to 8.30 (July) in males and 0.70 (August) to 10.46 (July) in females. GSI showed single peak in July. Fecundity ranged from 570 (TL - 240 mm; W - 30 g) to 10870 (TL - 601; W - 350g). Histology of the testes and ovaries of the eels were carried out to investigate the gonadal development stages during the reproductive months (August to November 2003). In case of male M. cuchia, the secondary primordial germ cells, primary spermatogonium, some spermatogonia A and clone of spermatogonium B in testis were observed in September. In October-males different sized lobules having spermatogonia, spermatocytes and spermatids were observed. In the ovary of M. cuchia, polygonal shaped oocytes were seen during September. The oogonia were reduced with dense and irregular shaped during October. Numerous pycnotic cells were visible during November. In male M. armatus numerous broken lobule walls were found in testes during September. In October, abundant primary germ cells, pycnotic nests of degenerating cells, spermatogonia and spermatids were observed. In females, ovaries had distinct yolk vesicles stage and yolk granules stages in August. In September, the follicular cells of the oogonia were ruptured, shrunk forming irregular shaped in October. Oogonia were also shrunk with thin, irregular shaped structure but broken parts of the ruptured follicular cells were scattered in case of M. armatus. Experimental attempts on artificial propagation indicated that both freshwater eels were difficult to breed using inducing agents like pituitary glands (PG) of 10, 20, 50, 100 and 150 mg per kg of body weight. Same doses were used for both sexes with equal sex-ratio. In both cases, brood fish died at higher doses of injection given at 100 and 150 mg PG/kg bodyweight. However, M. cuchia breed naturally in cisterns when provided with water hyacinths and tunnel in muddy bottom. M. cuchia fed with chopped cooked fish attained a mean weight of 18.75 ± 2.3 g and cent percent survival. While in case of M. armatus best growth by weight (12.0 ± 2.48 g) and cent percent survival were achieved using chopped raw fish. Car tyre was observed as best shelter for attaining the mean weight gain 22.53 ± 2.24 g and cent percent survival of M. cuchia. While PVC pipe was found to be the best shelter for M. armatus, where it attained the mean weight of 12.73 ± 1.88 g and cent percent survival.

Suggestive role of eye-stalk of Macrobrachium gangeticum Bate, 1868 in regulation of its reproductive behaviour

Role of eye-stalk of Macrobrachium gangeticum Bate, 1868 in its reproductive behaviour has been examined by conducting deletion and addition experiments. Eye-stalk ablation induced gonadal maturity in both sexes, leading to change in colour and size of ovaries and increase in GSI and oocyte diameter in females and increased length of testes and diameters of seminiferous tubules in males. Injection of eye-stalk extracts tended to at least partly restrict the effects in both sexes. The experiments thus suggested that the eye-stalk of M. gangeticum released some gonad inhibiting factors.

A nerve growth factor from the venom of Chinese cobra (Naja naja atra) and its effects on male reproductive system in rats

A nerve growth factor (NGF) was isolated from the venom of Chinese cobra (Naja naja ntr a) by ion exchange chromatography, gel filtration and fast protein liquid chromatography (FPLC). The N-terminal sequence of 22 amino acid residues was identical with other NGFs previously purified from the venom of the same genus. The NGF monomer molecular weight was estimated to be 13 500 by reducing SDS-PAGE and the isoelectric point was determined to be 7.2 by isoelectric focusing electrophoresis. NGF improved the epididymal sperm motility of male rats and increased the pregnancy rate and fetus number of mated female rats. The serum levels of luteinizing hormone (LH) and follicle stimulating hormone (FSH) of male rats administrated NGF + gossypol was lower than that of male rats administrated gossypol. Histological sections of testes and epididymides showed that NGF reduced the destructive effects of gossypol on rat testes. (C) 1999 Elsevier Science Inc. All rights reserved.

Plerchis heterorchis N. sp. (Digenea: Acanthocolpidae Luhe, 1909) from fishes Lutjanus johnii and Otolithus argenteus of Karachi coast, Pakistan

A new species of trematodes Pleorchis heterorchis is described from the fishes Lutjanus johnii and Otolithus argenteus of Karachi coast. The new species is characterized by having a lanceolate body with a notch at the middle of the posterior end of the body. Body surface is smooth, ventral sucker rounded, situated at the anterior middle region of the body, pre-pharynx is well developed, widened posteriorly, pharynx muscular, oesophagus short, intestine H-shaped with anterior arms much shorter than the posterior, intestinal bifurcation almost in the middle of fore body, anterior caeca wide and short extending as far as anterior limit of pharynx. Posteriorly caeca reach to posterior end of the body with no lateral out pocketing. Testes 44 in number, intercecal arranged in 2 parallel rows, sub-globular, entire to slightly irregular, almost of same sizes extending immediately from posterior of the ovary to anterior of excretory vesicle. Cirrus pouch overlaps the ventral sucker, extends into hind body, terminating above the ovary, containing bipartite seminal vesicle, pars prostatica and ejaculatory duct. Genital pore behind the intestinal bifurcation and pre-acetabular. Ovary pre-testicular, consists of 16 follicles of varying sizes. Vitellaria lateral, follicular, extending from post bifurcal to posterior extremity. Excretory vesicle reaches to the posterior level of last pair of testes.

The study of maturational progress in the whitespotted rabbitfish (Siganus sutor) under different photoperiod and light intensity conditions

The rule of light on the timing of maturation and spawning in tropical and subtropical regions is not clear well, because the reproductive cycle in these systems is lunar synchronized. In this study, thus, the effects of different light regimes were investigated on maturational progress of whitespotted rabbitfish, Siganus sutor, the commercial species in Persian Gulf and Oman Sea. During prespawning season, 50 adult fish were randomly divided into ten 300-l tanks (n=5). The fish in control tank received indoor light condition and the fish in each other tanks were exposed to nine different combinations of photoperiod (8L: 16D, 12L: 12D, 16L: 8D) and light intensity (1000, 2000, 3000 lux). After 60 days, GSI and HSI values, serum levels of estradiol (E2), testosterone (T), 17-α hydroxyprogestrone (OHP), calcium (Ca2+) and gonad histology were evaluated for females and males. In females the GSI mean values of exposed fish increased in comparison with control except for fish were kept under 8L, 2000 lux (tank 8). These differences were significant only for fishes in tank 7 (8L, 3000 lux). In the cases of HSI, the results were converse, so that, the most of thanks showed significant decreasing than fishes reared under indoor condition. Morphology and histology study of Ovaries showed three developmental phases including 3, 4 and chiefly 5 that were parallel with GSI values. Fortheremore the serum levels of E2 was recorded between 0.54-15.8 ng/ml in different fish and their mean values were lower than control in all treatmants (P> 0.05). In males, the similar results were obtained. The GSI and HSI mean values in experimental regimes compared with control were upper and lower, respectively, except for fishes were reared in tank 1 (16L, 3000 lux). Testes histology of fishes were reared under different regimes showed signs of stage 5, since no blood vessels observed and thick milt exuded on slight pressure. The mean values of testosterone consentration in fishes were kept in tanks 1 and 6 (12L, 1000 lux) were higher and in other ones were lower than control group. It is also noted that the OHP and Ca2+ had diverse results including increasing and decreasing mean values than control. So, these factors contrary to E2 and T were not suitable to evaluation of maturity in both sexes. On the basis of ovarian structure in stage 5, oocyte development pattern in this species was group synchronous. So, increased mean of GSI versus decreased values of HSI, E2 and perhaps Ca2+ were the signs of improved maturation. But in males, reduced levels of T and similarity of testes morphology in all samples caused that GSI mean value was the only indicator for differentiating among treatments. These findings suggest that alternations were used in light regimes have been the reason of improved maturity in all treatments except fishes reared in tank 8. The ١٠٧ rule of light intensity on induction of maturity was cleared by comparision between fishes in tanks 4, 5, 6 and control group. Because day length was the same whereas fishes in tanks 4, 5 and 6 were exposed to increased light intensity compared with control. This fact verified by results was obtained from fishes in tanks 9 and especially 7, since photoperiod in this group was lower than control. So, higher intensity was considered as the reason of alternations. Contrasting with indoor condition, Induced maturity was also cleared for fishes were kept in tanks 1, 2 and 3, where both of light duration and intensity were increased. But, the rule of photoperiod was individually demonstrated when obtained results were compared with similar light intensities in other treatments. In conclusion, with comparison among different light regimes it is declared that siganids were kept under light condition of tank 2 including 16h light duration combined with 2000 lux intensity showed the best signs of sexual maturation in both sexes. On the basis of this study, setting up the spring light condition during prespawning season induces maturation in withspotted rabbitfish. This improvement not only by influence of photoperiod or light intensity, separately, but obtained through interaction between them. Thus, determination of threshold and resistance to light be recommended before exposure, although using proper regime during suitable time are necessary to achieve purposes considerably.

Study on macroscopic, microscopic and hormonal changes in gonads of the crayfish Astacus leptodactylus in Aras Dam

This study was carried out to seasonal determination of some morphological characteristics, Seasonal fecundity, Seasonal fluctuations of vertebrate-type steroids and seasonal analysis of gonadal histology in both female and male sexes of freshwater crayfish (Astacus leptodactylus Eschscholtz 1823) in the area of Aras dam Lake. Crayfish were collected respectively in June, August, November (2011) and January (2012). The average length and weight of male crayfish was higher than that of females. %GSI of females fluctuated within an extended range (between 0.6 and 13.5% from June to January). Both of synchronous and asynchronous ovaries were seen in August sampled ovaries; however asynchronous form was higher than another. The annual reproductive cycle of male A. leptodactylus was surveyed by study on the seasonal changes of the external appearance of the testes and vasa deferentia, fluctuations in the gonadosomatic index (GSI%) and the histological analysis of the male reproductive system. Based on the histological differentiation of testis, spermatogenisis devided to 5 separated stages. The findings suggested asynchronous testis in the species A.leptodactylus. The presence of primary spermatophore layer may help keeping spermatozoa alive while the secondary spermatophore layer may produces spermatophore or synthesize of acellular material which forms spermatophre. Pleopodal fecundity was 37.3%lower than ovarian fecundity observed. The significantly higher number of eggs attached to 3rd and 4th pairs of pleopods. The egg number and gonadosomatic index increased with female size while egg weight and egg diameter didn’t increase with female size. Hemolymph levels of 17β-estradiol and progesterone followed a similar fluctuation pattern with % GSI in females, while testosterone didn’t follow the mentioned pattern. The testis of November sampled crayfish presented significantly higher gonadosomatic (%GSI) index (P < 0.05).The most observed gonadosomaticindices were 13.5%(forfemales) and 1.21% (for males, in autumn. Althogh the lowest GSI was (0.50%) formales in spring and (0.26%0 for spent females in January. Testosterone which followed a similar pattern with %GSI in males increased remarkably in November. 17β-estradiol increased strictly in January. The strictly enhancement of the three estroid hormones in January in both male and female sexes could bedue totheir stimulating role in in spermatophre and egg lying in the mating season (In January). Most of the ovaries followed the asynchoronous growth pattern. Also the testes presented asynchoronous growth pattern in autumn.

Study on biological characteristics, sexual maturation and reproduction of Liza abu in the water of Khozestan Province

As the most of the fish resources are known and exploited, protecting their generation is of the greatest importance. Aquaculture is one of the efficient procedures in protecting and reviving fish resources and knowing about the reproductive cycle and gonads development has an important role in approaching this aim. Liza abu belongs to the family Mugilidae that according to its resistance to the environmental condition and its fast growth , can be introduced as a fish with economical value. As there is no scientific data on the reproductive biology of this species , study on the reproductive biology and gonad development is considered as the aim of this research . For this purpose , 360 samples of this species were investigated during the period from February 2007 to January 2008 in Khozestan Province . After studing morphological and histological characteristics of gonad specimen , they were prepared through histological method. Samples were prepared through usual histological method and studied under light microscope. According to the results, the maturity stages of male and female Liza abu were separated to six different successive stages. In ovaries , these stages were as follow : In stage І, the oocytes were small , this stage was observed from July to October . In stage ІІ, considerable growth was observed in the oocytes . This stage was observed from October to January . In stage III, due to vitellogenesis, the maximum growth was observed and three layers of theca, granullosa and follicle cells were visible. This stage was observed during January and February . In stage IV, migration of germinal vesicle was observed and due to hydration of the oocytes , their diameter was increased. The ovaries were yellowish and in maximum size and ovules could be easily observed with naked-eye . This stage was observed in February and March . In stage V, spawning occured. This stage was observed in April . In stage VI, ovaries consisted of immature and atretic oocytes and also empty follicles. This stage was observed in May and June. In testes , these stages were as follow : In stage I , the testes were small in size and contained the spermatogonia which were the only cellular components.This stage was observed in August and September . In stage II (maturing virgin ) , the spermatogonia and the primary spermatocytes were visible. This stage was observed in October . In stage III (developing), intensive spermatogenesis was occured and the primary and the secondary spermatocytes were the most visible cells during this stage .This stage was observed from November to January. In stage IV(developed), cells of all stages of spermatogenesis could be seen but the secondary spermatocytes and spermatids were in large number. This stage was observed from January to March. In stage V , the testes were filled with sperms. This stage was observed in March and April .In stage VI, residual spermatozoa and the spermatogonia were visible in the testes. This stage was observed from May to August. According to cyclic changes in GSI, sexual maturation in breeding begins in January and spawning occurs in April. The ova diameter ranged from 30.75 μ in stage I to 472.19 μ in stage IV. In this study , the sex ratio was 1:2.7, and male and female percentage were 27.02% and 72.98% respectively. This means that females predominate males. In this study absolute fecundity was calculated and changing between 30805.44 to 431247.3 was observed and absolute fecundity was calculated 111275.3 in average.

HORMONAL-REGULATION OF TISSUE-TYPE PLASMINOGEN-ACTIVATOR AND PLASMINOGEN-ACTIVATOR INHIBITOR TYPE-1 IN CULTURED MONKEY SERTOLI CELLS

Sertoli cells play a central role in the control and maintenance of spermatogenesis. Isolated Sertoli cells of mouse and rat testes have been shown to secrete plasminogen activator (PA) and a plasminogen activator inhibitor type-1 (PAI-1) in culture. In this study, we have investigated the hormonal regulation of PA and PAI-1 activities in cultured monkey Sertoli cells. Sertoli cells (5x10(5) cells/well) isolated from infant rhesus monkey testes were preincubated at 35 degrees C for 16 h in 24-well plates precoated with poly(D-lysine) (5 mu g/cm(2)) in 0.5 mi McCoy's 5a medium containing 5% of fetal calf serum and further incubated for 48 h in 0.5 mi serum-free medium with or without various hormones or other compounds, PA as well as PAI-1 activities in the conditioned media were assayed by fibrin overlay and reverse fibrin autography techniques respectively. The Sertoli cells in vitro secreted only tissue-type PA (tPA), no detectable amount of urokinase-type PA (uPA) could be observed, Monkey Sertoli cells were also capable of secreting PAI-1, Immunocytochemical studies indicated that both tPA and PAI-1 positive staining localized in the Sertoli cells, spermatids and residual bodies of the seminiferous epithelium; Northern blot analysis further confirmed the presence of both tPA and PAI-1 mRNA in monkey Sertoli cells. Addition of follicle-stimulating hormone (FSH) or cyclic adenosine monophosphate (cAMP) derivatives or cAMP-generating agents and gonadotrophin-releasing hormone (GnRH) agonist or phorbol ester (PMA) to the cell culture significantly increased tPA activity. PAI-1 activity in the culture was also enhanced by these reagents except 8-bromo-dibutyryl-cAMP, forskolin and 3-isobutyl-1-methylxanthin (MIX) which greatly stimulated tPA activity, whereas decreased PAI-1 activity, implying that neutralization of PAI-1 activity by tile high level of tPA in the conditioned media may occur. These data suggest that increased intracellular signals which activate protein kinase A (PKA), or protein kinase C (PKC) can modulate Sertoli cell tPA and PAI-1 activities, The concomitant induction of PA and PAI-1 by the same reagents in the Sertoli cells may reflect a finely tuned regulatory mechanism in which PAI-1 could limit the excession of the proteolysis.

Expression and regulation of orphan receptor TR2 mRNA in germ cells of cryptorchid testis in rat and rhesus monkey

Expression and cellular localization of orphan receptor TR2 mRNA in relation to germ cell apoptosis in cryptorchid testes of rat and rhesus monkey have been studied by using in situ hybridization and in situ 3'-end labeling of DNA fragments (TUNEL). The results show that: (i) TR2 mRNA is specifically expressed in the germ cells, mainly in the spermatocytes, round and elongated spermatids. The expression level of TR2 mRNA varies with the seminiferous cycle, (ii) In the rat cryptorchid testes on days 3 and 5 after the surgery, the germ cells began to undergo apoptosis with no evident decrease in TR2 mRNA level. On day 7.5, however, most germ cells underwent apoptosis, while the expression level of TR2 mRNA declined markedly, and TR2 mRNA was rarely expressed on day 10 thereafter. (iii) On days 15 and 20 of the cryptorchid testes of rhesus monkey, TR2 mRNA was only expressed in a few of primary spermatocytes and the mRNA was almost undetectable on days 30, 45, 60. These results suggest that TR2 mRNA probably plays an important role in spermatogenesis and germ cell apoptosis.

KHAWIA SAUROGOBII N. SP (CESTODA: CARYOPHYLLIDEA) FROM FRESHWATER FISH SAUROGOBIO SPP. (CYPRINIDAE) IN CHINA

Khawia saurogobii n. sp. is described from the intestine of the cyprinid fishes Saurogobio dabryi and Saurogobio dumerili in China. The new species belongs to the Lytocestidae and is placed in Khawia because of its afossate scolex, vitelline follicles in the cortical parenchyma, uterus not looping anterior to the cirrus sac, gonopores separate but close together in distinct genital atrium, external seminal vesicle absent, and postovarian follicles present. The new species differs from other congeneric species by the shape of the body markedly tapering posteriorly from the end of the anterior third, the shape of the scolex that is very short and markedly wider than the neck, spatulate without incisions, but shallow superficial grooves, vitelline follicles and testes starting immediately posterior to the scolex, and an ovary with long, posterior arms bent medially, thus giving the shape of an inverted A.

Discovery of a male-biased mutant family and identification of a male-specific SCAR marker in gynogenetic gibel carp Carassius auratus gibelio

Gibel carp ( Carassius auratus gibelio) is a uniquely gynogenetic species with a minor ratio of males in natural habitats, but its male origin and sex determination mechanisms have been unknown. In this study, a male-biased mutant family was discovered from the gynogenetic gibel carp, and a male-specific SCAR marker was identified from the mutant family. Normal spermatogenesis was observed in the male testes by immuno. fluorescence histochemistry. Nearly identical AFLP profiles were observed between males and females, but a male-specific 86 bp AFLP fragment was screened by sex-pool bulked segregant analysis and individual screening. Based on the male-specific AFLP fragment, a total of 579 bp sequences were cloned by genome walking. Subsequently, a male-specific SCAR marker was designed, and the male-specific DNA fragment was confirmed to be steadily transmitted to the next generation and consistently detected only in males. (C) 2009 National Natural Science Foundation of China and Chinese Academy of Sciences. Published by Elsevier Limited and Science in China Press. All rights reserved.

Involvement of Fas/FasL system in apoptotic signaling in testicular germ cells of male Wistar rats injected i.v. with microcystins

Previous studies have shown that gonads were the second target organ of microcystins (MCs), and that MCs exposure exerted obvious toxic effects on male reproductive system of mammals. However, relevant molecular evidences are still lacking. Fas-signaling pathway plays a key role in toxicant-induced germ cell apoptosis. This study was to evaluate the responses of Fas/FasL system related genes and proteins in testes of rats injected intravenously with MCs. Enhanced apoptosis of germ cells in the testes of MCs-treated rats was detected by the terminal deoxynucleotidyl transferase-mediated deoxy-UTP nick end labeling (TUNEL) associated with up-regulation of the Fas/FasL system. Both Fas and FasL protein expression were induced evidently from I h post-injection, and this high expression level maintained throughout the experiment. In addition, the activation of caspase-8 and caspase-3 protein was also observed, which were indicators of apoptosis. These results suggested the likely involvement of Fas/FasL system in the MCs-induced germ cell apoptosis. It is also suggested that MCs can cause damage to Sertoli cells directly. (C) 2009 Elsevier Ltd. All rights reserved.