Mariano Elisio

Effects of global warming on fish reproductive endocrine axis, with special emphasis in pejerrey Odontesthes bonariensis

The ongoing of global warming trend has led to an increase in temperature of several water bodies. Reproduction in fish, compared with other physiological processes, only occurs in a bounded temperature range; therefore, small changes in water temperature could significantly affect this process. This review provides evidence that fish reproduction may be directly affected by further global warming and that abnormal high water temperature impairs the expression of important genes throughout the brain-pituitary-gonad axis. In all fishes studied, gonads seem to be the organ more readily damaged by heat treatments through the inhibition of the gene expression and subsequent synthesis of different gonadal steroidogenic enzymes. In view of the feedback role of sex steroids upon the synthesis and release of GnRH and GtHs in fish, it is possible that the inhibition observed at brain and pituitary levels in treated fish is consequence of the sharp decrease in plasma steroids levels. Results of in vitro studies on the inhibition of pejerrey gonad aromatase expression by high temperature corroborate that ovary functions are directly disrupted by high temperature independently of the brain-pituitary axis. For the reproductive responses obtained in laboratory fish studies, it is plausible to predict changes in the timing and magnitude of reproductive activity or even the total failure of spawning season may occur in warm years, reducing annual reproductive output and affecting future populations.

Seasonal changes and endocrine regulation of pejerrey (Odontesthes bonariensis) oogenesis in the wild

The goal of this study was to evaluate the essential components controlling the brain-pituitary-gonad axis during pejerrey (Odontesthes bonariensis) oogenesis in the wild. Ovarian developmental stages from vitellogenesis up to ovulation were associated with increasing day length and water temperatures below 21°C (winter and beginning of spring). Gonadal regression was observed when water temperature exceeded this value or when photoperiod decreased. Most females were arrested at primary growth stage during summer (high temperature) or at cortical alveoli stage between autumn and beginning of winter (short photoperiod). Plasma E2 and transcript levels of fshr, cyp19a1b and cyp19a1a increased during vitellogenesis, while fshb remained high at all vitellogenic stages. A significant correlation between plasma sex steroids (T and E2) and cyp19a1b as well as lhcgr transcript levels was observed during vitellogenesis, suggesting a steroid positive feedback. Gnrh-I, Gth subunits and lhcgr transcript levels increased significantly during late vitellogenesis and final maturation. Present results suggest that pejerrey vitellogenesis is controlled by Fsh/Fshr, stimulating gonadal aromatase and estradiol synthesis. Moreover, the increase of testosterone and estradiol during final vitellogenesis could induce coordinately the functioning of the Gnrh/Lh system (perhaps through brain P450 aromatase stimulation and brain estradiol increase) and the gonadal Lhcgr synthesis to promote the final maturation of oocytes. All these stimulation mechanisms of gonadal development would be possible only under permissive environmental conditions.

Effects of short periods of warm water fluctuations on reproductive endocrine axis of the pejerrey (Odontesthes bonariensis) spawning

The aim of this study was to assess fluctuations in daily water temperature in Chascomús Lagoon during one year, and to evaluate whether the highest temperature recorded during pejerrey spawning season can produce an endocrine disruption on brain-pituitary-gonads axis. Fish were subjected to daily temperature fluctuations: 17 °C to 19 °C (reproductive control), 19 °C to 25 °C, and 19 °C to 27 °C. After 8 days, ten fish per treatment were sacrificed and gene expression of gonadotropin-releasing hormone (GnRH-I, GnRH-II, GnRH-III), gonadotropin subunits-β (FSH-β, LH-β), glycoprotein hormone-α (GPH-α), gonadotropin receptors (FSH-R, LH-R), and gonadal aromatase (cyp19a1a) was analyzed. Also, plasma levels of sexual steroids and gonadal reproductive status were studied. Fish exposed to high temperature fluctuations quit spawning, presenting clear signs of gonadal regression. Fish recovered its spawning activity 11 weeks after heat treatment. At endocrine level, GnRH-I and FSH-β in both sexes, LH-β and GPH-α in males and FSH-R, LH-R and cyp19a1a in females decreased significantly in treated fish. Also, a strong reduction in plasma sex steroid levels was found for both sexes. This study demonstrated that pulses of warm water in natural environment during pejerrey spawning season can disrupt all levels of the reproductive axis, impairing reproduction.

Extrahypophyseal expression of gonadotropin subunits in pejerrey Odontesthes bonariensis and effects of high water temperatures on their expression

It has been traditionally accepted that the gonadotropins (GtHs), follicle stimulating hormone (FSH) and luteinizing hormone (LH), are synthesized and secreted only by the pituitary. However, the presence of theses hormones in extrapituitary tissues has been demonstrated in mammals, and more recently also in fish. In this study, we cloned the cDNAs and characterized the expression of FSH-β, LH-β, and glycoprotein hormone α (GPH-α) subunits from brain and gonads of male and female pejerrey Odontesthes bonariensis at different stages of gonadal maturation. In situ hybridization revealed that, in addition to their classical location in pituitary cells, the three GtH transcripts were also located in the gonads. FSH-β and GPH-α subunits were found in the cytoplasm of oogonia, previtellogenic and vitellogenic oocytes in ovaries. LH-β expression was detected in previtellogenic and vitellogenic oocytes but not in oogonia. In males, the three subunits were expressed in spermatogonia and to a lesser extent in spermatocytes. Exposure of fish to high water temperatures that impair pejerrey reproduction also induced a decrease of extrahypophyseal expression of GtH subunits.

Seasonal changes and endocrine regulation of pejerrey (Odontesthes bonariensis) spermatogenesis in the wild

The endocrine mechanisms that regulate spermatogenesis and their interaction with environmental cues have been poorly studied compared with oogenesis in fish. The aim of this work was to study the spermatogenesis in pejerrey under the influence of photoperiod and water temperature fluctuation in the wild, evaluating the transcript levels of brain Gnrh variants and cyp19a1b, pituitary Gth subunits, gonadal Gth receptors, 11β-hsd, and 11-KT plasma levels. Males at spermiogenic stage were observed during spring and autumn, under a photoperiod above 11h of light and a water temperature below 23 °C. Most arrested males were observed in summer when water temperatures increased above 23 °C. Males at spermatogonial stage were mainly observed in autumn, while most males at spermatocytary stage were caught in winter. An increase of gnrh-I, cyp19a1b, fshb, gpha and 11β-hsd transcripts and 11-KT plasma levels was observed during spermatogonial and/or spermatocytary stage (early spermatogenesis). The spermiogenic stage was associated to the maximum gnrh-I gene expression level and a significant increase of Gth receptors transcripts, being this fact more evident for lhcgr. During this last gonadal stage, cyp19a1b transcript level remained high, while fshb mRNA and 11-KT plasma levels showed a significant decreased compared to that occurred at the spermatocytary stage. Also, gphα and 11β-hsd gene expression levels fell during spermiation up to similar values to those observed in arrested males. A significant correlation between 11-KT and gnrh-I, cyp19a1b, gphα, fshb, 11β-hsd transcripts, and the number of spermatocytes was observed during spermatogenesis. All these findings suggested that in pejerrey, the spermatocyte proliferation occurs mainly during winter under the stimulation of 11-KT induced by FSH through the stimulation of specific enzymes, including the 11β-hsd while spermiation occurs after photoperiod increase and with temperatures of the water below 23 °C, through the stimulation of gnrh-I, cyp19a1b and lhcgr.

Quality of pejerrey (Odontesthes bonariensis) eggs and larvae in captivity throughout spawning season

The aim of this work was to assess the quality of pejerrey eggs and larvae throughout its spawning season. Fertilized eggs were taken on September, October, November, and December from a captive broodstock. The egg diameter, yolk diameter, and oil droplets area decreased along the spawning season, with higher values in September. Fertilization and hatching rates decreased throughout this period, with highest values in September (88.0%; 55.2%) and the lowest values on December (43.0%; 25.2%). The larvae hatched from eggs obtained on October were the heaviest and longest (1.57 mg; 8.24 mm). The survival rate at 30 days post hatching (dph) was similar in larvae from September and October eggs (66.1%; 62.9%) with a sharp decrease in larvae from November and December eggs (22.4%; 23.3%). Furthermore, the highest body weight (15.1 mg) and total length (15.25 mm) at 30 dph were obtained in larvae from October eggs. The results obtained showed that overall eggs quality was better at the beginning of the spawning period, influencing the larvae performance.

Kisspeptins and their receptors in the brain-pituitary-gonadal axis of Odonthestes bonariensis: Their relationship with gametogenesis along the reproductive cycle

In vertebrates, the reproduction is controlled by the brain-pituitary-gonadal (BPG) axis and kisspeptin has emerged as a key player of this axis. In this study, we analyzed changes in the expression levels of kiss1, kiss2, and their receptors, kissr2 and kissr3 during gametogenesis in the BPG axis of feral Odontesthes bonariensis. In females, levels of brain kiss1 showed an increase at final maturation (Fm), while kiss2 levels were shown to be high at primary growth (Pg) stage, with no differences in the expression of their receptors. In the pituitary, kiss1 and kiss2 peaked at the cortical alveoli (Ca) stage, and kissr3 at initial vitellogenesis. In parallel, there was an increase of kiss1, kissr2 and kissr3 in the ovary during the Ca stage and both receptors again at Fm stage. In males, the four genes were highly expressed in the brain at the arrested (A) stage. In the pituitary, kiss2 peaked at spermatogonial (SG) and spermatocytary (SC) stages; while kissr3 reached a peak at the spermiogenic stage (SP). In testes, kiss1 and kiss2 significantly increased during the SG and SC stages; meanwhile, kissr2 increased at SG and SC, whereas kissr3 levels were significantly high at SC and SP stages. Taken together these results showed that the kisspeptin system in pejerrey is expressed in the three levels of the BPG axis with different expression profiles during the gonadal cycle. These findings pointed that kisspeptins have different roles in gametogenesis in this species.

Aggregations and reproductive events of the narrownose smooth-hound shark (Mustelus schmitti) in relation to temperature and depth in coastal waters of the south-western Atlantic Ocean (38–42°S)

Knowledge of environmental influence on reproductive dynamics in chondrichthyans is critical for effective management. The present study assessed the potential influences of temperature and depth on aggregations and reproductive events of the narrownose smooth-hound shark (Mustelus schmitti), using an analysis criterion based on the co-occurrence of different reproductive stages. Data were collected from 363 sample sites during six research surveys, covering an area of ~47744km2 along the coast (38–42°S, depth of <50m). Aggregations of M. schmitti adults were mostly associated with the warmer spring surveys, with the aggregation sites being significantly warmer than those of non-aggregation. Evidence of pupping and ovulation was observed, almost simultaneously, during all spring surveys. However, the occurrence of these reproductive events showed a clear interannual variability pattern that was significantly explained by water temperature. Particularly, the occurrence of ovulation was associated with temperatures above 16–17°C. These results suggest that M. schmitti adults may follow the warming gradient along the environment and, thus, they aggregate in shallow and productive waters during the course of spring. Their reproductive events could be induced by temperature increase. This scenario suggests that changes in the patterns of bottom temperature may be important drivers of the reproductive dynamics of M. schmitti.