Seasonal correlative changes between sex steroids and lipid levels in the freshwater female catfish, Heteropneustes fossilis (Bloch)

Circannual variation in plasma levels of testosterone (T), oestradiol-17β(E₂) and 17α-hydroxy-progesterone (17α-OHPg) were measured in female, Heteropneustes fossilis . T and E2 levels increased during the preparatory phase, reached their peak in the early prespawning phase and fell during the late prespawning phase to reach their lowest levels post-spawning. 17a-OHPg was detected from the late preparatory to the late spawning phase showing its peak during the early spawning phase. The levels of free fatty acids (FFA), monoglycerides (MG), diglycerides (DG), triglycerides (TG), phospholipids (PL), free cholesterol (CF)and esterified cholesterol (CE) were estimated in liver, plasma and ovaries. The preparatory phase showed hepatic lipogenic activity while during the prespawning phase TG lipolysis was increased by FFA. Ovarian CF was depleted by enhancing plasma E, levels during the prespawning phase. The gonadosomatic index reached its peak during spawning.     

Diurnal and seasonal profiles of melatonin in the liver and their correlates of ovarian functions under natural photo-thermal conditions in adult carp Catla catla

Present study aimed to demonstrate daily rhythm features of hepatic melatonin concentrations in relation to ovarian functions during four reproductive phases of an annual cycle by measuring the levels of melatonin, 17-β estradiol (E2), vitellogenin (Vg) and maturation inducing hormone (MIH) in the liver and/or serum of adult carp Catla catla. Melatonin titres in liver, irrespective of reproductive phase, underwent daily variations with a peak in early dark phase and nadir at midday. However, the acrophase (Ø) of serum melatonin varied from late night in preparatory phase to midnight in the remaining parts of annual cycle. Their amplitude was highest during post-spawning phase and lowest during spawning phase. Hepatic E2 levels showed daily peak at midday and seasonal peak during pre-spawning phase. Though levels of serum Vg proteins and MIH did not exhibit daily variations, underwent seasonal changes with the highest and lowest values during spawning and post-spawning phases respectively. Hepatic melatonin titres always displayed significant negative correlation with the levels of both E2 and Vg. In essence, our study presented the first data on the daily and seasonal rhythm features of hepatic melatonin in carp and underlined their temporal relationship with the functions of ovary in any fish species.     

Effect of long photoperiod on the maturation of ovary of the catfish, Mystus tengara (Ham.)

A study has been made of the effect of photoperiodism on the ovarian maturation of fresh water teleost, Mystus tengara (Ham.). The magnitude of ovarian recrudescence is more during maturing virgin phase as compared to during immature virgin and preparatory virgin phases in long photoperiod (14 hours/day) treated fish. Thus, the response of the ovary to long photoperiod increases with the approach of natural spawning period. The mature eggs are more in long photoperiod treated fish than in controls exposed to 12 hours/day photoperiod. In long photoperiod treated fish, prespawning period is reduced considerably. During spawning and post-spawning periods, the ovaries are refractory, since long photoperiod is ineffective in maintaining gravid ovaries beyond spawning period and the ovarian regression during post spawning period sets in regardless of nature of photoperiod treatment.     

Seasonal migrations and reproductive patterns in the lake sturgeon, Acipenser fulvescens, in the vicinity of hydroelectric stations in northern Ontario

This study was conducted in order to evaluate seasonal migratory behaviour and reproductive pattern of lake sturgeon in a confined region of the Mattagami River system in northern Ontario where river flow is regulated by hydroelectric works. Radio tracking and the systematic sampling of lake sturgeon using gill nets indicated that the distribution of fish throughout the study site varied on a seasonal basis. This distribution was related to the migration of individuals to potential spawning sites in the spring, a post-spawning dispersal to feeding areas and late summer migration to an area of concentration on the Groundhog River which is a tributary of the Mattagami River. There was a high proportion of fish (about 50%), within the size range of reproductively active fish, found in the vicinity of suitable spawning habitat during early May. Measurement of the gonadosomatic index (GSI) and plasma sex steroid hormone levels revealed a divergent pattern of reproductive development between the sexes. Female sturgeon exhibited a prolonged period of ovarian regression following spawning. Resumption of ovarian development was not evident until September and was characterized by an increased GSI and plasma levels of testosterone and 17β-estradiol. In contrast, male lake sturgeon began testicular recrudescence within one month of spawning with the GSI reaching prespawning levels by September; reproductive hormones were at prespawning levels by the end of June. It seems that hydroelectric works has complex effects on sturgeon in the Mattagami system. The extensive migratory behaviour of lake sturgeon within the study area make it prone to impingement or entrainment whereas the altered river flow appears to enhance reproductive development. This revised version was published online in August 2006 with corrections to the Cover Date.     

Seasonal changes in the hypothalamo-hypophyseal-ovarian system in the catfish,Heteropneustes fossilis (Bloch)*

The annual reproductive cycle of the catfish, H. fossilis (Bloch) is divided into the preparatory period (February-April), the prespawning period (May-June), the spawning period (July-August) and the postspawning period (September-January). During the early postspawning period (September-November), the hypothalamo-hypophyseal-ovarian system shows a gradual regression. In January, the hypothalamic nuclei, the pars magnocellularis (PMC), the pars parvocellularis (PPC) of the nucleus preopticus (NPO), and the nucleus lateralis tuberis (NLT) show renewed activity, as shown by a significant increase in their nuclear diameters and an accumulation of neurosecretory material (NSM) in their cell bodies. The hypophysis and the ovary remain quiescent. During the preparatory period, all the hypothalamic neurons studied indicate decreased activity but simultaneously show an accumulation of NSM in their cell bodies. The number of granulated basophils in the proximal pars distalis (PPD) of the hypophysis remains low but ovarian weights increase, presumably due to the multiplication of oogonia. In the prespawning period, there is a marked accumulation of NSM in the cell bodies of the hypothalamic neurons and at the same time the number of granulated basophils in the PPD of the hypophysis dramatically increases with concomitant increase in vitellogenic activity in the ovary. During the spawning period, the hypothalamic neurons continue to store NSM in their cell bodies and simultaneously there is a tremendous increase in the number of granulated basophils in the PPD of the hypophysis and the ovary has a large proportion of yolky primary oocytes. Spawning is associated with a significant degranulation of the granulated basophils in the PPD of the hypophysis. The significance of the results is discussed in relation to the environmental and hormonal regulation of seasonal ovarian activity.     

Sexual cycle and seasonal changes in the ovary of the red mullet, Mullus surmuletus, from the southern coast of Brittany

The reproductive cycle of the red mullet is described on a macroscopic scale in terms of the GSI, HSI and K , and on a microscopic scale in terms of histological changes in the ovary and changes in the oocyte size frequency distribution. On the southern coast of Brittany the red mullet breeds in May and June. During oogenesis, the previtellogenic period lasts 6 months and the secondary phase of vitellogenesis no more than 3 months. When spawning commences the process of vitellogenesis ceases and up to 20% of the vitellogenic oocytes become atretic. Prior to spawning a single batch of oocytes can be seen to be entering secondary vitellogenesis. During the immediate prespawning and spawning periods the existing vitellogenic oocytes mature but there is no recruitment from the stock of previteilogenic oocytes. This results in a gap or hiatus in the oocyte size frequency distribution between previtellogenic and vitellogenic oocytes within which there are very few resting or maturing oocytes. The red mullet appears to be a determinate spawner, in which egg loss through atresia considerably reduces the potential fecundity.     

Circadian Variation in Circulating Levels of Melatonin,Testosterone and Estradiol-17β in a Tropical Freshwater Catfish,Clarias batrachus,during Different Phases of the Annual Reproductive Cycle

In the present investigation, daily variations in plasma levels of melatonin, testosterone and estradiol-17β were analysed in the tropical freshwater catfish, Clarias batrachus during early and late-preparatory, pre-spawning, late-spawning and post-spawning phases of its annual reproductive cycle. Plasma melatonin levels exhibited a significant circadian rhythm during all the reproductive phases but peaks were invariably at night. The amplitude of melatonin was high during the early-preparatory phase and low in the pre-spawning phase. Testosterone also displayed significant rhythm during all the studied phases except post spawning phase. Estradiol-17β revealed diurnal rhythm only during preparatory and pre-spawning phases. The peak in the levels of plasma testosterone and estradiol-17β were recorded during the photophase. During the late-preparatory phase, the rhythm of testosterone coincided with the rhythm of estradiol-17β.     

Circadian Variation in Circulating Levels of Melatonin, Testosterone and Estradiol-17β in a Tropical Freshwater Catfish, Clarias batrachus, during Different Phases of the Annual Reproductive Cycle

In the present investigation, daily variations in plasma levels of melatonin, testosterone and estradiol-17β were analysed in the tropical freshwater catfish, Clarias batrachus during early and late-preparatory, pre-spawning, late-spawning and post-spawning phases of its annual reproductive cycle. Plasma melatonin levels exhibited a significant circadian rhythm during all the reproductive phases but peaks were invariably at night. The amplitude of melatonin was high during the early-preparatory phase and low in the pre-spawning phase. Testosterone also displayed significant rhythm during all the studied phases except post spawning phase. Estradiol-17β revealed diurnal rhythm only during preparatory and pre-spawning phases. The peak in the levels of plasma testosterone and estradiol-17β were recorded during the photophase. During the late-preparatory phase, the rhythm of testosterone coincided with the rhythm of estradiol-17β.     

Annual Variations of Gonadotropin Content and Ovarian Development of Feral Female Catfish, Silurus asotus, in Central China

The Japanese catfish Silurus asotus is widely distributed throughout the freshwaters reservoirs, lakes and rivers of China, Korea and Japan, and is a commercially valuable aquaculture fish in some regions of China and Japan. We studied seasonal variations of gonadotropin (GtH) content of the plasma and pituitary as well as ovarian development. Plasma GtH titres increase in April in the Pearl River and in May in the Liao River in central China. Annual cycles of plasma GtH levels in both rivers are the same, with a gradual decrease after ovulation with the lowest values observed in September (in the Liao River) or October (in the Pearl River). Plasma GtH levels increase gradually in the early stages of ovarian development, with the highest values observed during the late stages of development, and with the lowest GtH titres observed during ovarian regression in both rivers. GtH content of the pituitary remained very high through the spawning period, with the highest titres observed in March in the Liao River, and in July in the Pearl River. As with plasma GtH, the lowest concentrations of pituitary GtH were observed during stages when the ovary was regressed. We analyzed the relationships between plasma and pituitary GtH and reproductive conditions, such as gonadosomatic index (GSI), hepatosomatic index (HSI), oocyte diameter (OD) and oocyte weight (OW). The results indicate that annual changes of GtH content of the plasma and pituitary paralleled GSI, OD and OW, but were negatively correlated with HSI in feral female catfish.     

Plasma levels of gonadotropin and 17α,20β-dihydroxy-4-pregnen-3-one in relation to spawning behaviour of rainbow trout, Oncorhynchus mykiss(Walbaum)

Manipulation of the opportunity to spawn was used to investigate the relationship between endocrine events, egg viability and spawning behaviour in female rainbow trout. Females were prevented from spawning by isolating them from males and gravel for up to 21 days after ovula- tion. Blood samples were taken before pairing with a male, at the onset of nesting activity, and at the completion of spawning. Plasma hormone levels of gonadotropin (GtH) and 17α,20β-dihydroxy-4-pregnen-3-one (17,2OP) were measured by specific radioimmunoassays. There were no qualitative or quantitative differences in the spawning behaviour of females paired on the day of ovulation or 7. 14, or 21 days after ovulation. There was a general decrease in the viability of eggs with increasing retention times. In females paired on the day of ovulation, or after 7 or 14 days, GtH levels increased with the onset of nesting behaviour and declined as fish reached the post-spawning condition. By day 21, GtH levels before pairing were significantly higher than prepairing levels in the other three treatment groups, and did not increase at the onset of nesting, or decrease in post-spawning fish. Plasma 17,20P remained high in prepairing and nesting samples of all four groups and declined to low levels in fish in post-spawning condition. In females paired on the day of ovulation there was a significant increase in 17,20P from the prepairing to the nesting stage. These results suggest that 17,20P plays a key role in the synchronization of behavioural and maturational events at the time of spawning.     

Seasonal fluctuation in lipid and cholesterol content of ovary, liver and blood serum in relation to annual sexual cycle in Heteropneustes fossilis (Bloch).

Seasonal fluctuation in lipid content of ovary and liver and cholesterol content of ovary, liver and serum in relation to annual reproductive cycle in H. fossilis has been worked out. Reduction in lipid content of liver was associated with decrease in hepatosomatic index (HSI) in prespawning and spawning phases. But there was an obvious increase in the lipid level of ovary in the above phases which coincided with enhanced gonadosomatic index (GSI) records. Serum and ovary showed a subsidence in their cholesterol contents in prespawning followed by increased level during spawning period. Substraction in liver cholesterol started in prespawning and continued till spawning phase and thereafter regained upward trend. Lipid and cholesterol levels in all organs studied along with GSI and HSI returned to initial values by the end of postspawning stage. Administration of clomid, sexovid, PGE1 and PGF2 alpha for a week did not induce any change in HSI, GSI, lipid and cholesterol values of liver and serum. However, these drugs were effective in suppressing the ovarian cholesterol within a week. The above drugs seem to affect cholesterol level indirectly since they trigger gonadotrophin secretion which in turn probably accelerated cholesterol utilization in ovary for increased steroid hormone output.     

Gonadal morphology, enzyme histochemistry and plasma steroid levels during the annual reproductive cycle of male and female brown bullhead catfish, Ictalurus nebulosus Lesueur

The annual reproductive cycle of the brown bullhead catfish, Ictalurus nebulosus Lesueur, was investigated over a two-year period. In females, GSI increased in the spring as follicles enlarged and the granulosa became hypertrophied, dropped during spawning in August, then rose in the autumn as follicles enlarged slightly. 3β-Hydroxysteroid dehydrogenase (3β-HSD) activity was limited to thecal nests of large, vitellogenic follicles. Plasma testosterone and estradiol-17β levels increased in parallel with GSI. Levels of both steroids dropped prior to the spawning period, although a peak in estradiol-17β was evident during the spawning period. No 11-ketotestosterone was detected in female plasma. In males, GSI increased in the spring as spermatogenesis proceeded, and dropped during spawning. 3β-HSD activity was confined to Leydig cells and was most intense prior to spawning. Plasma testosterone and 11-ketotestosterone peaked during the pre-spawning period, dropped prior to spawning, then rose slowly during the autumn. A peak in estradiol-17β occurred during the spawning period. Significant differences in GSI and plasma steroid levels during the pre-spawning and spawning periods were observed between the two yearly cycles; they may be related to differences in rainfall during these periods.     

Localization and dynamics of Mel1a melatonin receptor in the ovary of carp Catla catla in relation to serum melatonin levels

We studied the localization, sub-cellular distribution and daily rhythms of a 37 kDa melatonin receptor (Mel_1aR) in the ovary to assess its temporal relationship with the serum melatonin levels in four different reproductive phases in carp Catla catla. Our immunocytochemical study accompanied by Western blot analysis of Mel_1aR in the ovary revealed that the expression of this 37-kDa protein was greater in the membrane fraction than in the cytosol. Ovarian Mel_1aR protein peaked at midnight and fell at midday in each reproductive phase. Conversely, serum melatonin levels in the same fish demonstrated a minimum diurnal value at midday in all seasons, but a peak at midnight (during pre-spawning, spawning, and post-spawning phases) or at late dark phase (during preparatory phase). In an annual cycle, band intensity of Mel_1aR protein showed a maximum at night in the spawning phase and a minimum in the post-spawning phase, demonstrating an inverse relationship with the levels of serum melatonin. Our data provide first evidence of the presence of Mel_1a melatonin receptor in carp ovary and offer interesting perspectives especially for the study of the mechanisms of the control of its rhythmicity and its response to external factors.     

Seasonal Variations and Diurnal Rhythms of Gonadotropin (GtH2) Levels in Immature and Mature Female Carp (Cyprinus carpio L)

Daily changes of gonadotropin (GtH2) blood levels of immature and mature carp females were investigated in four seasons during the year: spring, summer (spawning period) and in autumn and winter (“resting” period). GtH2 levels were measured using ELISA method. The maturity stage of oocytes was determined histologically and on the base of germinal vesicle position in the oocyte. It appeared shown that GtH2 levels changed daily during the year in the range of 3.56 to 6.61 ng/ml, regardless of the state of ovarian maturity. Cosinor analysis and ellipse of errors revealed the presence of a circadian rhythm in GtH2 levels only in mature females during the reproduction period (L:D=16:8, water temp. 20 to 22°C).     

Dynamics of vitellogenin synthesis in juvenile giant freshwater prawn Macrobrachium rosenbergii

The dynamics of vitellogenin (Vg) mRNA expression and patterns of Vg and vitellin distribution in the hepatopancreas and ovary of juvenile Macrobrachium rosenbergii were examined using real-time RT-PCR and immunohistochemical methods. Eyestalk ablation was seen to induce rapid development of the gonads and Vg synthesis in females. In the female hepatopancreas, Vg mRNA expression was observed several days following ablation, after which levels increased gradually with increasing gonadosomatic index (GSI). Vitellin accumulation in the oocytes also increased with increasing Vg mRNA synthesis; expression was however negligible in the ovary. Hemolymph Vg levels in females ranged from 0.04 to 2.2 mg/ml. SDS PAGE/Western blotting analysis of hemolymph samples revealed that juvenile Vg was composed of 199 and 90 kDa subunits; the 102 kDa subunit present in adult female Vg (Okuno et al., 2002. J Exp Zool 292:417-429) could not be detected at any stage of vitellogenesis in juveniles. Vg was not detectable in non-ablated juveniles. The results of this study confirmed that the mode of involvement of eyestalk factors in regulating vitellogenesis is intrinsic to both juveniles and adults, and that a basic pattern of Vg synthesis and processing is conserved. However, the fact that juveniles are not able to produce the same Vg levels observed in adult females, and do not reach high GSI levels culminating in spawning suggests that other factors and physiological conditions specific to adult females are necessary to demonstrate full reproductive ability.     

Study of the sexual maturity of female bluefin tuna: purification and partial characterization of vitellogenin and its use in an enzyme-linked immunosorbent assay

Plasma steroid levels of female bluefin tuna (BFT) Thunnus thynnus rose from c. 1·5 ng ml⁻¹ during the quiescent period (March) to c. 7 ng ml⁻¹ during the ripening period (May). Testosterone (T) increased further to c. 8 ng ml⁻¹ during the pre-spawning period (June) while 17β-oestradiol (E₂) began to decrease. In the post-spawning period (August) steroid levels decreased to < 1 ng ml⁻¹. Vitellogenin (Vtg) plasma levels seemed to follow changes in E₂, showing an increase from the quiescent period to the ripening period of c. 18 mg ml⁻¹, decreasing slightly before spawning, and then decreasing after spawning. The Vtg content in plasma showed a good correlation both with the plasma levels of E₂ and T and with the percentage of vitellogenic oocytes at different periods of the reproductive cycle. Thus the ELISA could be taken as validated. Immunohistochemical staining of ovaries with anti BFT-Vtg serum demonstrated a high cross-reactivity with yolk proteins allowing the identification of vitellogenic oocytes.     

Comparative study of reproductive biology in single- and multiple-spawner cyprinid fish. I. Morphological and histological features

To clarify the dynamics and regulation of oogenesis in single- and multiple-spawning cyprinid fish with group-synchronous oocyte development, a multidisciplinary approach to their reproduction was undertaken using three species from the River Meuse (Belgium): the roach Rutilus rutilus as a single spawner, and the bleak Alburnus alburnus and the white bream Blicca bjoerkna as multiple spawners. The gonadosomatic index (GSI) and histomorphometric changes (distribution of oocyte size, relative proportion of the various oocyte stages) in the ovary are compared. Different patterns of GSI and oocyte growth were observed both between the single- and multiple-spawner fish and between the two multiple spawners. Maximum GSIs were higher in roach (21%) than in bleak and white bream (17.7 and 14.5%, respectively), and compared to the rapid decline of GSI in the roach population, the GSI of multiple spawners decreased progressively during the spawning season. In roach, a short gonadal quiescent period and an early onset of vitellogenesis was recorded from late summer onwards whereas, in bleak and white bream, exogenous vitellogenesis was not systematically observed before winter. A protracted spawning season and/or a low water temperature in autumn are hypothesized to explain this long period of gonadal quiescence. In bleak, during the spawning season, the oocytes recruited arose from the stock of endogenous vitellogenesis and attained the final maturation stage very rapidly. This recruitment occurred during the whole spawning season. In white bream, the differentiation of vitellogenic oocytes from smaller oocytes was completed before the onset of the spawning season. During the spawning period, the proportion of vitellogenic oocytes decreased progressively whereas the percentage of oocytes in the final maturation stage remained approximately constant.     

Melatonin in the regulation of annual testicular events in carp Catla catla: evidence from the studies on the effects of exogenous melatonin, continuous light, and continuous darkness

The physiological significance of melatonin in the regulation of annual testicular events in a major carp Catla catla was evaluated through studies on the effects of graded dose (25, 50, or 100 µg/100 g body wt.) of melatonin exogenously administered for different durations (1, 15, or 30 days) and manipulation of the endogenous melatonin system by exposing the fish to constant darkness (DD) or constant light (LL) for 30 days. An identical experimental schedule was followed during the preparatory (February-March), pre-spawning (April-May), spawning (July-August), and post-spawning (September-October) phases of the annual cycle. Irrespective of the reproductive status of the carp, LL suppressed while DD increased the mid-day and mid-night values of melatonin compared to respective controls. Influences of exogenous melatonin varied in relation to the dose and duration of treatment and the reproductive status of the carp. However, testicular response to exogenous melatonin (at 100 µg, for 30 days) and DD in each reproductive phase was almost identical. Notably, precocious testicular maturation occurred in both DD and melatonin-injected fish during the preparatory phase and in LL carps during the pre-spawning phase. In contrast, testicular functions in both the melatonin-treated and DD fish were inhibited during the pre-spawning and spawning phases, while the testes did not respond to any treatment during the post-spawning phase. In conclusion, this study provided the first experimental evidence that melatonin plays a significant role in the regulation of annual testicular events in a sub-tropical surface-dwelling carp Catla catla, but the influence of this pineal hormone on the seasonal activity of testis varies in relation to the reproductive status of the concerned fish.     

Enzyme activities in the blood plasma of grayling, Thymallus thymallus (Linn.), in the breeding season

Activities of alanine aminotransferase, aspartate aminotransferase, glutamate dehydrogenase and creatine kinase were determined in the blood plasma of grayling, Thymallus thymallus , in the pre-spawning, spawning and post-spawning periods. Enzyme activities increased to a maximum in the spawning period and then successively decreased, after spawning, to values close to the initial ones. Only in the case of aspartate aminotransferase did the increased activity level persist after spawning. The temporary but large increase in enzyme activity levels in the plasma of grayling appeared to be a natural and physiological phenomenon accompanying the spawning process.     

Seasonal Variations and Diurnal Rhythms of Gonadotropin (GtH2) Levels in Immature and Mature Female Carp (Cyprinus carpio L)

Daily changes of gonadotropin (GtH2) blood levels of immature and mature carp females were investigated in four seasons during the year: spring, summer (spawning period) and in autumn and winter (“resting” period). GtH2 levels were measured using ELISA method. The maturity stage of oocytes was determined histologically and on the base of germinal vesicle position in the oocyte. It appeared shown that GtH2 levels changed daily during the year in the range of 3.56 to 6.61 ng/ml, regardless of the state of ovarian maturity. Cosinor analysis and ellipse of errors revealed the presence of a circadian rhythm in GtH2 levels only in mature females during the reproduction period (L:D=16:8, water temp. 20 to 22°C).