Effects of Estradiol and Testosterone in the Hagfish Eptatretus burgeri

Effects of exogenous estradiol and testosterone on the gonad and pituitary were examined in the hagfish. Estradiol caused degeneration of the ovary. Testosterone had no effect on the ovary. Neither steroid had a significant effect on the testis. Neither estradiol nor testosterone provoked a conspicuous change in adenohypophysial cells. Therefore, a feedback mechanism between gonadal steroids and gonadotropin either does not exist or plays a very minor role. The degeneration of the ovary may be due to a pharmacological effect of estradiol.     

Ovulation and Eggs of the Hagfish Eptatretus burgeri

Abstract Ripe females and males of Eptatretus burgeri were kept in cages both in the sea and in the aquaria for about 2 months of the normal spawning period around October. All the 9 ripe females in the sea ovulated all their eggs (200) and deposited 178 of them. Only 5 eggs were ovulated and deposited in the aquaria, although there were 17 ripe females and some of them were injected with various hormones in an effort to induce ovulation. No eggs were fertilized. A peculiar surface pattern of hexagonal ridges is described around the entrance of the micropylar canal on the bottom of the micropylar funnel in the ovulated eggs.     

Spermiogenesis in the Hagfish Eptatretus burgeri (Agnatha)

The fine structure of spermatid differentiation in a primitive vertebrate, the hagfish, whose spermatozoa bear acrosomes, was investigated. In early round spermatids, the acrosomal vesicles were spherical and located in a shallow nuclear indentation, flanked by the plasma and the nuclear membranes. The vesicle underwent a transition through lens-shaped and cap-shaped stages until it attained the shape of a bell in mature spermatozoa. Electron-dense acrosomal material that appeared as deposits in three portions of the vesicle finally joined in the center region at a late stage. Condensation of chromatin occurred in the anterior region of the nucleus. During transformation of the spermatids, many regularly spaced microtubules appeared beneath the plasma membrane except in the anteriormost region of the cell. The microtubules in a single alignment lay parallel to one another and encased the nucleus diagonally. During an early stage, the centrioles changed their orientation from perpendicular to longitudinal and rotated to become parallel to the long axis of the nucleus. Thus, the flagellum lay nearly straight along the cell axis. A cytoplasmic canal appeared transiently during the early stage. A droplet of cytoplasm was eliminated after descending along the flagella. The features of spermiogenesis in hagfish, which lies between invertebrates and vertebrates, are compared with those of other animals.     

Cyclical Changes in the Testis of the Hagfish Eptatretus burgeri (Cyclostomata)

The development of the spermatogenetic stages in the testis of the hagfish Eptatretus burgeri was studied by means of histological techniques. These animals have an annual reproductive cycle with a spawning season in October. Spermatocytes begin to appear in January and reach a maximum in March. The development of spermatids of stage I and stage II starts in April and June, respectively. In August about 35% of the testicular follicles contain spermatids of stage II. The gonadosomatic index shows a more or less constant increase from November until August. During the spawning season, no animals with well developed testis could be trapped. About 10% of male hagfish bear undeveloped gonads.     

Cyclical Changes in the Ovary of the Hagfish Eptatretus burgeri (Cyclostomata)

Abstract The development of eggs and the degeneration of ovulated follicles in Eptatretus burgeri was studied over a period of one year. It was found that small resting eggs with a diameter of about 1.5 mm continue to grow in November. By the spawning season in October of the following year, the follicles attain a length of about 24 mm. Only one fertilized egg containing an embryo of 7.5 mm length was obtained. The ratio of males to females was found to vary throughout the year. The annual increase of body length in correlation to ovarian development in the hagfish is calculated to be 4 to 5 cm.     

Circadian Rhythms of Locomotor Activity in the Nile Tilapia Oreochromis niloticus

Behavioral rhythms of the Nile tilapia were investigated to better characterize its circadian system. To do so, the locomotor activity patterns of both male and female tilapia reared under a 12:12 h light-dark (LD) cycle were studied, as well as in males the existence of endogenous rhythmicity under free-running conditions (DD and 45 min LD pulses). When exposed to an LD cycle, the daily pattern of activity differed between individuals: some fish were diurnal, some nocturnal, and a few displayed an arrhythmic pattern. This variability would be typical of the plastic circadian system of fish. Moreover, reproductive events clearly affected the behavioral rhythms of female tilapia, a mouth-brooder teleost species. Under DD, 50% (6 of 12) of male fish showed circadian rhythms with an average period (τ) of 24.1±0.2 h, whereas under the 45 min LD pulses, 58% (7 of 12) of the fish exhibited free-running activity rhythms with an average τ of 23.9±0.5 h. However, interestingly in this case, activity was always confined to the dark phase. Furthermore, when the LD cycle was reversed, a third of the fish showed gradual resynchronization to the new phase, taking 7–10 days to be completely re-entrained. Taken together, these results suggest the existence of an endogenous circadian oscillator that controls the expression of locomotor activity rhythms in the Nile tilapia, although its anatomical localization remains unknown.     

A Scanning Electron Microscopic Study on the Thyroid Follicle of the Hagfish Eptatretus burgeri

The external form of the follicles was mostly spherical or oval and the large oval follicles reached more than 300 times 180 μm in size. Cytoplasmic inclusions of various sizes were observed in the follicular epithelium. Follicles were composed of hexagonal epithelial cells, and on the apical surface of the cells many microvilli were found (380–500 microvilli per cell) with globular or amorphic cytoplasmic protrusions. In a few cells a central cilium was also observed. From these results a relationship between thyroid structure and function in the hagfish is discussed.     

Sex Difference in the Cloacal Gland in the Hagfish,Eptatretus burgeri,and its Possible Significance in Reproduction

The cloacal gland of the hagfish, Eptatretus burgeri, was studied histologically in four seasons. Before the breeding season (spring and summer) the male cloacal gland was larger than the female cloacal gland. The gland was largest in the males with a more mature testis. After the breeding season (autumn and winter) there was no sex difference in the size of the cloacal gland. The cloacal gland consists of mucus and thread cells, as do the lateral slime glands. The sperm may be entangled within slime in the cloacal gland and be shed outside as a slimy sperm mass. Such a slimy sperm mass may play an important role in reproduction of the hagfish.     

Stocking Density Affects Circadian Rhythms of Locomotor Activity in African Catfish,Clarias gariepinus

The effect of stocking density on the locomotor activity of African catfish C. gariepinus under different light regimes was investigated. C. gariepinus were stocked under different densities (1, 5, or 10 fish/tank), and their locomotor activity recorded under light-dark (LD), constant light (LL), constant darkness (DD), and LD-reversed (DL) regimens. Under the LD cycle, catfish showed a crepuscular activity pattern, irrespective of stocking density, with most of the daily activity concentrated around the light-onset and light-offset times. When fish were subjected to DD, all 4 tanks with medium (5 fish) and high (10 fish) stocking densities showed circadian rhythmicity, with an average period (τ) of 23.3?±?0.5 and 24.6?±?0.5?h, respectively. In contrast, only 2 low (1 fish) density tanks showed free-running rhythms. Under LL, activity levels decreased significantly in comparison with levels observed under LD and DD. Moreover, fish of 1, 2, and 3 out of the 4 tanks with low, medium, and high densities, respectively, showed free-running rhythms under these conditions. When the photocycle was reversed (DL), fish of 3, 2, and 4 out of the 4 tanks with low, medium, and high stocking densities, respectively, showed gradual resynchronization to the new phase, and transient cycles of activity were observed. These results suggest that stocking density of fish affected the display of circadian rhythmicity and the intensity of activity levels. Thus, fish kept in higher densities showed more robust rhythmicity and higher levels of daily activity, indicating that social interactions may have an influence on behavioral patterns in the African catfish. (Author correspondence: lmvera@um.es)     

Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila

Most life forms exhibit daily rhythms in cellular, physiological and behavioral phenomena that are driven by endogenous circadian (≡24 hr) pacemakers or clocks. Malfunctions in the human circadian system are associated with numerous diseases or disorders. Much progress towards our understanding of the mechanisms underlying circadian rhythms has emerged from genetic screens whereby an easily measured behavioral rhythm is used as a read-out of clock function. Studies using Drosophila have made seminal contributions to our understanding of the cellular and biochemical bases underlying circadian rhythms. The standard circadian behavioral read-out measured in Drosophila is locomotor activity. In general, the monitoring system involves specially designed devices that can measure the locomotor movement of Drosophila. These devices are housed in environmentally controlled incubators located in a darkroom and are based on using the interruption of a beam of infrared light to record the locomotor activity of individual flies contained inside small tubes. When measured over many days, Drosophila exhibit daily cycles of activity and inactivity, a behavioral rhythm that is governed by the animal''s endogenous circadian system. The overall procedure has been simplified with the advent of commercially available locomotor activity monitoring devices and the development of software programs for data analysis. We use the system from Trikinetics Inc., which is the procedure described here and is currently the most popular system used worldwide. More recently, the same monitoring devices have been used to study sleep behavior in Drosophila. Because the daily wake-sleep cycles of many flies can be measured simultaneously and only 1 to 2 weeks worth of continuous locomotor activity data is usually sufficient, this system is ideal for large-scale screens to identify Drosophila manifesting altered circadian or sleep properties.     

Ependymal Absorption of Peroxidase into the Third Ventricle of the Hagfish Eptatretus burgeri (Girard)

Abstract The ependymal cells of both dorsal and ventral walls lining the lumen of the neurohypophysis of the hagfish, Eptatretus burgeri, absorb peroxidase injected into the third ventricle. Peroxidase diffuses rapidly into the connective tissue separating the neurohypophysis from the adenohypophysis, and also into the connective tissue located between the cell nests of the adenohypophysis.     

Circadian Rhythms of Self-feeding and Locomotor Activity in Zebrafish (Danio Rerio)

To investigate daily feeding rhythms in zebrafish, the authors have developed a new self-feeding system with an infrared photocell acting as a food-demand sensor, which lets small-size fish such as zebrafish trigger a self-feeder. In this paper, the authors used eight groups of 20 fish. Locomotor activity rhythms were also investigated by means of infrared sensors. Under a 12?h:12?h light (L)-dark (D) cycle, zebrafish showed a clear nocturnal feeding pattern (88.0% of the total daily food-demands occurring in the dark phase), concentrated during the last 4?h of the dark phase. In contrast, locomotor activity was mostly diurnal (88.2% of total daily activity occurring in the light phase). Moreover, both feeding and locomotor rhythms were endogenously driven, as they persisted under free-running conditions. The average period length (τ) of the locomotor and feeding rhythms was shorter (τ?=?22.9?h) and longer (τ?=?24.6?h) than 24?h, respectively. During the time that food availability was restricted, fish could only feed during ZT0–ZT12 or ZT12–ZT16. This resulted in feeding activity being significantly modified according to feeding time, whereas the locomotor activity pattern remained synchronized to the LD cycle and did not change during this trial. These findings revealed an independent phasing between locomotor and feeding activities (which were mostly nocturnal or diurnal, respectively), thus supporting the concept of multioscillatory control of circadian rhythmicity in zebrafish. (Author correspondence: javisan@um.es)     

Effect of Thyrotropin-releasing Hormone on the Thyroid of a Teleost,Chasmichthys dolichognathus,and a Hagfish,Eptatretus burgeri

Abstract Intraperitoneal injection of mammalian thyrotropin-releasing hormone (TRH) caused hypertrophy of possible thyrotropic cells in the pituitary gland of the teleost, Chasmichthys dolichognathus. The epithelial cells of the thyroid follicles also appeared hypertrophic after the TRH injection. On the other hand, in the hagfish, Eptatretus burgeri, TRH injection had no effect either on the cytological appearance of the adenohypophysis or on the thyroidal activity in terms of serum thyroxine concentration and epithelial cell height.     

Circadian Locomotor Activity Rhythms in the African Clawed Frog,Xenopus laevis: The Role of the Eye and the Hypothalamus

The effects of hypothalamic lesioning and removal of the eyes on locomotor activity rhythms of African clawed frog, Xenopus laevis were examined under light-dark cycles (LD12:12) and constant conditions. Frogs were kept individually and the activity rhythms at the bottom layer of water tank were recorded by means of the infrared photocells. Intact frogs displayed clear entrained nocturnal activity and expressed freerunning activity rhythms in constant darkness (DD), while some frogs did not freerun under co nstant dim light (dimLL) and constant light (LL). Freerunning periods in intact frogs were significantly shorter in dimLL than in DD. Although freerunning periods were shortened after blinding in same individuals, no significant changes in the freerunning periods were observed after blinding under dimLL and LL. When electrolytic lesions to the hypothalamus were performed, all frogs with more than 70% damage of the SCN abolished freerunning rhythms and in frogs with less than 70% damage, 57% of the animals became arrhythmic. In conclusion, (1) There is a circadian pacemaker somewhere outside the eyes, and it is probably situated in the hypothalamusincluding the SCN. (2) Both the eyes and the SCN are involved in the circadian system of the frogs.     

Circadian Locomotor Activity Rhythms in the African Clawed Frog, Xenopus laevis: The Role of the Eye and the Hypothalamus

The effects of hypothalamic lesioning and removal of the eyes on locomotor activity rhythms of African clawed frog, Xenopus laevis were examined under light-dark cycles (LD12:12) and constant conditions. Frogs were kept individually and the activity rhythms at the bottom layer of water tank were recorded by means of the infrared photocells. Intact frogs displayed clear entrained nocturnal activity and expressed freerunning activity rhythms in constant darkness (DD), while some frogs did not freerun under co nstant dim light (dimLL) and constant light (LL). Freerunning periods in intact frogs were significantly shorter in dimLL than in DD. Although freerunning periods were shortened after blinding in same individuals, no significant changes in the freerunning periods were observed after blinding under dimLL and LL. When electrolytic lesions to the hypothalamus were performed, all frogs with more than 70% damage of the SCN abolished freerunning rhythms and in frogs with less than 70% damage, 57% of the animals became arrhythmic. In conclusion, (1) There is a circadian pacemaker somewhere outside the eyes, and it is probably situated in the hypothalamusincluding the SCN. (2) Both the eyes and the SCN are involved in the circadian system of the frogs.     

Circadian Rhythms and time Measurement in Locomotor Activity of the Scorpion Leiurus Quinquestriatus (Buthidae)

The circadian rhythms of locomotor activity of the scorpion Leiurus quinqueslriatus were examined under different light-dark cycles and in free-running conditions. The circadian rhythm is bimodal in LD 12:12 with alternating cycles of temperature (35°-25°C) with high intensity (1300 lux) or in LD 12: 12 with constant temperature 35° C with 300 lux. In LD 12:12 (1300 lux), in long or in short light spans with constant temperature, the bimodal pattern is slightly changed with the appearance of a third minor peak of activity. In free-running conditions, the bimodal rhythm of locomotor activity persists in DD with T about 24 hr, but in LL the rhythm becomes unimodal with T about 24 hr. Cosinor and power spectrum analysis showed the presence of more than one periodic component. It seems that there is a correlation between the range of light regimens, temperature, light intensity and the coincidence of these components. These components are independently entrained by the environmental light cycle. The mechanism of entrainment of components is discussed.     

Circadian Rhythms of Locomotor Activity in Lycosa tarentula (Araneae, Lycosidae) and the Pathways of Ocular Entrainment

Lycosa tarentula is a ground-living spider that inhabits a burrow where it awaits the appearance of prey or conspecifics. In this study, circadian rhythms of locomotor activity were examined as well as the ocular pathway of entrainment. Thirty-three adult virgin females were examined under constant darkness (DD); all of them exhibited robust circadian rhythms of locomotor activity with a period averaging 24.1h. Fourteen of these spiders were studied afterwards under an LD 12:12 cycle; they usually entrained to in the first or second day, even when the light intensity was as low as 1 lx. During the LD cycle, locomotor activity was generally restrained to the darkness phase, although several animals showed a small amount of diurnal activity. Ten males were also examined under LD; they were also nocturnal, but were much more active than the females. Seven females were examined under constant light (LL); under this they became arrhythmic. Except for the anterior median eyes (OMAs), all the eyes were capable of entraining the locomotor activity to an LD cycle. These results demonstrate that under laboratory conditions and low light intensities locomotor activity of Lycosa tarentula is circadian and in accordance with Aschoff's 'rule'. Only OMAs are unable to entrain the rhythm; the possible localization of circadian clock is therefore discussed.     

Circadian Rhythms of Locomotor Activity in Lycosa tarentula (Araneae,Lycosidae) and the Pathways of Ocular Entrainment

Lycosa tarentula is a ground-living spider that inhabits a burrow where it awaits the appearance of prey or conspecifics. In this study, circadian rhythms of locomotor activity were examined as well as the ocular pathway of entrainment. Thirty-three adult virgin females were examined under constant darkness (DD); all of them exhibited robust circadian rhythms of locomotor activity with a period averaging 24.1h. Fourteen of these spiders were studied afterwards under an LD 12:12 cycle; they usually entrained to in the first or second day, even when the light intensity was as low as 1 lx. During the LD cycle, locomotor activity was generally restrained to the darkness phase, although several animals showed a small amount of diurnal activity. Ten males were also examined under LD; they were also nocturnal, but were much more active than the females. Seven females were examined under constant light (LL); under this they became arrhythmic. Except for the anterior median eyes (OMAs), all the eyes were capable of entraining the locomotor activity to an LD cycle. These results demonstrate that under laboratory conditions and low light intensities locomotor activity of Lycosa tarentula is circadian and in accordance with Aschoff's 'rule'. Only OMAs are unable to entrain the rhythm; the possible localization of circadian clock is therefore discussed.     

Cyclical Changes in Weight and Fat Content of the Liver and their Relationship to Reproduction in the Hagfish Eptatretus burgeri (Cyclostomata)

Patzner, R. A. 1980. Cyclical changes in weight and fat content of the liver and their relationship to reproduction in the hagfish Eptatretus burgeri (Cyclostomata). (Zoological Institute, University of Salzburg, Austria and Misaki Marine Biological Station, University of Tokyo, Japan.) — Acta zool. (Stockh.) 61(3): 157–160. The changes in weight and fat content of the liver in the West Pacific hagfish Eptatretus burgeri were studied over a period of one year. The liver somatic index shows a high increase from the beginning of December until migration in deeper water in July. From the beginning of April to the end of August the difference between male and female liver somatic index is statistically significant. This fact is brought in connection with the production of vitellogenin in the female liver.