CONTEMPORARY ISOLATION-BY-DISTANCE, BUT NOT ISOLATION-BY-TIME, AMONG DEMES OF EUROPEAN GRAYLING (THYMALLUS THYMALLUS, LINNAEUS) WITH RECENT COMMON ANCESTORS

The development of isolation by distance (IBD) and isolation by time (IBT) was contrasted among demes of European grayling ( Thymallus thymallus ) that have diverged within the last 25 generations following colonization of a lake (Lesjaskogsvatnet). We find low but significant levels of genetic differentiation among spawning tributaries and a pattern of IBD among them. We do not, however, find evidence for IBT despite an up to four-week difference in spawning date between "warm/early" and "cold/late" spawning demes and differences in the incubation temperatures experienced by offspring. It appears that IBD has developed more rapidly than IBT in this system and that adaptive divergence has been initiated in the absence of IBT. Although analysis of selected loci could reveal reduced recombination in parts of the genome associated with temporal divergence, our analysis of neutral genetic data suggests that IBD is a more important isolating mechanism in the early stages of adaptive divergence in European grayling.     

The migration of spawning stocks of graylingThymallus thymallus, in Lake Mjøsa, Norway

Synopsis The migration of grayling was studied in Lake Mjøsa and 13 of its tributaries in which grayling spawn. The study demonstrate that grayling spawn in creeks in May/June and leave for Lake Mjøsa shortly after spawning to return the next spring. Once mature, the grayling spawn every year. Grayling mix in the lake and are found at a mean distance of 11.9 km from the estuary of their spawning creek in September. Grayling embryos hatch in June and the juveniles stay in the tributaries until September/October. Grayling spend the winter months in the lake. Out of 1599 grayling tagged in tributaries during the spawning season, 240 of 284 recaptures (84.5%) were made in subsequent spawning seasons in the same tributary in which they were first tagged. Forty-four grayling were recaptured in adjacent tributaries. These results demonstrate that, despite the fact that all grayling leave the tributaries and mix in the lake, there is a high precision of homing. It is suggested that the grayling population in lake Mjøsa is composed of stocks using specific tributaries for spawning.     

Annual changes in fine structure of inner epithelial lining of the ovary of a marine sculpin,Alcichthys alcicornis (Teleostei: Scorpaeniformes), with internal gametic association

Alcichthys alciocornis has a viscous ovarian fluid in the ovarian cavity, which plays an important role in its unique mode of reproduction called internal gametic association (i.e., internal insemination and sperm-egg association but a delay in the physiological fertilization until spawning). Seasonal changes in fine structure of the inner epithelial lining and capillary endothelium of the ovary revealed that ovarian fluid originated as a result of the secretory activity of the tissues. The ovarian cavity of A. alcicornis is lined with an ovigerous lamella epithelium and an ovarian wall epithelium. During the spawning period, both epithelia actively secreted proteinaceous substances which seemed to constitute the ovarian fluid. The substances appear to be synthesized in the rough-surfaced endoplasmic reticulum from the material which was transported from the blood capillary, taken into the epithelial cells by endocytosis, accumulated in secretory vesicles via Golgi apparatus in the cells, and finally released into the ovarian cavity by exocytosis. Microapocrine secretion was also observed to occur in both epithelia. Secretory activity of both epithelia by exocytosis and microapocrine secretion showed distinct seasonal changes. Active exocytosis and microapocrine secretion were observed during the spawning period (April–May). These activities slightly declined during the degeneration period (May–June) and were lost during the early recovery period (July). During the mid to late recovery period (October–March), there was some exocytosis but no microapocrine secretion. © 1995 Wiley-Liss, Inc.     

Cytology of the interstitial tissue in scrotal and abdominal testes of post-puberal boars

The interstitial tissue of the testes from healthy boars, and unilateral and bilateral abdominal cryptorchid boars was examined by light and transmission electron microscopy. The left and right testes of healthy boars, and the left (scrotal) testis of unilateral cryptorchid boars had abundant mature Leydig cells, few fibroblasts and mast cells, scarce and small blood vessels, and little lymphatic areas. The right (abdominal) testis of unilateral cryptorchid boars contained abundant Leydig cells, fibroblasts and erythrocytes, scarce mast cells, and frequent blood vessels; Leydig cells exhibited either a mature but degenerative appearance or an immature appearance, and fibroblasts displayed immaturity signs. The interstitial tissue of the left (abdominal) testes of bilateral cryptorchid boars had small blood vessels surrounded by erythrocytes, lymphocytes, and few plasma cells, and abundant mature and immature Leydig cells, immature fibroblasts, and mast cells. Mature Leydig cells showed mid or advanced degeneration, and immature Leydig cells displayed either non-degenerative or degenerative patterns. The right (abdominal) testes of bilateral cryptorchid boars contained scarce immature Leydig cells in advanced degeneration, large fibrous and adipose areas, and blood vessels. These results indicated that unilateral abdominal cryptorchidism affect neither the structural nor the cytologic features of the interstitial tissue in scrotal testes. Unilateral and bilateral cryptorchidism induced abnormal differentiation of Leydig cells and fibroblasts leading to decreased steroid production and increased collagenization in abdominal testes.     

A preliminary investigation of spawning migrations of grayling in a small stream as determined by radio-tracking

Adult grayling Thymallus thymallus migrated from 230 to 4980 m up the Aisne stream, Belgium, to spawn between 18 and 29 March, under decreasing floods, increasing temperature and low turbidity. Males ( n =4) arrived on spawning grounds several days earlier than females ( n =2), stayed there longer (10–19 v. 2–3 days), and occupied a single ground each, whereas females moved between several places. After spawning, all grayling homed precisely into the pool-riffle sequences where they were tagged in late February, and remained here until late June. These observations indicate that resident grayling are far less mobile than autumn-spawning salmonids, and that the environmental factors triggering spawning migrations resemble more closely those of spring-spawning cyprinids than of other salmonids. The implications of these restricted mobility patterns are discussed within the scope of population structure, and impact of river management.     

Morphologic study of the testes from spontaneous unilateral and bilateral abdominal cryptorchid boars

Macroscopical and histological characteristics were examined in both testes from three healthy boars, three boars with unilateral abdominal cryptorchidism on the right side, and three boars with bilateral abdominal cryptorchidism. Abdominal cryptorchidism, unilateral and bilateral, provoked a significant decrease of the weight and volume of the ectopic testes. The scrotal testis of the unilateral cryptorchid boars showed an increase in its volume and weight. Cryptorchidism also induced abnormalities in the histological structure of seminiferous tubules, lamina propria, and interstitial tissue of the abdominal testes. The number of seminiferous tubules decreased; the seminiferous epithelium was constituted by few spermatogonia with an atypical pattern and by abnormal Sertoli cells. The lamina propria showed a variable degree of thickening and collagenization. The interstitial tissue was very developed but displayed a decrease in the Leydig cell population. These abnormalities were more critical in bilateral cryptorchidism than in unilateral cryptorchidism. The scrotal testis of the unilateral cryptorchid boars showed normal appearance, but a decrease of the number of seminiferous tubules was observed. Moreover, the seminiferous tubules showed impaired spermatid maturation. The alterations observed in the abdominal testes of the unilateral and bilateral cryptorchid boars were attributed to defective proliferation and differentiation of Sertoli cells and Leydig cells. The anomalies in the scrotal testis of the unilateral cryptorchid boars were due to disturbances in the Sertoli cell activity.     

The testicular cycle of a percoid teleost--Nandus nandus (Ham.).

Small paired testes of Nandus nandus are situated posteriorly in the body cavity. They open posteriorly in a common sperm duct. A urinogenital sinus is present. Each testis consists of a large number of seminiferous tubules extending from the periphery towards the centre. The seminiferous tubules are separated from each other by a layer of interstitial tissue. 6 different stages of spermatogenesis are recognised. On the basis of morphological and histological changes in the testes during different months of the year, the reproduvtice cycle has been divided into post-spawning (October to December), pre-spawning (January to March) and spawning (April to August) periods. The monthly volume of testes is in direct correlation with the monthly changes in water temperature. Statistical observations indicate that the process of spermatogenesis is very active during pre-spawning period. The relative number of spermatozoa is maximum in July (69.89%), suddenly decreases in August (54.28%) and continues to decrease upto October (49.66%) indicating the maximum spawning in July and August.     

Testis transplantation in male rainbow trout (Oncorhynchus mykiss)

The objective of the present study was to establish a procedure for the transplantation of an intact testis from one male rainbow trout (Oncorhynchus mykiss) to another individual and evaluate the reproductive function of the transplanted testis at sexual maturity. Isogenic (cloned) male rainbow trout were produced by crossing a completely homozygous male (YY) with a homozygous female (XX) to eliminate any problem of tissue rejection. Transplantation was performed on four pairs of sexually immature animals (n = 8); each served both as a donor and recipient. The left testis was removed by making a ventral midline incision to expose the body cavity and gonads. The left testis was disconnected at the anterior and posterior points of attachment and transferred to the recipient fish where it was placed in position adjacent to the pyloric cecae. The right testis was left intact. After 4 wk, the fish were injected (i.p.) twice weekly for 8 or 9 wk with salmon pituitary extract (1.5 mg/kg) to induce precocious sexual maturation. A similar number of untreated fish were maintained as controls. Following this treatment, all the fish were killed, and the right (intact) and left (transplanted) testes were removed, weighed, and sampled for sperm. Although the mean weights of the left, transplanted testes were significantly (P: < 0.05) smaller than the intact testes (transplants = 1.2 g; intact = 3.9 g), transplanted testes were present in all animals, had increased in mass, and were sexually mature containing sperm. The mean fertility, as measured by the proportion of eggs completing first cleavage, of sperm derived from transplanted testes (92%) was no different from the sperm obtained from intact testes (84%). Similarly, there was no difference in the number of embryos attaining the eyed stage of development, after 18 days of incubation, that were derived from transplanted (84%) or intact testes (85%).     

Synchronous reproduction of corals in the Red Sea

Multi-species synchronous spawning was first described on reefs off the east and west coast of Australia. In contrast, locally abundant species in the northern Red Sea and the central Pacific have little overlap in the time of reproduction. Consequently, the idea developed that high levels of spawning synchrony both within and among species was largely confined to Australian reefs. Here, we show that gamete maturity in colonies of the genus Acropora was highly synchronous in the Red Sea. In early April 2008, at two locations separated by 300 km, 13 of 24 species sampled had mature colonies, and a further 9 species had immature colonies. In late April–early May 2008, all colonies sampled had no oocytes, indicating colonies had spawned a few days after the full moon of 20 April 2008. Similarly, in 2009, 99% of colonies from 17 species at Hurghada were mature in late April, and all were empty in early May. Spawn slicks suggested many of these colonies had released gametes three night prior to the full moon on 8 May 2009. This level of synchrony in gamete maturity is among the highest ever recorded and similar to that typically recorded in Acropora assemblages on Australian reefs. While further work is required to document the night of gamete release, these data strongly suggest that high levels of spawning synchrony are a regular feature of these Red Sea coral assemblages and that multi-species spawning occurs on or around the full moon in April and/or May.     

Characterization of high mobility group protein levels during spermatogenesis in the rat

The distribution, quantitation, and synthesis of high mobility group (HMG) proteins during spermatogenesis in the rat have been determined. HMG1, -2, -14, and -17 were isolated from rat testes by Bio-Rex 70 chromatography combined with preparative gel electrophoresis. Amino acid analysis revealed that each rat testis HMG protein was similar to its calf thymus analogue. Tryptic peptide maps of somatic and testis HMG2 showed no differences and, therefore, failed to detect an HMG2 variant. Testis levels of HMG proteins, relative to DNA content, were equivalent to other tissues for HMG1 (13 micrograms/mg of DNA), HMG14 (3 micrograms/mg of DNA), and HMG17 (5 micrograms/mg of DNA). The testis was distinguished in that it contained a substantially higher level of HMG2 than any other rat tissue (32 micrograms/mg of DNA). HMG protein levels were determined from purified or enriched populations of testis cells representing the major stages of spermatogenesis; spermatogonia and early primary spermatocytes, pachytene spermatocytes, early spermatids, and late spermatids; and testicular somatic cells. High levels of HMG2 in the testis were due to pachytene spermatocytes and early spermatids (56 +/- 4 and 47 +/- 6 micrograms/mg of DNA, respectively). Mixtures of spermatogonia and early primary spermatocytes showed lower levels of HMG2 (12 +/- 3 micrograms/mg of DNA) similar to proliferating somatic tissues, whereas late spermatids had no detectable HMG proteins. The somatic cells of the testis, including isolated populations of Sertoli and Leydig cells, showed very low levels of HMG2 (2 micrograms/mg of DNA), similar to those in nonproliferating somatic tissues. HMG proteins were synthesized in spermatogonia and primary spermatocytes, but not in spermatids. Rat testis HMG2 exhibited two bands on acid-urea gels. A "slow" form comigrated with somatic cell HMG2, while the other "fast" band migrated ahead of the somatic form and appeared to be testis-specific. The "fast" form of HMG2 accounted for the large increase of HMG2 levels in rat testes. These results show that the very high level of HMG2 in testis is not associated with proliferative activity as previously hypothesized.     

Cytological evaluation of spermatogenesis within the germinal epithelium of the male European wall lizard, Podarcis muralis

The annual cytological changes to the male germinal epithelium were investigated in an introduced population of European wall lizards (Podarcis muralis). Testicular tissues were collected, embedded, sectioned by an ultramicrotome, and stained with the PAS procedure followed by a toluidine counterstain. Spermatogenesis in the lizard is divided into the proliferative, meiotic, and maturational phases. Wall lizards have a prenuptial pattern of spermatogenesis, where sperm development begins immediately prior to and continues through the months of breeding (April-June). The testis then involutes, undergoes a short period of quiescence, and recrudescence commences in mid-July. Germ cells undergo proliferation, meiosis, and the early stages of spermiogenesis (maturation) from late July through December. However, the late stages of spermiogenesis are retarded from December through February. Spermiogenesis continues at an accelerated pace from March through May, leading to a single massive spermiation event through the month of June. Although spatial relationships are seen between germ cells within the seminiferous epithelium, accumulation of spermatids during winter and acceleration of elongation in spring prevents determination of consistent cellular associations between early and late developing germ cells within the wall lizard testis. This temporal germ cell development is different from the consistent spatial development seen within seasonally breeding birds and mammals and may represent an evolutionary intermediate in terms of amniotic germ cell development.     

Expression of p57 in mouse and human testes

The expression of cyclin-dependent kinases inhibitors, p57kip2, was investigated during the postnatal development of mouse testis, and in adult human testis. Expression of p57kip2 mRNA was higher in immature than pubertal or adult mouse testes. In postnatal day 7 (PND7) testes, moderate p57kip2 immunoreactivity was found in spermatogonia, but signal was heterogeneous among the spermatogonia. In PND14 testes onward, strong immunoreactivity of p57kip2 was found in the nuclei of early spermatocytes but not in the late pachytene stage onward. In PND28 and PND50 testes, p57kip2 immunoreactivity was varying among the seminiferous tubules. There was no visible signal in late pachytene stage onward. In Leydig cells, heterogeneous immunoreactivity of p57kip2 was found in immature testis and the signal intensity was higher in adult testis than immature ones. In Sertoli cells, weak or negligible immunoreactivity of p57kip2 was found. In human seminiferous tubule, strong immunoreactivity of p57kip2 was found in the nucleus of early spermatocytes, but not in the late pachytene spermatocytes onward and Sertoli cells. These results suggest the possible role of p57kip2 in the regulation of early spermatogonial proliferation, meiotic progression of early spermatocytes and differentiation of Leydig cells in testis.     

Annual reproductive cycle of a bitterling, Tanakia tanago, reared in an outdoor tank

From June 2000 to September 2001, we investigated the presence of eggs spawned in Margaritifera laevis and the seasonal changes in the gonads of Tanakia tanago. Eggs were observed from mid-March to mid-September. In females with a shrunken ovipositor, as the GSI gradually increased, most ovaries were in the prespawning phase (Oct-Mar). As the GSI increased further, most ovaries were in the early spawning phase (Mar-Jun). As the GSI gradually deceased, ovaries in the late spawning phase appeared (Jun-Sep). When the GSI was very low, most ovaries were in the postspawning phase (Sep-Oct). In males, when the GSI was low, most testes were in the early prespawning phase from Oct-Dec. As the GSI gradually increased, most testes were in the late prespawning phase (Dec-Jan). As the GSI increased further, testes were in the early spawning phase (Jan-Jun). As the GSI gradually decreased, amost testes were in the late spawning phase (Jun-Sep). When the GSI was very low, most testes were in the postspawning phase (Sep-Oct). These results indicate that T. tanago has a distinct annual reproductive cycle and is a spring-autumn spawner. Based on the relationship between reproductive activity and environmental factors, the spawning season of T. tanago appears to be initiated by increasing temperature and / or longer days in spring and to be terminated by shorter days in autumn.     

Feeding migration of roach, Rutilus rutilus (L.), in Lake Arungen, Norway

In the period April–July 1980 we studied the feeding migration and food of roach in a small tributary of the eutrophic Lake Årungen, south-eastern Norway. Tagging experiments revealed a tendency in roach to utilize a specific tributary both for feeding and for spawning. The mean size of roach ascending the tributary in late June and July was significantly larger than the mean size of roach spawning 1–2 months earlier, probably due to higher water discharge in July than in May and June. The experiment indicates two separately motivated migrations involving homing. The roach fed more profitably in the tributary than in the lake, both in terms of food availability and predator avoidance.     

Stage-specific expression of three cell surface carbohydrate antigens during murine spermatogenesis detected with monoclonal antibodies

We have identified three germ cell surface carbohydrate antigens that exhibit a common, stage-specific pattern of expression during spermatogenesis in the mouse. IgM-class monoclonal antibodies designated "J1," "C6," and "A5" were absorbed by adult testis, but not by any adult somatic tissue tested. In indirect immunofluorescence assays using collagenase-dissociated prepuberal and adult testicular cells, these antibodies labeled the surfaces of early and late pachytene spermatocytes and round spermatids. Gonocytes from fetal and neonatal testes were not labeled. In paraffin sections of prepuberal and adult testes, sialidase treatment exposed antigens recognized by antibodies C6 and A5 on preleptotene, leptotene, and zygotene spermatocytes located near the perimeter of seminiferous tubules. The determinants recognized by antibodies J1, C6, and A5 were characterized partially using a sugar hapten inhibition assay. The binding of J1 to adult testicular cells was inhibited specifically by N-acetylglucosamine and the binding of both C6 and A5 was inhibited by N-acetyllactosamine. The glycoconjugates recognized by J1, C6, and A5 eluted from gel filtration columns with an apparent molecular weight greater than 1 X 10(6) and were sensitive to endo-beta-galactosidase (keratanase) treatment. The apparent high molecular weight of these glycoconjugates was confirmed by immunolabeling Western blots of testis extracts separated by SDS-polyacrylamide gel electrophoresis. The results suggest that polylactosamine (keratan) glycoconjugates of high molecular weight are associated with the plasma membranes of meiotic and haploid male germ cells. The effects of sialidase on antibody labeling patterns suggest that changes in cell surface sialylation accompany the transition of early meiotic germ cells to pachytene spermatocytes during spermatogenesis.     

The biology of the twaite shad, Alosa fallax fallax (Lacépède), in the Severn Estuary

The estuarine biology of the twaite shad was studied in the Severn Estuary. Adults enter the estuary at the start of the freshwater phase of their spawning migration between April and June. Peak immigration generally occurs in May and is associated with temperatures in the range 10.6–12.3°C. The mean (± s.d. ) instantaneous mortality rate for the mature population was 0.53±0.18. The effect of additional mortality on the spawning population was modelled assuming constant recruitment and no density-dependent effects.
Juvenile twaite shad are present in the estuary from July until they emigrate seaward during the autumn. A portion of these fish re-enter the estuary the following April–May and remain until late summer/early autumn before once more migrating seaward.
The 0 + age group feed mainly on harpacticoid and calanoid copepods and mysids, the relative preponderance of these in the diet being apparently related to tidal conditions. The possible implications of the proposed tidal power barrage in the Severn Estuary on the twaite shad population are discussed in relation to movement, diet and additional mortality of the mature population.     

Intratubular localization of 4-ene-5 beta-reductase and 17 beta-hydroxy-dehydrogenase in immature golden hamster testis

We have reported [1,2] in immature golden hamster testis that 5 beta-reductase is localized in the seminiferous tubules, while 5a-reductase is present in the interstitial tissue and that the 17 beta-ol-dehydrogenase activity is found predominantly in the seminiferous tubules. In the present study, we show the intratubular localization of these enzymes. The left testis of golden hamster was irradiated with 2000R or 8000R of X-rays at 22 days of age. The hamsters were killed at 28 days of age. Homogenates of the left irradiated and right intact testes were incubated with [14C]-4-androstone-3,17-dione and NADPH, and enzyme activity was estimated. Both testes were also examined histologically. The X-irradiation of the testis resulted in an almost complete disappearance of germ cells with a significant decrease in testis weight, but the interstitial tissue and tubular nongerm cells including Sertoli cells remained almost unchanged. However, the activities of 5 beta-reductase and 17 beta-ol-dehydrogenase expressed as nmol formed/testis/h did not decrease at all. These results show that 5 beta-reductase is localized in the tubular nongerm cells including the Sertoli cells and 17 beta-ol-dehydrogenase is present in the tubular nongerm cells and interstitial tissue in immature golden hamster testis.     

Spawning of walleye pollock (<Emphasis Type="Italic">Theragra chalcogramma</Emphasis>) off the Southwestern coast of Kamchatka

The results of ichthyoplankton surveys conducted in 2007–2008 showed that waters off Southwest Kamchatka and North Kuril Islands were areas of mass spawning of walleye pollock. The peak spawning occurred during the last 10 days of April and in early May, which was much later than the peak at the main spawning site off West Kamchatka. The spawning activity of walleye pollock near the southwestern shores of Kamchatka is a regular event, as the analysis of archive materials shows. This gave us grounds to suspect the existence of a southern site that coincided well with the mass spawning in the Shelikhov Gulf in its timing and scale, but was missed during standard ichthyoplankton surveys conducted in early April. After analyzing the growth rates of spawners, the assumption was made that the southern spawning was performed by the East Kamchatkan walleye pollock population, whose mass spawning usually occurs in late April-early May. According to the data of 2008, the estimated biomass of walleye pollock spawning in the area of the Ozernovskaya basin in late April was nearly 600000 tons. Regular monitoring of the southern spawning is proposed by means of additional ichthyoplankton surveys south of 53° N, including the Okhotsk Sea waters of the North Kuril Islands, in late April—early May.