Stereological study of gonadotropes in the frog,Rana pipiens,after GnRH stimulation in vitro

Summary Previous physiological results have indicated the existence of two releasable pools of gonadotropins in amphibian pituitaries: an acute releasable pool that appears independent of protein synthesis, and a storage pool involved in chronic release that depends on protein synthesis. To elucidate the ultrastructural localization of these pools and the morphological changes induced in gonadotrope cells after treatment with gonadotropin-releasing hormone, we carried out a morphometric study of immuno-identified gonadotrope cells using an in vitro superfusion system. Treatment with gonadotropin-releasing hormone induced a degranulation of small (110–255 nm) and medium (236–360 nm) secretory granules as well as hypertrophy of the endoplasmic reticulum and Golgi complex. Simultaneous incubation with gonadotropin-releasing hormone and cycloheximide inhibited the release of secretory granules although the endoplasmic reticulum and Golgi complex were hypertrophied. These morphological results strongly suggest: (1) that gonadotropin-releasing hormone induces degranulation and hypertrophy of the biosynthetic machinery in gonadotrope cells; and (2) that the activation of the endoplasmic reticulum and Golgi complex by stimulation with gonadotropin-releasing hormone is independent of protein synthesis, while the release of secretory granules is protein synthesis-dependent. In addition, the second or storage pool of gonadotropin is associated mainly with the small and medium secretory granules.     

Sexual isolation between Drosophila melanogaster females and D. simulans males: sexual maturation of males

Isofemale lines of D. simulans were examined to determine the age of sexual maturity of males with conspecific females, and for the frequency of hybridization with D. melanogaster females. Males started to mature sexually on the first day after eclosion but their ability to mate slowly increased during the following day. The estimates of both the age sexual maturation started and the switch from immature to mature males were strongly dependent on the female genotypes used in the tests. No clear differences in speed of maturation were apparent between male lines. In contrast, differences in frequency of hybridization with D. melanogaster females did occur. From the above results it is concluded that the differential hybridization success of male D. simulans lines is not related to the speed at which males mature sexually.

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Résumé Des lignées isofemelles de D. simulans ont été examinées pour déterminer l'âge de la maturité sexuelle des mâles avec des femelles conspécifiques et pour établir la fréquence de l'hybridation avec des femelles de D. melanogaster. Les mâles ont commencé à être sexuellement mûrs le premier jour après l'émergence, mais leur aptitude à la copulation a augmenté lentement pendant le jour suivant. Les estimations, tant de l'âge du début de la maturation sexuelle que de l'âge du passage de mâle immature à mâle sexuellement mûr dépendaient étroitement des génotypes des femelles utilisées dans les expériences. Il n'y avait pas de différences nettes entre les lignées de mâles. Par contre, des différences dans les fréquences d'hybridation avec les femelles de D. melanogaster ont été observées. De ces résultats, on peut conclure que les différences dans la réussite des hybridations des lignées de mâles de D. simulans n'étaient pas dues à la vitesse de maturation sexuelle des mâles.

     

Miltpain, a cysteine proteinase, from milt of Pacific cod (Gadus macrocephalus): purification and characterization

Miltpain (EC.3.4.22.-) is a cysteine proteinase that preferentially hydrolyzes basic proteins, previously found in the milt of chum salmon. Here we report a similar cysteine proteinase in the milt of the marine Pacific cod. The enzyme was isolated and purified 6900-fold and with an estimated mass of 63 kDa by gel filtration chromatography and 72 kDa by SDS/PAGE. Cod miltpain has an optimum pH of 6.0 for Z-Arg-Arg-MCA hydrolysis, and K_m of 11.5 μM and k_cat of 19.0 s⁻¹ with Z-Arg-Arg-MCA. It requires a thiol-inducing reagent for activation and is inhibited by E-64, iodoacetamide, CA-074, PCMB, NEM, TLCK, TPCK, ZPCK and o-phenanthroline. This proteinase strongly hydrolyzes basic proteins such as salmine, clupeine and histone, and exhibits unique substrate specificity toward paired basic residues such as Lys-Arg, Arg-Arg on the substrates of P2-P1. The isoelectric point is 5.2 by isoelectric focusing. N-Terminal sequencing gave a sequence of <EVPVEVVRXYVTSAPEK. The cysteine proteinase from Pacific cod very closely matches the previously reported miltpain from chum salmon.     

Sexuality, sex change and maturation patterns in the yellowfin seabream, Acanthopagrus latus (Teleostei: Sparidae) (Houttuyn, 1782)

Macroscopic and histological studies were carried out to describe the reproductive styles and sex reversal and to follow gonadal changes in captive yellowfin seabream during the second year of life. Four reproductive styles are found in Acanthopagrus latus (Houttuyn, 1782): (i) males and females (gonochorism), (ii) functional males, (iii) transitionals and (iv) functional females. The species is a protandric hermaphrodite and begins life as a functional male with testicular zone undergoing active spermatogenesis, while the ovarian zone is arrested at the primary growth (perinucleolar) phase. Males and females were encountered in virtually all size‐classes. Functional males outnumbered the functional females in all size‐classes in which they were encountered. Sex reversal begins in the transitionals from July to August, after spawning in the functional males, at 14.9–20.2 cm standard length (SL) and, by November, maturation of the ovarian tissue begins. A. latus in cages in Kuwait waters spawns from January to April with a peak in February for males, and a peak in March for females and transitionals. Spawning begins in the 18.3–20.2 cm size‐range fish, peaking in the 20.3–22.2 cm size‐range in both males and females and also in the transitionals, although a few of the latter spawn from 14.9 cm SL. In relation to age, spawning begins at 20 months in males and peaks at 21 months. Females begin to spawn at 21 months with a peak at 22 months, while transitionals generally begin to spawn at 20 months, although a few 14–15‐month‐old sex‐changing individuals were encountered. Temperature, either alone or in combination with other unknown factors, triggers spawning in A. latus.     

Sexual maturation,serum steroid concentrations,and mRNA expression of IGF-1, luteinizing and progesterone hormone receptors and survivin gene in Japanese quail hens

In avian species, sexual maturation represents the evidence of start laying, which is a consequence of the development of ovarian follicles. These follicles are the functional reproductive unit whose maturation and viability critically depends on endocrine, paracrine, and autocrine factors beyond the signals from the central nervous system. The present study was undertaken to investigate the correlation of sexual maturity with tissue growth, mRNA expression of certain genes, and serum steroid concentrations in Japanese quail hens. To carry out the present study, a total of forty Japanese quail hens (5 weeks) were housed individually under uniform husbandry condition with ad libitum quail layer ration and water at 14-hour photo schedule. On sixth week onwards, four birds were sacrificed at each time on 1, 3, 7, 10, 13, 16, 19, 22, 25, and 28 days. Serum was extracted aseptically to analyze the gonadal steroid hormones (estrogen and progesterone) and corticosterone to investigate the liaison with sexual maturation of the species. Expression analyses of four genes i.e., insulin-like growth factor-1, luteinizing hormone receptor, progesterone receptor, and survivin were carried out in the three largest ovarian yellow follicles. A significant (P < 0.05) increase in body weight gain and oviduct weight was recorded during the phase of sexual maturation. Smaller follicles revealed higher insulin-like growth factor-1 and survivin gene expression, whereas the reverse result was manifested in both the luteinizing and progesterone hormone receptors. In biochemical study, the gonadal steroids (estrogen and progesterone) were recorded higher at the first half of the experiment when a gradual decrease in corticosterone concentration was confirmed from the very beginning of this study. This result substantiated that sexual maturation in Japanese quail may be completed by the time of 8 weeks after its birth in support of the analyzed information studied in the current investigation.     

Aspects of the fecundity of the feral catfish,Clarias macrocephalus (Gunther), population obtained from the rice fields used for rice-fish farming,in Malaysia

The catfish, Clarias macrocephalus, harvested from the irrigated rice fields of Malaysia, exhibited spawning activities that are influenced by water level management. The multiple spawning potential of the fish is negated by declining water level and the fish spawned only once per growing season. Although oocyte development started in conjunction with water level increase, subsequent oocyte development did not take place due to low water level. Absolute and relative fecundity also decreased with declining water level, however, mean relative fecundity is relatively constant (17.6–53.7 oocytes g^–1 body weight) and not related to body size. Absolute fecundity ranged between 785 to 14066 oocytes female^–1, whereas relative fecundity is between 7.1 to 83.5 oocytes g^–1 body weight, respectively.     

Sexual size dimorphism,growth, and maturity of the fluvial eight-barbel loach in the Kako River,Japan

The maturation and growth pattern of the fluvial eight-barbel loach Lefua sp. (Japanese name: nagare-hotoke-dojo), an endangered species, was investigated using an individual identification-recapture method from 1995 to 1998 in an upper reach of a headwater tributary of the Kako River, Hyogo Prefecture, Japan. Based on observations of the gonads through the abdominal skin, the loach was estimated to breed mostly from May to July. All the males matured by age 1⁺, and all the females matured by age 2⁺. Gamete release in all individuals of both males and females was predicted from recaptured loaches during each breeding season. The standard length of mature females was significantly larger than that of males, showing sexual size dimorphism (SSD). The maximum sizes recorded were 75.4 mm SL for females and 61.2 mm SL for males. Both males and females of immature specimens grew mainly from May to November, including the breeding season, with no significant differences in growth rates between them. After sexual maturity, both males and females grew mainly from July to October (or November), after the breeding season, and the females exhibited higher growth rates than males. Therefore, SSD of the species seems to be attributable to the different growth rates after maturity. The longevity of the loach was estimated to exceed ten years based on individual growth patterns of various sizes during the survey period. It is likely that the loach has an iteroparous life history, breeding every year, and moderate growth rates after maturity.     

Cardicola aurata sp. n. (Digenea: Sanguinicolidae) from Mediterranean Sparus aurata L. (Teleostei: Sparidae) and its unexpected phylogenetic relationship with Paradeontacylix McIntosh, 1934

A new sanguinicolid trematode, Cardicola aurata sp. n., is described from gilthead seabream Sparus aurata L., from off the Spanish Mediterranean coast. The morphology of C. aurata sp. n. generally agrees with the diagnosis of the genus, however, in contrast to all other reported Cardicola spp. the male pore is located sub-medially at the posterior end of the body instead of sinistrally before the posterior end of the body. Based on a comparison of the morphology as well as partial 28S and ITS2 rDNA sequence data from the present species with that from closely related species, it was decided to emend the diagnosis of Cardicola rather than create a new genus, as the aberrant position of the male pore is likely to be an autapomorphy. The phylogenetic analyses revealed a close relationship between Cardicola and Paradeontacylix, two genera with considerable morphological differences; C. aurata sp. n. occupies a position intermediate to these genera. Thus, a morphological comparison of Cardicola, Paradeontacylix and Braya, a genus which is morphologically similar to Cardicola but clusters basal to the Cardicola/Paradeontacylix clade, was conducted. The results of this comparison showed that despite large differences with regard to body shape, the organisation of the internal organs is very similar in species of Cardicola and Paradeontacylix. The synopsis of morphological data and molecular phylogeny allows for interpretations regarding the importance of different morphological features for the phylogenetic inference of the Sanguinicolidae.     

Somatometrical features of Japanese monkeys in the Koshima islet: In viewpoint of somatometry,growth, and sexual maturation

The Japanese monkeys inhabiting Koshima islet were investigated morphologically, and compared with those living in other areas of Japan. The morphological features of the Koshima monkeys are thought to reflect many interrelated factors. Their body size is the smallest inMacaca fuscata fuscata. The lower latitude and the warmer winter of the islet represent the major factors in this small body size. Another physical characteristic is their longer limbs, which may be related to the same environmental factors. The adult males of the Koshima troop show the widest variation, while the variation in the adult females is the narrowest, among the troops ofMacaca fuscata examined. This may be explained either by sexually differentiated growth, that is, males continue to grow considerably after 8 years of age, whereas females do not; or by isolation, that is, males sometimes immigrate into or emigrate from the islet, whereas almost of all the females remain there. It was found that the influence of food limitations on body weight depends on sex and age; babies and juveniles, and adult males did not lose their body weight, but monkeys aged from 3 to 7 years and adult females had lost much weight compared to those of the same ages and sexes measured in 1963 (feeding was not limited). This fact may support the idea of “sex-age-class selection” proposed byMori (1979), but the weight loss pattern could be explained by changes of body composition with growth. The growth of the anterior trunk length, too, was restrained. Sexual maturation in males was delayed by at least one year.     

Potassium fluxes in the rat reticulocyte Ouabain sensitivity and changes during maturation

K⁺ turnover is markedly enhanced in the rat reticulocyte, both influx and efflux rates being increased by factors of approximately 3 over the corresponding rates in adult cells. These accelerated fluxes are observed despite the absence of any appreciable change in intracellular K⁺ concentration during the course of maturation. Qualitative characteristics of the active transport process for K⁺ influx appear to be identical in reticulocytes and mature erythrocytes with regard both to K⁺ sensitivity, and to ouabain sensitivity as a function of external K⁺ concentration. The number of ouabain binding sites per unit volume of cells, however, is increased by a factor of approximately three in the reticulocyte and thus correlates well with the observed degree of enhancement of active K⁺ influx in these cells. Half-maximal rates of ouabain-sensitive K⁺ influx are observed at external K⁺ concentrations well below 1 mM for both reticulocytes and mature erythrocytes. It is concluded that the enhanced rate of K⁺ accumulation in the reticulocyte can be quantitatively attributed to an increased number of pump units which are qualitatively identical to those in the mature cell, and which function at a near-maximal rate at the ambient K⁺ concentration present in normal rat plasma.     

Resorption of unemitted gametes in Lithognathus mormyrus (Sparidae,Teleostei): a possible synergic action of somatic and immune cells

The resorption of unemitted gametes during the post-spawning period of the male and female reproductive cycles in Lithognathus mormyrus was studied by histochemical, histological and cytological methods. The resorption of residual spermatozoa involved the phagocytotic activity of Sertoli cells bounding the seminiferous cysts of spermatozoa, and those associated with spermatogonia lining the lobular lumen. Spermatozoa remaining in the sperm duct were phagocytozed by the lining epithelial cells. Eosinophilic granulocytes and macrophages were identified in the vicinity of residual spermatozoa. The remnants of oocytes underwent an atretic phenomenon in which follicle cells were firstly involved, inducing a progressive fragmentation of the oocyte cytoplasm. Subsequently, eosinophilic granulocytes invaded oocyte degenerative areas and clung to the remaining vitelline inclusions ensuring their biotransformation into waste products (brown bodies). The analogy of the resorption processes of both male and female unemitted gametes during the post-spawning period of natural reproductive cycle, involving first the enveloping somatic cells and then immune cells, is emphasized.     

Sexual segregation in ungulates: individual behaviour and the missing link

Previous "explanations" of sexual segregation in ungulates establish no more than a prerequisite for habitat segregation because they do not include a model of competitive habitat selection. Here we provide one based on the ideal free distributions of mutually competing, optimally foraging, individual deer. We parameterised our model using field data collected from a population of fallow deer (Dama dama) in a Mediterranean forest. The predictions of the inter-sex competition model were in full agreement with observational data, but those of single sex distributions (conventional theory) were not. The "conventional" hypothesis, that segregation arises simply from sex differences, predicted no more than moderate (20–40%) levels of segregation, even in optimal conditions. By introducing inter-sex resource competition, the predicted segregation can generally more than double and full segregation becomes possible in some circumstances. The modelling showed segregation to be density-dependent, varying in complicated ways with season and animal density. Sensitivity analysis showed the vulnerability of the "conventional" understanding of environmental variation and uncertainty. Using our competition model we show that as diet difference increases, direct competition between the sexes declines, so that as males increasingly differ from females, segregation declines and the two sexes are more likely to be found mixed (as long as the chosen food is available to both in the same area). Conversely, small differences among male and female deer are amplified by both food depletion and inter-sex competition to give substantial levels of segregation. The theoretical framework on which our model is built strongly suggests that sexual dimorphism in the context of scramble competition may be the fundamental cause of sexual habitat-segregation among ungulates.     

Long-term cryopreservation of Greek fir embryogenic cell lines: recovery, maturation and genetic fidelity

In coniferous species, including Greek fir (Abies cephalonica Loud), the involvement of somatic embryo plants in breeding and reforestation programs is dependent on the success of long-term cryostorage of embryogenic cultures during clonal field testing. In the present study on Greek fir, we assayed the recovery, morphological characteristics and genetic fidelity of embryogenic cell lines 6 and 8 during proliferation and maturation after long-term cryostorage. Our results indicate successful recovery of both cell lines after 6 years in cryostorage. In the maturation phase, both cell lines were capable of producing somatic embryos although some differences were detected among experiments. However, these changes were more dependent on the differences in the components of the maturation media or in the experimental set-up than on the long-term cryostorage. During both proliferation and maturation phases, the morphological fidelity of the embryogenic cultures as well as of the somatic embryos were alike before and after cryopreservation. The genetic fidelity of the cryopreserved cell line 6 that was assayed by random amplified polymorphic DNA (i.e. RAPD) markers demonstrated some changes in the RAPD profiles. The results indicate possible genetic aberrations caused by long-term cryopreservation or somaclonal variation during the proliferation stage. However, in spite of these changes the embryogenic cultures did not lose their proliferation or maturation abilities.     

Nucleotide pool changes in coelomic cells (eleocytes) of the polychaete Nereis virens during sexual maturation

Eleocytes (a type of coelomic cell) of the polychaete Nereis virens can store large amounts of adenine nucleotides at certain times. Since eleocytes have specific functions related to gametogenesis, we tested whether the presence of these large nucleotide stores in eleocytes is specific to gender or related to specific events during gametogenesis. Nucleotide pools in eleocytes isolated at different stages of sexual maturation from N. virens were analysed using high-performance liquid chromatography. Eleocytes from immature and male animals had extremely high concentrations of both AMP and ADP (each > 10 μmol/ml of packed cell volume). In eleocytes from male animals, the high nucleotide stores were maintained throughout the maturation phase and decreased at a late stage, while in female animals the nucleotides were degraded at an early stage of maturation. In male eleocytes, the decrease in the adenine nucleotide pool may be the result of its conversion to inosine which is then released by the eleocytes and reutilized by male germ cells for nucleic acid biosynthesis, as has been suggested previously. Our study shows that the time of degradation of the adenine nucleotide pool coincides with the period of spermatogonia proliferation which involves intense nucleic acid synthesis. ATP levels (0.4–1.5 μmol/ml packed cell volume) and the guanine nucleotide pool (GTP+GDP+GMP; 0.08–0.18 μmol/ml packed cell volume) were similar in both sexes, did not change during germ cell development and were decreased only in eleocytes from prespawning females. The GTP/GDP ratios were initially higher (up to 14) in eleocytes from females compared to ratios in eleocytes from immature (4–9) and male animals (up to 8), and decreased during the maturation phase of the animals. GTP levels were correlated with those of ATP; this correlation was much closer in eleocytes from females than from males. The results further support the hypothesis that the adenine nucleotide stores in the eleocytes are maintained as a supply of purine precursors for the growing germ cells.     

Autumnal changes in tissue nitrogen of autumn olive,black alder and eastern cottonwood

Two experiments were conducted to determine patterns of N change in tissues of autumn olive (Elaeagnus umbellata Thunb.) and black alder (Alnus glutinosa [L.] Gaertn.) during autumn in central Illinois, U.S.A. In the first study leaf nitrogen concentrations of autumn olive decreased 40% at an infertile minespoil site and 39% at a fertile prairie site throughout autumn whereas nitrogen concentrations in respective bark samples increased by 39% and 37%. Salt-extractable protein concentrations increased in bark and decreased in leaves over the sampling period. Free amino acid concentrations of autumn olive leaves decreased over the course of the experiment from peak concentrations in August. Asparagine, glutamic acid and proline were major constituents of the free amino acid pools in leaves. Total phosphorus concentrations of autumn olive leaves declined by 40–46% during autumn while bark concentrations of P did not significantly change.In the second experiment non-nodulated seedlings of alder receiving a low level of N-fertilization did not exhibit net resorption of leaf N during autumn whereas foliar N concentration of contrasting nonactinorhizal cottonwood plants (Populus deltoides Bartr. ex. Marsh) under the same fertilization regime decreased by 27% after the first frost. A gradual but significant decrease of 38% in foliar N concentration of nodulated alder seedlings grown under a low N-fertilization regime was associated with the cessation of nitrogenase activity during autumn in nodules. Compared with the low N fertilization regime, the higher level of N-fertilization resulted in smaller autumnal decreases of foliar N concentration in nodulated alder (17%) and in cottonwood (20%); but there was no decrease in foliar N concentration in non-nodulated alder. The higher level of N-fertilization promoted a greater accumulation of N in the roots than in the bark of both tree species after the first frost.Our results suggest that black alder lackingFrankia symbionts does not exhibit net leaf N resorption and that autumnal decreases in leaf N ofFrankia-nodulated black alder result primarily from declining foliar N import relative to export due to low temperature inhibition of N₂ fixation. In contrast, autumn olive exhibited greater and more precipitous autumnal declines in foliar N concentration than those of alder, and the pattern of N decline was unaffected by site fertility.     

Correlated changes in the structure of the anterior pituitary gland,testes and interrenal tissue during sexual maturation of male lizards

Summary The anterior pituitary gland, testes and interrenal glands of a series of young males of the teiid Cnemidophorus l. lemniscatus (L.) have been studied by light microscopy in order to correlate the changes occurring during sexual maturation. In the testes of the smallest animals, spermatogenesis does not progress beyond primary spermatocytes and there is no differentiated interstitial tissue. In medium-sized animals, spermatids and some interstitial cells appear, and in the largest lizards, spermatogenesis is completely established and Leydig cells abound. Simultaneously with the development of the testes, interrenal glands undergo great hypertrophy and hyperplasia, especially in the peripheral reactive zone. Starting in animals of intermediate size, the anterior hypophysis exhibits a considerable hypertrophy of two rostral cell types: the chromophobic corticotrophs and the acidophilic PAS-positive cells considered to be interstitiotrophs. These cells show large, vesicular nuclei and prominent nucleoli, signs of enhanced cellular activity. The hypertrophy begins in the dorso-rostral region of the gland close to the median eminence, at the site of entry of the portal vessels. This suggests a hypothalamic influence on the function of these pars distalis cells. The scattered basophilic gonadotrophs or folliculotrophs are scarce, small, and do not vary appreciably among the animals studied. The hyperactivity of corticotrophs may account for enlargement of the interrenal glands. Testicular development is apparently related to an increased activity of interstitiotrophs but to a stable level of activity in folliculotrophs.This research forms part of project No. 31.26.S1-0244 supported by the Consejo Nacional de Investigaciones Científicas y Tecnológicas.     

Sexual Dimorphism and Mortality Bias in a Small Miocene North American Rhino, <Emphasis Type="Italic">Menoceras arikarense</Emphasis>: Insights into the Coevolution of Sexual Dimorphism and Sociality in Rhinos

Rhinos are the only modern perissodactyls that possess cranial weapons similar to the horns, antlers and ossicones of modern ruminants. Yet, unlike ruminants, there is no clear relationship between sexual dimorphism and sociality. It is possible to extend the study of the coevolution of sociality and sexual dimorphism into extinct rhinos by examining the demographic patterns in large fossil assemblages. An assemblage of the North American early Miocene (∼22 million years ago) rhino, Menoceras arikarense, from Agate Springs National Monument, Nebraska, exhibits dimorphism in incisor size and nasal bone size, but there is no detectible dimorphism in body size. The degree of dimorphism of the nasal horn is greater than the degree of sexual dimorphism of any living rhino and more like that of modern horned ruminants. The greater degree of sexual dimorphism in Menoceras horns may relate to its relatively small body size and suggests that the horn had a more sex-specific function. It could be hypothesized that Menoceras evolved a more gregarious type of sociality in which a fewer number of males were capable of monopolizing a larger number of females. Demographic patterns in the Menoceras assemblage indicate that males suffered from a localized risk of elevated mortality at an age equivalent to the years of early adulthood. This mortality pattern is typical of living rhinos and indicates that young males were susceptible to the aggressive behaviors of dominant individuals in areas conducive to fossilization (e.g., ponds, lakes, rivers). Menoceras mortality patterns do not suggest a type of sociality different from modern rhinos although a group forming type of sociality remains possible. Among both living and extinct rhinos, the severity of socially mediated mortality seems unrelated to the degree of sexual dimorphism. Thus, sexual dimorphism in rhinos is not consistent with traditional theories about the co-evolution of sexual dimorphism and sociality.     

Co-existence of gonadotrophins (FSH,LH) and thyrotrophin (TSH) in single anterior pituitary cells of the musk shrew,Suncus murinus

According to recent immunocytochemical studies of anterior pituitary cells, it is obvious that the one cell-one hormone theory must be modified. Many pituitary morphologists have demonstrated that there are some cells that contain two hormones. In this study, we demonstrate by means of immuno-electronmicroscopy the co-existence of gonadotrophins (FSH and LH) and thyrotrophin (TSH) in the same anterior pituitary cells of the musk shrew. These cells were remarkably altered in their ultrastructural features by either gonadectomy or thyroidectomy. Double labeling for gonadotrophins and thyrotrophin was present not only in the same cells but also in the same secretory granules. Our ability to demonstrate co-existence of gonadotrophins and thyrotrophin in the same cell may be due to our selection of fixative and embedding media for electron-microscopic immunocytochemistry. Our conclusion that gonadotrophins and thyrotrophin are produced in a single cell type of the anterior pituitary gland in the musk shrew, i.e., thyrogonadotrophs, suggests the need to consider a modification of the classic scheme for classification of anterior pituitary cells.     

Efficient synthesis of a novel m-phenylene derivative as a selective EP4 agonist inducing follicular growth and maturation in the ovary

An efficient and straightforward synthesis of a novel m-phenylene derivative has been developed. The optically pure dibromo compound was selected as a starting material. Through a protocol involving the Prins reaction and two steps of the Horner–Wadsworth–Emmons reaction, the basic skeleton was constructed with appropriate alpha and omega side chains. The compound proved to be a highly selective EP₄ agonist and a possible drug candidate for maturation of the uterine cervix.     

Immunocytochemical identification of prolactin cells in the pituitary gland of tilapia larvae (Oreochromis mossambicus: Teleostei)

Summary Using an antiserum to highly purified chum salmon prolactin, prolactin cells were identified in the putative rostral pars distalis of newly hatched tilapia larvae (Oreochromis mossambicus) by the immunogold method for the electron microscope. In the putative rostral pars distalis, some cells had another kind of secretory granule which was much less numerous, much smaller in size, and without immunoreactivity to salmon prolactin antiserum. Controls incubated with salmon prolactin-preabsorbed antiserum or normal serum showed no immunoreactive cells, confirming the specificity of the antiserum. The possible role of prolactin in the osmoregulation of tilapia larvae is discussed.