Influences of the environment on the endocrine and paracrine fish growth hormone–insulin‐like growth factor‐I system

Insulin‐like growth factor‐I (IGF‐I) is a key component of the complex system that regulates differentiation, development, growth and reproduction of fishes. The IGF‐I gene is mainly expressed in the liver that represents the principal source of endocrine IGF‐I but also in numerous other organs where the hormone most probably acts in an autocrine–paracrine manner. The primary stimulus for synthesis and release of IGF‐I is growth hormone (GH) from the anterior pituitary. Thus, in analogy to mammals, it is usual to speak of a fish ‘GH–IGF‐I axis'. The GH–IGF‐I system is affected by changes in the environment and probably represents a target of endocrine disrupting compounds (EDC) that impair many physiological processes in fishes. Thus, the review deals with the influences of changes in different environmental factors, such as food availability, temperature, photoperiod, season, salinity and EDCs, on GH gene expression in pituitary, IGF‐I gene expression in liver and extrahepatic sites and the physiological effects resulting from the evoked alterations in endocrine and local IGF‐I. Environmental influences certainly interact with each other but for convenience of the reader they will be dealt with in separate sections. Current trends in GH–IGF‐I research are analysed and future focuses are suggested at the end of the sections.     

Plant volatiles in the sexual communication of Melolontha hippocastani: response towards time-dependent bouquets and novel function of (Z)-3-hexen-1-ol as a sexual kairomone

1. Swarming males of Melolontha hippocastani are known to locate females that stay feeding within the host trees by orienting towards damage‐induced plant volatiles (green leaf volatiles) and a sex pheromone. Thus, volatiles emitted by freshly damaged leaves might indicate to a male the presence of currently feeding females. 2. The hypothesis was studied that volatiles from freshly damaged leaves are more attractive to males than volatiles from old damaged leaves. The odour bouquets of damaged leaves from three plant species that have been shown to attract male M. hippocastani in the field were analysed 10 min (fresh damage) and 1.5 h (old damage) after damaging, using coupled gas chromatography–mass spectrometry. The results showed clear differences between the bouquets: the bouquet of freshly damaged leaves of all species consisted of typical leaf aldehydes, i.e. hexanal, (Z)‐3‐hexenal, (Z)‐2‐hexenal, (E)‐2‐hexenal, the leaf alcohol (Z)‐3‐hexen‐1‐ol, and the corresponding acetate. One and a half hours after damage, aldehydes had almost vanished and (Z)‐3‐hexen‐1‐ol and (Z)‐3‐hexenyl acetate predominated; however males of M. hippocastani were equally attracted to traps baited with volatiles from old and freshly damaged leaves in field experiments. When traps were baited with synthetic volatile mixtures mimicking the bouquets of old and freshly damaged leaves, M. hippocastani males even preferred the old damage mixture. 3. Experiments addressing the role of individual green leaf volatiles revealed that only (Z)‐3‐hexen‐1‐ol was highly attractive while the other compounds tested individually were behaviourally inactive, however all tested compounds elicited comparable electrophysiological responses on male M. hippocastani antennae. 4. In analogy to the term aggregation kairomone used for feeding‐induced plant volatiles that attract both sexes of an insect, the term sexual kairomone is suggested to describe the novel function of (Z)‐3‐hexen‐1‐ol in the sexual communication of M. hippocastani.     

Electromechanical coupling between skeletal and cardiac muscle. Implications for infarct repair

Skeletal myoblasts form grafts of mature muscle in injured hearts, and these grafts contract when exogenously stimulated. It is not known, however, whether cardiac muscle can form electromechanical junctions with skeletal muscle and induce its synchronous contraction. Here, we report that undifferentiated rat skeletal myoblasts expressed N-cadherin and connexin43, major adhesion and gap junction proteins of the intercalated disk, yet both proteins were markedly downregulated after differentiation into myo-tubes. Similarly, differentiated skeletal muscle grafts in injured hearts had no detectable N-cadherin or connexin43; hence, electromechanical coupling did not occur after in vivo grafting. In contrast, when neonatal or adult cardiomyocytes were cocultured with skeletal muscle, approximately 10% of the skeletal myotubes contracted in synchrony with adjacent cardiomyocytes. Isoproterenol increased myotube contraction rates by 25% in coculture without affecting myotubes in monoculture, indicating the cardiomyocytes were the pacemakers. The gap junction inhibitor heptanol aborted myotube contractions but left spontaneous contractions of individual cardiomyocytes intact, suggesting myotubes were activated via gap junctions. Confocal microscopy revealed the expression of cadherin and connexin43 at junctions between myotubes and neonatal or adult cardiomyocytes in vitro. After microinjection, myotubes transferred dye to neonatal cardiomyocytes via gap junctions. Calcium imaging revealed synchronous calcium transients in cardiomyocytes and myotubes. Thus, cardiomyocytes can form electromechanical junctions with some skeletal myotubes in coculture and induce their synchronous contraction via gap junctions. Although the mechanism remains to be determined, if similar junctions could be induced in vivo, they might be sufficient to make skeletal muscle grafts beat synchronously with host myocardium.     

Hormonal Interactions in Fruit Development

Fruit development involves a complex interplay of cell division, differentiation and expansion of sporophytic and gametophytic tissues that is carefully coordinated temporally and spatially. Plant hormones are signal molecules that regulate many processes of plant development, including fruit development leading to mature fruit and viable mature seed. Auxins, gibberellins, cytokinins, abscisic acid, and ethylene have been implicated at various stages of fruit development. In the past, hormone application studies and hormone analysis studies have supported the hypothesis that fruit development is in part regulated by hormonal interaction. More recently, biochemical, genetic, and molecular studies are beginning to unravel the complexities of how hormones affect fruit development. In the current work, we review selected studies that show the interplay between hormones during fruit development, with an emphasis on the interaction between auxin and gibberellin in pea fruit development.     

Hormone and seed-specific regulation of pea fruit growth

Growth of young pea (Pisum sativum) fruit (pericarp) requires developing seeds or, in the absence of seeds, treatment with gibberellin (GA) or auxin (4-chloroindole-3-acetic acid). This study examined the role of seeds and hormones in the regulation of cell division and elongation in early pea fruit development. Profiling histone H2A and gamma-tonoplast intrinsic protein (TIP) gene expression during early fruit development identified the relative contributions of cell division and elongation to fruit growth, whereas histological studies identified specific zones of cell division and elongation in exocarp, mesocarp, and endocarp tissues. Molecular and histological studies showed that maximal cell division was from -2 to 2 d after anthesis (DAA) and elongation from 2 to 5 DAA in pea pericarp. Maximal increase in pericarp gamma-TIP message level preceded the maximal rate of fruit growth and, in general, gamma-TIP mRNA level was useful as a qualitative marker for expanding tissue, but not as a quantitative marker for cell expansion. Seed removal resulted in rapid decreases in pericarp growth and in gamma-TIP and histone H2A message levels. In general, GA and 4-chloroindole-3-acetic acid maintained these processes in deseeded pericarp similarly to pericarps with seeds, and both hormones were required to obtain mesocarp cell sizes equivalent to intact fruit. However, GA treatment to deseeded pericarps resulted in elevated levels of gamma-TIP mRNA (6 and 7 DAA) when pericarp growth and cell enlargement were minimal. Our data support the theory that cell division and elongation are developmentally regulated during early pea fruit growth and are maintained by the hormonal interaction of GA and auxin.     

Neurohormonal peptides, serotonin, and nitric oxide synthase in the enteric nervous system and endocrine cells of the gastrointestinal tract of neotenic and thyroid hormone-treated axolotls (Ambystoma mexicanum)

Immunoreactivity against vasoactive intestinal polypeptide (VIP), neurotensin (NT), substance P (SP), calcitonin gene-related peptide (CGRP), gastrin/cholecystokinin (GAS/CCK), somatostatin (SOM), serotonin (SER), and nitric oxide synthase (NOS) was investigated in the gastrointestinal tract of the urodele Ambystoma mexicanum, the axolotl, by the use of immunohistochemical techniques. The study also compares the distribution patterns and frequencies of the neurohormones, and NOS in neotenic and thyroxine-treated (metamorphosed) individuals. GAS/CCK, SP, NT, SOM, and SER immunoreactivities occurred in endocrine mucosal cells and VIP, SP, CGRP, NTSER, SER, and NOS immunoreactivities in the enteric nervous system. The GAS/CCK-immunoreactive (-IR) cells were restricted to the upper small intestine. NT-IR and SP-IR endocrine cells were found in the entire gastrointestinal tract and were most prominent in the distal large intestine. The density of the SOM-IR cells decreased from the stomach toward the large intestine. SER-IR endocrine cells were found throughout the gastrointestinal tract, with particularly high densities in the stomach and distal large intestine. The VIP-IR enteric nerve fibers were the most prominent ones, present in all layers of the entire gastrointestinal tract, and supplied the smooth muscle and the vasculature. The SER-IR fibers exhibited similar distribution patterns but were less numerous. Very few NT-IR but many SP-IR fibers were found in the muscle and submucosal layers. The NT-IR fibers mainly supplied blood vessels, while the SP-IR fibers were also in contact with the smooth muscle. In the muscle and submucosal layers, CGRP-IR fibers were associated to the vasculature; CGRP immunoreactivity occurred also in a minority of SP-IR fibers. NOS-IR nerve fibers were in contact with submucosal arteries but were the least frequent . After metamorphosis provoked by exogenous thyroxine, the number of SOM-IR endocrine cells in the stomach mucosa was increased as well as the density of VIP-IR, SER-IR, and SP-IR nerve fibers in the gastrointestinal tract. It is proposed that the observed increases may reflect refinements of the neurohormonal system after metamorphosis.     

The scent of food and defence: green leaf volatiles and toluquinone as sex attractant mediate mate finding in the European cockchafer Melolontha melolontha

Mate finding in the European cockchafer, Melolontha melolontha , has been investigated in the field. Our data suggest that mainly males show flight activity at dusk. In a landing cage bioassay, male cockchafers preferred cages baited with females over cages baited with males. Gas chromatographic analysis of beetle extracts with electronantennographic detection revealed the presence of electrophysiologically active compounds, among them toluquinone. In funnel trap bioassays none of these compounds was attractive towards males per se . A mixture of green leaf volatiles (GLV) mimicking the bouquet of mechanically damaged leaves was highly attractive to M. melolontha males. The attractiveness of the same GLV mixture was synergistically enhanced when toluquinone was added to the lure. GLV attract males to damaged leaves. Toluquinone as a sex pheromone indicates the presence of conspecific females and synergistically enhances the attractiveness of GLV.     

Immunocytochemistry of cardiac polypeptide hormones (cardiodilatin/atrial natriuretic polypeptide) in brain and hearts of Myxine glutinosa (Cyclostomata)

With the use of different region-specific antisera against partial sequences of porcine cardiodilatin (CDD)-126 and the peroxidase-antiperoxidase (PAP) technique, the central nervous system as well as the systemic and the portal vein heart of the cyclostomian species Myxine glutinosa were investigated for a possible existence of cardiac polypeptides. In contrast to mammals, CDD-immunoreactions were obtained only with antisera directed against the C-terminus of CDD (CDD 99-126) which is identical to human atrial natriuretic polypeptide (alpha hANP). CDD-immunoreactive myocardiocytes were found in high densities in the atrium of the systemic heart and in the portal vein heart. In the ventricle of the systemic heart, CDD-immunoreactive cells were extremely scarce. In agreement with the immunohistochemical results, myoendocrine cells analyzed by electronmicroscopy exhibited specific granules of an average diameter of 0.21 + 0.02 micron in equivalent localizations. Furthermore, with the use of the protein A-gold (PAG) technique, CDD-immunoreactivity was ultrastructurally localized within the specific granules of atrial myocardiocytes. In the central nervous system of Myxine glutinosa, CDD-immunoreactive perikarya and/or fibers were present on all levels from the telencephalon to the spinal cord. The results of the present study are compared with those obtained in mammals and their possible functional relevance and their meaning in phylogeny are discussed as well.     

Nematode parasites from Burchell's zebras in South Africa

Twenty-five Burchell's zebras (Equus burchelli antiquorum) which were culled at monthly intervals in the Kruger National Park were examined for helminths. Twenty-nine species of nematodes belonging to the families Atractidae, Habronematidae, Onchocercidae, Oxyuridae, Strongylidae, Strongyloididae and Trichostrongylidae were recovered. The cyathostomes (small strongyles) most abundant were Cyathostomum tetracanthum, Cylicostephanus calicatus, Cylindropharynx sp. (? C. intermedia Theiler, 1923) and Cylicocyclus auriculatus. Cyathostomum alveatum, Cyathostomum montgomeryi, Cylicostephanus calicatus and Cylindropharynx sp. (? C. intermedia Theiler, 1923) were the most prevalent cyathostomes (small strongyles) while Craterostomum acuticaudatum was the most prevalent of the large strongyles. Of all the species recovered those most abundant were Crossocephalus viviparus and Probstmayria vivipara with intensities of 100 to 3,857,772 and 18,400 to 104,120,467, respectively. Four new species, two Triodontophorus spp. (Strongylidae) and two Habronema spp. (Habronematidae) were identified. Furthermore, this study furnishes a first report of Triodontophorus minor in zebras. The fourth stage cyathostomes as well as the adults of 11 of the 14 species were present in significantly greater intensities in autumn and winter.