Bihormonal control of oogenesis in the freshwater prawn, Macrobrachium kistnensis

The hormonal control of oogenesis has been investigated in the freshwater prawn, Macrobrachium kistnensis. Experiments using bilateral eyestalk ablation and injection of eyestalk extract supported the presence of a gonad inhibiting factor in the eyestalk of female prawns. Injections of brain and thoracic ganglion extract in normal female prawns and in those subjected to bilateral eyestalk ablation revealed the presence of a gonad stimulating factor in the central nervous tissue.     

Induction of ovarian growth in the red claw crayfish,Cherax quadricarinatus,by the enkephalinergic antagonist naloxone: in vivo and in vitro studies

Summary

The effects of spiperone and domperidone (dopaminergic antagonists), as well as naloxone (enkephalinergic antagonist) on ovarian growth of the crayfish Cherax quadricarinatus, were evaluated during the non-reproductive period. In vivo assays were carried out by administering these compounds by adding them to the crayfish food, and subsequently measuring the oocyte diameter (oocytes in secondary vitellogenesis). Only naloxone produced a significantly (p <0.05) higher oocyte diameter than that of the controls, suggesting that this antagonist was blocking the effects of endogenous enkcephalins on the secretion of one or more neurohormones involved in crustacean reproduction. To test this hypothesis, an in vitro incubation of ovarian pieces with thoracic ganglion (TG), a source of the gonad stimulating hormone (GSH), or eyestalk tissue (ET), the source of the gonad inhibiting hormone (GIH), with or without naloxone added to the incubation medium, was done. A significant increase of ovarian growth was observed when TG was present. Moreover, the addition of naloxone to this preparation was able to further increase the ovarian growth, presumably by preventing enkephalinergic inhibition of GSH secretion.     

Species-specific effects on hemolymph glucose control by serotonin, dopamine, and L-enkephalin and their inhibitors in Squilla mantis and Astacus leptodactylus (crustacea)

Hemolymph glucose level is controlled by crustacean Hyperglycemic Hormone (cHH) released from the eyestalk neuroendocrine centers under conditions of both physiological and environmental stress. Biogenic amines and enkephalin have been found to mediate the release of several neurohormones from crustacean neuroendocrine tissue. We investigated the effect of serotonin, dopamine, and Leucine-enkephalin in vivo--injected into the stomatopod Squilla mantis and the decapod Astacus leptodactylus--whether increasing or depressing glycemia. Serotonin had a marked effect in elevating glucose level compared with initial values in both species. 5-HT1-like receptors are more involved in mediating serotonin action as co-injected cyproheptadine was a more effective antagonist than ketanserin (5-HT2-like receptor inhibitor). Dopamine injection in intact animals produced a decrease below initial levels of hemolymph glucose. This effect was significantly antagonized by domperidone. No significant effect of both amines occurred in eyestalkless animals. L-enkephalin shows a differential effect: in S. mantis it induced hypoglycemia while in A. leptodactylus it caused an increase of glucose level. Co-injected antagonist naloxone affected the direction of the response. Serotonin appears to provide a major control on glucose mobilization, whereas dopamine and L-enkephalin act as modulators whose plasticity in use or action varies among species.     

A trematode metacercaria encysted in the nerves of the dungeness crab, Cancer magister

Of 188 Dungeness crabs, Cancer magister, 8 contained unidentified trematode metacercariae encysted in various components of the nervous system, including the thoracic ganglion, brain, lamina ganglionaris of the eyestalk, and major nerves arising from the thoracic ganglion. A single cyst was present in the available tissue sections of 7 of the crabs and no behavioral abnormalities were exhibited. One crab with multiple cysts in major nerves near the thoracic ganglion was markedly ataxic. The cysts and included worms distorted, compacted, and destroyed nervous tissue, and occupied most of the nerves where present. Host response was minimal, but some cysts invoked massive hemocytic accumulations near the infection site. Infections probably seriously affect nerve impulse transmission and accounts for the lethargic behavior of the crab with multiple cysts in major nerves. The present report is the first record of digenetic trematode infections in the Dungeness crab and the apparent restriction of the worm to nervous tissue is unusual if not unique in the Digenea. Because of the absence of grossly recognizable lesions and the small samples excised at necropsy, both the incidence and intensity of infection in Dungeness crab populations are almost certainly higher than indicted by our data.     

The distribution of APGWamide and RFamides in the central nervous system and ovary of the giant freshwater prawn, Macrobrachium rosenbergii

Immunohistochemistry was used to identify the distribution of both APGWamide-like and RFamide-like peptides in the central nervous system (CNS) and ovary of the mature female giant freshwater prawn, Macrobrachium rosenbergii. APGWamide-like immunoreactivity (ALP-ir) was found only within the sinus gland (SG) of the eyestalk, in small- and medium-sized neurons of cluster 4, as well as their varicosed axons. RFamide-like immunoreactivity (RF-ir) was detected in neurons of all neuronal clusters of the eyestalk and CNS, except clusters 1 and 5 of the eyestalk, and dorsal clusters of the subesophageal, thoracic, and abdominal ganglia. The RF-ir was also found in all neuropils of the CNS and SG, except the lamina ganglionaris. These immunohistochemical locations of the APGWamide-like and RF-like peptides in the eyestalk indicate that these neuropeptides could modulate the release of the neurohormones in the sinus gland. The presence of RFamide-like peptides in the thoracic and abdominal ganglia suggests that it may act as a neurotransmitter which controls muscular contractions. In the ovary, RF-ir was found predominantly in late previtellogenic and early vitellogenic oocytes, and to a lesser degree in late vitellogenic oocytes. These RFs may be involved with oocyte development, but may also act with other neurohormones and/or neurotransmitters within the oocyte in an autocrine or paracrine manner.     

Black mat syndrome,an invasive mycotic disease of the tanner crab, Chionoecetes bairdi

Black mat syndrome, caused by an encrusting fungus on the exterior of the carapace of tanner crabs, has been known for many years. Although it is a nuisance when processing crabs for the commercial market, it has been thought to be restricted to the external surfaces of the crab and, therefore, nonpathogenic. In the present study, 11 tanner crabs from the Kodiak area of Alaska with and 9 without grossly recognizable masses of the fungus on the carapace were necropsied and examined histologically utilizing special stains selective for fungi. In all individuals with the syndrome, hyphae of the fungus, previously identified as Phoma fimeti, penetrated the carapace and virtually replaced the underlying epidermis. In the more advanced cases, the eyestalk was invaded and the epidermis destroyed, and hyphae extended into the eyestalk musculature and nervous tissue. To date, infections of the connective tissue sheaths surrounding the esophagus, stomach, heart, hemopoetic tissue, thoracic ganglion, antennal gland, and ovary have also been observed. None of the crabs without the syndrome contained internal hyphae. Although data on the lethality of the disease are not yet available, the ease with which the hyphae penetrate the chitinous exoskeleton, their extensive proliferation in the epidermis, and their ability to invade deep tissues causing obvious pathological effects, are highly suggestive that it is a virulent, probably fatal, disease that may have a significant impact on tanner crab population dynamics.     

Effects of Neuronal Activity on Inositol Phospholipid Metabolism in the Rat Autonomic Nervous System

The effect of nerve stimulation on inositol phospholipid hydrolysis in autonomic tissue was assessed by direct measurement of [3H]inositol phosphate production in ganglia that had been preincubated with [3H]inositol. Within minutes, stimulation of the preganglionic nerve increased the [3H]inositol phosphate content of the superior cervical sympathetic ganglion indicating increased hydrolysis of inositol phospholipids. This effect was blocked in a low Ca2+, high Mg2+ medium. It was also greatly reduced when nicotinic and muscarinic antagonists were present together in normal medium. However, neither the nicotinic antagonist nor the muscarinic antagonist alone appeared to be as effective as both in combination. In other experiments, stimulation of the vagus nerve caused dramatic increases in [3H]inositol phosphate in the nodose ganglion but did not increase [3H]inositol phosphate in the nerve itself. This effect was insensitive to the cholinergic antagonists. Thus, neuronal activity increased inositol phospholipid hydrolysis in a sympathetic ganglion rich in synapses, as well as in a sensory ganglion that contains few synapses. In the sympathetic ganglion, synaptic stimulation activated inositol phospholipid hydrolysis and this was primarily due to cholinergic transmission; both nicotinic and muscarinic pathways appeared to be involved.     

The influence of optic nerve section on blood metabolite levels in Carcinus maenas (Crustacea: Brachyura)

• 1.1. Blood metabolite levels were assayed in Carcinus maenas as an indicator of the functioning of the hyperglycemic hormone, HGH, secreted by the crab's eyestalk neuroendocrine tissue.
• 2.2. Bilateral eyestalk ablation eventually resulted in a hypoglycemic response after 2–3 days.
• 3.3. Bilateral optic nerve section produced a significant, long-term hypoglycemic response suggesting that release of HGH from the eyestalk sinus gland is controlled, via a promotive neural pathway, by the CNS and probably by the cerebral ganglia.
• 4.4. Injection of eyestalk extract into operated crabs consistently produced significant, short-term hyperglycemia.

     

Neurosecretion and Salt and Water Balance in the Annelida and Crustacea

The possible role of the neurosecretory system in regulationof salt and water has been studied in the annelids and crustaceans.In the earthworm, Lumbricus terrestris, a brain factor influences'the salt and osmotic concentration of the blood and coelomicfluid. Removal of the brain results in the increase of waterinflux with a decrease in the salt and osmotic concentrationsof the body fluids. The decreases in salt and osmotic concentrationscan be prevented by the implantation of the brain or the injectionof brain homogenates. In the freshwater crayfish, Procambarus clarkii, a factor, presumablysecreted in the brain and released in the eyestalk, seems tomaintain the normal permeability of the body surfaces to water.Eyestalk removal, which eliminates the release site, resultsin the increased influx of water with a decrease in the saltconcentration of the blood. A brain factor also seems to beinvolved in maintaining the sodium and osmotic concentrationsof the blood. In the semi-terrestrial grapsid crab, Metopograpsus messor,the thoracic ganglion, under the control of an eyestalk element,secretes a factor involved in increasing the permeability ofthe body surfaces to water. The removal of the eyestalks, theimplantation of the thoracic ganglion, or the injection of extractsof thoracic ganglia, results in changes in the osmotic concentrationof the blood tending toward that of the medium. In all threespecies studied, the neuroendocrine factors seem to be involvedprimarily in the regulation of the permeability of the bodysurfaces to water.     

Current progress in shrimp endocrinology-a review

One problem in aquaculture is obtaining brood because many commercially important species are incapable of spontaneous maturation under artificial conditions. Commercial shrimp hatcheries commonly use eyestalk ablation to stimulate gonadal maturation in shrimps. Research has been conducted on the inhibition of reproductive maturation by hormones originating in the eyestalk glands and on other endocrine sources (e.g.,brain, thoracic ganglion, ovary, mandibular organ, androgenic gland and Y-organs) to determine their roles. Alternate techniques for acceleration of gonad maturation through the use of synthetic hormones or neurotransmitters may benefit aquaculture. Neurohormones and neuroregulators have been shown to accelerate gonadal maturation but an effective delivery technique must be developed for use in a large-scale aquaculture operation.     

Octopamine distribution in the tsetse fly Glossina morsitans

Tissues of Glossina morsitans were assayed for octopamine using an enzymatic technique. Octopamine was detected at the highest concentration in the brain (7.06-7.99 ng mg-1 tissue protein) and thoracic ganglion (10.9-13.89 ng mg-1 tissue protein). Octopamine was present in haemolymph at a concentration of 1.0-1.27 X 10(-7) M. This was not found to vary when insects were flown or mechanically stressed. Nervous tissue, flight muscle and haemolymph showed a significant ability to metabolize octopamine. The greatest enzyme activity was present in the haemolymph.     

Tissue-Specific Expressed Sequence Tags from the Black Tiger Shrimp Penaeus monodon

Expressed sequence tag data were generated from complementary DNA libraries created from cephalothorax, eyestalk, and pleopod tissue of the black tiger shrimp (Penaeus monodon). Significant database matches were found for 48 of 83 nuclear genes sequenced from the cephalothorax library, 22 of 55 nuclear genes from the eyestalk library, and 6 of 13 nuclear genes from the pleopod library. The putative identities of these genes reflected the expected tissue specificity. For example, genes for digestive enzymes were identified from the cephalothorax library and genes involved in the visual and neuroendocrine system from the eyestalk library. A few sequences matched anonymous EST or genomic sequences, and others contained mini-satellite or microsatellite repeat sequences. The remainder, 31 from the cephalothorax library, 25 from the eyestalk library, and 5 from the pleopod library, were sequences of high nucleotide complexity with no matches in any database searched and thus may represent novel genes. Received February 12, 1999; accepted April 13, 1999     

Neuroendocrinology of Osmoregulation in Decapod Crustacea

Experimental evidences for neuroendocrine control of osmoreregulationin decapod crustaceans are mounting. The eyestalk system, brainand thoracic ganglionic centers, and the pericardial organ appearto be involved in this control. Evidences based on experimentationwith eyestalk removal and the injection of extracts of neuroendocrinecenters are presented. Neuroendocrine extracts affect the movementsof salts and water in the gills, stomach, intestine and antennalglands. The pericardial organ material may affect osmoregulationby increasing rate of circulation and influencing salt movement. Three factors have been partially separated from CNS tissues.A factor of the freshwater crayfish, and other freshwater decapods,increases the influx of salt. Two factors from an estuarinecrab influence the movement of water, an acetone-soluble factorincreasing its influx and a water-soluble factor decreasinginflux and increasing efflux. The factors may be involved inthe adaptation of the animals to their osmotic environments. None of the factors have yet been identified in circulationnor in effector tissues. Future research must place specialemphasis on the identification of the CNS factors in circulationas well as in secretory cells and effector tissues to establishtheir true hormonal nature.     

Leptin receptors,NPY, and tyrosine hydroxylase in autonomic neurons supplying fat depots in a pig

The goal of this study was to determine the immunohistochemical characteristics of peripheral adrenergic OBR-immunoreactive (OBR-IR) neurons innervating adipose tissue in a pig. The retrograde tracer, Fast Blue (FB), was injected into either the subcutaneous, perirenal, or mesentery fat tissue depots of three male and three female pigs each with approximately 50 kg body weight. Sections containing FB(+) neurons were stained for OBR, tyrosine hydroxylase (TH) or neuropeptide Y (NPY) using a double labeling immunofluorescence method. OBR, TH, and NPY immunoreactivities were present in the thoracic (T) and lumbar (L) ganglia of the sympathetic chain, as well as in the coeliac superior mesenteric ganglion (CSMG), inferior mesenteric ganglion (IMG), intermesenteric ganglia (adrenal-ADG, aorticorenal-ARG, and ovarian-OG or testicular-TG ganglion). These results indicate that, in addition to neuroendocrine functions, leptin may affect peripheral tissues by acting on receptors located in sympathetic ganglion neurons.     

Differences in cardiac but not in neuroendocrine responses in healthy volunteers after administration of two antimuscarinic agents, atropine and pirenzepine

Neuroendocrine and cardiac responses were studied in healthy volunteers with the classical muscarinic antagonist, atropine and the new antimuscarinic agent, pirenzepine. The secretion of prolactin (PRL) and growth hormone (GH) was increased after metoclopramide. Typically, an antidopaminergic drug such as metoclopramide decreases rather than increases GH concentrations in serum. Pretreatment with both atropine and pirenzepine abolished the increase of GH secretion, which suggests an important role of cholinergic mechanisms in the regulation of GH secretion. The increase of PRL secretion was not inhibited by the two muscarinic antagonists. With the doses used, antimuscarinic activities in serum were comparable after atropine and pirenzepine treatments for the most part of the study. Heart rate was, however, significantly increased during atropine and higher than during saline or pirenzepine treatments throughout the study period. When compared to placebo, pirenzepine lowered heart rate slightly but significantly. The exact mechanism of this effect is unclear. We conclude that in contrast to the identical neuroendocrine effects, the cardiac responses clearly differ during atropine and pirenzepine treatments which confirms the ability of pirenzepine to distinguish muscarinic receptor sites in the central nervous system from those of the heart.     

Two type I crustacean hyperglycemic hormone (CHH) genes in Morotoge shrimp (Pandalopsis japonica): cloning and expression of eyestalk and pericardial organ isoforms produced by alternative splicing and a novel type I CHH with predicted structure shared with type II CHH peptides

Crustacean hyperglycemic hormone (CHH) peptide family members play critical roles in growth and reproduction in decapods. Three cDNAs encoding CHH family members (Pj-CHH1ES, Pj-CHH1PO, and Pj-CHH2) were isolated by a combination of bioinformatic analysis and conventional cloning strategies. Pj-CHH1ES and Pj-CHH1PO were products of the same gene that were generated by alternative mRNA splicing, whereas Pj-CHH2 was the product of a second gene. The Pj-CHH1 and Pj-CHH2 genes had four exons and three introns, suggesting the two genes arose from gene duplication. The three cDNAs were classified in the type I CHH subfamily, as the deduced amino acid sequences had a CHH precursor-related peptide sequence positioned between the N-terminal signal sequence and C-terminal mature peptide sequence. The Pj-CHH1ES isoform was expressed at a higher level in the eyestalk X-organ/sinus gland (XO/SG) complex and at a lower level in the gill. The Pj-CHH1PO isoform was expressed at higher levels in the XO/SG complex, brain, abdominal ganglion, and thoracic ganglion and at a lower level in the epidermis. Pj-CHH2 was expressed at a higher level in the thoracic ganglion and at a lower level in the gill. Real-time polymerase chain reaction was used to quantify the effects of eyestalk ablation on the mRNA levels of the three Pj-CHHs in the brain, thoracic ganglion, and gill. Eyestalk ablation reduced expression of Pj-CHH1ES in the brain and Pj-CHH1PO and Pj-CHH2 in the thoracic ganglion. Sequence alignment of the Pj-CHHs with CHHs from other species indicated that Pj-CHH2 had an additional alanine at position #9 of the mature peptide. Molecular modeling showed that the Pj-CHH2 mature peptide had a short alpha helix (α1) in the N-terminal region, which is characteristic of type II CHHs. This suggests that Pj-CHH2 differs in function from other type I CHHs.     

Behavioral and neuroendocrine actions of the Met-enkephalin-related peptide MERF

The effects and the mediation of the action of the proenkephalin derivative Met(5)-enkephalin-Arg(6)-Phe(7) (MERF) on the hypothalamo-pituitary-adrenal (HPA) system and open-field behavior were investigated in mice. Intracerebroventricular injection of the heptapeptide increased square crossing, rearing, and plasma corticosterone level. To characterize the receptors involved in these neuroendocrine processes, animals were pretreated either with the nonselective opioid antagonist naloxone or the kappa-antagonist nor-binaltorphimine (nor-BNI). Both antagonists dose-dependently attenuated the HPA activation elicited by MERF. Naloxone also blocked the behavioral responses, but nor-binaltorphimine did not elicit a significant inhibition. The dopamine antagonist haloperidol and a corticotropin-releasing hormone (CRH) antagonist were also preadministered to shed light on the transmission of the actions of MERF. Both the motor responses and the HPA activation were diminished by the preadministration of the CRH antagonist, while haloperidol attenuated only square crossing and rearing. To investigate the direct effect of MERF on the dopaminergic system, dopamine release of striatal slices was measured in a superfusion system. Neither the basal nor the electric impulse-evoked dopamine release was modified by MERF. The results suggest that opioid-mediation predominate in the neuroendocrine actions of MERF, and the effect of the heptapeptide on the HPA system seems to be mediated by kappa-receptors. In the behavioral responses evoked by MERF, both CRH release and the action of the dopaminergic neurons of the subcortical motor system might be involved. MERF also appears to activate the paraventricular CRH neurons, but dopaminergic transmission does not seem to play a significant role in its hypothalamic action.