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PubMed1996

Distribution of aromatase-immunoreactive cells in the forebrain of zebra finches (Taeniopygia guttata): implications for the neural action of steroids and nuclear definition in the avian hypothalamus.

Cells immunoreactive for the enzyme aromatase were localized in the forebrain of male zebra finches with the use of an immunocytochemistry procedure. Two polyclonal antibodies, one directed against human placental aromatase and the other directed against quail recombinant aromatase, revealed a heterogeneous distribution of the enzyme in the telencephalon, diencephalon, and mesencephalon. Staining was enhanced in some birds by the administration of the nonsteroidal aromatase inhibitor, R76713 racemic Vorozole) prior to the perfusion of the birds as previously described in Japanese quail. Large numbers of cells immunoreactive for aromatase were found in nuclei in the preoptic region and in the tuberal hypothalamus. A nucleus was identified in the preoptic region based on the high density of aromatase immunoreactive cells within its boundaries that appears to be homologous to the preoptic medial nucleus (POM) described previously in Japanese quail. In several birds alternate sections were stained for immunoreactive vasotocin, a marker of the paraventricular nucleus (PVN). This information facilitated the clear separation of the POM in zebra finches from nuclei that are adjacent to the POM in the preoptic area-hypothalamus, such as the PVN and the ventromedial nucleus of the hypothalamus. Positively staining cells were also detected widely throughout the telencephalon. Cells were discerned in the medial parts of the ventral hyperstriatum and neostriatum near the lateral ventricle and in dorsal and medial parts of the hippocampus. They were most abundant in the caudal neostriatum where they clustered in the dorsomedial neostriatum, and as a band of cells coursing along the dorsal edge of the lamina archistriatalis dorsalis. They were also present in high numbers in the ventrolateral aspect of the neostriatum and in the nucleus taeniae. None of the telencephalic vocal control nuclei had appreciable numbers of cells immunoreactive for aromatase within their boundaries, with the possible exception of a group of cells that may correspond to the medial part of the magnocellular nucleus of the neostriatum. The distribution of immunoreactive aromatase cells in the zebra finch brain is in excellent agreement with the distribution of cells expressing the mRNA for aromatase recently described in the finch telencephalon. This widespread telencephalic distribution of cells immunoreactive for aromatase has not been described in non-songbird species such as the Japanese quail, the ring dove, and the domestic fowl.

Animals↗

PubMed1977

The influence of the fetal hypothalamus and pituitary on the onset and course of parturition.

The possibility that the fetal brain or pituitary either initiates parturition or influences the course of labour was studied in human and rat. The results when corticotropin or neurohypophysial hormones were injected directly into human anencephalic fetuses in utero, and data obtained from 147 clinical records of such fetuses, seemed to show that the fetal brain does not trigger the onset of parturition. On the other hand, the course of labour was seriously protracted in anencephalic fetuses. Gestation length of brain-aspirated rat fetuses was not significantly longer than in sham-operated controls. However, the course of labour was protracted in the brain-aspirated fetuses. A similarly protracted expulsion pattern was observed in Brattleboro rats homozygous for a hypothalamic form of diabetes insipidus. These data all pointed to the likelihood that fetal neurohypophysial hormones stimulate the course of labour. Neither oxytocin nor vasopressin could be demonstrated in the rat fetus on the last day of pregnancy, when specific immunofluorescence was used. However, a closely related compound was found that was identified as most probably being vasotocin. The hypothesis is put forward that this fetal hormone normally stimulates the course of labour.

Anencephaly↗

PubMed2002

Distribution and quantification of corticotropin-releasing hormone (CRH) in the brain of the teleost fish Oreochromis mossambicus (tilapia).

The recent characterization of the corticotropin-releasing hormone (CRH) prehormone of the fish tilapia (Oreochromis mossambicus) showed that more variation exists between vertebrate CRH amino acid sequences than recognized before. The present study investigates whether the deviating composition of tilapia CRH coincides with an atypical distribution of CRH in the brain. For this purpose we applied immunohistochemistry, as well as radioimmunoassay (RIA) quantification in brain slices. The results are plotted in a new atlas and reconstruction of the tilapia brain. The largest population of CRH-immunoreactive (ir) neurons is present in the lateral part of the ventral telencephalon (Vl). Approximately tenfold less CRH-ir neurons are observed in the preoptic and tuberal region. The CRH-ir neurons observed in the preoptic region are parvocellular and do not, or hardly, display arginine-vasotocin (AVT) immunoreactivity. CRH-ir neurons are also present in the glomerular layer of the olfactory bulb, in the periventricular layer of the optic tectum, and caudal to the glomerular nucleus. A very dense plexus of CRH-ir terminals is located in the most rostral part of the dorsal telencephalon. This region has not been described in other teleosts and is in the present study subdivided into the anterior part of the dorsal telencephalon (Da) and the anterior part of the laterodorsal telencephalon (Dla). High densities of CRH-ir terminals were observed in and around Vl, in the tuberal region, around the rostral part of the lateral recess, and in the caudal part of the vagal lobe. In the pituitary, CRH-ir terminals are concentrated in the neuro-intermediate lobe. Overall, the immunohistochemical and quantitative data correlated well, as the RIA CRH profile in serial 160-microm slices revealed four peaks, which corresponded with major ir-cell groups and terminal fields. Our results strongly suggest that the CRH-ir cells of Vl project to the rostro-dorsal telencephalon. Consequently, they may not be primarily involved in regulation of pituitary cell types but may subserve other functions. The presence of a CRH-containing Vl-Da/Dla projection seems to be restricted to the most modern group of teleosts, i.e., the Acanthopterygians. Further anatomic indications for non-pituitary-related functions of CRH are found in the vagal lobe and the optic tectum of tilapia. Although the low CRH content of the preoptic region reported here for tilapia may be typical for unstressed fish, the fact remains that remarkably few CRH-ir neurons are involved in regulating the pituitary. Overall, the CRH distribution in the brain of tilapia is more widespread than previously reported for other teleosts.

Animals↗

PubMed1980

Functional and morphological studies of peptide-containing neuroendocrine cells in goldfish hypothalamus.

We report the development of a double-label method which combines intracellular recording, dye-marking, and immunocytochemistry for the study of functional and morphological aspects of peptide-containing neurons in the magnocellular preoptic nucleus of the goldfish hypothalamus. Using multiple techniques, we distinguish three types of peptide-containing neurons: enkephalin (27%), isotocin (49%), and vasotocin (24%). Drawings of serial sections containing the dye-injected cells were the basis for subsequent reconstruction. These peptide-containing cells measure 14-56 micrometer in somata diameter, are unipolar, bipolar, or multipolar in somata shape and lie 6-244 micrometer from the ependymal lining of the preoptic recess of the third ventricle. Their electrophysiological properties match those of other mammalian and fish magnocellular neuroendocrine cells. This report confirms the one neuron, one hormone (peptide) hypothesis, supports synaptic over electrotonic coupling between peptidergic and adjacent hypothalamic neurons, and suggests that chemical and functional classification of single electrophysiologically and neuroanatomically studied central neurons is feasible technologically.

Animals↗

PubMed1985

The distribution of hypothalamic and extrahypothalamic vasotocinergic cells and fibers in the brain of a lizard, Gekko gecko: presence of a sex difference.

The distribution of vasotocin in the brain of the lizard Gekko gecko was studied with immunocytochemical methods. Vasotocinergic cells were found in the nucleus supraopticus, the nucleus paraventricularis, the bed nucleus of the stria terminalis, and in the rhombencephalon. Vasotocinergic fibers were found in the preoptic area, the lateral and ventral hypothalamus, and in many extrahypothalamic brain areas. Furthermore, evidence was obtained of a conspicuous sex difference with regard to vasotocinergic innervation of the lateral septum, the ventrocaudal telencephalon (nucleus sphericus), and the periaqueductal gray; in these areas vastocinergic innervation is much denser in males than in females. The results are discussed in relation to the sexually dimorphic vasopressinergic innervation of the rat brain. It is suggested that the vasotocinergic fiber system in the sexually dimorphic brain areas in Gekko gecko is related to the bed nucleus of the stria terminalis.

Animals↗

PubMed1992

Comparative analysis of the vasotocinergic and mesotocinergic cells and fibers in the brain of two amphibians, the anuran Rana ridibunda and the urodele Pleurodeles waltlii.

To obtain more insight into the vasotocinergic and mesotocinergic systems of amphibians and the evolution of these neuropeptidergic systems in vertebrates in general, the distribution of vasotocin (AVT) and mesotocin (MST) was studied immunohistochemically in the brains of the anuran Rana ridibunda and the urodele Pleurodeles waltlii. In Rana, AVT-immunoreactive cell bodies are located in the nucleus accumbens, the dorsal striatum, the lateral and medial part of the amygdala, an area adjacent to the anterior commissure, the magnocellular preoptic nucleus, the hypothalamus, the mesencephalic tegmentum, and in an area adjacent to the solitary tract. In Pleurodeles, AVT-immunoreactive somata are confined to the medial amygdala, the preoptic area, and an area lateral to the presumed locus coeruleus. In both species, the distribution of MST-immunoreactive cell bodies is more restricted: in the frog, MST-immunoreactive somata are present in the medial amygdala and the preoptic area, whereas, in the urodele, cell bodies are found only in the preoptic area. Both in Rana and Pleurodeles, AVT- and MST-immunoreactive fibers are distributed throughout the brain and spinal cord. A major difference is that in Rana the number of MST-immunoreactive fibers is evidently higher than that of AVT-immunoreactive fibers, whereas the opposite is found in Pleurodeles. This holds, in particular, for the forebrain and the brainstem. The presence of several extrahypothalamic AVT-immunoreactive cell groups and the existence of well-developed extrahypothalamic networks of AVT- and MST-immunoreactive fibers are features that amphibians share with amniotes. However, this study has revealed that major differences exist not only between species of different classes of vertebrates, but also within a single class. In order to determine whether features of these neuropeptidergic systems are primitive or derived, a broad selection of species of each class of vertebrates is needed.

Animals↗

PubMed2002

Cell signaling and ion transport across the fish gill epithelium.

A large array of circulating and local signaling agents modulate transport of ions across the gill epithelium of fishes by either affecting transport directly or by altering the size and distribution of transporting cells in the epithelium. In some cases, these transport effects are in addition to cardiovascular effects of the same agents, which may affect the perfusion pathways in the gill vasculature and, in turn, affect epithelial transport indirectly. Prolactin is generally considered to function in freshwater, because it is the only agent that allows survival of some hypophysectomized fish species in freshwater. It appears to function by either reducing branchial permeability, Na,K-activated ATPase activity, or reducing the density of chloride cells. Cortisol was initially considered to produce virtually opposite effects (e.g., stimulation of Na,K-activated ATPase and of chloride cell size and density), but more recent studies have found that this steroid stimulates ionic uptake in freshwater fishes, as well as the activity of H-ATPase, an enzyme thought to be central to ionic uptake. Thus, cortisol may function in both high and low salinities. Growth hormone and insulin-like growth factor appear to act synergistically to affect ion regulation in seawater fishes, stimulating both Na,K-activated ATPase and Na-K-2Cl co-transporter activity, and chloride cell size, independent of their effects on growth. Some of the effects of the GH-IGF axis may be via stimulation of the number of cortisol receptors. Thyroid hormones appear to affect seawater ion regulation indirectly, by stimulating the GH-IGF axis. Natriuretic peptides were initially thought to stimulate gill ionic extrusion, but recent studies have not corroborated this finding, so it appears that the major mode of action of these peptides may be reduction of salt loading by inhibition of oral ingestion and intestinal ionic uptake. Receptors for both arginine vasotocin and angiotensin have been described in the gill epithelium, but their respective roles and importance in fish ion regulation remains unknown. The gill epithelium may be affected by both circulating and local adrenergic agents, and a variety of studies have demonstrated that stimulation of alpha-adrenergic versus beta-adrenergic receptors produces inhibition or stimulation of active salt extrusion, respectively. Local effectors, such as prostaglandins, nitric oxide, and endothelin, may affect active salt extrusion as well as gill perfusion. Recent studies have suggested that the endothelin inhibition of salt extrusion is actually mediated by the release of both NO and prostaglandins. It is hoped that modern molecular techniques, combined with physiological measurements, will allow the dissection of the relative roles in ion transport across the fish gill epithelium of this surprisingly large array of putative signaling agents.

Adaptation, Physiological↗

PubMed1984

Endocrine regulation of avian renal function.

The avian kidney contains reptilian-type (RT) nephrons that do not function in concert and mammalian-type (MT) nephrons that do function in concert to permit production of concentrated urine. Significant data on endocrine regulation of renal function are available only for antidiuretic hormone (arginine vasotocin, AVT) and parathyroid hormone (PTH). Plasma AVT increases and number of filtering RT nephrons decrease when plasma osmolality increases. Small doses of AVT reduce the number of filtering RT nephrons, and all RT nephrons filter following acute neurohypophysectomy. In addition to changing tubule permeability to water, AVT affects the concentrating mechanism by reducing the number of filtering RT nephrons and flow through collecting ducts. Both net tubular reabsorption and net tubular secretion of PO4 occur. Parathyroidectomy (PTX) stimulates PO4 reabsorption, and PTH replacement decreases reabsorption and stimulates secretion, apparently in proximal tubules of RT nephrons. PTX stimulates and PTH replacement inhibits Na-dependent PO4 transport by renal brush border membrane vesicles. PTH also stimulates Ca2+ reabsorption but apparently not long the proximal tubule of RT nephrons.

Animals↗

PubMed1986

Changes in body water and plasma constituents during bullfrog development: effects of temperature and hormones.

The osmoregulatory responses to warmer temperatures and hormone treatment in cold-adapted (5 degrees C) Rana catesbeiana tadpoles and newly metamorphosed frogs were examined. Tadpoles transferred to 11 degrees C and 18 degrees C and left for 5 days lost 7% and 10% of their body weight. Plasma [Na+] was elevated 28% and 21%, respectively. Control (5 degrees C) animals maintained their body weight and plasma [Na+] constant. Daily treatment with either ovine prolactin (oPRL) or ovine growth hormone (oGH) prevented the weight loss and the increase in extracellular [Na+] that occurred when tadpoles were transferred to 18 degrees C. Neither propylthiouracil (PTU) nor arginine vasotocin (AVT) were effective in countering temperature-induced weight loss in tadpoles. Newly metamorphosed frogs transferred to 18 degrees C also lost weight; this was not prevented by daily treatment with saline, oPRL, oGH or PTU. However, in frogs treated daily with AVT, initial BW was regained by day 6. When warm-adapted (18 degrees C) tadpoles were treated daily for 18 days with saline, bPRL, bGH, thyroxine (T4), ergocornine, cortisol, or cortisol + T4, bPRL was most effective in retarding weight loss and maintaining body water content, whereas T4 + cortisol caused the greatest loss of weight and body water. By day 20, the correlations between weight loss and both body water content and hematocrit were highly significant. These data suggest that reported increases in plasma solute concentrations in larval amphibians may actually reflect decreases in extracellular fluid volume, rather than increased amounts of solutes, per se.

Animals↗

PubMed2002

Induction of parturition in snow skinks: can low temperatures inhibit the actions of AVT?

The influence of environmental factors on the timing of parturition has not been investigated in viviparous squamates. We investigated the interaction between temperature and parturition in two viviparous skink species, the southern snow skink (Niveoscincus microlepidotus) and the metallic skink (N. metallicus). In these species, the timing of parturition is separated from the completion of embryonic development; the delay is attributed to their cool and variable habitats. We examined whether the neurohypophyseal hormone arginine vasotocin (AVT) stimulated parturition in southern snow skinks with late stage embryos in autumn (approximately 6-7 months prior to parturition) and in metallic skinks with late stage embryos in summer (approximately 2-3 weeks prior to parturition). The experiments were conducted at a range of environmentally relevant temperatures (6 degrees C, 15 degrees C, 22 degrees C, and 28 degrees C). AVT induced parturition in both species at all temperatures; time until birth, however, occurred more quickly at warmer temperatures (22 degrees and 28 degrees C), whereas cooler temperatures delayed parturition. We hypothesize that if cool temperatures are preventing parturition, then temperature must act at some level within the brain to prevent or slow the secretion of AVT. Future experiments will need to determine how temperature influences AVT production. Further research is also required to determine how the timing of parturition is influenced by interactions between temperature, photoperiod, and seasonal hormone patterns.

Animals↗

PubMed2001

Design of oxytocin antagonists, which are more selective than atosiban.

We report the solid phase synthesis of four pairs of L- and D-thienylalanine (Thi/D-Thi) position two modified analogues of the following four oxytocin (OT) antagonists: des-9-glycinamide [1-(beta-mercapto-beta,beta-pentamethylene propionic acid), 2-O-methyltyrosine, 4-threonine]ornithine-vasotocin (desGly(NH2)9,d (CH2)5[Tyr(Me)2,Thr4]OVT) (A); the Tyr-(NH2)9 analogue of (A), d(CH2)5[Tyr(Me)2,Thr4,Tyr-(NH2)9]OVT (B); the Eda9 analogue (where Eda = ethylenediamine) of (A), d(CH2)5[Tyr(Me)2, Thr4, Eda9]OVT (C); and the retro Tyr10 modified analogue of (C), d(CH2)5[Tyr(Me)2, Thr4, Eda9<--Tyr10]OVT (D). The eight new analogues of A-D are (1) desGly(NH2),d(CH2)5[Thi2,Thr4]OVT, (2) desGly(NH2),d(CH2)5[D-Thi2,Thr4]OVT, (3) d(CH2)5[Thi2, Thr4,Tyr-(NH2)9]OVT, (4) d(CH2)5[D-Thi2,Thr4,Tyr-(NH2)9]OVT (5) d(CH2)5[Thi2,Thr4Eda9]OVT, (6) d(CH2)5[D-Thi2,Thr4,Eda9]OVT, (7) d(CH2) [Thi2,Thr4,Eda9<--Tyr10]OVT, (8) d(CH2),[D-Thi2,Thr4,Eda9<--Tyr10]OVT. We also report the synthesis of (C). Peptides 1-8 and C were evaluated for agonistic and antagonistic activities in in vitro and in vivo OT assays, in in vivo vasopressor (V1a receptor) assays and in in vivo antidiuretic (V2 receptor) assays. None of the eight peptides nor C exhibit oxytocic or vasopressor agonism. Peptides 1-8 are extremely weak V2 agonists (antidiuretic activities range from < 0.0005 to 0.20 U/mg). Peptide C is a weak mixed V2 agonist/antagonist. Peptides 1-8 and C exhibit potent in intro (no Mg2+) OT antagonism (anti-OT pA2 values range from 7.76 to 8.05). Peptides 1-8 are all OT antagonists in vivo (estimated in vivo anti-OT pA2 values range from 6.54-7.19). With anti-V1a pA2 values of approximately 5-5.80, peptides 1-8 exhibit marked reductions in anti-V1a potencies relative to those of the parent peptides A-D (anti-V1a pA2 range from 6.48 to 7.10) and to l-deamino[D-Tyr(Et)2, Thr4]OVT (Atosiban, trade name Tractocile) (anti-V1a pA2-6.14). Atosiban has recently been approved in Europe for clinical use for the prevention of premature labour (Pharm. J. 264(7-100): 871). Peptides 1-8 exhibit striking gains in in vitro anti-OT/anti-V1a selectivities with respect to the parent peptides A, B, C and D and to Atosiban. Peptides 1-8 exhibit anti-OT (in vitro)/anti-V1a selectivities of 450, 525, 550, 450, approximately 1080, 116, 355, 227 respectively. The corresponding values for A-D and Atosiban are 30, 4.2, 4.3, 2.6 and 37. With the exception of peptide 6, the remaining seven peptides exhibit 3-18-fold gains in anti-OT (in vivo)/anti-V1a selectivity with respect to Atosiban, peptides 1-8 exhibit anti-OT (in vivo)/anti-V1a selectivities of 22, approximately 82, approximately 82, 147, approximately 83, 11, 31 and 42. By comparison, Atosiban exhibits an anti-OT (in vivo)/anti-V1a selectivity = 8. With an estimated in vivo anti-OT pA2 value = 7.19+/-0.06, peptide 4 is equipotent with Atosiban (pA2 = 7.05+/-0.05). However, with its significantly reduced anti-vasopressor potency, pA2 = approximately 5, it is approximately 18 times more selective for OT receptors with respect to VP V1a receptors than Atosiban. Since we have shown that V1a antagonism could be an unwanted side-effect in tocolytics, peptide 4 and some of the OT antagonists reported here have advantages over Atosiban and thus may be suitable candidates for evaluation as potential tocolytic agents for the treatment of preterm labour.

Biological Assay↗

PubMed2005

Design of peptide oxytocin antagonists with strikingly higher affinities and selectivities for the human oxytocin receptor than atosiban.

The peptide oxytocin (OT) antagonist atosiban, approved for tocolytic use in Europe (under the tradename Tractocile), represents an important new therapeutic advance for the treatment of premature labor. This paper presents some new peptide OT antagonists which offer promise as superior tocolytics. The solid phase synthesis is reported of four pairs of L and D-2-naphthylalanine (L/D-2Nal) position-2 modified analogs of the following four oxytocin (OT) antagonists: des-9-glycinamide [1-(beta-mercapto-beta,beta-pentamethylene propionic acid), 2-O-methyltyrosine, 4-threonine]ornithine-vasotocin (desGly-NH(2),d(CH(2))(5)[Tyr(Me)(2),Thr(4)]OVT) (A); the Tyr-NH(2) (9) analog of (A), d(CH(2))(5)[Tyr(Me)(2),Thr(4),Tyr-NH(2) (9)]OVT (B); the Eda(9) analog of (A), d(CH(2))(5)[Tyr(Me)(2),Thr(4),Eda(9)]OVT (C); and the retro COCH(2)Ph(4-0H)(10) modified analog of (C), d(CH(2))(5)[Tyr(Me)(2),Thr(4),Eda(9)<-- COCH(2)Ph(4-0H)(10)]OVT (D). The eight new analogs of A-D are (1) desGly-NH(2),d(CH(2))(5)[D-2Nal(2),Thr(4)]OVT, (2) desGly-NH(2),d(CH(2))(5)[2-Nal(2),Thr(4)]OVT, (3) d(CH(2))(5)[D-2Nal(2),Thr(4),Tyr-NH(2) (9)]OVT, (4) d(CH(2))(5)[2Nal(2),Thr(4),Tyr-NH(2) (9)]OVT, (5) d(CH(2))(5)[D-2Nal(2),Thr(4),Eda(9)]OVT, (6) d(CH(2))(5)[2Nal(2),Thr(4),Eda(9)]OVT, (7) d(CH(2))(5)[D-2Nal(2),Thr(4),Eda(9)<-- COCH(2)Ph(4-0H)(10)]OVT, (8) d(CH(2))(5)[2Nal(2),Thr(4),Eda(9)<-- COCH(2)Ph(4-OH)(10)]OVT. Peptides 1-8 were evaluated for agonistic and antagonistic activities in in vitro and in vivo rat bioassays, in rat OT receptor (rOTR) binding assays and in human OT receptor (hOTR) and human vasopressin (VP) vasopressor (V(1a)) receptor (hV(1a)R) binding assays. Also reported are the hOTR and hV(1a)R affinity data for atosiban and for B. None of the eight peptides exhibit oxytocic or vasopressor agonism. Peptides 1-8 exhibit weak antidiuretic agonism (activities in the range 0.014-0.21 U/mg). Peptides 1-6 exhibit potent in vitro (no Mg(2+)) OT antagonism (anti-OT pA(2) values range from 7.63 to 8.08). Peptides 7 and 8 are weaker OT antagonists. Peptides 1-6 are all OT antagonists in vivo (estimated in vivo anti-OT pA(2) values in the range 6.94-7.23). Peptides 1-8 exhibit vasopressor antagonism, anti-V(1a) pA(2) values in the range 5.1-7.65. Peptides 1-8 exhibit high affinities for the rOTR (K(i) values = 0.3-7.8 nM). Peptides 1-4 and B exhibit surprisingly very high affinities for the hOTR; their K(i) values are 0.17, 0.29, 0.07, 0.14 and 0.59 nM, respectively. Peptides 1-4 and B exhibit respectively 449, 263, 1091, 546 and 129 times greater affinity for the hOTR than atosiban (K(i) = 76.4 nM). Peptides 1-4 exhibit high affinities for the hV(1a)R (K(i)s = 1.1 nM, 1.3 nM, 0.19 nM and 0.54 nM, all higher than the hV1(a)R affinities exhibited by atosiban (K(i) = 5.1 nM) and by B (K(i) = 5.26 nM). Because of their strikingly higher affinities for the hOTR than atosiban, peptides 1-4 and B exhibit gains in anti hOT/anti hV(1a) receptor selectivity compared with atosiban of 93, 64, 39, 56 and 127, respectively. These OT antagonists are thus promising candidates for development as potential new tocolytic agents.

Animals↗

PubMed1993

Peptide hormone-stimulated second messenger production in the teleostean nephron.

Renal actions of arginine vasotocin (AVT) and atrial natriuretic peptide (ANP) in teleosts are poorly defined. Administration of these hormones changes renal function, largely explained by systemic vascular effects. The present experiments examine the potential direct action of these hormones on the teleost nephron. This has been assessed from the hormones' abilities to stimulate their second messenger systems within isolated nephroi of the rainbow trout Onchorhynchus mykiss. Following initial experiments to define the experimental conditions, AVT was found to induce a dose-dependent increase in cAMP content of a suspension of nephroi incubated for 10 min at 10 degrees; 10(-5) M AVT provoking an increase of 846% (SW) and 829% (FW) above basal cAMP accumulation of 88 and 125 pmol/mg protein, respectively. This V2-type receptor response to AVT (or AVP) has previously been considered to be present only in the tetrapod kidney. ANP evoked a dose-dependent stimulation of cGMP accumulation from 1.3 (SW) or 1.9 (FW) pmol/mg protein basal levels to 5806 and 4405% of these levels, respectively, at the highest concentration used of 3.3 x 10(-7) M. Further experiments using isolated glomeruli from the eel Anguilla anguilla localized at least a part of the response to ANP to a glomerular site.

Anguilla↗

PubMed1998

The hypothalamus and adrenal regulate modulation of corticosterone release in redpolls (Carduelis flammea--an arctic-breeding song bird).

Free-living redpolls (Carduelis flammea-a species that breeds in the Alaskan Arctic), modulate corticosterone release in response to capture and restraint depending upon the breeding site. We extended these findings to adults undergoing a prebasic molt (the energetically costly replacement of feathers) and to juveniles. Results indicate not only that the stress response is dramatically reduced at one breeding site, but that the stress response during molt and in juveniles is lower still. In fact, juveniles failed to secrete any corticosterone in response to capture and handling, suggesting a stress hyporesponsive period. We also examined possible mechanisms underlying stress modulation. Corticosterone binding protein capacity does not change, and the stress response is only correlated with the overall condition of the bird (assessed by fat storage) at one site, suggesting that neither can explain the different corticosterone responses. Adrenal insensitivity also does not appear to fully explain reduced maximal output since exogenous ACTH enhanced corticosterone release. Exogenous ACTH, however, cannot stimulate corticosterone to stress-induced levels at the high-response site, implying reduced adrenal capacity. Redpoll pituitaries responded to exogenous corticotrophin-releasing factor and arginine vasotocin, suggesting a mechanism upstream from the pituitary blunts corticosterone release. Taken together, these results indicate that corticosterone release in this species is modulated depending upon the ecological and physiological state of the animal, and that the maximal corticosterone response is controlled at multiple sites in the hypothalamic-pituitary-adrenal axis.

Adrenal Glands↗

PubMed1998

Preoptic AVT immunoreactive neurons of a teleost fish with alternative reproductive tactics.

Recent reports have implicated an important role for arginine vasotocin (AVT) in the socially mediated sexual differentiation of fishes. This study focuses on the plainfin midshipman (Porichthys notatus) which exhibits two male morphs, type I and type II, differing in a suite of behavioral, neurobiological, and endocrine traits. Immunocytochemical techniques were used to detect neurons containing AVT-like peptide in the forebrain of juveniles, adult females, and type I and type II males. AVT immunoreactive (ir) somata were localized to three regions: the terminal nerve ganglion, the preoptic area (POA), and the pineal stalk. The profile area, or size, of AVT-ir POA neurons differed across the four classes of midshipman and was strongly correlated to differences in body size among the groups. By contrast, the number of AVT-ir cells in the POA exhibited no difference across the classes of midshipman. The number of POA cells containing AVT is therefore likely to be set early in development and not to change with the growth of the animal. An analysis of AVT-ir cell number normalized by body mass revealed that the larger morphs, type I males and females, have fewer cells per gram body mass than type II males and juveniles. Therefore, type II males have a juvenile-like AVT POA phenotype with smaller cells and more numerous cells per unit body mass than type I males. Type II males also exhibit more variability in the number of AVT-ir cells found in the POA compared to type I males.

3-Hydroxysteroid Dehydrogenases↗

PubMed1999

Effects of naloxone on neurohypophyseal peptide release by hypertonic stimulation in chicks.

The effects of opioid peptides on the osmotic release of neurohypophyseal hormones, arginine vasotocin (AVT) and mesotocin (MT), were determined in 2-day-old chicks. Experiment 1 examined the effect of a variety of doses of naloxone, an opioid antagonist, on chicks administered isotonic or hypertonic solution. Plasma osmolality in chicks administered hypertonic solution was significantly higher than that in groups administered isotonic solution. None of the doses of naloxone affected plasma osmolality in response to isotonic and hypertonic solution. Plasma levels of AVT increased in hypertonic solution and this response was further enhanced by naloxone injection as the doses increased. The hypertonic solution alone did not affect plasma levels of MT, but additional treatment with naloxone slightly increased plasma levels of MT. Experiment 2 examined the effect of DAMGO ([d-Ala(2), N-Me-Phe(4),Gly-ol]-enkepha lin), a specific mu receptor agonist. Relatively high plasma osmolality caused by hypertonic solution was not affected by additional treatment with DAMGO. Plasma levels of AVT in response to hypertonic solution and to additional treatment with naloxone were reduced by higher doses of DAMGO. Experiment 3 examined the effect of naloxone on chicks administered different concentrations of NaCl. Administration of hypertonic solution resulted in an increase in plasma osmolality and plasma levels of AVT. Naloxone administration enhanced the increase in plasma AVT levels in response to hypertonic solution. Experiment 4 examined the effect of naloxone on different kinds of hypertonic solution, 0.15 M NaCl, 1.5 M NaCl, 2.55 M urea, and 1.95 M sucrose. The increases in plasma osmolality resulting from the administration of the urea and sucrose solutions were the same as those in the chicks injected with 1.5 M NaCl. In sucrose-treated chicks, plasma levels of AVT increased in chicks administered naloxone but not in chicks injected with normal saline. In contrast, no significant changes in plasma levels of AVT were observed in urea treatment with or without naloxone. In Experiments 3 and 4, plasma levels of MT after administration of hypertonic solutions did not change. However, naloxone administration enhanced plasma levels of MT in osmotically stimulated chicks. The results of the present study suggest that opioid peptides attenuate the increase in plasma AVT and MT in hypertonic states.

Analgesics, Opioid↗

PubMed1999

Seasonal changes in expression of neurohypophysial hormone genes in the preoptic nucleus of immature female masu salmon.

In relevance to osmoregulatory and reproductive functions, activity of the hypothalamic magnocellular neurosecretory system may vary seasonally in teleosts. The changes in the expression of vasotocin (VT) and isotocin (IT) genes were thus studied by an in situ hybridization technique and an immunohistochemical avidin-biotin complex method in immature female masu salmon (Oncorhynchus masou). The plasma levels of testosterone and estradiol were also measured by enzyme immunoassay. Fish were sampled in March, May, August, and November 1994 and January 1995. The intensity of autoradiographic hybridization signals and immunoreactivity were determined in individual neurosecretory cells (NSC) in the rostroventral, middle, and dorsocaudal regions of the magnocellular part of the preoptic nucleus (PM). The VT hybridization signals and immunoreactivity were high in November, along with the elevation of plasma levels of testosterone and estradiol. These results suggest that sex steroid hormones are involved in seasonal regulation of VT gene expression. The hybridization signals for IT mRNA were increased in May and decreased in November, whereas IT immunoreactivity was low in March and high in November. NSCs thus showed seasonal variations in the intensity of hybridization signals for VT and IT mRNAs and immunoreactivity of VT and IT, although the patterns of changes were different between VT and IT. VT and IT genes may be seasonally expressed under different regulatory mechanisms.

Animals↗

PubMed1999

Differences in seasonal expression of neurohypophysial hormone genes in ordinary and precocious male masu salmon.

Our previous study showed the seasonal variations in expression of vasotocin (VT) and isotocin (IT) genes in preoptic magnocellular neurons of female masu salmon (Oncorhynchus masou). The changes in the level of VT mRNA were coincident with those in plasma testosterone and estradiol levels. In the present study, generality of this phenomenon in salmonid was verified in males. We examined changes in expression of VT and IT genes by an in situ hybridization technique and an immunohistochemical avidin-biotin complex method in the preoptic nuclei of ordinary and precocious male masu salmon. Plasma levels of testosterone and estradiol were measured by enzyme immunoassay. Fish were sampled in March, May, August, and November 1994 and January 1995. The intensities of hybridization signals for VT and IT mRNAs, as well as immunoreactivity of VT and IT, showed seasonal variations, although the profiles were different between the ordinary and precocious males. In the ordinary males, the intensities of hybridization signals for VT and IT mRNAs were high in January. These strong hybridization signals, representing elevation of VT and IT gene expression, were accompanied by increases in plasma levels of testosterone and estradiol. However, in precocious males, changes in VT and IT mRNA levels were not coincident with variation of plasma levels of sex steroid hormones. The sensitivity to sex steroid hormones of VT and IT gene expression may be different between the ordinary and precocious male masu salmon.

Animals↗