Phylogenetic study of the oxytocin-like immunoreactive system in invertebrates

1. A phylogenetic study of oxytocin (OXT)-like immunoreactive cells was performed by the PAP method in the central nervous system of invertebrates. 2. The immunoreactivity was detected in the nerve cells of Hydra magnipapillata of the Coelenterata; Neanthes japonica and Pheretima communissima of the Annelida; Oncidium verrucosum, Limax marginatus and Meretrix lamarckii of the Mollusca; and Baratha brassica of the Arthropoda. 3. No immunoreactive cells were found in Bipalium sp. of the Platyhelminthes; Pomacea canaliculata, Aplysia kurodai, Bradybaena similaris and Achatina fulica of the Mollusca; and Gnorimosphaeroma rayi, Procambarus clarkii, Hemigrapsus sanguineus, Helice tridens and Gryllus bimaculatus of the Arthropoda; Asterina pectinifera of the Echinodermata; and Halocynthia roretzi of the Protochordata. 4. These results demonstrate that an OXT-immunoreactive substance is widely present not only in vertebrates but also in invertebrates. 5. OXT seems to have been introduced into these invertebrates at an early stage of their phylogenetic history.     

Arginine vasotocin mRNA revealed by in situ hybridization in bovine pineal gland cells

Arginine vasopressin (AVP) is the main antidiuretic hormone in mammals and arginine vasotocin (AVT) in submammalian vertebrates. The possibility that the genetic material encoding AVT is maintained in mammals is controversial. In this study, we investigated by radioactive in situ hybridization the possible presence of the mRNA encoding AVP and AVT, and using immunocytochemistry the presence of structures immunoreactive for AVP and AVT in the bovine pineal gland. In situ hybridization was performed by use of ³⁵S-labelled oligoprobes. Immunocytochemistry was performed using specific polyclonal rabbit antibodies and the avidin-biotin-complex method. In situ hybridization revealed positive signals for both AVP mRNA and AVT mRNA in a few cells scattered throughout the pineal body. Immunocytochemistry revealed thin AVP-immunoreactive fibres in the pineal stalk and the pineal gland. It also revealed staining of several AVT-immunoreactive nerve fibres in both the pineal stalk and the gland. In addition, polyhedral, neuron-like cell bodies from which two to three processes emerged were also AVT-immunoreactive. Thus, our investigation shows the presence of AVP/AVT-immunoreactive cellular structures in the bovine pineal gland. Our data further show the presence of mRNAs encoding both AVT and AVP. We therefore suggest that AVT mRNA is translated into an AVT-like peptide in the bovine pineal.     

Oxytocin antagonist blocks the vasodepressor but not the vasopressor effect of neurohypophysial peptides in chickens

Cockerels with permanent cannulas in the brachial artery and vein were put into isolated slings. Arterial pressure and heart rate were continuously recorded. Following habituation, tests were initiated. In each cockerel 2 nmol/kg of the tested neurohypophysial peptide (NPs) or analogue was IV injected six times at 6-min intervals. Arginine vasotocin (AVT) caused an immediate vasodepressor (VDP) effect and tachycardia. These subsided within 20–30 s and were followed by a vasopressor (VP) response and bradycardia. On repeated injections of AVT, the VDP response declined and bradycardia intensified. Arginine vasopressin (AVP), oxytocin (OT), and mesotocin (MT) had short-lasting VDP effect in the following order of potency: OT = MT > AVT > AVP. Only AVT and, more effectively, AVP, caused a VP response. The VDP effect of MT and OT declined on repeated injections. When AVT was injected after three injections of MT, it had mostly an immediate VP effect. Although the V₁ agonist is VP in chickens, at the dose used the V₁ antagonist, [d(CH₂)₅,O-Me-Tyr²]AVP, had no effect on cardiovascular responses to AVT. Pretreatment with OT antagonist, [d(CH₂)₅-O-Me-Tyr^2,Thr^4,Tyr^9,Orn⁸]VT, abolished the VDP effect of all NPs. Thus, MT had no effect on blood pressure, whereas AVP and, more effectively, AVT, had a marked immediate VP action. In chickens the VDP effect of NPs is probably mediated by an OT/MT-like receptor, wherein the peptide's ring structure, shared by AVT, OT, and MT, is important. The VP effect is mediated by a receptor only partially similar to the mammalian V₁ receptor, where arginine in position 8, shared only by AVT and AVP, is necessary for action, and the native AVT is more effective than the mammalian AVP. This receptor reacts to the V₁ agonist but probably not to the V₁ antagonist.     

Antipyretic effect of arginine vasotocin in toads

Arginine vasotocin (AVT) is a nonmammalian analog of the mammalian hormone arginine vasopressin (AVP). These peptides are known for their antidiuretic and pressor effects. More recently, AVP has been recognized as an important antipyretic molecule in mammals. However, no information exists about the role of AVT in febrile ectotherms. We tested the hypothesis that AVT is an antipyretic molecule in the toad Bufo paracnemis. Toads equipped with a temperature probe were placed in a thermal gradient, and preferred body temperature was recorded continuously. A behavioral fever was observed after lipopolysaccharide (LPS) was injected systemically (200 microg/kg). Systemically injected AVT (300 pmol/kg) alone caused no significant change in body temperature, but abolished LPS-induced fever. Moreover, a smaller dose of AVT (10 pmol/kg), which did not affect LPS-induced fever when injected peripherally, abolished fever when injected intracerebroventricularly. We therefore conclude that AVT plays an antipyretic role in the central nervous system, by means of behavior, in an ectotherm, a fact consistent with the notion that AVT/AVP elicits antipyresis by reducing the thermoregulatory set point.     

Immunocytochemical evidence for the presence of an albumin-like substance in the rat pineal gland

Summary Two rabbits and two guinea pigs were immunized with arginine vasotocin (AVT) conjugated to bovine albumin with glutaraldehyde. Only one preparation of antiserum (anti-G 7) was of value. Anti-G 7 immunochemically defined the same rat pineal cells previously reported as presumptive AVT cells. However, absorption of anti-G 7 with bovine albumin inhibited the staining of the pineal cells demonstrating that they contained an albumin-like substance. Positive immunochemical staining of the rat pars nervosa suggested that anti-G 7 contained antibodies able to react with arginine vasopressin (AVP). Loss of a positive reaction in the posterior lobe on absorption of anti-G 7 with AVT or AVP confirmed this. However, the addition of AVT to anti-G 7 failed to inhibit the immunochemical staining of the pineal cells. This study reports the presence of an albumin-like substance in pineal cells previously described as presumptive AVT cells, and discusses possible explanations for the inability of anti-G 7 to recognize immunocytochemically the native AVT molecule. Confirmation of AVT in the pineal gland by immunocytochemistry must await the availability of more specific antisera.     

A comparison of grooming behavior potencies of neurohypophyseal nonapeptides

We have previously demonstrated that intracerebroventricular (ICV) administration of oxytocin (OXY) enhanced grooming behaviors in male and female rats at a 1 microgram dose. In the present study female rats were injected ICV with 1 microgram OXY or equimolar doses of other peptides. At this dose arginine-vasopressin (AVP), arginine-vasotocin (AVT) and lysine-vasopressin (LVP), as well as alpha-MSH, were as effective as OXY in increasing grooming behavior. At equimolar doses, ACTH1-10, tocinoic acid (the ring structure of OXY) and Pro-Leu-Gly-NH2 (the tail structure of OXY) had no significant effect on grooming behavior. The potency of AVP and AVT was determined across a 0.05-5 microgram dose range. Grooming scores increased in an apparent linear manner across a similar OXY dose range. Both AVP and AVT, however, manifested an inverted U grooming response curve. Maximum grooming scores resulted from a 0.1 microgram dose of AVT or a 0.5 microgram AVP dose. Analyses of the aspects of grooming separately found that nonapeptides OXY, AVP and AVT all elevated body grooming, washing, and scratching. Because AVT and AVP administration resulted in grooming scores significantly higher than OXY at lower doses, we concluded that the CNS is more sensitive to the effects of AVT and AVP on grooming behavior than OXY.     

Developmental and Evolutionary Aspects of the Neurohypophysis

Although phylogenetic studies of neurohypophysial principlesare well advanced, studies of their development are rare. Fetalmammals have been shown to have unusually high vasopressor/oxytocicratios in their pituitaries. Paper chromatography of extractsfrom pituitaries of fetal sheep and fetal seals (Callorhinusursinus) indicated the presence of AVT, in addition to AVP andoxytocin. This was confirmed by the pharmacological and chemicalanalysis of AVT, purified from fetal seal pituitaries. The presenceof AVT suggested a "recapitulation" of a principle previouslyknown only in sub-mammalian vertebrates. Studies of ammocoetelarvae and adult lampreys (Lampetra nchardsoni) showed the presenceof AVT, but there was no evidence for any "recapitulation" ofan "early" neutral principle. It seemed possible that an apparentrecapitulation might occur only when the principle had a usein fetal life. Therefore the possible actions of AVT (and AVP)in the mammalian fetus (guinea pig) were studied. AVT and AVPwere shown to act on the isolated amnion to slow, and even reversefetal-maternal water movement. AVP acted on the yolk sac, thefetal skin and the fetal bladder to increase water flow inwards,towards the fetus. These actions suggested an extraplacentalmechanism for supplying water to the fetus and its environment,through the extraembryonic membranes. These actions were reminiscentof those known in the anurans. The apparent success of the conceptof recapitulation, in both the molecular and functional fields,may have been fortuitous. However, the results suggest thatphylogenetic considerations may well be useful in understandingthe physiology of the mammal during fetal life.     

Do circulating plasma AVT and/or cortisol levels control pulsatile urea excretion in the gulf toadfish (Opsanus beta)?

Previous work has shown that pulsatile urea excretion at the gills of the gulf toadfish is due to periodic activation of a facilitated diffusion transport system with molecular and pharmacological similarity to the UT-A transport system of the mammalian kidney. In mammals, AVP and glucocorticoids are two important endocrine regulators of this system. The present study focused on the potential role of circulating AVT (the teleost homologue of AVP) and cortisol levels as possible triggers for urea pulses. Long-term (34–84 h) monitoring of plasma levels by repetitive sampling at 2-h intervals from chronic cannulae in individual toadfish demonstrated that circulating AVT concentrations are low (10⁻¹²–10⁻¹¹ M), and show no relationship to the occurrence of natural urea pulses. In contrast, plasma cortisol levels decline greatly prior to natural pulses and rise rapidly thereafter. AVT injections into the caudal artery or ventral aorta elicited pulse events, but these were extremely small (1–10%) relative to natural pulses, and occurred only at unphysiological dose levels (10⁻⁹ M in the plasma). AVP was a partial agonist, but isotocin, insulin-like growth factor-1, and atrial natriuretic peptide were without effect at the same concentration. Artificially raising plasma cortisol levels by cortisol injection tended to reduce responsiveness to AVT. Pharmacological reduction of plasma cortisol levels by metyrapone injection elicited small pulses similar to those caused by AVT. Following such pulse events, AVT was ineffective in inducing pulses. We conclude that decreases in circulating cortisol play an important permissive role in urea pulsing, but that circulating AVT levels are not involved.     

Neurohypophyseal hormones protect against pentylenetetrazole-induced seizures in zebrafish: Role of oxytocin-like and V1a-like receptor

Oxytocin (OT) and arginine-vasopressin (AVP) are involved in the physiological response to different stressors like the occurrence of seizures which is regarded as a severe stress factor. Zebrafish (Danio rerio) is recently featured as a model of epilepsy but the role of neurohypophyseal hormones on this teleost is still unknown. We attempted to determine whether non-mammalian homologues like isotocin (IT) and vasotocin (AVT) affected pentylenetetrazole (PTZ)-induced seizures in adult zebrafish in comparison with OT/AVP. The mechanism was studied using the most selective OT and AVP receptor antagonists. Zebrafish were injected i.m. with increasing doses (0.1-40ng/kg) of the neuropeptides 10min before PTZ exposure. DesGly-NH2-d(CH2)5-[D-Tyr2,Thr4]OVT (desglyDTyrOVT) for OT receptor and SR49059 for V1a subtype receptor, were injected together with each agonist 20min before PTZ exposure. All the peptides significantly decreased the number of seizures, increased the mean latency time to the first seizure and decreased lethality. This protective effect led to a dose-response curve following a U-shaped form. IT was approximately 40 times more active than OT while AVT was 20 times more potent than AVP in reducing the number of seizures. DesglyDTyrOVT was more effective in antagonizing OT/IT, while SR49059 mainly blocked AVP/AVT-induced protection against PTZ-induced seizures. The present findings provide direct evidence of an important involvement of IT/OT and AVP/AVT as anticonvulsant agents against PTZ-induced seizures with a receptor-mediated mechanism in zebrafish. These data reinforce zebrafish as an emerging experimental model to study and identify new antiepileptic drugs.     

Effects of oxytocin,vasopressin and vasotocin and their agonists on steroid secretion by bovine granulosa cells

The effects of oxytocin and vasopressin and their agonists on the secretion of progesterone and oestradiol-17β by bovine luteinised granulosa cells cultured in a serum-supplemented medium were analysed. The effects of oxytocin (OT), its long-acting agonist 2-0-methyl-tyrosin (deamino-karba)-oxytocin (DK-OT), arginine-8-vasopressin (AVP), 1-desamino-arginine-8-vasopressin (D-AVP, a vasopressin analogue with high antidiuretic and without vasopressor properties) and arginine-8-vasotocin (AVT) were investigated. It was found that OT and DK-OT had a stimulatory effect on progesterone release, while AVP, D-AVP and AVT had an inhibitory effect. All peptide hormones investigated significantly increased oestradiol-17β secretion. The results suggest the involvement of nonapeptide hormones of both oxytocin and vasopressin groups in the regulation of steroidogenesis by granulosa cells from bovine ovarian follicles.     

Arginine Vasotocin: Structure-Activity Relationships and Influence on Gonadal Growth and Function

When AVT (arginine vasotocin) was given neonatally during theperiod when the brain is undergoing sexual differentiation,increased growth of the reproductive organs was observed inadulthood. Injection of AVT after this neonatal period in immatureanimals led to diminished growth of the accessory organs andin some cases the gonads themselves. The hypertrophic responseof the in situ ovary in adult mice following unilateral ovariectomy(UO) was inhibited in a dose-related manner by a single intraperitonealinjection of freshly prepared AVT. Much less AVT was requiredfor this response when injected into the third ventricle. Afterintraperitoneal injection, arginine vasopressin (AVP), lysinevasopressin (LVP), and 4-leucine vasotocin (4-leu-AVT) alsoinhibited compensatory ovarian hypertrophy whereas oxytocindid not. The commonality in die structure of these antigonadotrophicpeptides include a closed ring and a basic amino acid in position8. After opening the disulfide bond of these nonapeptides withmercaptoethanol, a single injection of the reduced AVT, AVP,LVP, or 4-leu-AVT into UO mice causes exaggerated hypertrophyof the remaining ovary. When added with leuteinizing hormone-releasinghormone (LRH) to culture medium containing hemipituitaries fromcastrated estrogen-progesterone primed female rats, AVT significantlyincreased the release of radioimmunoassayable LH above thatdue to LRH alone. AVT might interact at all levels of the hypothalamo-hypophysealgonadalaxis.     

Galanin immunoreactivity increased in chicken supraoptic neurons after activation of the vasotocin system at oviposition

The avian arginine vasotocin (AVT) synthesized in the hypothalamic magnocellular neurons and released from the posterior pituitary is known to be involved in the regulation of uterine contractions for oviposition in chickens. However, regulation of AVT synthesis and release within the magnocellular hypothalamus has not been elucidated. Galanin, the oviposition inducing factor in the oviduct of the hen, has been demonstrated to have sexually dimorphic stimulatory action in oxytocin- and vasopressin neurons in the mammalian hypothalamus. In this study, galanin and AVT immunoreactivity was investigated around the time of oviposition in the supraoptic nucleus (SON) to determine if galanin modulates AVT synthesis and/or release. Within SON neurons increased AVT immunoreactivity before oviposition and the decreased AVT immunoreactivity after oviposition implied function-related peptide release. The significantly increased number of galanin neurons co-localizing with AVT immediately after oviposition suggests that galanin is involved in the negative feedback to limit AVT release in the SON. Thus, these data support the idea that AVT in the SON is involved in central regulation of oviposition and that AVT release could be modulated by the neuropeptide galanin.     

ACTH-releasing activity of urotensin I and ovine CRF: interactions with arginine vasotocin, isotocin and arginine vasopressin

The release of ACTH from superfused dispersed goldfish anterior pituitary cells was examined to determine if the neurohypophyseal peptides arginine vasotocin (AVT), isotocin (IST) or arginine vasopressin (AVP) potentiate the ACTH-releasing activities of the structurally homologous peptides urotensin I (UI) or ovine corticotropin-releasing factor (CRF). The ACTH-releasing activities of the neurohypophyseal peptides and UI or CRF were additive. AVT, IST or AVP failed to potentiate the ACTH-releasing activity of UI or CRF. These results suggest that in teleost fishes neurohypophyseal peptides have intrinsic ACTH-releasing activity but, unlike mammals, do not potentiate the release of ACTH evoked by CRF, or by the piscian CRF-like peptide, UI.     

Galanin immunoreactivity increased in chicken supraoptic neurons after activation of the vasotocin system at oviposition.

The avian arginine vasotocin (AVT) synthesized in the hypothalamic magnocellular neurons and released from the posterior pituitary is known to be involved in the regulation of uterine contractions for oviposition in chickens. However, regulation of AVT synthesis and release within the magnocellular hypothalamus has not been elucidated. Galanin, the oviposition inducing factor in the oviduct of the hen, has been demonstrated to have sexually dimorphic stimulatory action in oxytocin- and vasopressin neurons in the mammalian hypothalamus. In this study, galanin and AVT immunoreactivity was investigated around the time of oviposition in the supraoptic nucleus (SON) to determine if galanin modulates AVT synthesis and/or release. Within SON neurons increased AVT immunoreactivity before oviposition and the decreased AVT immunoreactivity after oviposition implied function-related peptide release. The significantly increased number of galanin neurons co-localizing with AVT immediately after oviposition suggests that galanin is involved in the negative feedback to limit AVT release in the SON. Thus, these data support the idea that AVT in the SON is involved in central regulation of oviposition and that AVT release could be modulated by the neuropeptide galanin.     

Presence of neurophysins I and II in the human pineal gland: Comparison with the content of neurohypophyseal hormones

We have previously measured the individual content of immunoreactive vasopressin (AVP), oxytocin (OT) and vasotocin (AVT) in 155 human pineal glands, and report here identification and measurement of the neurophysin (Np) content of the same glands, using specific homologous human neurophysin I (HNp I) and neurophysin II (HNp II) radioimmunoassays. Median values for HNp I were for men 47 ng/gland (range, 5 to 1360) and for women 24 ng/gland (range, 5 to 1000); median values for HNp II were respectively 7 ng/gland (range, 2 to 191) and 15 ng/gland (range, 2 to 356) with no significant difference between men and women for HNp I and HNp II but a significant difference (p<0.001) between HNp I and HNp II for both sexes. Gel filtration showed that pineal neurophysins were eluted at the same volume as both standard Np and Np from human posthypophyses used as controls. HNp I correlated both with AVP (r_s=0.54 for men and 0.55 for women) and OT (r_s=0.86 for men and 0.57 for women) but not with AVT, while HNp II correlated with AVP (r_s=0.52 for men and 0.53 for women) and OT (r_s=0.92 for men and 0.50 for women) but not with AVT. This study thus confirms the presence of two neurophysins in the human pineal gland and further indicates that they are related to AVP and OT concentrations in the same gland. The results also imply, however, that the presence of immunoreactive AVT (more probably a closely related peptide) is independent of the neurophysins.     

Vasopressin gene expression in the normal and Brattleboro rat: a histological analysis in semi-thin sections with biotinylated oligonucleotide probes

We analyzed expression of the vasopressin (AVP) gene in semi-thin sections in normal and Brattleboro rats by using in situ hybridization and immunohistochemistry. AVP mRNA was detected as follows: vibratome sections of rat hypothalamus were hybridized with a biotinylated oligonucleotide probe, embedded in Araldite, and cut into semi-thin sections which were reacted with streptavidin-alkaline phosphatase and the appropriate substrate. Adjacent serial sections were treated by immunohistochemistry to detect AVP or oxytocin immunoreactivity. In normal rat, AVP mRNA can be detected in magnocellular neurons of the supraoptic and paraventricular nuclei and in parvocellular neurons of the suprachiasmatic nucleus. AVP mRNA was present throughout the cytoplasm of the cell bodies, their processes, and in punctate structures in the vicinity of the AVP cell bodies. Most neurons containing AVP mRNA also contain AVP immunoreactivity, but the staining intensity was not consistently correlated for each reaction. A few neurons contained AVP mRNA without detectable AVP immunoreactivity. In the Brattleboro rat, staining intensity of the reaction was lower than in normal rat and the AVP mRNA was restricted mostly to the periphery of the cytoplasm. In this strain, the neurons containing the AVP mRNA did not contain AVP or oxytocin immunoreactivity. These results demonstrate that neuropeptide mRNA can be detected in semi-thin sections with a biotinylated oligonucleotide probe, and that AVP gene deletion provokes modification of the intracellular localization of the AVP mRNA.     

Do circulating plasma AVT and/or cortisol levels control pulsatile urea excretion in the gulf toadfish (Opsanus beta)?

Previous work has shown that pulsatile urea excretion at the gills of the gulf toadfish is due to periodic activation of a facilitated diffusion transport system with molecular and pharmacological similarity to the UT-A transport system of the mammalian kidney. In mammals, AVP and glucocorticoids are two important endocrine regulators of this system. The present study focused on the potential role of circulating AVT (the teleost homologue of AVP) and cortisol levels as possible triggers for urea pulses. Long-term (34-84 h) monitoring of plasma levels by repetitive sampling at 2-h intervals from chronic cannulae in individual toadfish demonstrated that circulating AVT concentrations are low (10(-12)-10(-11) M), and show no relationship to the occurrence of natural urea pulses. In contrast, plasma cortisol levels decline greatly prior to natural pulses and rise rapidly thereafter. AVT injections into the caudal artery or ventral aorta elicited pulse events, but these were extremely small (1-10%) relative to natural pulses, and occurred only at unphysiological dose levels (10(-9) M in the plasma). AVP was a partial agonist, but isotocin, insulin-like growth factor-1, and atrial natriuretic peptide were without effect at the same concentration. Artificially raising plasma cortisol levels by cortisol injection tended to reduce responsiveness to AVT. Pharmacological reduction of plasma cortisol levels by metyrapone injection elicited small pulses similar to those caused by AVT. Following such pulse events, AVT was ineffective in inducing pulses. We conclude that decreases in circulating cortisol play an important permissive role in urea pulsing, but that circulating AVT levels are not involved.     

Identification and characterization of arginine vasopressin-like substances in the rat testis

We have previously demonstrated the suppression of Leydig cell steroidogenesis by arginine vasopressin (AVP) in vitro. Since the circulating level of AVP is too low to mediate a testicular action of this peptide, we have conducted studies to identify testicular AVP-like substances. The supernatant of homogenized, acid-extracted rat testes was found to contain AVP immunoreactivity which showed parallelism with synthetic AVP in a specific radioimmunoassay for AVP. Chromatography of this extract on a Sephadex G-25 column produced three peaks of AVP immunoreactivity. The largest peak eluted close to the column void volume, a second smaller peak eluted at the total column volume, while a third peak co-eluted with synthetic AVP. Following acetone precipitation, ether extraction, and octadecylsilica (C18) adsorption chromatography of the acid extract, the third peak of AVP immunoreactivity (about 600 pg/testis) was fully retained by C18 chromatography and showed parallelism with synthetic AVP in both radioimmuno- and radioreceptor assays. This substance also co-migrated with synthetic AVP on both Sephadex G-25 and reverse-phase thin layer chromatography (RPTLC). The second peak was only partially retained by C18 adsorption chromatography, dilution curves were not parallel with synthetic AVP in radioimmuno- or radioreceptor assay, and this material failed to co-migrate with synthetic AVP on Sephadex G-25 and RPTLC. The first peak of apparent AVP immunoreactivity was associated with an enzyme(s) that degraded labeled AVP. This enzymatic activity, as well as the immunoreactivity, could be eliminated by heating the extract to 90 degrees C. These results demonstrate, by a number of independent criteria, that rat testes contain a substance which behaves like authentic AVP. Due to its high concentration, the AVP-like peptide may be synthesized or concentrated by testis cells. In addition, testis tissue contains enzymatic activity which degrades AVP and could represent a site of regulation of AVP action. Coupled with the previously demonstrated testis action of AVP, these results suggest a paracrine or autocrine role of the neurohypophysial hormone at the testis level.     

Arginine vasotocin neuronal phenotypes, telencephalic fiber varicosities, and social behavior in butterflyfishes (Chaetodontidae): potential similarities to birds and mammals

The neuropeptide arginine vasopressin (AVP) influences many social behaviors through its action in the forebrain of mammals. However, the function of the homologous arginine vasotocin (AVT) in the forebrain of fishes, specifically the telencephalon remains unresolved. We tested whether the density of AVT-immunoreactive (-ir) fiber varicosities, somata size or number of AVT-ir neuronal phenotypes within the forebrain were predictive of social behavior in reproductive males of seven species of butterflyfishes (family Chaetodontidae) in four phylogenetic clades. Similar to other fishes, the aggressive (often territorial) species in most cases had larger AVT-ir cells within the gigantocellular preoptic cell group. Linear discriminant function analyses demonstrated that the density of AVT-ir varicosities within homologous telencephalic nuclei to those important for social behavior in mammals and birds were predictive of aggressive behavior, social affiliations, and mating system. Of note, the density of AVT-ir varicosities within the ventral nucleus of the ventral telencephalon, thought to be homologous to the septum of other vertebrates, was the strongest predictor of aggressive behavior, social affiliation, and mating system. These results are consistent with the postulate that AVT within the telencephalon of fishes plays an important role in social behavior and may function in a similar manner to that of AVT / AVP in birds and mammals despite having cell populations solely within the preoptic area.     

Effect of vasotocin on locomotor activity in bullfrogs varies with developmental stage and sex

Although neurohypophysial peptides are present in many regions of the developing and adult bullfrog (Rana catesbeiana) brain, the function of these peptides remains unclear. To investigate possible behavioral actions, we examined locomotor activity following peptide injection in bullfrogs at various developmental stages. An intraperitoneal (ip) injection of arginine vasotocin (AVT) in tadpoles (stages V, X, or XVII) produced an immediate and dose-dependent inhibition of locomotor activity. On the other hand, AVT stimulated activity when administered ip to juvenile or adult female bullfrogs, but did not influence activity in juvenile or adult males. The minimum effective dose of AVT, when injected directly into the brain of tadpoles, was 100-fold less than that observed when injected ip, suggesting a central nervous system site of action for this peptide. A vasopressin receptor antagonist (d(CH2)5[Tyr(Me)2]AVP administered ip or icv) significantly increased locomotor activity in tadpoles, compared to controls. Oxytocin, vasopressin, and AVP4-9 inhibited activity in tadpoles while mesotocin, des Gly(NH2)AVP, and pressinoic acid had no significant effect. Injection of PGF2 alpha also significantly decreased activity levels in tadpoles. However, pretreatment of tadpoles with indomethacin, a prostaglandin synthesis inhibitor, did not prevent the behavioral effects of AVT, suggesting that prostaglandin synthesis is not required for this response. In summary, AVT influenced locomotor activity in bullfrog tadpoles and female frogs. This effect shifted during development from an inhibitory action in tadpoles to a stimulatory effect in metamorphosed female frogs. The effect of AVT on juvenile and adult frog locomotion was sexually dimorphic, as this peptide altered female behavior but not male behavior.