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[Morphologic and biochemical changes in the hypothalamus in hormonally stimulated sheep].

The effect of exogenous hormonal preparations (PMSG, SG, PMSG + PGF2 alpha and FSH) was studied as exerted on neurosecretion and on changes in the nuclear volume of nucleus paraventricularis, nucleus ventromedialis, nucleus infundibularis and nucleus tuberomamillaris, and on histological changes in the third cerebral ventricle. Changes in catecholamine concentrations were determined in the rostral, medial, and caudal hypothalamus of the ewes. The study was performed with 28 ewes of the Slovak Merino breed during the oestrus period. The oestrus was synchronized by the Agelín vaginal tampons (Spofa). The ewes of four experimental groups were stimulated by the administration of 1000 I.U. PMSG (Antex-Leo), 1000 I. U. Serum Gonadotropin (Bioveta Ivanovice na Hané), 1000 I. U. PMSG with 250 micrograms Oestrophan inj. (Léciva Praha), and 450 I. U. Folistiman (VEB Arzneimittelwerk Dresden, GDR). The brain samples were processed by common histological methods. A caryometric analysis was performed at 3000-fold magnification and by measurement of 200 cells of one sample. The data were subjected to mathematical processing after Fischer and Inke (1956). The amount of neurosecretory material was evaluated by light microscopy (Nakahara, 1963). The multiplication of neurosecretion in the hypothalamic nuclei testifies to the fact that the administration of hormonal preparations stimulates the function of hypothalamic structures, which corresponds with the results of the caryometric analysis. It also ensues from the results that the administration of serum gonadotropins (PMSG and SG) at the dose of 1000 I. U. results in a statistically significant decline (P less than 0.001) of the concentrations of norepinephrine in the rostral, medial and caudal hypothalami of the ewes, although the PMSG preparation has a more pronounced effect when compared with SG. The hormonal stimulation with PGF2 alpha and with FSH causes no significant changes in the concentration of norepinephrine in the hypothalamus of the ewes. As norepinephrine is considered as a neurosecretion inhibitor, the multiplication of neurosecretion in the hypothalamus nuclei after stimulation with serum gonadotropins correlates with the decline of hypothalamic norepinephrine concentrations.

Animals↗

The effect of X-radiation on the hypothalamo-hypophyseal neurosecretory system of the rat.

The aim of the study was to reveal alterations in the macrocellular neurosecretory system of the rat which occur under the influence of X-radiation. The experiment was carried out on 15 male Wistar rats, divided into 1 control and 4 experimental groups. The animals were subjected to total-body-X-irradiation in a dose of 300R. Brains and cerebral pituitaries were collected after decapitation and fixed in Bouine's fluid. Staining with haematoxylin & eosin and paraldehyde fuchsin was applied. A reduction in the neurosecrete content was found after 7 days following irradiation both in the area of perikarya and nerve processes of SON and PVN. In animals dissected on the 14th and 21st day following irradiation, a shift of neurosecrete along the neurosecretory path was observed (via nerve processes to the posterior lobe of cerebral pituitary). On the 28th day, the morphological picture of the system examined was similar to the control one. The above results indicate that initially X-radiation causes a decrease in the production of neurosecrete within SON and PVN; later, however, the amount of neurosecrete is increased, like in control nuclei.

Animals↗

Monovalent cation dependency for the inhibition of outward transport of [3H]norepinephrine.

Rat hearts were labelled with [3H]norepinephrine in vivo. Slices of ventricles were prepared, preincubated in Krebs-HCO3 medium (KRB), and then incubated in a Na+-deficient, choline+-Krebs HCO3 (Ch+-Ca2+). The choline+-Krebs HCO3 medium induced a delayed neurosecretion which could be inhibited by either one of the blockers of the uptake of NE, cocaine or desipramine, when included in both the Krebs and the Ch+-Ca2+. When either desipramine or cocaine was present in the Na+-rich preincubation medium only, the duration of action of desipramine was more prolonged than that of cocaine. When desipramine was first added to the Ch+-Ca2+ medium at various times after incubation, the inhibitory response became smaller as the duration of preliminary Na+-deprivation was increased. After incubation for 80 min neither desipramine nor cocaine inhibited secretion. Sodium (added to Ch+-Ca2+ medium or in a Krebs replacement medium) then slowed the rate of release stimulated by Ch+-Ca2+ and facilitated an inhibitory action for both inhibitors. The inhibition did not require the continued presence of Na+. Depolarizing concentrations of K+ in the Ch+-Ca2+ medium with Na+ and either one of the inhibitors, prevented the inhibition of neurosecretion. However, K+ which was added to the preincubation medium with either inhibitor did not prevent the prolonged inhibition of neurosecretion. By contrast with the secretory response to Ch+-Ca2+, the secretory response to K+ in Na+-enriched Ch+-Ca2+ medium was weakly inhibited by the inhibitors of transport.

Animals↗

[Qualitative and quantitative changes in the hypothalamus and the ependyma of the 3d cerebral ventricle in sheep after irradiation and hormonal stimulation].

In the submitted work we concentrated our attention on the study of the changes in neurosecretion. PAS positive mucopolysaccharides in the hypothalamus and ependyma of the IIIrd cerebral ventricle and on studying the changes in the volume of the cell nuclei of the neurons of nucl. supraopticus (NSO), nucl. hypothalamicus centromedialis (NVM) and nucl. tuberomamillaris (NTM) of sheep after irradiation and after hormonal stimulation. We made our observations on 28 sheep of the Slovak Merino breed, of average live weight 35 to 40 kg, two- and three-years old in the period of physiological anoestrum. The first group of four ewes and the second group of four yearling lambs were controls. The third group of six ewes and the fourth group of six yearlings were exposed to irradiation all over with a dosage of 2.46 Gy (250 R) for a period of five days. To synchronize the oestrum of all the sheep we used agelline sponges which we fitted five days before irradiation. After the irradiation and removal of the sponges we gave the test sheep hormonal stimulation with 3 X 500 i. u. of serum gonadotropin. The fifth group of four ewes and the sixth group of four yearlings were only stimulated without irradiation. The fifth day after stimulation we slaughtered the sheep. We processed the brain samples by the usual histological methods. We carried out karyometric analysis with 3000X magnification and the measurement of 200 cells from one sample. We processed the values obtained mathematically according to Fischer and Inke (1956). We evaluated the quantity of neurosecretion material with a light microscope according to Nakahara (1962). We found that the multiplication of neurosecretion and the increased activity of the PAS reaction in the hypothalamic nuclei studied show that the irradiation and hormonal treatment stimulate the function of the hypothalamic structures. The results of the karyometric analysis in the yearlings also confirm this opinion. In the ewes the inhibitive influence of irradiation was probably manifest. The giving of hormones in combination with irradiation causes the multiplication of ependyma cells and the desquamation of surface layers.

Animals↗

[Changes in the hypothalamus in continuously irradiated sheep].

Neurosecretion, PAS-positive mucopolysaccharides and the Nissl substance were studied in the neurons of the rostral, medial and caudal hypothalamus of continuously irradiated ewes. The study was performed on 21 ewes of the Slovak Merino breed at the live weight of 34 kg. The animals were in the period of physiological anoestrus and their age was two to three years. The first group of six ewes was the control. The second group included 15 sheep irradiated with a total dose of 6.7 Gy (700 R) for seven days. Co60 was used as the source of irradiation. The animals of this group were killed seven days from treatment. The ewes in the third group were left for the study of mortality. The brains were perfused with 2% buffered paraformaldehyde immediately after the bleeding of the sheep; then the brains were removed from skulls and fixed in buffered picroformol. Paraffin slices were stained with haematoxylin-eosine, aldehyde-fuchsine and alcian blue for neurosecretion, with PAS-reaction for mucopolysaccharides and with cresyl violet for the Nissl substance. As found, the irradiation of the whole body inhibits the activity of neurosecretory cells in rostral and medial hypothalamus, thus reducing neurosecretion. These regions also show a reduced activity of PAS reaction used for the demonstration of mucopolysaccharides. The observed changes also included damage of the endothelium of blood vessels with the occurrence of erythrocyte extravasates and with haemorrhages.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

[The influence of digitalis glycosides on certain neural and hormonal functions in patients with chronic heart failure].

Digitalis works not only as a positive agent in congestive heart but as a modulator of neurosecretion as well. Opposite to its positive inotropic activity the latter refers to small doses. It leads to restoring of the autonomic balance. Its effect is resulted from the increased vagal activity that suppresses the adrenergic tone. Disregulating of the neurosecreting system may occur in patients with left ventricular impairment even without symptoms of left ventricular failure. Digitalis then besides stimulating contractility stops the neurosecretion and may be used not only to improve hemodynamics but also to slow the process of progressive deterioration of cardiac function.

Atrial Natriuretic Factor↗

Some effects of mating in female tsetse, Glossina austeni Newst.

Virgin females of G. austeni were mated at carefully chosen physiological states to determine the effect of mating on ovulation, neurosecretion and blood meal size. The results suggest that ovulation and larviposition are controlled by neurosecretion, and that the release of neurosecretion requires both a mating stimulus and the presence of a mature egg in one of the ovarioles. The mating stimulus was also found to lead to an increase in blood meal size. Unmated female flies do not ovulate and their mature eggs eventually disintegrate. Virgin blood meal sizes remain relatively low.

Animals↗

Membrane potential has no direct role in evoking neurotransmitter release.

Neurons communicate by secreting a transmitter that excites or inhibits other neurons at synapses. The role of presynaptic membrane potential in triggering transmitter release is still controversial. In one view, presynaptic action potentials trigger the release by the entry of calcium ions into presynaptic terminals through voltage-dependent calcium channels. Calcium acts at high local concentrations at release sites near channel mouths to cause neurosecretion. An opposing view is that, in addition to elevating presynaptic calcium, presynaptic potential stimulates transmitter release by a distinct direct action. The relative importance of depolarization and calcium entry in neurosecretion cannot be determined because the two events are tightly linked. To delineate the roles of presynaptic potential and calcium entry in transmitter release, we have used nitr-5, a photolabile calcium chelator, and a voltage-clamp technique to control intracellular calcium and membrane potential independently at a synapse formed between cell bodies of cultured neurons of the fresh water snail Helisoma trivolvis. We found transmitter release occurred when presynaptic calcium levels were elevated to concentrations of a few micromolar, and that presynaptic voltage had no direct effect on neurosecretion.

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An opioid-neuropeptide-Y transmission line to luteinizing hormone (LH)-releasing hormone neurons: a role in the induction of LH surge.

We tested the hypothesis that a decrease in hypothalamic inhibitory opioid tone produced by naloxone (NAL) will activate neuropeptide-Y (NPY) neurosecretion in 17 beta-estradiol (E2)-primed ovariectomized (ovx) rats. NPY neurosecretion was assessed in two ways. First, we studied the effects of iv saline (controls) or NAL infusion (2 mg/h) between 1100-1400 h on NPY concentrations in seven microdissected sites in the medial basal hypothalamus (MBH) and preoptic area in association with the increase in the rate of LH secretion, and then we examined the effects of NAL on the in vitro release of NPY and LHRH from the MBH of E2-primed ovx rats. We observed that in control rats, NPY concentrations in selected hypothalmic sites (median eminence, medial preoptic area, and arcuate nucleus) increased either just before LH rise at 1400 h or in association with the moderate LH surge in the afternoon. NAL infusion advanced the onset and amplified the magnitude of LH surge in the afternoon. In association with this augmentation of LH response, NAL infusion significantly increased NPY concentrations selectively in the median eminence, medial preoptic area, and arcuate nucleus. During NAL infusion at 1300 h and at the end of infusion at 1400 h, NPY concentrations in these sites increased compared to preinfusion levels at 1100 h and corresponding control levels at 1300 h. During the post-NAL infusion period until 1800 h, NPY levels remained elevated in these sites, but were not significantly different from those in control rats, which also displayed increments at this time. Further, NAL increased the in vitro efflux of both NPY and LHRH from the MBH; the increased release of two neuropeptides was dose related between 0.01-0.5 mg/ml, with the maximal increase occurring at 1.0 mg/ml NAL. Cumulatively, these studies show that 1) in association with the spontaneous LH surge in E2-primed rats, NPY concentration increased only in sites confined to the preoptic-tuberal pathway, which previously has been shown to mediate the induction of the LH surge; and 2) a decrease in the inhibitory opioid tone imposed by NAL readily augmented the hypothalamic NPY neurosecretion concomitant with an increase in duration and magnitude of the LH surge. These findings are in accord with the thesis that a decrease in the inhibitory opioid tone by the neural clock, postulated to occur before the LH surge, initiates a chain of neurosecretory events that may include a site-specific activation of NPYergic neurons.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Secretory activity and endocrine regulation of male accessory glands in the blood-sucking bug Panstrongylus megistus (Hemiptera: Reduviidae).

The epithelial cells of Panstrongylus megistus male accessory glands (MAG) present ultrastructural characteristics of a secretory cell. Their secretory products are accumulated in the lumen of the four MAG lobes. During the first 8 days of adult life a strong secretion activity occurs, accumulating enough material to produce the first spermatophore. Cerebral neurosecretions as well as juvenile hormone are both involved in MAG secretory activity regulation. Juvenile hormone seems to be the responsible for the stimulation of most protein synthesis in male accessory glands. Cerebral neurosecretion seems to be necessary to stimulate juvenile hormone production and release by the corpus allatum. Furthermore, neurosecretion is required for some polypeptides synthesis by MAG. Although topic application of precocene II to adult males does not reproduce the same effects on MAG as does allatectomy, this compound causes strong reduction on male reproductive capacity.

Animals↗

Coordinated release of ATP and ACh from cholinergic synaptosomes and its inhibition by calmodulin antagonists.

Isolated cholinergic synaptosomes from elasmobranch electric organ release both ACh and ATP when depolarized in the presence of calcium. The conditions that trigger ATP release are the same as those known to stimulate neurosecretion. The ratio of ACh to ATP released is the same as that found in purified elasmobranch synaptic vesicles. Both ACh and ATP release are inhibited either by the removal of extracellular calcium or the addition of the "calmodulin antagonist" trifluoperazine (TFP). Taken together, these data suggest that both ACh and ATP are released by exocytosis from synaptic vesicles within nerve terminals. A number of drugs, collectively known as calmodulin antagonists, were examined for their effect on neurosecretion. TFP completely inhibited secretion in a rapid and reversible manner. Other related drugs had similar effects, though with relative efficacies different from those reported for inhibiting calmodulin. It therefore seems likely that a calmodulin-like component of the nerve terminal, distinct from calmodulin itself, is essential for the process of neurosecretion.

Acetylcholine↗

[Development of the ultrastructural organization of the rat neurohypophysis during postnatal ontogenesis].

The development of structural components of the posterior hypopysis lobe of rats, the development of the neurosecretory activity of the hypothalamo-hypophyseal nervous system and the establishment of its function as a single system within the first three weeks after birth (1, 7, 14 and 21 days) as well as embryos of the 20th and 21st days of development were studied electron-microscopically. It has been established that the pituicytes of the posterior lobe of the hypophysis of newborn rats are represented by a single type of cells and are completely differentiated. On the 21st day they are in different morpho-functional states and consist of dark and light cells. By the 14th day of development a considerably increased amount of the neurosecretion is observed. By the 21st day the density of neurosecretion granules per a preparation unit does not change. The basal layer of the capillaries is formed not definitely by the birth time; on the 14th day the organization of the vessel wall is almost similar to that of mature animals. The Herring bodies make their appearance on the 21st day after birth. The hypothalamus and neurohypophysis of rats are functionally active before birth. After birth their neurosecretion activity is well pronounced while the hypothalamo-hypophyseal nervous system is not mature yet. By the time of getting sight this system is almost formed and sufficiently active.

Animals↗

Effect of lithium chloride on the neurosecretory system of the rat hypothalamus.

The response of the hypothalamic-pituitary neurosecretory system (HPNS) of rats to a single and repeated injection (200 mg/kg each time) of lithium chloride was studied by quantitative cytochemical analysis. The response of the HPNS was found to depend directly on the dose of lithium given and to consist of activation of synthesis and liberation of neurosecretion after a single dose of LiCl or inhibition of hormone formation in the hypothalamus and exhaustion of the reserves of neurosecretion in the neurohypophysis after a course of injections. In the recovery period (7--30 days after stopping the course of injections of LiCl) the previous state of the HPNS was gradually restored.

Animals↗

Electron-microscopic study of the secretion of the ependymal cells in the domestic cat (ependymin-beta cells).

We have studied, by electron microscopy, the ultrastructural aspects of secretion (neurosecretion) of the ependyma of the third ventricle of the domestic cat. We have found cytoplasmic protrusions and isolated masses of cytoplasm, some with homogeneous cytoplasm and others with very dense granulation (protein-beta?). Axons, synaptic terminals and free secretory granules in the ventricular lumen were also seen. The existence of ependymin-beta cells (ependymocyte-beta) and axohormonal buttons is suggested. The ependymal cells are classified into seven types: (1) covering ependymocytes, (2) tanycyt ependymocytes, (3) secretory ependymocytes, (4) ependymocytes-beta, (5) neurosecretory ependymocytes, (6) neurosensorial ependymocytes (crown-like) and (7) supraependymal microgial ependymocytes. A neurohormonal hypothesis and the possible existence of one or more cerebral hormones (neurohormones) are suggested. These hormones would flow into the CSF through some of the ependymal cells (by microapocrine secretion, liberation of neurosecretion granules, or by axohormonal buttons): this could be the most important link in the endocrine system, assuring the functional unity throughout the ventricular system of the cerebrospinal axis which it winds through, although its basic influence is exercised) on the hypophysis level as a vertex of the classical endocrine system.

Animals↗

[Effect of synaptosomal cytosolic [3H]GABA pool depletion on secretory ability of alpha-latrotoxin].

alpha-Latrotoxin, a presynaptic neurotoxin from the venom of Latrodectus mactans tredecimguttatus, induces massive [3H]GABA release from rat brain synaptosomes as a result of interaction with either Ca(2+)-dependent (neurexin 1 alpha or Ca(2+)-independent (latrophilin) membrane receptor. The main aim of the study was to elucidate whether the binding of alpha-latrotoxin to different types of receptors led to [3H]GABA secretion from one pool or in each case the source of neurotransmitter differs: in the presence of Ca2+ exocytosis is induced, while in the absence of Ca(2+)--outflow by mobile membrane GABA transporter from cytoplasm. We examined the effect of the depletion of cytosolic [3H]GABA pool by competitive inhibitors of the GABA transporter (nipecotic acid and 2,4-diaminobutyric acid) on the alpha-latrotoxin-stimulated neurotransmitter release. We also compared the influence of these agents on neurosecretion, evoked by depolarization with that evoked by alpha-latrotoxin. Depolarization was stimulated by 4-aminopyridine in the Ca(2+)-containing saline and high KCl in Ca(2+)-free medium. In synaptosomes treated with nipecotic acid unstimulated [3H]GABA release was significantly augmented and high KCl-evoked Ca(2+)-independent [3H]GABA release was essentially inhibited. But under the same conditions neurosecretion stimulated by alpha-latrotoxin greatly raised with respect to the control response. The similar results were obtained with the synaptosomes treated with 2,4-diaminobutyric acid. Another way to determine which of GABA pool is the target of alpha-latrotoxin action lay in analysis of the toxin effects on the preliminary depolarized synaptosomes. alpha-Latrotoxin influence was diminished by the preceding depolarization by 4-aminopyridine in Ca2+ presence. But after the high KCl stimulation effect of alpha-latrotoxin didn't change. These data suggest that alpha-latrotoxin triggers neurotransmitter release from synaptic vesicles via exocytosis. We suppose that the type of membrane receptor does not determine the mechanism of GABA release evoked by the toxin.

4-Aminopyridine↗

[Reactive characteristics of the neurohypophysis in the early postnatal period].

The maturity of the hypothalamic-neurohypophyseal system in the first 10 days of the postnatal life of rats was analysed through the potential inhibitory effect of ethanol on oxytocin and vasopressin secretion. Experimental animals were intraperitoneally injected with ethanol, and the control ones with the corresponding amount of physiological solution. Hypophyses were extracted 30 min later to be histologically analysed. Although the morphological features of neurohypophysis with functionally active pituicytes and dense vascular net were present even from the first natal day, neurosecretory grains were noticed not before the 8 th day. But the inhibitory effect of ethanol on neurosecretion did not manifest itself until the 10 th day, so it can be concluded that in this period the hypothalamic-neurohypophyseal system had not yet reached its level of complete maturity. Fatty acids in pituicytes being elements of neurohypophyseal neuroglia are pronunced even in animals younger than 10 days, which indicates that their origin could not be necessarily linked to the process of neurosecretion. In acutely alcoholized offsprings the amount of fatty acids in the neurohypophysis was reduced. This is substantiated by the engagement of the alcohol dehydrogenase (ADH) and the microsomal ethanol-oxidizing system (MEOS) in alcohol metabolism with which follows retarded oxidation process of fatty acids.

Animals↗

[Changes in neurosecretory activity in the hypothalamus of sheep after administration of PMSG].

By means of histological methods of histochemical nature we examined the qualitative alterations in neurosecretory cells of anterior (nucl. paraventricularis - NPV. nucl. preopticus medialis - NPM), medial (nucl. arcuatus s. infundibularis - NARC), and posterior (nucl. tuberomamillaris - NTM) hypothalamus of sheep with a simultaneous caryometric analysis of neurons of the same nuclei after administration of various PMSG doses. The examination was performed on the brain samples of 23 two- to three-year-old sheep, of the Slovak Merino breed, at the live weight of 30 to 40 kg. in the period of physiological anoestrus (June-July). The animals were divided into three experimental groups and were instilled agellin sponges (20 mg). On the 13th day the sponges were removed and the first group was applied 1000 i. u. PMSG, the second group 750 i. u. PMSG. The sheep were killed within 30 to 36 hours after the oestrus determination with a teaser ram. After bleeding of animals the brains were perfused with 2% buffered paraformaldehyde and after taking out the brains from the crania their fixation was finished in buffered picroformol. Paraffin slices were stained with haematoxylin-eosine and crotonaldehyde-fuchsin to neurosecretions. Caryometric analysis was carried out with a 1000 fold magnifying and with a measuring of 200 cells from one sample. We recorded the neurosecretion increase in all hypothalamic nuclei under study. The caryometric analysis has shown a moderate shift to the right, it means the volume increase of nuclei of neurosecretory cells, which demonstrates a stimulation of the function of hypothalamic structures.

Animals↗

Vasopressin-immunoreactive neurons and neurohemal systems in cockroaches and mantids.

Vasopressin-like neuropeptides of insects are of special interest because of their possible function as hormones and neuromodulators. Therefore, this study was undertaken by using whole-mount immunofluorescent staining by two antisera that recognize different types of vasopressin-like immunoreactive groups of neurons in the cockroaches Periplaneta americana, Leucophaea maderae, Nauphoeta cinerea, Diploptera punctata, and Blaberus discoidalis and in the mantids Litaneuria minor and Tenodera aridifolia sinensis. Using an antiserum to Arg/vasopressin, only two cells, the paired ventral paramedian (PVP) neurons, were immunostained in the central nervous system (CNS) of the cockroaches. These cells are located in the subesophageal ganglion, project throughout the CNS, and appear to be neurosecretory. Their varicose collaterals extend into the dorsal (motor) neuropil of the segmental ganglia, and this neuropil may be the principal site of the release of their neurosecretion. The PVP neurons were also stained by an antiserum to Lys/vasopressin; in addition, this antiserum stained several other groups of neurons, most of which appeared to be neurosecretory. Two pairs of Lys/vasopressin-immunoreactive cells are similar to the PVP neurons in that they are located in the subesophageal ganglion, extend through the ventral nerve cord, have collaterals in the dorsal neuropil of the segmental ganglia, and appear to be neurosecretory within the CNS. In addition, midventral and anteroventral clusters of Lys/vasopressin-immunoreactive neurosecretory neurons in the subesophageal ganglion project neurohemal release sites on the corpora allata. Other types of Lys/vasopressin-immunoreactive neurons include median and lateral neurosecretory cells of the protocerebrum and neurosecretory cells in the tritocerebrum, all of which project to the corpora cardiaca. In the abdominal ganglia there are posterolateral clusters of Lys/vasopressin neurosecretory neurons, and these cells extend to neurohemal release sites on the transverse and lateral cardiac nerves. In mantids the anti-Arg/vasopressin and anti-Lys/vasopressin antisera stained most of the same groups of neurons that these antisera recognized in cockroaches. The results of this study suggest that there are two or more vasopressin-like peptides in cockroaches and mantids and that these peptides may be released either as hormones in the blood or as neurosecretions within the CNS. Their function(s) in these insects remains to be determined.

Animals↗