High K+-induced release of somatostatin from the cortical preparation of rat brain

Summary Release of endogenous somatostatin (SRIF) from the rat cerebral cortical slices incubated in Krebs-bicarbonate buffer was increased from the basal rate of 3.4±0.6% of the total SRIF content in 15 min at [K⁺]_o=5.6 mM, to 13.1±1.6% upon raising the [K⁺]_o to 56.6 mM. The high-K⁺ evoked SRIF release was absent when Ca⁺⁺ in the medium was replaced by Mn⁺⁺. The isolated synaptosomes from rat cerebral cortex contain 13.2±3.1 ng SRIF/mg protein compared to 0.33±0.01 ng/mg protein in the cortical tissue as a whole, suggesting that nerve terminals are the main source of the peptide released upon membrane depolarization.The study was supported by a grant from the Medical Research Council of Canada. Results of this work have been published in part as abstracts: Can. Physiol.9, 45 (1978), and Fedn Proc.37, 665 (1978).The authors are greatly indebted to Dr M. Gotz and the Ayerst Research Laboratories for the most generous supply of the synthetic somatostatin.     

The effect of anesthetics on the basal secretion of immunoreactive growth hormone in rats]

The authors investigated basal levels of plasma immunoreactive growth-hormone in the rat (R-GH) after administration of 3 different anesthetic drugs: urethan, chloral hydrate and gamma-hdroxy-butyrate (GHB). Lowest R-GH concentrations (5 +/- 3 ng/ml) are observed after urethan; they are significantly higher (15 +/- 4 ng/ml) after chloral hydrate but this anesthetic also causes hyperglycemia (210 +/- 30 mg/100 ml). Normal blood glucose levels are observed under GHB narco-analgesia which elicits a clear-cut R-GH secretory episode (70 +/- 5 ng/ml); basal values (12 +/- 3 ng/ml) are maintained for several hours thereafter.     

4-Proline and 4-hydroxyproline analogs of arginine vasopressin: role of the proline substitution in the two beta-turns of vasopressin

[4-L-Proline]arginine vasopressin, [4-D-proline]arginine vasopressin, [4-hydroxyproline]arginine vasopressin and [4-proline, 7-hydroxyproline] arginine vasopressin were synthesized and found to have antidiuretic activities of 91 +/- 4, 1.7 +/- 0.2, 1.0 +/- 0.1 and 4.4 +/- 1.0 units/mg, respectively. None of these analogs exhibited a significant level of rat pressor activity. The observed activities of these and other analogs with substitutions at position 4 and/or 7 are discussed on the basis of hypotheses and data bearing on the solution conformation of vasopressins.     

Increase in the oxygen available to the cerebral cortex after the administration of carbonic anhydrase inhibitors]

Oxygen tension (pO2) in cerebral cortex was measured by polarographic method in unanesthetized rabbits. Intravenous administration (25 mg/kg) of carbonic anhydrase inhibitors (acetazolamide, methazolamide, dichlorphenamide, sulthiame) induced an early important rise of cortical p O2, which is not dependent on increase of p O2 and p CO2 and decrease of pH in arterial blood. High dosage of acetazolamide (250 mg/kg) produced the same effect and did not suppress the increase of cortical p O2 under air-CO2 inhalation. This result suggests that CO2 might act specifically upon cerebral vessels.     

Opioids and cytosolic calcium in rat anterior pituitary: dynorphin preparation showed LHRH-like action due to contamination

The effect of dynorphin A-(1-13) (Dyn A-(1-13] and other opioids on the cytosolic free calcium concentration [(Ca2+]i) in rat anterior pituitary cells was examined using the fluorescent indicator fura-2. A commercial synthetic Dyn A-(1-13) preparation elevated [Ca2+]i. Results, which were obtained with receptor antagonists, and in LHRH receptor radioligand binding studies as well as by HPLC combined with LHRH radioimmunoassay, strongly suggest that this effect of the dynorphin preparation was due to contamination with a LHRH-like compound. Dyn A-(1-13), purified by HPLC, as well as Dyn A-(2-13), [Leu5]enkephalin, beta-endorphin, morphine, or U50,488H had no effect on [Ca2+]i. LHRH caused a rapid increase in [Ca2+]i by about 50 nM which was blocked by the LHRH antagonist, [D-pGlu1,D-Phe2,D-Trp3,6] LHRH.     

Dissociation of digoxin-like immunoreactivity and Na+,K+-ATPase inhibitory activity in rat plasma

We measured endogenous digitalis-like factor (EDF) in rat plasma during acute saline infusion by two different procedures. Na+,K+-ATPase inhibitory activity in the rat plasma significantly increased during saline loading (7.8 +/- 2.2 vs 2.5 +/- 0.9%, with and without acute saline loading, respectively, p less than 0.05). On the other hand, the plasma digoxin-like immunoreactivity significantly decreased during acute saline loading (16.9 +/- 1.6 vs 32.0 +/- 2.8 pg digoxin equivalents/ml, with and without acute saline loading, respectively, p less than 0.01). These results indicate that the major substances detected by digoxin-like immunoreactivity and direct Na+,K+-ATPase inhibitory activity are completely different, at least in rat plasma.     

Differential inhibitory effects of noradrenaline upon insulin secretion by the isolated perfused pancreas of the rat

Summary Responses of basal and glucose-induced secretion phases of the isolated perfused pancreas of the rat to graded doses of noradrenaline (NA) are described. Maximal inhibition of basal secretion was achieved at [NA]=40, ng/100 ml; at [NA]=80 ng/100 ml, the 1st phase of glucose induced secretion was abolished; at [NA]=100 ng/100 ml the 2nd phase was abolished. At [NA]40 ng/100 ml basal secretion was less sensitive to NA inhibition than glucose induced output.Acknowledgment. The authors are most grateful to Mr J. Stevenson for advice upon and assistance with statistical analysis of data.     

Spectral changes resulting from the interaction of some N-alkyl nitrosamines and rat liver microsomes

Summary Dimethyl (DMN) and diethyl nitrosamine (DEN) do not give characteristic spectral changes upon interaction with rat liver microsomes, while dipropyl (DPN) and dibutyl (DBN) nitrosamine cause type I spectral changes. The spectral binding constant is 100 mM for DPN and 1.17 mM for DBN. The maximal spectral change is 3.2×10⁶ and 1.0×10⁶ absorbance units per milligram protein for DPN and DBN respectively.Acknowledgment. This work was supported by Grants AM 13195-07 from the National Institute of Health (USA) and from the Consejo Nacional de Investigaciones Cientificas y Técnicas (Argentina).     

Human growth hormone in urine: development of an ultrasensitive radiometric assay

An immunoradiometric assay for human growth hormone (HGH) has been developed which has a detection limit of 1 ng/l and can measure HGH in unextracted urine from normal children and adults. The assay is based on a two-step procedure, using a solid-phase goat-anti-HGH immunosorbent for immunoextraction and [125I]-labeled monoclonal HGH-antibody for detection and quantification. The assay is not affected by urea, NaCl or changes of pH from 5-8. The mean urine HGH concentration in normal children is 6.78 +/- 7.6 (SD) pg/ml, in patients with HGH-deficiency 1.3 +/- 0.9 pg/ml which increases to 11.7 +/- 13.4 pg/ml on the day of growth hormone injection.     

Influence of flupirtine, a novel nonopioid analgesic agent on somatosensory evoked potentials in rats.

The effect of flupirtine, a novel nonopioid analgesic, on somatosensory evoked potentials (SEP) was investigated in anesthetized rats. Primary somatosensory potentials were evoked in the cerebral cortex by stimulation of the skin of the whiskery part of the face. Flupirtine injected i.p. dose-dependently prolonged the latency and reduced the amplitude of SEP with ID50-values of 5.4 mg/kg (2.6-9.3 mg/kg) and 7.9 mg/kg (3.9-13.8 mg/kg), respectively. This effect of flupirtine (10 mg/kg, i.p.) on the latency and the amplitude of SEP, did not change when naloxone (1 mg/kg, i.p.) was given before flupirtine. The results indicate that the analgesic flupirtine decreases the primary somatosensory evoked potential by diminishing the excitability of cortical neurons. Opioid mechanisms are not involved.     

Manganese action potentials in mammalian cardiac muscle.

The membrane potential in guinea-pig's papillary muscles from right ventricle was recorded by glass microelectrodes and stimulation was effected by current pulses applied through a sucrose-gap. Action potentials with overshoot were recorded in the solution lacking Na+ and Ca++ but containing 2-95 mM Mn++. The overshoot was increased with the increase of [Mn++]o by about 30 mV/decade. Similar Mn++ dependent action potentials were also obtained in Na-free solution containing 0.6 mM Ca++. The results indicate that Mn inward current is sufficient to generate action potentials in cardiac muscle.     

Transplantation into the mammalian adult spinal cord

Summary Embryonic cerebral cortical tissue obtained from rat embryos of 15-day gestation was transplanted into the cervical spinal cord of adult rats. The cortical transplants survived, grew, and established connections with the host animal's spinal cord. In other animals, knife lesions were first made in the host's spinal cord, and then embryonic cortical tissue was transplanted into the site of the lesion. The cortical transplants in these animals were observed to become an integral part of the host animal's spinal cord.     

Cytosolic free Ca2+ in insulin secreting cells and its regulation by isolated organelles

The role of Ca2+ in secretagogue-induced insulin release is documented not only by the measurements of 45Ca fluxes in pancreatic islets, but also, by direct monitoring of cytosolic free Ca2+, [Ca2+]i. As demonstrated, using the fluorescent indicator quin 2, glyceraldehyde, carbamylcholine and alanine raise [Ca2+]i in the insulin secreting cell line RINm5F, whereas glucose has a similar effect in pancreatic islet cells. The regulation of cellular Ca2+ homeostasis by organelles from a rat insulinoma, was investigated with a Ca2+ selective electrode. The results suggest that both the endoplasmic reticulum and the mitochondria participate in this regulation, albeit at different Ca2+ concentrations. By contrast, the secretory granules do not appear to be involved in the short-term regulation of [Ca2+]i. Evidence is presented that inositol 1,4,5-trisphosphate, which is shown to mobilize Ca2+ from the endoplasmic reticulum, is acting as an intracellular mediator in the stimulation of insulin release.     

Characterization of the (Na+-K+)-ATPase from endometrial plasma membranes of immature mammals]

The (Na+-K+)-ATPase in plasma membrane from Mammiferous endometrium is characterized by the Mg/ATP ratio equal to one, and by a distinct affinity for Na+ (1.3 mM) and K+ (2 mM). The activity is maximum for pH 7.4-7.5 in presence of Mg++ 2mM and ATP 2 mM, Na+ 140 mM and K+ 10 mM.     

Intravenous thyrotropin releasing hormone (TRH) enhances the excitatory actions of acetylcholine (ACh) on rat cortical neurons

Summary I. v. administered TRH (1.25-10 mg/kg) enhanced the excitatory actions of iontophoretically applied ACh on spontaneously active cerebral cortical neurons of pentobarbital-anaesthetized rats. These observations are consistent with the hypothesis of a cholinergic-link in the anti-anaesthetic actions of exogenously administered TRH.     

Synthesis and pharmacological properties of [1-N4-dimethyl-asparagine]-angiotensin II.

[1-N4-Dimethyl-asparagine]-angiotensin II was synthesized by Merrifield's solid-phase procedure. The analogue gave rat blood pressure about 70% relative potency to Hypertension (Ciba). Rabbit aorta strips gave intrinsic activity alpha E = 1, a PD2 of 6.92 +/- 0.09 and an affinity relative to [Asn1]-angiotensin II of 6.5%.     

Plasma estrogens in Pleurodeles waltlii. Comparison between diploid and triploid females]

Estrogens have been examined in the plasma of diploid and triploid newts Pleurodeles waltlii. Estradiol-17 beta (E2) and estrone (E1) were determined by radioimmunoassay, before and after enzymatic hydrolysis of the conjugates. Total (t) and unconjugated (u) E2 levels were positively correlated (E2u = 0.478 35 E2t + 0.579 98; r = 0.883), but no correlation was detected between E1 levels. No statistical difference was found for the estrogen levels between the different experimental lots of diploid newts (E2t = 7.5 +/- 0.37 ng/ml, E2u = 4.3 +/- 0.20 ng/ml, E1t = 2.19 +/- 0.08 ng/ml, E1t = 0.41 +/- 0.02 ng/ml) but every estrogen level was lower in the triploid group (E2t = 1.8 +/- 0.60, E2u = 1.0 +/- 0.18, E1t = 1.4 +/- 0.13, E1u = 0.3 +/- 0.04 ng/ml). This difference is discussed in relation to lower fertility of the triploid females.     

Net ATP synthesis by running the red cell calcium pump backwards.

Ca2+ loaded inside-out vesicles from human red blood cells, yielding C2+ into a Ca2+ free medium with 4 mM EGTA, 2 mM ADP and 10 mM phosphate, produced an excess of 14.9 pmoles . min-1 . (mg protein)-1 of ATP compared to controls in which the transmembrane Ca2+ gradient was abolished by the ionophore A 23 187.     

Estimation of systemic catecholamine levels in the frog, using the technic of radio-enzymatic assay]

We have developed a radio-enzymatic assay for systemic catecholamines in the frog. Such are its specificity and sensibility that adrenaline and noradrenaline may be measured in 50 microliter of plasma samples, the withdrawal of which strongly influenced the results. The smaller values were obtained in plasma withdrawn from canulated animals. In this case, adrenaline was the major catecholamine in the plasma: 190 +/- 55 ng/100 ml versus 35 +/- 18 ng/100 ml for noradrenaline.     

Characteristics of the partial derivative of hemoglobin saturation with respect ot oxygen pressure : delta S 02/delta P 02]

The first (delta S/delta P) and second (delta 2S/delta [H+] delta P) partial derivatives of hemoglobin saturation with respect to oxygen pressure and [H+] were numerically calculated for the blood of non-smoking young adult at different pH, PCO2 and [2.3-DPG]. Their functional value in respiratory physiology was stressed.