Decreased number of asialoglycoprotein receptors in diabetic BB Wistar rat

The binding of asialoglycoproteins by hepatic binding protein was studied in freshly isolated hepatocytes from genetically diabetic BB Wistar rats. The number of cell surface asialoglycoprotein receptors was dramatically decreased (58,000 +/- 38,000 for diabetic rats compared to 267,000 +/- 70,000 for normal rats), while the association equilibrium constant was not changed. These results parallel those obtained with streptozotocin-diabetic rats and support the hypothesis that insulin deprivation is responsible for the decrease in the receptor number.     

Effects of blood sampling,anesthesia and surgery on plasma vasopressin concentration in rats

The influence of blood sampling, anesthesia and surgery on plasma vasopressin concentration was assessed in rats. Mean plasma concentration in conscious, chronically catheterized rats was 1.4±0.1 pg/ml (n=6). This value remained constant over repeated plasma samplings in the same animals. On the other hand, decapitation increased the plasma vasopressin concentration to 6.0±2.4 (in pg/ml) (n=6), inactin anesthesia to 2.9±0.6 (n=6), anesthesia and femoral cannulation to 13.3±5.8 (n=6) and surgery for renal micropuncture to 81.3±35.0 (n=6). It is concluded that the level of circulating plasma vasopressin is highly dependent on the sampling technique and is closely related to the extent of surgery.     

Effect of diabetes on the vertebral column of rats and its modification by estradiol

Summary In vertebrae of genetically selected sucrose-fed diabetic rats a statistically significant bone deficit was found after diabetes had been present for about 8 months. No osteopenia was observed in diabetic rats following treatment with estrogenic hormone for 5–7 months. The development of osseous centers in the end plates of the vertebrae was retarded in diabetic rats, but was about normal in diabetic rats given estrogen.—No differences were noted in the growth zones or in the tendency to develop articular lesions in rats of the various groups. Possible differences in the amount of GAG in intervertebral discs of diabetic and non-diabetic rats respectively await further confirmation.These investigations were aided by grant No. 2 RO1 EYO 1837-03, of the Institute of Arthritis and Metabolic Disease, National Institutes of Health, Bethesda, Maryland, USA.We are indebted to Professor Dr J. Rüttner, Institute of Pathology, University of Zürich, for his permission to use the calculator.     

C-peptide fragments stimulate glucose utilization in diabetic rats

Studies of C-peptide cellular effects show that not only the full-length native peptide but also specific C-terminal fragments are biologically active in in vitro systems. In the present study, the effect of five C-peptide fragments and the native peptide on whole-body glucose turnover was studied in streptozotocin diabetic rats using the insulin clamp technique. Insulin was infused intravenously at 18 pmol kg^–1 min^–1 for 90 min and blood glucose concentration was clamped at 8 and 4 mM in diabetic and non-diabetic animals. A steady state was reached during the last 30 min of the study period. Rat C-peptide II and fragments comprising residues 27–31 and 28–31 were effective in augmenting glucose turnover in diabetic rats (+100% to 150%), while no significant effects were seen for segments 1–26, 11–19 and 11–15. The metabolic clearance rate for glucose during infusion of C-peptide or fragments 27–31 and 28–31 in diabetic rats was similar to that seen in non-diabetic animals. We conclude that C-terminal tetra- and pentapeptides, but not fragments from the middle segment of C-peptide, are as effective as the full-length peptide in stimulating whole-body glucose turnover in diabetic rats.Received 18 December 2003; received after revision 19 January 2004; accepted 21 January 2004     

Änderungen des Elektrolytgehaltes von Erythrozyten und Plasma bei nephrektomierten Ratten

Summary After two-stage nephrectomy in rats the potassium concentration in the red blood corpuscles (RBC) decreases from 10.6 ± 0.3 to 5.5 ± 0.3 mEq per 100 ml of RBC within 48 h. The decrease is accompanied by a much smaller increase in the plasma potassium concentration; the hyperkaliemia in nephrectomized rats is less pronounced than in nephrectomized dogs or anuric humans. Na⁺ in RBC increases by about 44% after nephrectomy; while there is only a very slight decrease of Na⁺ in plasma. Plasma chlorides drop from 10.92 ± 0.08 mEq/100 ml of plasma to 6.00 ± 0.81 mEq % within 48 h after nephrectomy. RBC chlorides tend to increase again after an initial drop from 5.22 ± 0.07 to 3.82 ± 0.90 mEq% within the first 8 h.     

The influence of different nutrients on plasma cholecystokinin levels in the rat

Summary Isocaloric and isovolemic amounts of protein (casein), fat (intralipid) and carbohydrate (saccharose) and an isovolemic control solution of water were administered intragastrically to conscious rats. The plasma CCK levels, determined by a sensitive and specific radioimmunoassay, showed an increment of 6.3±0.6, 2.7±0.5, 1.7±0.4 and –0.9±0.4 pM, respectively (basal value 2.5±0.3 pM). The threshold increment of plasma CCK to stimulate pancreatic enzyme secretion by exogenous CCK was found to be 1.5 pM. It is therefore concluded that casein is a potent stimulus for CCK secretion and pancreatic secretion, but that fat and even carbohydrate, although less potent, also produce a CCK increment above the threshold for pancreatic secretion.Supported by Grant IKW 86-16 from the Netherlands Cancer Foundation KWF.     

Effect of experimental diabetes on estradiol binding by the anterior pituitary and hypothalamus in ovariectomized rats

Summary In diabetic rats,³H-estradiol binding by the cytosol and nuclei of the anterior pituitary was lower than normal. Changes in affinity and receptor numbers were demonstrated by Scatchard analysis. Protein synthesis in diabetic pituitaries, however, was in the normal range.     

Hemodynamic studies in a parabiotic model of portal hypertension

Summary Splanchnic and systemic hemodynamic studies were performed in a rat model of parabiosis and portal hypertension. A portal hypertensive and a normal rat were surgically united side to side. A hyperdynamic circulation, characterized by increased cardiac index (413±26 vs 318±23 ml·min^–1·kg^–1; p<0.05) and portal venous inflow (9.61±1.29 vs 6.33±0.36 ml·min^–1·100 g b.wt^–1; p<0.05), was found in all the portal hypertensive rats but not in the normal parabiotic partners. These results do not support the existence of a transferable humoral factor mediating the hyperdynamic circulatory state of chronic portal hypertension.Acknowledgment. The authors express their appreciation to Martha Shea for technical assistance.     

Copper metabolism in the LEC rat: Involvement of induction of metallothionein and disposition of zinc and iron

The Cu concentration was about 40 times higher in the liver of LEC (Long-Evans with a cinnamon-like coat color) rats aged 77 days (227.5±21.6 g/g liver) than in Fischer rats (5.2±0.1 g/g liver). However, in the kidney and brain of the LEC rats, Cu concentrations were lower than in these organs of the Fischer rats. Cu concentration in the hepatic metallothionein fraction was about 130 times higher in the LEC rats than in the Fischer rats. The LEC rats showed markedly low concentrations of Cu in the serum and bile. It seems likely that excretion of Cu from the liver into the bile and blood (as ceruloplasmin) is inherently lacking in the LEC rat.     

Influence of common bile duct cannula size on maximal secretory rate of taurocholate in the rat

Summary The common bile duct of male Sprague-Dawley rats was cannulated with either PE 10 or PE 50 tubing. Maximal secretory rate of taurocholate averaged 389±67 (SD) and 657±115 nmoles·min^–1·g liver^–1 in the PE 10 and PE 50 group, respectively (p<0.005). Maximal bile secretory pressure was significantly higher in the PE 10 group (240±28 vs 174±8 mm H20; p<0.005). When the maximal secretory rate was exceeded, bile flow decreased in both groups but this was accompanied with a decrease in maximal bile secretory pressure in the PE 10 group only. Maximal secretory rate of bile salts is markedly influenced by experimental technique. Use of small caliber common bile duct cannulae leads to partial obstruction and decreases the apparent maximal secretory rate for taurocholate.Acknowledgments. J. Reichen was the recipient of a Faculty Development Award in Clinical Pharmacology from The Pharmaceutical Manufacturer's Association Foundation, and is the recipient of a Research Career Development Award (KO4 AM 1189) from the National Institutes of Health. Supported by National Institutes of Health grant RO1 AM 27597.     

Capillary blood flow in the testes and testosterone secretion in the starved rat

Summary The average capillary blood flow in the testes was found to be 181 l/min/g testis tissue (n=19) in rats starved for 5 days and 273 l/min/g (n=18, p<0.01) in the control group. Plasma testosterone was significantly decreased in the starved animals (1.00±0.06 ng/ml vs 5.43±0.63 ng/ml). When starved and control rats were stimulated with human chorion gonadotropin, testosterone values in plasma were greatly increased in both groups. The capillary blood flow was not altered. The data indicate that human chorion gonadotropin can stimulate testosterone production in the starved rat without influencing the reduced capillary blood flow.     

Pineal N-acetyltransferase depression in rats exposed to heat

Summary Exposure of adult male rats to increased temperature of 33±1°C for 3 and 10 days brought about decreases in pineal N-acetyltransferase activity. These and previous findings of pineal HIOMT inhibition under similar conditions support the postulation of a possible thermoregulatory role for the pineal gland.     

HCN channels contribute to the intrinsic activity of cochlear pyramidal cells

A hyperpolarization-activated current recorded from the pyramidal cells of the dorsal cochlear nucleus was investigated in the present study by using 150- to 200-m-thick brain slices prepared from 6- to 14-day-old Wistar rats. The pyramidal neurones exhibited a slowly activating inward current on hyperpolarization. The reversal potential of this component was –32 ± 3 mV (mean ± SE, n = 6), while its half-activation voltage was –99 ± 1 mV with a slope factor of 10.9 ± 0.4 mV (n = 27). This current was highly sensitive to the extracellular application of both 1 mM Cs⁺ and 10 M ZD7288. The electrophysiological properties and the pharmacological sensitivity of this current indicated that it corresponded to a hyperpolarization-activated non-specific cationic current (I_h). Our experiments showed that there was a correlation between the availability of the h-current and the spontaneous activity of the pyramidal cells, suggesting that this conductance acts as a pacemaker current in these neurones. Immunocytochemical experiments were also conducted on freshly isolated pyramidal cells to demonstrate the possible subunit composition of the channels responsible for the genesis of the pyramidal h-current. These investigations indicated the presence of HCN1, HCN2 and HCN4 subunits in the pyramidal cells.Received 15 May 2003; received after revision 26 June 2003; accepted 21 July 2003     

Identification of calcium antagonist receptor binding sites using (3H)nitrendipine in bovine tracheal smooth muscle membranes

Summary (³H)Nitrendipine binding to the bovine tracheal muscle membrane at 25°C was rapid, saturable (B_max=14.8±3.9fmol/mg protein) and of high affinity (K_d=0.15±0.04 nM). The rank order of Ca²⁺ antagonists competing for airway (³H)nitrendipine binding was nitrendipine nisoldipine nifedipine » verapamil. Cromolyn, however, neither inhibited nor increased the binding.J.B.C. is a visiting associate professor at the NYMMC. We thank Ms. Pang-jang Chang for technical assistance. This work is supported by a grant (NSC 72-0412-BO10-R20) from the National Science Council, ROC. To whom reprint requests should be addressed: Allergic Disease Center, Creighton University School of Medicine, Omaha, Nebraska 68178.     

Chronic sensory denervation reduces thrombin-stimulated endothelin release from aortic endothelial cells

The long-term (trophic) influence of perivascular nerves on the endothelium was investigated by measuring changes in thrombin-stimulated release of the potent vasoconstrictor, endothelin, after selective chronic denervation. Rat pups were treated with either guanethidine or capsaicin to destroy sympathetic or sensory nerves, respectively. The abdominal aortas from the rats at three months of age (5 pooled per experiment) were incubated with 4U thrombin/ml in medium for 24 h at 37°C, and the amount of endothelin released from the preparation determined by immunoassay. After neonatal sensory denervation there was a significant reduction in the thrombinstimulated release of endothelin compared to the controls (0.012±0.012 (4) compared to 0.063±0.012 (6), pmol/cm²/24 h, p<0.02). There was no change in endothelin release after sympathetic denervation. In summary, sensory nerves play a trophic role in the expression of endothelin in endothelial cells of the intima.     

Beta-adrenergic receptors in rat myocardium: Direct detection by a new fluorescent beta-blocker

Summary A new fluorescent -blocker, 9-amino-acridin propranolol (9-AAP), was administered i.v. to rats. Multiple fluorescent 9-AAP binding sites were observed on cardiac muscle cells in frozen sections. Intensity and density of cardiac 9-AAP fluorescence were markedly reduced following pretreatment with (±)- and (–)-propranolol but not with (+)-propranolol. Our findings suggest that 9-AAP may label -adrenergic receptor sites in rat myocardium.D. A. is supported by a fellowship from the Lady Davis Fellowship Trust and by a Research Grant from the Bat-Sheva de Rothschild Fund for The Advancement of Science and Technology. The technical assistance ofA. Itzhak is gratefully acknowledged.     

The conduction velocity,number and diameter of unmyelinated fibers in Remak's nerve

Summary Remak's nerve in the chicken was examined ultrastructurally and electrophysiologically to determine the characteristics of fibers in the nerve trunk. The ratio of unmyelinated fibers to myelinated ones was 1111. The mean number of unmyelinated fibers was 3555±232 (SEM, n=5) and they had a mean diameter of 0.502±0.034 (SEM) m. The compound action potential consisted almost entirely of a large diphasic waveform which had a mean peak conduction velocity of 0.62±0.031 (SEM, n=5) m·s^–1 at 37°C.     

The regulatory peptide system of the large bowel in equine grass sickness

Summary In recent years, distinct changes in regulatory peptides have been found in a number of gastrointestinal diseases. Grass sickness is a fatal disease of horses for which the etiology has yet to be fully ascertained. In this study, the peptide-containing nerves and ganglionic and mucosal endocrine cells of the ileum, colon and rectum were investigated in horses with sub-acute or chronic grass sickness and compared with normal controls using immunocytochemistry, at both the light and electron microscopical levels, and radioimmunoassay. A substantial loss of both peptide-containing cells and nerves was found in all of the sick horses, particularly in the ileum. Electron microscopy revealed marked degeneration of nerves in the gut wall. fibers containing granules immunostained for substance P or VIP, using the immunogold staining technique, underwent extensive degranulation in grass sickness, with the formation of multiple vacuoles.Radioimmunoassay of peptide content also showed that the most drastic changes occurred in the ileum. For example, VIP content was significantly reduced from 109±19.8 (mean±SEM) pmoles/g in controls to 6.8±1.4 pmoles/g in grass sickness (p<0.001) and substance P from 65.9±8.1 to 31.3±9.5 (p<0.02). These results may have applications in the diagnosis and treatment of grass sickness.The authors gratefully acknowledge the financial support of the Wellcome Trust and the Grass Sickness Fund.     

Role of calcium in the histamine release induced by D-galactosamine from rat mast cells

Summary Rat peritoneal mast cells were isolated and purified by differential centrifugation in Ficoll. Cells pooled from three to four rats were suspended at approximately 10⁶ cells/ml in a buffered salt solution and incubated for 1 h at 37°C in 300 l volumes in the absence or presence (9×10^–4 M) of calcium chloride. Addition of D-galactosamine hydrochloride (DGM; 2.8×10^–4 M) caused (in addition to basal release) a mean ±SEM percent histamine release of 15.7±5.2 in the presence of Ca⁺⁺ and 19±4.9 in the absence of Ca⁺⁺ (p>0.05). It is suggested that D-galactosamine does not require extracellular Ca⁺⁺ for the release of histamine from the rat mast cell.A preliminary analysis of these results was presented at the International Symposium on calcium entry blockers and tissue protection, Rome, 15–16 March 1984.     

Evaluation of the ability of intact platelets to accumulate acridine organge

Summary A new approach to the evaluation of the uptake of fluorescent, probes by intact, cells is described. Acridine organe (AO) was used because it can be selectively accumulated by serotonin-containing granules of platelets. Analysis of the fluorescence signal allows the estimation of the relative volume of the granules and the equilibrium coefficients for AO transport across the cytoplasm and granule membranes. The following results were obtained for human and rabbit platelets: the relative volumes of the granules were 14±1% and 29±2%, the ratios of intragranular-extracellular probe concentration were 2260±382 and 30,000±5500, and the cytoplasm-extracellular medium concentration ratios were 375±60 and 225±60, respectively.