Embryonic chick muscle produces an FGF-like activity

Normal and pathological formation of blood vessels is of considerable interest both in terms of basic scientific processes and clinical applications. Angiogenic events in the adult are likely to represent persistence of developmental mechanisms, and embryos are therefore a suitable experimental model for these processes. Among embryonic tissues, muscle is particularly appropriate for investigation, since it is highly vascularised from early stages. There are a number of competing explanations of how this process is controlled. Bioassays offer advantages over conventional molecular localisation techniques, in that they reveal the presence of active processed forms of the molecules under study, rather than non-processed forms, or non-translated meassages. Using these techniques, we report here that embryonic chick muscle, taken from the stages at which blood vessels are forming, produces an angiogenic activity on the chick chorioallantoic membrane (CAM), and transforms NR6 cells in soft agar. Basic fibroblast growth factor (bFGF) is shown to be angiogenic on the CAM in the same way, and also transforms NR6 cells (NR6 cells lack functional epidermal growth factor/transforming growth factor-a receptors, and are believed to respond only to bFGF in this way). Anti-bFGF removes the transforming activity of the embryonic muscle. We conclude that this represents evidence that embryonic chick muscle is producing an FGF-like molecule which is capable of acting as an angiogenic agent at the appropriate times in development.     

Alpha 1-adrenergic stimulation of ketogenesis and fatty acid oxidation is associated with inhibition of lipogenesis in rat hepatocytes

The effect of norepinephrine on fatty acid synthesis (3H2O incorporation into fatty acids), on fatty acid oxidation to CO2 and on ketogenesis was studied in isolated hepatocytes of fed rats. After incubation with norepinephrine (50 microM), lipogenesis was lower (5.7 +/- 1.1 nmoles 3H2O incorporated into fatty acids/mg dry weight/30 min) than in controls (7.5 +/- 1.7; n = 6, p less than 0.02). In contrast, (1-14C) palmitate conversion into total ketone bodies was increased to 10.9 +/- 1.8 nmoles/mg/30 min with norepinephrine, vs 8.5 +/- 1.6 in controls (p less than 0.05), and more (1-14C) palmitate was converted to 14CO2 with norepinephrine than in controls (1.48 +/- 0.10 nmoles/mg/30 min vs 1.06 +/- 0.11, p less than 0.05). The inhibitory effect of norepinephrine on lipogenesis was abolished by addition of the alpha 1-receptor blocker prazosin, but not by alpha 2 or beta-blockers. The results demonstrate that the ketogenic effect of norepinephrine is coupled with an inhibitory effect on lipogenesis which may be explained by diminished activity of acetyl-CoA carboxylase, diminished formation of malonyl-CoA and decreased activity of carnitine palmitoyl transferase I.     

Concurrent occurrence of elevated ketone body and depressed carnitine level in underfed guinea-pigs

Summary Food ingestion of male guinea-pigs was restricted to 10 g/day. Total carnitine content of liver of the underfed animals fell to 3.75 moles per liver (46.1% of control). Serum level of total carnitine was 35.0 nmoles/ml (64.4%) in the underfed animals while they developed high ketonemia, 384.2 nmoles/ml (555.2%). In restricted animals the carnitine levels also decreased in the muscles.     

In situ hybridization analysis of globin mRNAs in the primitive erythroid cells of the chick embryo

The possibility that the minor embryonic chick hemoglobins might be present in a particular subgroup of primitive erythroid cells has been investigated by in situ hybridization. Probe to detect the mRNA for the ^A globin chain of the minor embryonic hemoglobin was used, and the results of the hybridization were compared with those obtained using as probes the cDNAs for total globin mRNAs. All erythroid cells circulating in a 4-day-old chick embryo gave positive signals with both probes at an approximately constant ratio. This shows that all cells contain a similar assortment of hemoglobin types, excluding the possibility that a subgroup might contain the minor primitive hemoglobins exclusively. However, the cells are not homogeneous, since about 10% of them show a distinctly higher concentration of mRNA of all globin types.     

Antidiuretic and thermogenic effects of intracerebroventricular prostaglandin H2 in ethanol-anaesthetized rats

Summary When PGH₂ was administered intracerebroventricularly at doses of 5 and 15 nmoles in ethanol-anaesthetized rats, alcohol diuresis was inhibited and rectal temperature, blood pressure and heart rate were all significantly increased.     

Distribution of type A and B monoamine oxidase activities in the central nervous system of rat and chick

Summary The distribution of type A and B monoamine oxidase (MAO) activities in the central nervous system (CNS) of rat and chick was investigated using 5-hydroxytryptamine and -phenylethylamine as specific substrates. The distribution of type A MAO was similar to that of type B MAO in rat CNS, but quite different in chick CNS. This may be ascribed to the difference in animal species. The major part of MAO activity in the spinal cord was found to be type A.     

Transport of the synthetic peptide DSIP through the blood-brain barrier in rabbit

Summary The synthetic delta sleep inducing peptide (DSIP) passes the blood-brain barrier, since i.v. injection in free moving rabbits (30 nmoles/kg) significantly increases the cortical delta activity and decreases the motor activity during 5 h.Acknowledgment. This work was supported by grants from the Swiss National Science Foundation (Nos 3.871-72, 3.525.75, 3.443.074, 3.780.076), Fonds für Lehre und Forschung der Universität Basel, Merck Sharp and Dohme (USA), Ciba-Stiftung Basel.     

N-cadherin detected in the membrane fraction of lens fiber cells

N-cadherin was identified as a glycoprotein present in the fiber cell membranes of frog, chick, bovine, rabbit and human lenses. The molecular size of N-cadherin varies with the species. Homogenization of the chick lens in the presence of Ca2+ resulted in a decrease in the concentration of N-cadherin. This suggests that the lens contains a Ca2(+)-activated protease which can act on N-cadherin.     

Comparison of X-prolyl dipeptidyl-aminopeptidase activity in human cerebrospinal fluid with that in serum.

X-Prolyl dipeptidyl-aminopeptidase activities in cerebrospinal fluid and serum from the same patients without neurological diseases, undergoing surgery under lumbar anesthesia, were assayed fluorometrically with a newly synthesized fluorogenic substrate, 7-glycylproline-4-methylcoumarinamide; the values were 129.1 +/- 19.5 nmoles/min/l and 54.17 +/- 3.11 mumoles/min/l (mean +/- SEM, n = 23), respectively, and there was no correlation between both activities (r = 0.0894).     

N-cadherin detected in the membrane fraction of lens fiber cells

Summary N-cadherin was identified as a glycoprotein present in the fiber cell membranes of frog, chick, bovine, rabbit and human lenses. The molecular size of N-cadherin varies with the species. Homogenization of the chick lens in the presence of Ca²⁺ resulted in a decrease in the concentration of N-cadherin. This suggests that the lens contains a Ca²⁺-activated protease which can act on N-cadherin.     

Complete replacement of serum in primary cultures of chick embryo brain cells by growth-promoting alpha-globulin

Summary Growth-promoting alpha-globulin (GPAG), a specific serum protein complex which induces mitotic activity in continually replicating metazoan cells in vitro, was shown in this study to support growth of astrocytes and mesenchymal cells as well as process formation of nerve cells isolated from cerebral hemispheres of chick embryos.     

Complete replacement of serum in primary cultures of chick embryo brain cells by growth-promoting alpha-globulin

Growth-promoting alpha-globulin (GPAG), a specific serum protein complex which induces mitotic activity in continually replicating metazoan cells in vitro, was shown in this study to support growth of astrocytes and mesenchymal cells as well as process formation of nerve cells isolated from cerebral hemispheres of chick embryos.     

Oxidation of synephrine by type A and type B monoamine oxidase.

Synephrine (SP) was found to be a substrate for monoamine oxidase (MAO) in rat brain mitochondria, showing the Km and Vmax values of 250 microM and 32.6 nmoles/mg of protein/30 min respectively. The inhibition studies showed that the SP oxidation was carried out by both type A and type B MAO and a major part of the activity was due to type A MAO.     

Transport of the synthetic peptide DSIP through the blood-brain barrier in rabbit

The synthetic delta sleep inducing peptide (DSIP) passes the blood-brain barrier, since i.v. injection in free moving rabbits (30 nmoles/kg) significantly increases the cortical delta activity and decreases the motor activity during 5 h.     

Single-cell recordings from chick pineal glands in vitro reveal ultradian and circadian oscillations

Evidence is clear that each melatonin-producing cell in the chick pineal gland contains a circadian oscillator that continues to function in vitro, resulting in a prominent day/night rhythm of melatonin secretion. The aim of the present investigation was to examine whether the circadian organization of the gland has an electrophysiological correlate. To this end, single-cell recordings were made from isolated chick pineal glands kept in vitro under a light/dark cycle of 12:12 h, identical to that of the donors, or under continuous light or darkness. In all the glands investigated, a very small percentage of cells exhibited sodium-dependent spontaneous spike activity with a mean frequency below 10 Hz. The cells revealed rhythms with periods in the 15- to 60-min range and, additionally, exhibited ultradian and circadian rhythms in firing, with periods of 10.75+/-1.06 h and 26.25+/-1.26 h (mean +/- standard deviation), respectively. Most of the cells exhibited circadian rhythms with higher activity during daytime than at night, showing that the electrical activity and melatonin rhythm were out of phase. Under constant light or darkness, the circadian rhythm persisted. When the light/dark cycle of the donors was phase-advanced by 5 h, the cells revealed complete entrainment. We discuss whether the cells, albeit small in number, could function as a secondary ultradian/circadian oscillator contributing to the ultradian/circadian organization of the gland.     

Comparison of X-prolyl dipeptidyl-aminopeptidase activity in human cerebrospinal fluid with that in serum

Summary X-Prolyl dipeptidyl-aminopeptidase activities in cerebrospinal fluid and serum from the same patients without neurological diseases, undergoing surgery under lumbar anesthesia, were assayed fluorometrically with a newly synthesized fluorogenic substrate, 7-glycylproline-4-methylcoumarinamide; the values were 129.1±19.5 nmoles/min/1 and 54.17±3.11 moles/min/1 (mean±SEM, n=23), respectively, and there was no correlation between both activities (r=0.0894).     

Receptors for polymerized albumin on liver cells.

By the use of rabbit polymerized albumin labelled with fluorescein isothiocyanate, or coated on sheep red blood cells, specific receptors on rabbit liver cells were demonstrated. The possible biological role of these receptors is discussed.     

Thermoregulatory effects on the sheep of intracerebroventricular injections of L-glutamic acid

L-glutamic acid injected in doses of 200--1000 n moles . kg-1 into the cerebral ventricles of sheep had dose-dependent thermoregulatory effects: an increase in heat production and/or a decrease in respiratory frequency, and a rise in rectal temperature. A dose of 800 nmoles . kg-1 had effects comparable with those of a similar injection of 3 nmoles . kg-1 carbamylcholine.     

The effect of levamisole on phosphodiesterase activity.

Phosphodiesterase activity of mouse liver homogenates was estimated in presence and absence of levamisole. The enzyme activity was 1394 and 1399 nmoles/mg protein/30 min respectively. Our data show that levamisole does not affect the phosphodiesterase activity.     

The effect of levamisole on phosphodiesterase activity

Summary Phosphodiesterase activity of mouse liver homogenates was estimated in presence and absence of levamisole. The enzyme activity was 1394 and 1399 nmoles/mg protein/30 min respectively. Our data show that levamisole does not affect the phosphodiesterase activity.