Radu Racotta

Food and water intake of rats after intraperitoneal and subcutaneous administration of glucose, glycerol and sodium lactate.

Abstract Glucose, glycerol and DL — sodium lactate were injected in various doses by intraperitoneal (IP) and subcutaneous (SC) routes in rats accustomed previously to a 24 hr-feeding, 24 hr-fasting schedule. Administration of isosmolar sodium chloride and measurement of water ingestion served as controls for osmotic effects. The three substances determined a dose-dependent feeding depression in the first half hour when administered IP. SC injections produced a much weaker and non dose-dependent feeding depression. Some doses prolonged the hypophagia up to three or even 24 hr. No relation can be established between the hypophagia and the concurrent hyperdipsia since the last effect does not depend on the route of administration. It is suggested that the hypophagia which followed IP administration was due to information of hepatic origin.

Hepatic chromaffin cells

Abstract Chromaffin cells were identified in the liver of mouse, rat, and rabbit. They are found in portal areas, sinusoid walls, or below the connective capsule. The histochemical technique which was used permited to differentiate them from enterochromaffin and mast cells. Liver chromaffin cells could be important for the adrenergic-induced hepatic glucose output.

Epinephrine and dopamine colocalization with norepinephrine in various peripheral tissues: Guanethidine effects

Chemical sympathectomy with guanethidine (Gnt) selectively destroys the postganglionic noradrenergic neurons, whereas dopaminergic fibers and nonneural catecholamine-secreting cells are spared. As a result, the relative proportions of norepinephrine (NE), epinephrine (E), and dopamine (DA) in tissues can be differentially affected. This study was done to show the possible differences in the relative amount of catecholamines in some organs and tissues that might indicate the nature of the secretory cells from which they originate. The contents of NE, E, and DA were assessed in rats neonatally treated with Gnt. Gnt-treated rats showed significantly lower levels of NE (P < 0.01) in all tissues except the adrenal gland and paraganglia. Epinephrine was present in all tissues with mean levels below 25 ng/g, with the exception of the adrenal gland (700 microg/gland) and paraganglia (100 ng/g). Only the heart showed lower values in Gnt-treated rats. Mean DA levels were also very high in paraganglia (530 ng/g). In the Gnt-treated rats, DA levels fell practically to zero except in the duodenum, mesentery, and adrenal, whereas there were high levels in the paraganglia, which were significantly different from controls. The results suggest that the three catecholamines are contained mainly in noradrenergic sympathetic fibers of muscle, white adipose tissue, heart, liver, pancreas, and spleen. The duodenum and mesentery may have dopaminergic fibers or E- and DA-containing nonneural cells. Hepatic-vagus paraganglia contain all the catecholamines in relatively high amounts in nonneural cells, and Gnt treatment raises DA levels without affecting the other amines.

Depletion and recovery of catecholamines in several organs of rats treated with reserpine

Chemical sympathectomy with reserpine depletes catecholamines in every neuronal or nonneuronal cell producing a nonspecific temporal sympathectomy. After reserpine administration, most of the drug is distributed to tissues based on their blood flow and would then either be metabolized or be reversibly bound in lipid depots from where it might be released. Consequently, reserpine concentration and the catecholamine-depleting effect in the various tissues are expected to differ according to the route of administration. This study was designed to compare the effects of intraperitoneal (i.p.) and subcutaneous (s.c.) administration of reserpine on catecholamine depletion and recovery in the liver, portal vein, and adrenal gland on days 1, 4, and 10 after reserpine dosage. Catecholamine determinations were extended to 25 days after the treatment only in s.c. reserpine-treated rats and adding samples of heart and brown adipose tissue to the testing. I.p. and s.c. reserpine administration had the same norepinephrine-depleting effect in the portal vein and liver but full recovery was present in both tissues only in i.p. reserpine-treated rats. In the adrenal gland, both routes of administration produced the same depleting and recovery effect of norepinephrine and epinephrine concentrations. A significant temporary overshoot in epinephrine levels was observed several days after s.c. reserpine treatment. Except for the liver, reserpine injected s.c. depleted norepinephrine concentrations significantly in all other tissues up to the end of the experiment. Our results suggest that chemical sympathectomy caused by reserpine administered s.c. produces a generalized and prolonged decrease in peripheral sympathetic activity that could be compensated by an increase in activity of the adrenal gland.

Catecholamines in paraganglia associated with the hepatic branch of the vagus nerve : effects of 6-hydroxydopamine and reserpine

Paraganglia are clusters of cells containing catecholamines (CA), mainly norepinephrine (NE) and dopamine (DA). The presence of epinephrine (E), on the other hand, has only been determined by indirect methods in retroperitoneal paraganglia of newborn and aged rats. Because their location, paraganglia associated with the hepatic branch of the vagus nerve may be a possible source of CA for the liver. The main purposes of the present study were to determine CA levels and whether E can be found in the omentum minus which includes paraganglia associated with the hepatic branch of the vagus nerve, and then to study the effects of 6-hydroxydopamine and reserpine on their CA content. Twenty-four female Wistar rats were randomly ascribed to three groups receiving two intraperitoneal injections of either 6-hydroxydopamine, reserpine or saline. Twenty-four hours after the last administration the rats were anesthetized and a portion of the omentum minus was obtained. Left adrenal medulla and a liver fragment were also collected as controls. The samples were processed to be analyzed by high performance liquid chromatography and catecholamine histofluorescence. The results confirm previous reports about the presence of considerable amounts of norepinephrine and dopamine in paraganglia. Norepinephrine and dopamine in the omentum like the adrenal medulla were significantly depleted by reserpine but not by 6-hydroxydopamine treatment, suggesting that some other sources in addition to sympathetic terminals are responsible for CA in the omentum. On the contrary, both drugs reduced liver NE, consistent with the localization of this amine mainly to hepatic sympathetic terminals. Histofluorescence of the omentum revealed 2-4 paraganglia per tissue fragment. Paraganglia associated with the hepatic branch of the vagus nerve contain also E. The presence of perihepatic sources of extra-adrenal CA, and more specifically E, could be of physiological significance.

Effect of lipectomy and long-term dexamethasone on visceral fat and metabolic variables in rats☆

Intraperitoneal (IP) fat accumulation in humans is a risk factor for a number of diseases. We tried to increase this particular adipose mass in rats by long-term administration of low-dose dexamethasone (Dex) and/or elimination of other fat depots. Male adult Wistar rats were lipectomized (Lip) or sham-operated (Sh). Bilateral lipectomy of retroperitoneal and inguinal fat pads was performed under anesthesia with Na pentobarbital 40 mg/kg supplemented with ether. After 8 days, half the animals of each group received Dex in their drinking water (0.1 microgram/mL) while the other half received water (W), for a total of four groups: Sh-W, Lip-W, Sh-Dex, and Lip-Dex. Body weight (BW) and food and water intake were measured throughout the treatment period. A glucose tolerance test was performed 34 days after starting Dex treatment, and then rats were killed, fat depots were weighed, and plasma and liver were obtained for metabolic determinations. Dex rats ate the same amount of food as W controls, but gained significantly less weight (2.02 +/- 0.18 v 3.82 +/- 0.10 g/d, P < .01). Mean daily W intake was approximately 40 mL/d in all groups, which means that Dex rats ingested approximately 4 micrograms/d Dex. Average glycemic values during the 180-minute glucose tolerance test were as follows: Sh-W, 162 +/- 13; Lip-W, 166 +/- 7; Sh-Dex, 118 +/- 6; and Lip-Dex, 229 +/- 27 mg/dL. These values show that glucose tolerance was improved by Dex treatment alone, but was impaired in Lip-Dex animals. The same trend was evident for the relative weights (percent of BW) of two intact adipose depots: IP and epididymal (EPI) (Sh-W, 2.08 +/- 0.13 and 1.35 +/- 0.11, respectively; Lip-W, 1.67 +/- 0.15 and 1.17 +/- 0.11; Sh-Dex, 1.66 +/- 0.10 and 1.28 +/- 0.07; Lip-Dex, 2.41 +/- 0.11 and 1.53 +/- 0.09). Average glycemia for all rats was significantly correlated with IP (r = .55, P < .01) but not with EPI; moreover it was correlated in the Sh-W control group (r = .81, P < .05), suggesting a normal relation between these variables. Liver triglycerides (LTG), which were elevated in Dex rats, were also correlated with IP (r = .51, P < .02 for all rats and r = .82, P < .05 for Sh-W rats). The results show that long-term administration of low-dose Dex has some different effects in normal versus Lip rats concerning mainly the IP fat depot, the relative mass of which seems to significantly affect glucose tolerance.

Effects of catecholamines on water intake in rats.

It is known that intraperitoneally (IP) injected adrenaline (A) inhibits food intake in otherwise hungry animals. In a recent work, Hinton et al. (6) showed that IP A also inhibits water intake in thirsty rats, concluding that As effect is unspecific. We administered A IP or intramuscularly (IM) in different doses in rats made thirsty either by 18-h water deprivation or by subcutaneous injection of hypertonic saline or polyethylene glycol. IP A reduced water intake in all experimental conditions. A dose-related inhibition was observed in water-deprived animals. On the other hand, IM A showed a small effect only at the highest dose (50 micrograms/100 g body weight). When some of these experiments were repeated using noradrenaline (NA) and isoproterenol (IS), IM administration of either substance showed no effect. IP administration reduced water intake significantly only at the highest dose of NA (50 micrograms/100 g). It is concluded that water intake inhibition by catecholamines in rats made thirsty either by osmotic or by volumetric challenges is of porto-hepatic origin and, in contrast with food intake inhibition, has no beta-adrenergic component.

Some metabolic effects on lactating rats of a low-energy diet restricted in good-quality protein

Adult female Sprague-Dawley rats were fed ad libitum during pregnancy and lactation a control diet (CD; 16.1 kJ/g) or a low-energy diet with wheat gluten as the main protein source (LED; 13.3 kJ/g). Body weight, food intake, resting energy expenditure, respiratory quotient and substrate use by the mammary gland were measured. After the animals had been killed, the parametrial and retroperitoneal fat pads were weighed. The mean food intake (g) of the two groups of rats was similar, resulting in a lower energy intake by the LED rats, significantly different during the last 2 weeks of lactation. The mean body weight of both dams and pups in the LED group was lower, starting at day 9 of lactation. The resting energy expenditure increased gradually during lactation in the control group, whereas this increase was not seen in rats of the LED group in the last week of lactation. Rats that had fasted overnight had a respiratory quotient of 0.7 or less, whereas for rats that had been fed, the mean respiratory quotient was over 1.0. Under both conditions, rats showed ketonuria. The arteriovenous difference in 3-hydroxybutyrate level was higher and those for glucose, lactate and triacylglycerol were lower across the mammary glands of LED rats. The parametrial fat depot weighed less in LED rats. Reducing the increase in resting energy expenditure and using ketone bodies to a greater extent as fuels may represent important mechanisms in the LED dams to cover the energy cost of milk production.

Specificity of alpha- and beta-adrenergic inhibition of water and food intake

In previous publications from our laboratory it was shown that catecholamines (CA) injected intraperitoneally (IP) to fasted rats induce a transient inhibition of food intake. This effect seems to be both alpha- and beta-adrenergic. According to more recent data (20), IP CA also reduced water intake in water-deprived rats, and the effect is exclusively alpha-adrenergic. In order to obtain more information on the adrenergic specificity of the two inhibitory effects we measured the amount of food and water ingested during 30 min by male and female rats previously deprived of both food and water for 18 h. Three adrenergic agonists (norepinephrine, isoproterenol, and salbutamol) were injected IP after the administration (IP) of the following adrenergic antagonists: phentolamine, prazosin, yohimbine, propranolol, or metoprolol. Results showed that, under these experimental conditions, water intake inhibition was due exclusively to an alpha 1 effect, whereas food intake inhibition seemed to depend on alpha 1 and beta 1 actions plus some beta 2 participation. It is also suggested that blocking one type of receptors may enhance the responsiveness of the other type.

Chemical sympathectomy alters food intake and thermogenic responses to catecholamines in rats.

It has been suggested that the sympathetic nervous system contributes to the short-term control of feeding. The adrenergic innervation of some splanchnic organs seems to be especially involved in such processes, since catecholamines reduce feeding only when injected intraperitoneally or intraportally. In this work, the effects of neonatal sympathetic denervation with guanethidine (Gnt) upon food intake were assessed in adult rats. Gnt-treated male rats had lower body weight gain. The hypophagic response to intraperitoneal (ip) norepinephrine was 70% higher in Gnt-treated animals as compared to controls (P < 0.05); that of epinephrine (E) by 33% (P < 0.05) and that of isoproterenol was not significantly modified. As in normal rats, the hypophagic effect was much stronger after ip than after intramuscular (im) administration (P < 0.05). On the other hand, resting oxygen consumption (VO2) was consistently lower in denervated animals. Ip E administration did not modify VO2, while im E caused increased motor activity and VO2 (P < 0.05). In contrast to control rats, the respiratory exchange ratio in ad libitum fed Gnt rats did not decrease after Ip E administration, suggesting a lack of effect upon lipid mobilization. The lower rate of body weight gain induced by neonatal Gnt sympathectomy might be due to lower daily food intake possibly related, in part, to the sensitization of the alpha-adrenergic porto-hepatic response to endogenous catecholamines. Compared with controls, Gnt-treated rats also showed a limited thermogenic capacity not related to feeding, and a greater degree of carbohydrate oxidation, possibly due to a defect in E-induced lipolysis, which is beta-adrenergic.