S 23521 Decreases Food Intake and Body Weight Gain in Diet‐Induced Obese Rats

Objective: To investigate the effect of S 23521, a new glucagon‐like peptide‐1‐(7‐36) amide analogue, on food intake and body weight gain in obese rats, as well as on gene expression of several proteins involved in energy homeostasis. Research Methods and Procedures: Lean and diet‐induced obese rats were treated with either S 23521 or vehicle. S 23521 was given either intraperitoneally (10 or 100 μg/kg) or subcutaneously (100 μg/kg) for 14 and 20 days, respectively. Because the low‐dose treatment did not affect food intake and body weight, the subcutaneous treatment at high dose was selected to test the effect on selected end‐points. Results: Treated obese rats significantly decreased their cumulative energy intake in relation to vehicle‐treated counterparts (3401 ± 65 vs. 3898 ± 72 kcal/kg per 20 days; p < 0.05). Moreover, their body weight gain was reduced by 110%, adiposity was reduced by 20%, and plasma triglyceride levels were reduced by 38%. The treatment also improved glucose tolerance and insulin sensitivity of obese rats. Regarding gene expression, no changes in uncoupling protein‐1, uncoupling protein‐3, leptin, resistin, and peroxisome proliferator‐activated receptor (PPAR)‐γ were observed. Discussion: S 23521 is an effective glucagon‐like peptide‐1‐(7‐36) amide analogue, which induced a decrease in energy intake, body weight, and adiposity in a rat model of diet‐induced obesity. In addition, the treatment also improved glucose tolerance and insulin sensitivity of obese rats. These results strongly support S 23521 as a putative molecule for the treatment of obesity.     

Differential Short-Term Distribution of Estrone and Oleoyl-Estrone Administered in Liposomes to Lean and Obese Zucker Rats

Thirteen-week-old female Zucker lean (Fa/Fa) and obese (fa/fa) rats were injected through a cannula inserted in the left jugular vein with 1 mL/kg of ³H-labeled oleoyl-estrone in liposomes (Merlin-2) (i.e., 670 fmol, 84 kBq). The rats were killed 10 minutes later and dissected. The presence of intact or hydrolyzed oleoyl-estrone was later determined in all samples. The pattern of distribution of estrone was quite different from that of oleoyl-estrone both in rats that were lean and in those that were obese. Estrone was better retained by white adipose tissue than oleoyl-estrone. Liver, spleen, and lungs accumulated more oleoyl-estrone and split part of it, from 4.7% (lung, obese) to 27% (liver, lean). The overall high retention of estrone by the rat tissues results in its very low circulating levels. The fast splitting of liposome-carried oleoyl-estrone by most tissues (up to more than 67% by intestine and skin of lean rats) may help explain the rise in blood free estrone. The differences between lean and obese Zucker rats are mainly quantitative in the case of estrone, the main differences being found in blood and adipose tissues. However, when we compare the data for oleoyl-estrone, the differences cannot be dismissed simply as due to differences in body size or the extent of fat deposits. A large portion of the label remained in the blood of the rats that were obese but not in those that were lean, the tissues of which took up more label. Brown adipose tissue shows a fair affinity for oleoyl-estrone in the rats that were lean but practically does not retain label in the rats that were obese, suggesting that oleoyl-estrone may have a direct effect on brown adipose tissue. The decreased uptake of oleoyl-estrone in rats that were obese shows that the mechanism regulating the turnover or disposal of this signal is altered in this type of genetic obesity.     

Retrograde Gastric Electrical Stimulation Reduces Food Intake and Weight in Obese Rats

Objective: To investigate the therapeutic potential of retrograde gastric electrical stimulation (RGES) for obesity in a rodent model of obesity. Research Methods and Procedures: The study was performed in 12 obese Zucker rats implanted with two pairs of gastric serosal electrodes, one pair for stimulation and the other for recording intrinsic gastric myoelectrical activity. It was composed of an acute study in three sessions to study the effect of RGES on intrinsic gastric myoelectrical activity and acute food intake and a chronic phase to study the short‐term effect of RGES on weight. RGES was performed through the distal stomach using long pulses at a frequency of tachygastria (known to induce gastric hypomotility). Results: RGES completely entrained intrinsic gastric myoelectrical activity and turned it into tachygastria at a certain strength. RGES reduced acute food intake compared with the control (p < 0.01). A 2‐week treatment of RGES resulted in a significant reduction in food intake (p = 0.002) and a significantly greater weight loss than sham stimulation (p = 0.004). Discussion: RGES at a tachygastrial frequency reduces food intake and results in weight loss in obese Zucker rats, and its effect is probably attributed to the introduction of tachygastria in the stomach.     

Fiber Intake of Normal Weight,Moderately Obese and Severely Obese Subjects

ALFEERI, MARGARET AH, JOCELINE POMERLEAU, D MICHAEL GRACE AND LORRAINE ANDERSON. Fiber intake of normal weight, moderately obese and severely obese subjects. Obes Res. The lack of dietary fiber may be a contributing factor in obesity. This study examined the fiber intake of three weight groups: normal (20.0≤BMI≤27.0), moderately obese (27.1≤BMI≤39.9) and severely obese (BMI≥40.0). Each group contained 50 subjects. Detailed 3-day food records were used to gather the nutritional data. Fiber intake in the normal weight group was 18.8 ± 9.3 grams, the moderately obese consumed 13.3 ± 5.8 grams of fiber and the severely obese 13.7 ± 5.7 grams. Total fiber intake in grams was found to be significantly higher in the lean group (p<0.05) and was positively associated with sex and education level with men and more highly educated individuals consuming more fiber. Using regression analysis total fiber in grams and fiber in g/1000 kcalories was inversely associated with BMI after adjusting for sex, age, education level and income (p<0.01). A high fiber diet may help to promote a negative energy balance by causing early satiety secondary to gastric distention. Dietitians and physicians need to emphasize the importance of a high fiber diet to their obese patients.     

Food Viscosity Influences Caloric Intake Compensation and Body Weight in Rats

Objective: To determine the effects of food viscosity on the ability of rats to compensate for calories in a dietary supplement. Research Methods and Procedures: In a series of four experiments, rats consumed dietary supplements equated for caloric and nutritive content but differing in viscosity. Experiments 1 to 3 examined the ability of the rats to compensate for the calories consumed in low‐ compared with high‐viscosity premeals by reducing intake of a subsequent test meal. Caloric compensation was assessed with a wide range of premeal viscosity levels and with two different non‐nutritive thickening agents. Experiment 4 assessed the effects of consuming daily a low‐viscosity compared with an equicaloric high‐viscosity dietary supplement on longer term body weight gain. Results: Consuming a lower viscosity premeal was followed by significantly more caloric intake (i.e., less caloric compensation) compared with consuming premeals with higher viscosity levels. This effect was not specific to one thickening agent. Furthermore, rats given a low‐viscosity supplement daily gained significantly more weight over a 10‐week period compared with rats given a high‐viscosity supplement. Discussion: The results of these experiments suggest that food viscosity may be an important determinant of short‐term caloric intake and longer term body weight gain.     

Parabrachial Interleukin-6 Reduces Body Weight and Food Intake and Increases Thermogenesis to Regulate Energy Metabolism

1. Download high-res image (179KB)
2. Download full-size image

     

Opioidergic Modulation of Ventilatory Response to Sustained Hypoxia in Obese Zucker Rats

Objective: To determine whether altered central and/or peripheral opioidergic mechanisms contribute to the altered ventilatory response to sustained hypoxia in obese Zucker rats. Research Methods and Procedures: Eight lean (176 ± 8 [SEM] g) and eight obese (225 ± 12 g) Zucker rats were studied at 6 weeks of age. Pulmonary ventilation (V?_E), tidal volume (V_T), and breathing frequency (f) at rest and in response to sustained (30 minutes) hypoxic (10% O₂) challenges were measured on three separate occasions by the barometric method after the randomized, blinded administration of equal volumes of saline (control), naloxone methiodide (N_M; 5 mg/kg, peripheral opioid antagonist), or naloxone hydrochloride (N_HCl; 5 mg/kg, peripheral and central opioid antagonist). Results: Administration of N_M and N_HCl in lean animals had no effect on V?_E either at rest or during 30 minutes of sustained exposure to hypoxia. Similarly, N_M failed to alter V?_E in obese rats. In contrast, N_HCl significantly (p < 0.05) increased V?_E and V_T both at rest and during 2 to 10 minutes of hypoxic exposure in obese rats. After 20 to 30 minutes of hypoxic exposure, V_T remained elevated with N_HCl, but the earlier elevation of V?_E seemed to be attenuated due to a decrease in f at 20 minutes of exposure to hypoxia. Discussion: Thus, endogenous opioids modulate both resting V?_E and the ventilatory response to sustained hypoxia in obese, but not in lean, Zucker rats by acting specifically on opioid receptors located within the central nervous system.     

Effects of a Fat Substitute,Sucrose Polyester,on Food Intake,Body Composition,and Serum Factors in Lean and Obese Zucker Rats

Sucrose polyester, a fat substitute, has shown promise in reducing blood cholesterol and body weight of obese individuals. Effects of this compound in the Zucker rat, a genetic model of obesity, are unknown. Thus, we examined food intake, body weight, body composition, and several metabolic parameters in sera of lean and obese female Zucker rats. Eight-week-old lean and obese animals were given a choice between a control diet (15% corn oil) and fat substitute diet (5% corn oil and 10% sucrose polyester) for 2 days. Next, one-half of the lean and obese groups received control diet; the remaining lean and obese rats received fat substitute diet for 18 days. Cumulative food intake was depressed in fat substitute groups relative to control-fed animals; however, this effect was more predominant in obese animals. Obese rats consuming fat substitute diet (O-FS) gained less weight as compared to obese control-fed animals (O-C). Lean rats given fat substitute (L-FS) did not have significantly different body weights as compared to the L-C group. Fat substitute groups, combined, had lower body fat and higher body water as compared to controls. The O-FS group had lower serum glucose and insulin and higher fatty acid levels compared to the O-C group. There were no differences in serum cholesterol, HDL, or triglyceride levels due to fat substitute diet. These data suggest that the obese Zucker rat is unable to defend its body weight when dietary fat is replaced with sucrose polyester.     

Regional Fat Pad Growth and Cellularity in Obese Zucker Rats: Modulation by Caloric Restriction

Objective : To investigate, in young obese male Zucker rats, the effects of chronic food restriction and subsequent refeeding on: 1) parameters of nonadipose and adipose growth, 2) regional adipose depot cellularity [fat cell volume (FCV) and number], and 3) circulating leptin levels. Research Methods and Procedures : Obese (fa/fa) and lean (Fa/?) male Zucker rats were studied from age 5 to 19 weeks. After baseline food intake monitoring, 10 obese rats were subjected to 58 days of marked caloric restriction from ad libitum levels [obese‐restricted (OR)], followed by a return to ad libitum feeding for 22 days. Ten lean control rats and 10 obese control rats were fed ad libitum for the entire experiment. All rats were fed using a computer‐driven automated feeding system designed to mimic natural eating patterns. Results : After food restriction, OR rats weighed significantly less than did lean and obese rats and showed a significant diminution in body and adipose growth as compared with obese rats. Relative adiposity was not different between obese and OR rats and was significantly higher than that of lean rats. The limitation in growth of the adipose tissue mass in OR rats was due mostly to suppression of fat cell proliferation because the mean FCV in each of the four depots was not affected. Serum leptin levels of OR and obese rats were not different from each other but were significantly higher than those of lean rats. Discussion : Marked caloric restriction affects obese male Zucker rats in a manner different from that of nongenetic rodent models (i.e., Wistar rats). In comparison with the response to caloric deprivation of Wistar rats, these calorically restricted obese male Zucker rats appeared to defend their relative adiposity and mean FCV at the expense of fat cell number. These findings indicate that genetic and/or tissue‐specific controls override the general consequences of food restriction in this genetic model of obesity.     

Effect of Chronic Central Administration of Glucagon-Like Peptide-1 (7–36) Amide on Food Consumption and Body Weight in Normal and Obese Rats

Glucagon-like peptide (7–36) amide (GLP-1) acutely inhibits food and water consumption in rats after intrace-rebroventricular (icv) administration. To assess the potential for desensitization of these effects, we investigated the effects of chronic icv administration of GLP-1 on food consumption and body weight in Sprague-Dawley (SD) rats and Zucker (fa/fa) obese rats. In vitro functional densensitization of the GLP-1 receptor was not observed after overnight exposure of Rin m5F insulinoma cells to GLP-1 at concentrations up to 10 nM. Administration of GLP-1 to SD rats (30 ug icv twice a day for 6 days) resulted in significant reductions in 24-hour food consumption each day (25 ±1%). Continuous icv infusion of GLP-1 for 7 and 14 days significantly inhibited cumulative food consumption and reduced body weight in SD rats. In the genetically obese Zucker rat, chronic dosing with GLP-1 (30 ug icv) once a day for 6 days caused significant reductions in food consumption each day and a reduction in body weight. These results indicate that the GLP-1 pathways in the central nervous system controlling food consumption do not desensitize after chronic exposure to GLP-1 and suggest that agonists of the central GLP-1 receptor may be effective agents for the treatment of obesity.     

The Relationship Between Daytime and Nighttime Food Intake in an Obese Night-Eater

Some obese individuals consume food during awakenings from nighttime sleep. Three studies were conducted on a 28-year-old morbidly obese male with chronic sleeping complaints and insignificant weight loss, despite self-reported daily caloric restriction: I. For 3 mo, the subject recorded food intake for 24-h periods. Mean daytime intake was 1286 kcal ± 386 (SD), and mean nighttime intake was 1036 kcal ± 487 (SD). Caloric values of daytime and nighttime intake were negatively correlated, r = ?0.22, df= 82, p<.05. II. Seven consecutive 24-h food intake recordings were obtained with an automated formula dispenser when the subject was an inpatient on a metabolic ward and received ad libitum formula as his sole food source. Mean daytime intake was 1245 ± 662 (SD), and mean nighttime intake was 231 ± 236 (SD). There was a non-significant negative correlation between daytime and nighttime intake, r = -0.32, df = 5, NS. III. The subject underwent polysomnographic studies on 2 non-consecutive nights, following the administration of either a low (600 kcal) or high (1800 kcal) daytime caloric condition. The subject, upon awakening from nighttime sleep, could eat from a platter of sandwich quarters placed at his bedside. The addition of 1200 kcal to daytime intake decreased nighttime intake by 654 kcal, or by 55% of the additional calories delivered during the day. The three studies (I, II, and III) show that daytime food intake can be negatively correlated with nighttime intake, and that daytime intake can influence nighttime intake in a documented obese night-eater.     

Normal Distribution of Body Weight Gain in Male Sprague‐Dawley Rats Fed a High‐Energy Diet

Objective: To investigate the effect of a high‐energy (HE) diet on caloric intake, body weight, and related parameters in outbred male Sprague‐Dawley (SD) rats. Research Methods and Procedures: Twenty‐eight SD rats were fed either chow (C) for 19 weeks or HE diet for 14 weeks and then C for 5 weeks. Blood hormones and metabolites were assayed, and expression of uncoupling protein‐1 and hypothalamic energy‐balance‐related genes were determined by Northern blotting and in situ hybridization, respectively. Results: HE rats gained body weight more rapidly than C animals with a range of weight gains, but there was no evidence that weight gain was bimodally distributed. Caloric intake was transiently elevated after introduction of the HE diet. Transfer of HE rats back to C resulted in a drop in caloric intake, but a stable body weight. In terminal analysis, two of four dissected adipose tissue depots were heavier in rats that had previously been fed HE diet. Blood leptin, insulin, glucose, and nonesterified fatty acids were not different between the groups. Uncoupling protein‐1 mRNA was elevated in interscapular brown adipose tissue from HE rats. There was a trend for agouti‐related peptide mRNA in the hypothalamic arcuate nucleus to be higher in HE rats. Discussion: Contrary to other studies of the SD rat on HE diet, body weight and other measured parameters were normally distributed. There was no segregation into two distinct populations on the basis of susceptibility to diet‐induced obesity. This characteristic may be dependent on the breeding colony from which animals were sourced.     

Time-Dependent Effects of Progressive Gamma -Linolenate Feeding on Hyperphagia,Weight Gain,and Erythrocyte Fatty Acid Composition During Growth of Zucker Obese Rats

Obese Zucker rats (fa/fa) have low levels of arachidonic acid (AA) in liver phospholipids (PL). We have previously shown that a 70% gamma-linolenate concentrate (GLA; an AA intermediate) fed at a fixed dose (0.07 g/day) normalized hepatic PL AA and reduced weight gain selectively in the obese animals. In a follow-up study, 16 obese (fa/fa) and 16 lean (Fa/Fa) 4-week-old male rats were randomized into 4 groups of 8 each and gavaged daily with soybean oil (SOY) containing 55% 18:2ω6 (an AA precursor) or GLA, using a progressive dose (≤ 5% of total calories) based on body weight. A defined diet with 11% of energy as SOY was fed ad libitum for 60 days. GLA obese had lower body weight (p<0.0001) and 60-day cumulative food intake (p<0.05) compared to SOY obese, but neither parameter differed between the lean groups. For the last twenty days cumulative food intake was identical for GLA obese and SOY lean, whereas SOY obese consumed 18% more (p<0.05). Thus the progressive dose of GLA selectively suppressed hyperphagia in obese Zucker rats. Erythrocytes collected at 15-day intervals showed parallel increases in AA in both genotypes over time, suggesting normal AA availability during rapid growth. Thus, the reduced PL AA in the livers from the obese rats probably reflects impaired distribution in selected tissues rather than reduced hepatic production. Due to the potential health risks of enriching tissue lipids with AA, great caution is advised in considering GLA as therapy for human obesity.     

Corticosterone Assimilation by a Voluntary Oral Administration in Palatable Food to Rats

Drug delivery in research on nonhuman animals in the laboratory is still challenging because it is usually invasive and stressful. Stress-free voluntary oral drug administration in water lacks precise control of dose and timing of substance ingestion. Voluntary oral consumption of corticosterone has been previously successfully applied in mice using oat flakes, but protocols for oral corticosterone administration in rats remain unavailable. This study assessed the effectiveness of voluntary oral administration to rats of a palatable piece of bread soaked with corticosterone that can be rapidly prepared and is reliably dose- and timing-controllable. After three familiarization days, all rats ate the bread within 120 seconds of presentation, irrespective of the presence or absence of corticosterone or vehicle. Corticosterone plasma levels remained at basal levels with consumption of vehicle-containing bread, and they were significantly increased with corticosterone-containing bread. Hence, the method enabled corticosterone bodily assimilation while avoiding stress, making it a possible alternative for invasive and stressful procedures. This article includes a methodological refinement that lessens unnecessary discomfort to laboratory animals and is potentially suitable for acute and chronic protocol studies.     

Effects of Exendin‐4 Alone and With Peptide YY3–36 on Food Intake and Body Weight in Diet‐Induced Obese Rats

Significant weight loss following Roux‐en‐Y gastric bypass surgery (RYGB) in obese humans correlates with enhanced secretion of anorexigenic gut hormones glucagon‐like peptide‐1 (GLP‐1) and peptide YY_3–36 (PYY_3–36). Our aim here was to identify a dosing strategy for intraperitoneal (IP) infusion of GLP‐1 homologue exendin‐4 alone and with PYY_3–36 that produces a sustained reduction in daily food intake and body weight in diet‐induced obese (DIO) rats. We tested 12 exendin‐4 strategies over 10 weeks. Exendin‐4 infused during the first and last 3 h of the dark period at 15–20 pmol/h (0.15 nmol/kg/day) produced a sustained 24 ± 1% reduction in daily food intake for 17 days, and decreased body weight by 7%. In a separate group of DIO rats, none of seven dosing strategies combining exendin‐4 and PYY_3–36 produced a similar reduction in daily food intake for >10 days. The subsequent decline in efficacies of exendin‐4 alone and with PYY_3–36 on food intake and body weight in each experiment suggested possible receptor downregulation and tolerance to treatments. However, when treatments were discontinued for 1 day following losses in efficacies, daily food intake significantly increased. Together, these results demonstrate that (i) intermittent IP infusion of a low dose of exendin‐4 can produce a relatively prolonged reduction in daily food intake and body weight in DIO rats, (ii) co‐infusion of exendin‐4 and PYY_3–36 does not further prolong this response, and (iii) activation of an orexigenic mechanism gradually occurs to counteract the inhibitory effects of exendin‐4 alone and with PYY_3–36 on food intake and body weight.     

硫胺素、核黄素和烟酰胺对大鼠体重增长及脂代谢影响的实验研究

目的：B族维生素以辅酶的形式参与糖、脂肪和蛋白质代谢,本文观察硫胺素、核黄素和烟酰胺补充对高脂饲料诱导大鼠肥胖的影响。方法：采用预防肥胖模型法,2×2×2析因设计分为8组：高脂对照组（F0组）,高脂＋硫胺素（F1组）,高脂＋核黄素（F2组）,高脂＋烟酰胺（F3组）,高脂＋硫胺素＋核黄素（F4组）,高脂＋硫胺素＋烟酰胺（F5组）,高脂＋核黄素＋烟酰胺（F6组）,高脂＋硫胺素＋核黄素＋烟酰胺（F7组）,每组12只大鼠,给予高脂饲料喂养,同时硫胺素（100mg／kgbw／d）、核黄素（100mg／kgbw／d）、烟酰胺（250mg／kgbw／d）灌胃,另设正常对照组（C组）12只,普通饲料喂养,自来水灌胃,15周后,分析其体重、摄食量、体脂重量、血脂等实验前后的变化情况及各组动物之间的差别。结果：经过15周喂养后,高脂喂养大鼠体重比正常对照组平均增加了15．7％;而补充烟酰胺（F3）及联合硫胺素（F5）或核黄素（F6）以及三者联合补充（F7）组高脂喂养大鼠与高脂对照组（F0）相比,体重分别降低了35．0％,30．0％,30．1％和30．6％（P值均小于0．05）;甚至比正常对照组大鼠平均体重分别下降了22．8％,17．0％,17．0％和17．7％（P值均小于0．05）;而补充核黄素或和硫胺素组大鼠体重没有明显增加或降低（P值均大于0．05）。血脂分析结果显示高脂喂养并联合补充核黄素或／和烟酰胺和／或硫胺素组大鼠血清CHOL和LDL水平明显低于高脂对照组;而高脂喂养并联合补充烟酰胺（F3）及联合硫胺素（F5）或核黄素（F6）以及三者联合补充（F7）组大鼠LDL／HDL比值分别0．29、0．26、0．25和0．26,明显低于F0组的0．37（P值均小于0．05）。结论：大剂量烟酰胺可有效地调节血脂水平和控制肥胖大鼠的体重增长,而核黄素及硫胺素对控制肥胖     

The Association of Body Weight,Dietary Intake,and Energy Expenditure with Dietary Restraint and Disinhibition

LAWSON, OLGA J, DONALD A WILLIAMSON, CATHERINE M CHAMPAGNE, JAMES P DELANY, ELLEN R BROOKS, PAULA M HOWAT, PATRICIA J WOZNIAK, GEORGE A BRAY AND DONNA H RYAN. The association of body weight, dietary intake, and energy expenditure with dietary restraint and disinhibition. Obes Res. 1995;3:153–161. The hypotheses that dieting and/or overeating are associated with adiposity, eating disturbances, and lowered energy expenditure were tested in this study. A sample of 44 premenopausal women scoring high and low on measures of dietary restraint and disinhibition of dietary control, as measured by the Three Factor Eating Questionnaire, was studied. A 2 × 2 factorial design was employed (High/Low Restraint x High/Low Disinhibition). Dependent variables were: body composition, dietary intake, activity, resting metabolic rate, and thermic effect of food. Unrestrained overeaters (Low Restraint/High Disinhibition group) were very obese. High Dietary Restraint was associated with intent to diet and controlled eating. High scores on the Disinhibition Scale were associated with episodic overeating. Groups did not differ in resting metabolic rate (controlled for fat-free mass). Lower thermic effect of food was found to be associated with the obesity found in High Disinhibition subjects. Thus, Dietary Restraint was not associated with significant adverse effects upon physical or psychological health. High Disinhibition, however, was associated with adiposity and significant disturbances of eating.     

Chronic Intracerebroventricular Infusion of Insulin Failed to Alter Brain Insulin-Binding Sites, Food Intake, and Body Weight

The present study was performed to explore the role of exogenous insulin in CSF in the control of energy balance in the rat. For this purpose, adult male Sprague-Dawley rats carrying an indwelling cannula in the right lateral cerebral ventricle were infused for a maximum of 10 days with insulin (Actrapid) at various rates (starting at 0, 45, 85, 170, and 600 ng/day) or anti-insulin antibody (IgG fraction; diluted 1:10 wt/vol) with an osmotic minipump. All those treatments did not modify the growing rates; neither total daily food intake nor the circadian rhythm of food intake was further modified. The chronic insulin infusion starting at 600 ng/day resulted in a chronic significant increase in CSF insulin levels without changing the plasma insulin level. It failed to alter specific insulin binding sites to Triton X-100 solubilized microsomal membranes from various brain areas (cerebral cortex, olfactory bulbs, and lateral and medial hypothalami) at the end of the 5- or 10-day period of insulin infusion. Purification of insulin receptors on a wheat germ agglutinin did not reveal any further effect of insulin. From these results, it seems unlikely that the input to the brain insulin-effector systems could arise from CSF insulin.