The effects of lactation on impulsive behavior in vasopressin-deficient Brattleboro rats

Vasopressin (AVP)-deficient Brattleboro rats develop a specific behavioral profile, which—among other things—include altered cognitive performance. This profile is markedly affected by alterations in neuroendocrine state of the animal such as during lactation. Given the links between AVP and cognition we hypothesized that AVP deficiency may lead to changes in impulsivity that is under cognitive control and the changes might be altered by lactation. Comparing virgin and lactating AVP-deficient female Brattleboro rats to their respective controls, we assessed the putative lactation-dependent effects of AVP deficiency on impulsivity in the delay discounting paradigm. Furthermore, to investigate the basis of such effects, we assessed possible interactions of AVP deficiency with GABAergic and serotonergic signaling and stress axis activity, systems playing important roles in impulse control. Our results showed that impulsivity was unaltered by AVP deficiency in virgin rats. In contrast a lactation-induced increase in impulsivity was abolished by AVP deficiency in lactating females. We also found that chlordiazepoxide-induced facilitation of GABAergic and imipramine-induced enhancement of serotonergic activity in virgins led to increased and decreased impulsivity, respectively. In contrast, during lactation these effects were visible only in AVP-deficient rats. These rats also exhibited increased stress axis activity compared to virgin animals, an effect that was abolished by AVP deficiency. Taken together, AVP appears to play a role in the regulation of impulsivity exclusively during lactation: it has an impulsivity increasing effect which is potentially mediated via stress axis-dependent mechanisms and fine-tuning of GABAergic and serotonergic function.     

Behavioral differences between vasopressin-deficient (Brattleboro) and normal Long-Evans rats

Behavioral comparisons were made between rats of the Brattleboro strain with hereditary hypothalamic diabetes insipidus (DI) and normal Long-Evans rats. Measurements were made of activity behavior in a lighted open field and in a darkened activity chamber. Subtle measurement specific differences in the activity behavior of DI rats were found which suggested altered emotion, motivation and/or attention in the DI rats. In terms of learned behavior, DI and normal rats displayed a similar degree of habituation to all within-session activity measures in both the open field and darkened activity chamber. In a passive avoidance test, DI rats exhibited a degree of avoidance behavior equivalent to that of normal animals. Thus, these studies provide evidence that the vasopressin-deficient rat is not defective in learning and memory processes. The data can be interpreted as suggesting that vasopressin may influence memory tasks by extrinsic modulation of related states of emotionality, motivation and/or attention rather than by direct involvement in the retrieval and consolidation of information.     

Effects of naloxone and cholecystokinin on food and water intake in vasopressin-deficient rats (Brattleboro strain)

Opioid peptides and cholecystokinin (CCK) have been shown to play a role in regulation of feeding behavior. Another neuropeptide that has recently been suggested to be involved in feeding is vasopressin. We explored possible interactions between opiates, CCK and vasopressin in feeding regulation by studying feeding suppression produced by naloxone and CCK in Brattleboro (DI) rats, which are homozygous for diabetes insipidus and lack the ability to synthesize vasopressin. Ten DI and 15 age-matched Long Evans (LE) rats were food deprived for 14 hours on two different days and then injected with naloxone (2.5 mg/kg) on one day or saline on the other. Thirty minutes later the food was returned and food and water consumption were measured after 1, 3 and 4 hr. Naloxone suppressed the food consumption of both DI and LE rats but the suppression was greater for the DI rats. This result was specific to feeding as water consumption was suppressed in LE more than in DI rats. Two weeks later, the same rats were food deprived for 6 hours on two different days and then injected with CCK-8 (2.5 micrograms/kg) on one day and with saline on the other. Food was returned one minute after the injection and food and water consumption were measured 30 and 60 minutes later. Food intake was reduced equally for both DI and LE rats. Water intake was not reduced. The results suggest that the suppression of feeding by CCK does not require an intact vasopressinergic system. The greater feeding suppression by naloxone in DI rats may suggest that opiates are interacting with vasopressin in producing their effects on food intake.     

Simvastatin enhances aquaporin-2 surface expression and urinary concentration in vasopressin-deficient Brattleboro rats through modulation of Rho GTPase

Statins are 3-hydroxyl-3-methyglutaryl-CoA reductase inhibitors that are commonly used to inhibit cholesterol biosynthesis. Emerging data have suggested that they also have "pleotropic effects," including modulating actin cytoskeleton reorganization. Here, we report an effect of simvastatin on the trafficking of aquaporin-2 (AQP2). Specifically, simvastatin induced the membrane accumulation of AQP2 in cell cultures and kidneys in situ. The effect of simvastatin was independent of protein kinase A activation and phosphorylation at AQP2-Ser(256), a critical event involved in vasopressin (VP)-regulated AQP2 trafficking. Further investigation showed that simvastatin inhibited endocytosis in parallel with downregulation of RhoA activity. Overexpression of active RhoA attenuated simvastatin's effect, suggesting the involvement of this small GTPase in simvastatin-mediated AQP2 trafficking. Finally, the effect of simvastatin on urinary concentration was investigated in VP-deficient Brattleboro rats. Simvastatin acutely (3-6 h) increased urinary concentration and decreased urine output in these animals. In summary, simvastatin regulates AQP2 trafficking in vitro and urinary concentration in vivo via events involving downregulation of Rho GTPase activity and inhibition of endocytosis. Our study provides an alternative mechanism to regulate AQP2 trafficking, bypassing the VP-vasopressin receptor signaling pathway.     

Binding protein of somatomedin A in serum from men and rats

After gel chromatography of human and rat serum at pH 7.4, all endogenous somatomedin A was recovered in the high molecular weight range. The largest peak was found in the gamma-globulin (II) region and the next largest peak found in the albumin region (III). The amounts of somatomedin A in the peak II region increased in serum from acromegalies and decreased in serum from growth hormone deficient patients. Four radioactive peaks were observed after gel chromatography of serum incubated with 125I-somatomedin A. Only the two peaks corresponding to peaks II and III out of the four peaks were displaced by adding 50 microgram of partially purified cold somatomedin A. The radioactivity of peak II decreased in sera from growth hormone deficient patients and increased after growth hormone administration. These observations support the hypothesis that the growth hormone regulates not only somatomedin A but also its carrier protein.     

Effect of vasopressin on open field and activity behavior of the vasopressin-deficient (Brattleboro) rat

The effect of 1 and 5 micrograms AVP injections on open field and photoactivity chamber behavior of D.I. and normal Long-Evans animals was studied. Administration of 5 micrograms AVP (SC) resulted in a statistically significant depression of both open field and photochamber activity in the D.I. rat, but had a less pronounced effect on normal animals. However, 1 microgram AVP resulted in only minor alterations of activity in both D.I. and normal animals. In terms of learned behavior, D.I. and normal animals displayed similar within-session habituation when comparisons were made following the same treatment conditions. Thus, this study supports the hypothesis that vasopressin may influence memory tasks by modulation of related states of emotionality, motivation, and/or attention rather than by direct involvement in the retrieval and/or consolidation of information.     

Effects of gonadotrophin-induced elevation of serum testosterone upon somatomedin C levels and serum thymidine activity in children

3H-thymidine uptake into lectin-activated human lymphocytes allows to measure a growth-stimulating activity of serum, the thymidine activity (TA), which is GH dependent in vivo and related to somatomedins (Sm). In this work, it is shown: that addition of human chorionic gonadotrophin (HCG) or testosterone in vitro does not increase the 3H-thymidine uptake into lymphocytes; that the gonadotrophin-induced elevation of testosterone in children is accompanied by a significant increase of TA and, at a lesser degree, of Sm C; that these two increases are significantly correlated, and that the age-related variation of TA and Sm C after HCG stimulation test are not parallel.     

Effect of ovine prolactin on serum somatomedin bioactivity in hypophysectomized female rats.

Ovine prolactin (oPRL) increased serum somatomedin (SM) bioactivity in hypophysectomized female rats. ACTH, in small but not large doses, augmented this oPRL effect. These results suggest that in the female rat PRL may regulate SM production. Adrenal factors may variably modulate SM production or serum SM bioactivity.     

Effects of testosterone and estradiol on serum somatomedin A and growth rate of rats

The mode of involvement of sex steroids in the growth spurt during adolescence was studied in Wistar rats with a special reference to the level of serum somatomedin A (SMA) determined by radioreceptor assay. Intramuscular administration of testosterone propionate (T; 1 mg/day, alternately for 10 days) to female or gonadectomized male rats provoked a small but significant increase in their body weight or body length without affecting the serum SMA level. In contrast, in hypophysectomized (hypox) male rats T caused a considerable increase in body weight and the serum SMA level only when administered concurrently with bovine growth hormone (bGH; 0.2 U/day, ip). T did not affect the sulfation activity in vitro. These results suggest that androgen participates in the growth spurt during adolescence by enhancing the SMA effect and/or potentiating the SMA production by GH. Estradiol benzoate (E2; 100 micrograms/day, alternately for 10 days) caused a decrease in the serum SMA level and the growth rate in normal male rats. However, E2 produced an increase in the SMA level when administered to hypox male rats, although the growth was retarded and sulfation potency of the serum was sharply reduced. These results indicate that E2 may suppress the growth by lowering SMA generation in normal rats and cause a production of biologically inactive SMA in hypox male rats.     

Gastric mucosal endogenous prostacyclin levels are different in Brattleboro rats compared with Wistar strain

Homozygous Brattleboro rats were investigated and compared to normal (physiological) Wistar strain rats regarding their gastric mucosal endogenous prostacyclin (PG-I(2)) level. It seems that the Brattleboro animals have a significantly lower level of this important protective material. Wistar rats having an artificial pituitary stalk lesion (which is the artificial equivalent of homozygous Brattleboro animals) showed no differences in endogenous mucosal prostacyclin level compared to normal Wistar rats. Therefore, we concluded that this hitherto unknown property of the homozygous Brattleboro rats is genetically determined.     

Production of corticosterone in male homozygous Brattleboro rats

Plasma concentration, metabolic clearance rate and in vitro adrenal production of corticosterone were measured in Brattleboro male rats homozygous for diabetes insipidus (DI) and in Long-Evans male rats (LE) as controls in resting conditions, under stress caused by pentobarbitone anesthesia and surgery and after three days water deprivation. In resting animals, plasma concentrations and in vitro adrenal production of corticosterone were higher in DI rats than in LE rats. Under pentobarbitone anesthesia and surgery, plasma concentrations and metabolic clearance rate of corticosterone were slightly but not statistically lower in DI rats; however, the in vivo production rate of corticosterone was significantly lower. After three days water deprivation, increasing plasma corticosterone level was consistently higher in DI than in Le rats. These results are not in favour of a reduced glucocorticoid activity of the adrenal of DI rats and of an important role played by vasopressin on the stimulation of the hypothalamopituitary adrenal activity at least in resting conditions; its role may depend upon stressful circumstances.     

Immunocytochemical identification of dynorphin-containing vesicles in Brattleboro rats

Vasopressin and its carrier protein, vasopressin-associated neurophysin, are co-packaged together with an opioid peptide, dynorphin, into 160 nm diameter neurosecretory vesicles in the normal rat hypothalamo-neurohypophysial system. The homozygous Brattleboro rat lacks vasopressin and vasopressin-associated neurophysin, but contains substantial amounts of dynorphin in the vasopressin-deficient neurosecretory cells. We used post-embedding electron microscopic immunocytochemistry to determine the subcellular location of dynorphin in Brattleboro rats. The results show that dynorphin is present within 100 nm neurosecretory vesicles in homozygous Brattleboro cell bodies and axons, and within 160 nm vesicles in heterozygous (control) neurosecretory cell bodies and axons. Oxytocin-associated neurophysin is present in a separate population of magnocellular neurons in both homozygous and heterozygous rats, and is contained within 160 nm vesicles in both cases. Therefore, the absence of synthesis of the vasopressin prohormone results in a dramatic reduction of neurosecretory vesicle size, despite the continued synthesis and packaging of dynorphin peptides.     

Transient enhancement of GLUT-4 levels in rat epitrochlearis muscle after exercise training.

The purpose of the present study was to examine the effect of detraining on the glucose transport system after short-term swim training (5 days), long-term swim training (5 wk), and treadmill run training (5 wk). Skeletal muscles were isolated from female Wistar rats at 24 or 48 h posttraining. SST produces a 48% increase in GLUT-4 mRNA, a 30% increase in GLUT-4 protein, and a 60% increase in insulin-stimulated glucose transport activity at 24 h posttraining but not at 48 h posttraining. Similar to SST, long-term swim training produces a 60% increase in GLUT-4 mRNA and a 30% increase in GLUT-4 protein content at 24 h posttraining but not at 48 h posttraining. Finally, treadmill run training produces a transient 35% increase in GLUT-4 protein content that is completely reversed at 48 h after the last bout of exercise. These results demonstrate that the increase in GLUT-4 mRNA and GLUT-4 protein occurs during the first week of exercise training and is rapidly lost after training cessation. We believe that the transient enhancement in GLUT-4 protein after exercise training is due to a short GLUT-4 half-life, a process that is primarily regulated by pretranslational mechanisms.     

Arginine vasopressin deficient Brattleboro rats fail to develop tolerance to the hypothermic effects of ethanol

We have tested the hypothesis that animals with reduced levels of arginine vasopressin (AVP) would show reduced tolerance to ethanol. Brattleboro rats either heterozygous or homozygous for the diabetes insipidus (DI) trait and normal Sprague-Dawley rats were exposed to ethanol vapor for 21 days. Two days later, tolerance was evaluated by monitoring body temperature reductions after intraperitoneal injection of 2 g/kg (20% w/v) ethanol. Under the same conditions of chronic ethanol exposure, Sprague-Dawley rats, but not Brattleboro rats, displayed tolerance to the hypothermic effects of intraperitoneal ethanol. This phenomenon did not appear to be related to differences in ethanol metabolism or blood alcohol levels in Brattleboro rats. These data support a possible role for AVP in the development or maintenance of tolerance.     

Phospholipid and glycolipid synthesis in brain structures of Brattleboro rats

The incorporation of labeled precursors in phospholipids and glycolipids was studied in discrete brain areas of rats with innate vasopressin deficiency (Brattleboro, DI) and intact Long Evans animals (LE). Tracer incorporation was found to be reduced in septal, hypothalamic and hippocampal phospholipids, but enhanced in the glycolipid fraction isolated from the hypothalamus and hippocampus of Brattleboro rats. The results indicate that inherited vasopressin deficit seems to be associated with altered lipid synthesis in some brain areas of the Brattleboro rat, suggesting a probability for impaired translation of chemical signals.     

Signs of attenuated depression-like behavior in vasopressin deficient Brattleboro rats

Vasopressin, a peptide hormone functioning also as a neurotransmitter, neuromodulator and regulator of the stress response is considered to be one of the factors related to the development and course of depression. In the present study, we have tested the hypothesis that congenital deficit of vasopressin in Brattleboro rats leads to attenuated depression-like behavior in tests modeling different symptoms of depression. In addition, hypothalamic-pituitary-adrenocortical axis activity was investigated. Vasopressin deficient rats showed signs of attenuated depression-like behavior in forced swimming and sucrose preference tests, while their behavior on elevated plus maze was unchanged. Vasopressin deficiency had no influence on basal levels of ACTH and corticosterone and had only mild impact on hormonal activation in response to forced swimming and plus-maze exposure. However, vasopressin deficient animals showed higher level of dexamethasone induced suppression of corticosterone response to restraint stress and higher basal levels of corticotropin-releasing hormone mRNA in the hypothalamic paraventricular nucleus. In conclusion, present data obtained in vasopressin deficient rats show that vasopressin is involved in the development of depression-like behavior, in particular of the coping style and anhedonia. Moreover, behavioral and endocrine responses were found to be dissociated. We suggest that brain vasopressinergic circuits distinct from those regulating the HPA axis are involved in generating depression-like behavior.