NGF decreases isolation-induced aggressive behavior, while increasing adrenal volume, in adult male mice.

Intravenous administration of highly purified murine nerve growth factor (NGF, either 15 or 30 micrograms/subject/day) for six consecutive days to adult male mice of the CD-1 strain markedly influenced a number of items of aggressive behavior induced by 5 weeks of individual housing. Control mice received the same doses of cytochrome c. During a 20-min fighting session on day 7, both NGF-treated groups showed longer Latency to the first Attack, while Total Attacking Time and Aggressive grooming were significantly lower when compared to control animals. Twenty-four h later, adrenals from the NGF- and cytochrome c-treated groups were dissected and analyzed by means of a Vidas Image-Analyzer. NGF administration produced a remarkable dose-dependent increase in both adrenal weight and volume (particularly in the medullary zone), supporting the hypothesis that adrenals represent a possible target for the NGF release from submaxillary salivary glands occurring upon intraspecific fighting.     

Serum FSH and LH in male mice following aggressive and nonaggressive interaction

Previously isolated, adult, male mice were maintained for four days either in continued isolation or as pairs which were either separated from each other or not by a wire mesh barrier. Males without the barrier in place were categorized as dominant or subordinate on the basis of wounding. In general, fighting resulted in decreased concentrations of circulating FSH and LH, as well as the expected increase in adrenal weight and decrease in weight of the seminal vesicles. Such changes were decidedly more exaggerated in subordinates than among dominant males. Body weight was not affected by any of the experimental treatments. Pituitary content of the gonadotropins also remained relatively unchanged except for a significant depression in LH among dominant males.     

Endocrine and lymphoproliferative response changes produced by social stress in mice

Daily dyadic resident-intruder encounters and uninterrupted cohabitation in pairs were used to assess the impact of different durations (5 and 15 days) of dominance and subordination experiences on splenic lymphoproliferative responses in male OF1 strain mice. HPA axis activity was assessed by measuring serum corticosterone levels, whereas splenic norepinephrine (NE) content provided a sympathetic activity index. Corticosterone levels in subordinate subjects were generally higher than in their control or dominant counterparts in both treatment paradigms. Corticosterone levels in dominant subjects were lower than in their control counterparts in both. Increasing the duration of treatments generally decreased such titers, especially so in subordinate subjects. No differences were detected in splenic NE content. Animals subjected to social interaction generally showed greater proliferation than their control counterparts. This effect was more pronounced in subordinates than dominants and after longer- rather than short-duration treatments. There was no inverse relation between proliferative responses and the subject's corticosterone levels. While corticosterone may have a general immunomodulating effect, other mediators apparently account for the effects produced by these social stress paradigms on splenic proliferative response.     

Lead exposure and agonistic behavior of adult mice of two ages

Adult male Heterogeneous stock (HET) mice were exposed to a 0.5% lead acetate solution when they were either 65 or 330 days of age. Fifteen weeks later they were paired with same age (young or old) water control HET mice and tested for aggression. All pairs of younger mice fought and six out of eleven pairs of older mice exhibited agonistic behavior. Although not all pairs of mice which fought achieved dominance, when dominant/subordinate relationships were established, the younger adults exposed to lead typically were subordinate. In contrast, older adults exposed to lead were always dominant. Differences in agonistic behavior patterns also were noted, with younger adults displaying more frequent and longer bouts of fighting than the older mice.     

Splenomegaly and other hematological parameters in the socially dominant mouse

Within a social dominance hierarchy, subordinate mice show hematological changes such as increased erythropoiesis and splenomegaly. The present experiment demonstrates similar findings for the unwounded dominant mouse. In addition, total serum protein, serum albumin and plasma fibrinogen were measured. Male DBA/2j mice were placed into social triads for three 24 hr periods. The resultant dominant and subordinate mice were compared with isolated control mice. Splenomegaly, thymus involution, decreased hematocrit, and increased fibrinogen levels were found in dominant mice. Subordinate mice demonstrated the same changes to a greater extent, as well as an increased reticulocyte count. Only dominant mice showed a reduction in total serum protein. Wounding-independent processes must be involved in these cellular and non-cellular hematological effects of psychosocial stress.     

Adrenal activity in isolated mice and mice of different social status

The individual housing of male mice increases the incidence of aggressive behaviour. This study examines two suggestions, that isolation is stressful, and that it produces mice similar to dominant animals. Adrenal catecholamine content, plasma corticosterone, the weights of seminal vesicles, preputial and adrenal glands were measured in dominant, subordinate, isolated and group-housed mice. There was no evidence to suggest that isolation is stressful. Grouped mice had heavier adrenals than individually housed mice, and there was no difference in plasma corticosterone titre. In terms of adrenal weight and indices of gonadal function, the isolated mice were similar to dominant males. However, the literature indicates that isolated and dominant mice differ in their reactivity to stress. It was concluded that isolated male mice have physiological characteristics of their own.     

Comparison of arginine vasotocin immunoreactivity differences in dominant and subordinate green anole lizards

The neuropeptide arginine vasotocin (AVT) and its mammalian homologue arginine vasopressin (AVP) are believed to be involved in many social behaviors including territorial aggression. Testosterone (T) is also important for controlling territorial aggression, and it is believed to be involved in modulating AVT/AVP levels in the brain. In this study, male Anolis carolinensis were paired (n = 11 pairs) in a neutral cage and were allowed to establish a dominant-subordinate relationship for 10 days (experimental groups) or housed in a neutral cage with or without a female (control groups; each n = 4). On 10th day animals were sacrificed and their brain sections were processed for AVT immunohistochemistry and their serum was analyzed for testosterone levels. AVT immunoreactive (AVT-ir) cell numbers were counted in the anterior hypothalamus (AH), paraventricular nucleus (PN), posterior hypothalamus (PH), preoptic area (POA), and supra optic nuclei (SON). 2-way randomized block design was conducted to assess AVT-ir cell number differences between dominant and subordinate animals and Pearson's correlations were used to determine if a relationship existed between T levels and AVT-ir cell numbers. Dominant animals had more AVT-ir cells in the POA compared to subordinate animals, and subordinate animals had fewer AVT-ir cells in the POA compared to males housed either singly or with a female. There were no differences in AVT-ir cell numbers between dominant and subordinate animals in other areas. T levels were not correlated with the AVT-ir cell numbers in any area. Thus dominant animals have increased AVT-ir cell numbers compared to subordinate animals in a brain region known to be important in male sexual behavior. However, this difference is not related to differences in T.     

Brain leptin and nerve growth factor are differently affected by stress in male and female mice: Possible neuroendocrine and cardio-metabolic implications

The aim of the present study was to investigate the variations in leptin and nerve growth factor (NGF) expression induced by immobilization stress in the brain of male and female adult CD1 mice. We found that 10 days of repeated immobilization stress induced an increase of hypothalamus and thalamus NGF that was more pronounced in female than in male mice. We also found that this type of stress induced an increase of leptin expression in the hypothalamus of female mice, and a decrease in the thalamus of both male and female mice, associated with enhanced expression of leptin receptors in the hypothalamus and thalamus, both in male and female mice. The observation that the brain leptin and NGF receptors were altered by stress suggests a functional role for these molecules in neuroendocrine and cardiovascular response to stress events.     

Modulation of aggression in male mice: influence of group size and cage size

Aggression in group-housed male mice is known to be influenced by both cage size and group size. However, the interdependency of these two parameters has not been studied yet. In this study, the level of aggression in groups of three, five, or eight male BALB/c mice housed in cages with a floor size of either 80 or 125 cm(2)/animal was estimated weekly after cage cleaning for a period of 14 weeks. Furthermore, urine corticosterone levels, food and water intake, body weight, and number of wounds were measured weekly. At the end of the experiment, tyrosine hydroxylase (TH) activity, testosterone levels, and weight of spleen, thymus, testes, and seminal vesicles were determined. Results indicate a moderate increase of intermale aggression in larger cages when compared to the smaller cages. Aggression in groups of eight animals was considerably higher than in groups of three animals. The increase of agonistic behavior was observed both in dominant and subordinate animals. Physiological parameters indicate differences in stress levels between dominant and subordinate animals. It is concluded that aggressive behavior in group-housed male BALB/c mice is best prevented by housing the animals in small groups of three to five animals, while decreasing floor size per animal may be used as a temporary solution to decrease high levels of aggression in an existing social group.     

Effects of nerve growth factor and glucocorticoid on cultured human pheochromocytoma cells

Primary cell cultures of two human pheochromocytomas (PC) that were associated with high serum levels of adrenaline and noradrenaline were developed to study the effects of nerve growth factor (NGF) and dexamethasone on the morphology and function of PC cells in vitro. By phase-contrast microscopy, cultured cells were small and hyperchromatic on the first day of culture; neurite-like processes that extended to other cells developed several days later and were maintained for more than 3 months. NGF (100ng/ml), dexamethasone (10^–5M), or NGF + dexamethasone were added to the culture media 2 weeks after the cultured cells had stabilized. Catecholamine concentrations in the medium were maintained at higher levels after addition of NGF, dexamethasone, or NGF + dexamethasone as compared to control cells. In the presence of NGF, extension of neurite-like processes was clearly accelerated, while high levels of dexamethasone inhibited growth of processes. These in vitro studies showed that the addition of NGF or the removal of dexamethasone induces differentiation of adrenal neurons present in pheochromocytomas, suggesting that adrenocortical steroid hormones influence the morphological control of adrenal medullary cells.     

Altered T-lymphocyte response following aggressive encounters in mice

Intermale aggression is a natural form of psychosocial stress that can alter a variety of physiological functions, including immune function. In Experiment 1, daily fighting between pairs of previously isolated male mice differentially altered immunological measures of T-cell responsiveness in dominant and submissive animals. Submissive mice had lower T-cell proliferation and IL-2 production, when compared to dominant, nonfought, or witness mice. Since the fighting behavior often results in wounding of the submissive animal, Experiment 2 used a relatively nonaggressive test to determine whether the immunological differences between dominant and submissive mice were due to wounding or due to the psychosocial state of dominance. Dominant mice had elevated T-cell proliferation and IL-2 production when compared to the other treatment groups. Therefore, it appears that in dominant/submissive pairs of mice a severe physical stress, such as intense fighting, influences the immune system in a different manner than psychological or mild aggressive encounters.     

Variations in behavior, innate immunity and host resistance to B16F10 melanoma growth in mice that present social stable hierarchical ranks

The present study analyzed the effect of social stable hierarchical dominance/submissive relationships in C57BL/6 mice on behavior, innate immunity, serum corticosterone levels and host resistance to B16F10 melanoma growth. Adult mice (90 days old) kept in pairs since weaning, were analyzed for dominant/submissive ranking in three consecutive days according to the presence or absence of fighting and/or anticipatory submissive responses. Only the pairs of mice where dominant/submissive relationships were clearly stated were employed. Results showed that submissive mice presented in relation to dominants: (1) decreased time spent in the central open-field area; (2) decreased number of entries into the open arms and decreased time spent in the exploration of the open arms of the plus maze; (3) increased time spent in exploration of the plus-maze closed arms; (4) decreased number of entries and in the time spent in the exploration of the third part of the plus-maze open arms; (5) increased number of B16F10 metastasis in the lungs; (6) decreased NK cell cytotoxicity measured in vitro in the peripheral blood and spleen; (7) decreased basal but not in S. aureus induced oxidative burst in both neutrophils and monocytes and (8) similar basal serum levels of corticosterone. The present behavioral findings show that submissive mice, within a stable social hierarchy, present anxiety like-responses a fact that would make than more prone to stressful stimuli. This condition would be responsible for the decreases presently observed on basal neutrophil oxidative burst, NK cell activity and resistance to B16F10 tumor growth. Together the obtained data show that mice that present stable hierarchical relationships display neuro-immune alterations comparable to those reported in mice under a situation of chronic social stress.     

Elevated concentrations of beta-nerve growth factor in selected tissues from senescence-accelerated mice (SAM-P/8)

Levels of the beta-subunit of nerve growth factor (beta-NGF) were determined in various tissues from senescence-accelerated mice (SAM-P/8) and compared with those from senescence-resistant control mice (SAM-R/1) at 4 months of age. (1) In SAM-P/8, the testis was 30% larger in terms of wet weight than that from SAM-R/1, whereas the adrenal glands from males and females were smaller than those from the respective controls by 45% and 20%, respectively. (2) About 70% of SAM-P/8 individuals had high concentrations of testosterone in serum (greater than 5ng/ml). (3) In SAM-P/8, endogenous levels of beta-NGF were significantly higher in the adrenal gland (20 and 7 times higher on average in males and females, respectively), in the thymus (100 and 5 times higher in males and females, respectively) and in the testis (500 times higher) than those in the control tissues. In other tissues there were little or no differences in terms of levels of beta-NGF. (4) Morphological changes in the adrenal gland, thymus and testis of SAM-P/8 mice were not as marked as expected from the elevated levels of beta-NGF in these tissues. (5) These results show that, in SAM-P/8 mice at 4 months of age, an elevation in the endogenous level of beta-NGF has already occurred in some peripheral tissues before senescence becomes accelerated.     

Age-related changes in brain RNA of noise stressed mice

Correlative data were obtained on RNA levels of cerebrocortical neurons, adrenal fasciculate cell RNA content and serum corticosterone levels in 10, 40 and 100 week old male C57BL/6J mice immediately following a 30 interval of continuous noise stimulation (100 dB re 2 x 10(-5) N/m2, 0.35-20 kHz) and at 1 hr and 4 hr recovery intervals. Quantification of cellular RNA levels entailed use of scanning-integrating microdensitometry of azure B stained tissue sections. Hormonal analyses were done by radioimmunoassay. Noise stimulation induced a mild to moderate adrenocortical activation and a moderate elevation of serum corticosterone levels in all age groups. Noise exposure also resulted in an increase in the RNA content of cerebrocortical neurons. However, maximal neuronal RNA levels proved to be considerably lower (32-36%) in 100 week old mice as compared to both 10 and 40 week old mice during the post-exposure intervals. The overall data support the existence of a lower neuronal capacity for sensory induced RNA synthesis in 100 week old mice. The data also suggest that age dependent differences in neuronal responsiveness are not directly related to the extent of adrenal RNA activation or to a lower availability of circulating glucocorticoids.     

Contribution of galanin to stress-induced impairment of insulin secretion in swimming mice.

This study examines the potential role of the neuropeptide, galanin, in stress-induced inhibition of insulin secretion in swimming mice. Firstly, the pancreatic and adrenal content of galanin-like immunoreactivity was determined in mice after swimming stress. It was found that pancreatic content was significantly lower in stressed mice than in resting controls, both after 2 (P less than 0.05) and 6 (P less than 0.025) minutes of swimming, suggesting partial release of pancreatic galanin during stress. In contrast, the adrenal content of galanin-like immunoreactivity did not change during the swimming stress. Gel filtration of tissue extracts indicated that (1) mouse pancreas contains two forms of galanin-like immunoreactivity; one co-eluting with synthetic porcine galanin (centered on Kav of 0.70) and another with a larger molecular weight (centered on Kav of 0.30), and (2) mouse adrenal contains a small void volume-peak and a larger peak of immunoreactivity, the latter co-eluting with synthetic galanin. Secondly, the effects of swimming stress on plasma glucose and insulin levels were compared in mice that received high titre rabbit anti-galanin serum with those in mice receiving normal rabbit serum. In normal rabbit serum-pretreated swimming mice, glucose-induced insulin levels were only 50% of resting controls (P less than 0.01). Immunoneutralization of galanin with specific antiserum abolished this swimming stress-induced inhibition of glucose-stimulated insulin levels. This was accompanied by a modestly enhanced rate of glucose disappearance. These findings suggest that pancreatic galanin is released during swimming stress in mice and that endogenous galanin makes a major contribution to stress-induced impairment of insulin secretion.     

Influence of aggression on erythropoiesis--the hypersympathetic syndrome

Male CF₁ mice were isolated for 3 weeks at weaning. Pairs were then selected for similarity in weight and placed together for two 30 min periods per day for 21 days. A dominant-subordinate relationship was established by fighting. The subordinate animals developed a marked splenomegaly, decreased hematocrit and increased reticulocyte count while dominant and control mice showed no change in these parameters. The experiment was repeated using splenectomized individuals. Again the subordinates showed decreased hematocrits and increased reticulocyte counts with no change observed in dominants and controls. An hypothesis is advanced which suggests that the behaviorally induced stimulus to erythropoiesis results from a series of events which we have termed a hypersympathetic syndrome.     

Role for cystic fibrosis transmembrane conductance regulator protein in a glutathione response to bronchopulmonary pseudomonas infection

The lung maintains an elevated level of glutathione (GSH) in epithelial lining fluid (ELF) compared to serum. The mechanism(s) by which the lung maintains high levels of ELF GSH and factors that modulate them are largely unexplored. We hypothesized that lung cystic fibrosis transmembrane conductance regulator protein (CFTR) modulates GSH efflux in response to extracellular stress, which occurs with lung infections. Mice were challenged intratracheally with Pseudomonas aeruginosa, and on the third day of infection bronchoalveolar lavage fluid was obtained and analyzed for cytokines and antioxidants. Lung tissue antioxidants and enzyme activities were also assessed. P. aeruginosa lung infection increased levels of inflammatory cytokines and neutrophils in the ELF. This corresponded with a marked threefold increase in GSH and a twofold increase in urate levels in the ELF of P. aeruginosa-infected wild-type mice. A twofold increase in urate levels was also observed among lung tissue antioxidants of P. aeruginosa-infected wild-type mice. There were no changes in markers of lung oxidative stress associated with the P. aeruginosa lung infection. In contrast with wild-type mice, the CFTR knockout mice lacked a significant increase in ELF GSH when challenged with P. aeruginosa, and this correlated with a decrease in the ratio of reduced to oxidized GSH in the ELF, a marker of oxidative stress. These data would suggest that the lung adapts to infectious agents with elevated ELF GSH and urate. Individuals with lung diseases associated with altered antioxidant transport, such as cystic fibrosis, might lack the ability to adapt to the infection and present with a more severe inflammatory response.     

Elevation of nerve growth factor synthesis by constitutive expression of v-src oncogene in cultured rat fibroblasts.

Rat fibroblast 3Y1 cells transformed by Rous sarcoma virus (RSV) or transfected with the v-src gene showed a highly constitutive v-src gene expression. Simultaneously, marked increases in the cellular level of nerve growth factor (NGF) mRNA and NGF content in the culture medium were observed. The levels of NGF mRNA and NGF secreted into the medium were correlated with the expression level of v-src mRNA gene in both transformants and control 3Y1 cells. These results suggest that v-src gene expression is relevant to regulation of NGF synthesis in rat 3Y1 fibroblasts.     

Revisiting the Koebner phenomenon: role of NGF and its receptor system in the pathogenesis of psoriasis

Nerve growth factor (NGF) influences the key pathological events of psoriasis: keratinocyte proliferation, angiogenesis, and T-cell activation. We have systematically examined the kinetics of NGF expression, keratinocyte proliferation, and migration of T lymphocytes in the epidermis in Koebner-induced developing psoriatic plaques. In skin traumatized by the tape-stripping method (n = 12), a marked up-regulation of NGF in Koebner-positive lesions (n = 7) was observed 24 hours after trauma. Synthesis of NGF reached its maximum level in the 2nd week. Furthermore, cultured keratinocytes from nonlesional skin of psoriasis patients produced 10 times higher levels of NGF compared with keratinocytes from healthy individuals. To substantiate the in vivo effect of NGF secreted by keratinocytes in psoriatic plaques, we studied psoriatic plaques and normal human skin in a SCID-human skin xenograft model. The transplanted psoriatic plaques demonstrated marked proliferation of NGF-R (p75)-positive nerve fibers compared with only a few nerves in the transplanted normal human skin. Our results demonstrate that 1) in a developing psoriatic lesion, up-regulation of NGF together with keratinocyte proliferation are early events and precede epidermotropism of T lymphocytes; 2) keratinocytes in patients with psoriasis are primed to produce elevated levels of NGF; and 3) NGF synthesized by these keratinocytes is functionally active.     

Scent marking and the maintenance of dominant/subordinate status in male golden hamsters

Since it is thought that flank marking communicates dominance status, experiments were designed to look at changes in aggression and flank marking behaviors in pairs of male hamsters with intact flank glands (Experiment One) or when one (Experiment Two) or both (Experiment Three) members of a pair had their flank glands surgically removed. In Experiment One the dominant members of twelve pairs of hamsters had a mean daily frequency of flank marks that was over two-fold greater than their subordinate partners, F(1,11) = 17.59, p less than 0.001. Over the course of five consecutive daily tests there was a significant decrease in the aggression index of both the dominant, t(44) = 4.49, p less than 0.01, and subordinate, t(44) = 3.33, p less than 0.01, hamsters. Accompanying the decrease in aggression was a significant increase in the flank marking of both dominant, t(44) = 7.8, p less than 0.01, and subordinate, t(44) = 3.59, p less than 0.01, hamsters. In Experiment Two, six out of eleven flank glandectomized hamsters were dominant over their sham operated partners while the remaining five were subordinate. Unlike Experiment One there was no significant difference in the flank marking between dominant and subordinate hamsters, in fact, in seven pairs the subordinate hamsters flank marked more than their dominant partners. In Experiment Three both hamsters had their flank glands removed, and as in Experiment Two, there was no significant difference in flank marking between dominant and subordinate hamsters, neither was there any significant change in their aggression and flank marking behaviors over the course of the five test periods.(ABSTRACT TRUNCATED AT 250 WORDS)