Ejaculatory abnormalities in mice with targeted disruption of the gene for heme oxygenase-2.

Nitric oxide (NO) is well established as a neurotransmitter in the central and peripheral nervous systems. More recently, another gas, carbon monoxide (CO) has also been implicated in neurotransmission. In the nervous system CO is formed by a subtype of heme oxygenase (HO) designated HO2. HO2 is localized to discrete neuronal populations in the brain resembling localizations of soluble guanylyl cyclase, which is activated by CO. CO may also function in the peripheral autonomic nervous system, in conjunction with NO. The majority of ganglia in the myenteric plexus possess both HO2 and neuronal NO synthase (NOS). Defects in myenteric plexus neurotransmission occur both in mice with targeted deletion of genes for HO2 and neuronal NOS. HO2 also occurs in other autonomic ganglia including the petrosal, superior cervical and nodose ganglia. Neuronal NOS is localized to neurons regulating male reproductive behavior, such as penile erection, and NOS inhibitors prevent erection. Because of the other parallels between NO and CO, we speculated that CO may play a role in male reproductive behavior. In the present study we describe HO2 localization in neuronal structures regulating copulatory reflexes. Reflex activity of the bulbospongiosus muscle, which mediates ejaculation and ejaculatory behavior, is markedly diminished in mice with targeted deletion of the gene for HO2 (HO2-).

Neurobehavioral deficits in mice lacking the erythrocyte membrane cytoskeletal protein 4.1.

The erythrocyte membrane cytoskeletal protein 4.1 (4.1R) is a structural protein that confers stability and flexibility to erythrocytes via interactions with the cytoskeletal proteins spectrin and F-actin and with the band 3 and glycophorin C membrane proteins. Mutations in 4.1R can cause hereditary elliptocytosis, a disease characterized by a loss of the normal discoid morphology of erythrocytes, resulting in hemolytic anemia [1]. Different isoforms of the 4.1 protein have been identified in a wide variety of nonerythroid tissues by immunological methods [2-5]. The variation in molecular weight of these different 4.1 isoforms, which range from 30 to 210 kDa [6], has been attributed to complex alternative splicing of the 4.1R gene [7]. We recently identified two new 4.1 genes: one is generally expressed throughout the body (4. 1G) [8] and the other is expressed in central and peripheral neurons (4.1N) [9]. Here, we examined 4.1R expression by in situ hybridization analysis and found that 4.1R was selectively expressed in hematopoietic tissues and in specific neuronal populations. In the brain, high levels of 4.1R were discretely localized to granule cells in the cerebellum and dentate gyrus. We generated mice that lacked 4.1R expression; these mice had deficits in movement, coordination, balance and learning, in addition to the predicted hematological abnormalities. The neurobehavioral findings are consistent with the distribution of 4.1R in the brain, suggesting that 4.1R performs specific functions in the central nervous system.

Elimination of aggressive behavior in male mice lacking endothelial nitric oxide synthase.

Male mice with targeted deletion of the gene encoding the neuronal isoform of nitric oxide synthase (nNOS(-/-)) display increased aggressive behavior compared with wild-type (WT) mice. Specific pharmacological inhibition of nNOS with 7-nitroindazole also augments aggressive behavior. We report here that male mice with targeted deletion of the gene encoding endothelial NOS (eNOS(-/-)) display dramatic reductions in aggression. The effects are selective, because an extensive battery of behavioral tests reveals no other deficits. In the resident-intruder model of aggression, resident eNOS(-/-) males show virtually no aggression. Latency for aggression onset is 25-30 times longer in eNOS(-/-) males compared with WT males in the rare instances of aggressive behaviors. Similarly, a striking lack of aggression is noted in tests of aggression among groups of four mice monitored in neutral cages. Although eNOS(-/-) mice are hypertensive ( approximately 14 mmHg blood pressure elevation), hypertension does not appear responsible for the diminished aggression. Reduction of hypertension with hydralazine does not change the prevalence of aggression in eNOS(-/-) mice. Extensive examination of brains from eNOS(-/-) male mice reveals no obvious neural damage from chronic hypertension. In situ hybridization in WT animals reveals eNOS mRNA in the brain associated exclusively with blood vessels and no neuronal localizations. Accordingly, vascular eNOS in the brain appears capable of influencing behavior with considerable selectivity.

Photoperiod and temperature interact to affect immune parameters in adult male deer mice (Peromyscus maniculatus).

Nontropical rodents often experience large seasonal fluctuations in both food availability and energy demands. The energy required for thermoregulation is highest during the winter when food availability may be at an annual minimum. Failure to cope with winter probably accounts, in part, for the increased prevalence of disease and death relative to that in summer. Winter conditions may elevate circulating glucocorticosteroid levels, which can compromise immune function. To increase the odds of surviving the energetic demands of winter, individuals of some rodent species appear to enhance immune function before conditions deteriorate. Previous laboratory studies suggest that immune enhancement can be induced by short days. These findings contrast with the results of several field studies reporting suppressed immune function during the winter. To resolve this conflict, the authors hypothesized that winter stressors present in field studies counteracted the short-day enhancement of immune function reported in laboratory studies. If true, then immune function of captive mice in short days should be compromised by low temperature or reduced food availability. Both ambient temperature and photoperiod were manipulated in the present study to assess their effects on immune parameters in male deer mice (Peromyscus maniculatus). Animals in short days regressed their reproductive systems and also displayed significantly higher immunoglobulin G (IgG) levels than did those in long days. Deer mice maintained in low temperatures had significantly reduced splenic masses and basal IgG levels independent of day length. Animals maintained in both short days and low temperatures displayed IgG levels comparable to those of mice in long-day/mild-temperature conditions. Animals maintained in long days and low temperatures had significantly higher serum corticosterone levels than did animals maintained in long days at mild temperatures. These data are consistent with the hypothesis that immune parameters are enhanced in short days to counteract stress-mediated immune suppression occurring during the winter.

Exogenous melatonin enhances cell-mediated, but not humoral, immune function in adult male deer mice (Peromyscus maniculatus).

Many nontropical rodent species display seasonal changes in reproductive physiology and metabolism, as well as in immune function. Field studies of seasonal changes in immune function typically report decreased immune function in the short days of winter compared to summer; presumably, reduced immunity in winter reflects increased glucocorticoid secretion in response to environmental stressors. In contrast, laboratory studies of photoperiodic changes in immunity invariably demonstrate increased immune function in short compared to long days. Although the precise mechanisms regulating short-day enhancement of immune function are not known, it is hypothesized that increased immunity is due to the increased duration of melatonin secretion in short compared to long days. However, melatonin can act both directly (i.e, via melatonin receptors located on lymphatic tissue) and indirectly (i.e., via alterations in gonadal steroids) to affect immune function. The present study examined the effects of exogenous melatonin administration on both cell-mediated and humoral immune function in adult male deer mice (Peromyscus maniculatus), as well as the role of gonadal steroid hormones in mediating these effects. Mice either were castrated to remove circulating androgens or received sham operations and were implanted with empty capsules or capsules containing melatonin. Individual mice implanted with melatonin underwent reproductive regression and displayed enhanced splenocyte proliferation to the T-cell mitogen concanavalin A; antigen-specific serum immunoglobulin M production was unaffected by melatonin treatment. Castration had no effect on either cell-mediated or humoral immune function. Taken together, these results suggest that exogenous melatonin enhances cell-mediated, but not humoral, immune function in adult male deer mice and that this effect is independent of gonadal steroid hormones. These results are consistent with a direct effect of melatonin on immunity.

Photoperiod, ambient temperature, and food availability interact to affect reproductive and immune function in adult male deer mice (Peromyscus maniculatus).

Winter is often stressful. Increased energetic demands in winter and concurrent reductions in energy availability can lead to an energetic imbalance and compromise survival. To increase the odds of surviving winter, individuals of some nontropical rodent species have evolved mechanisms to enhance immune function in advance of harsh winter conditions. Short day lengths provide a proximate cue for enhancement of immune function, an adaptive functional response to counter environmental stress-induced reduction in immune function. In the present study, photoperiod, ambient temperature, and food availability were manipulated and reproductive function and cell-mediated immunity were assessed in adult male deer mice (Peromyscus maniculatus). Mice maintained in short days regressed their reproductive systems and displayed enhanced immune function compared to long-day animals. Reduced food availability elevated corticosterone concentrations and suppressed reproductive and immune function, whereas ambient temperature alone had no effect on cell-mediated immunity. The suppressive effect of food restriction on reproductive and immune function was overcome by maintaining animals in short days. However, short-day, food-restricted mice maintained at low ambient temperatures displayed reduced reproductive and immune function compared to animals maintained at mild temperatures. Taken together, these results suggest that short-day enhancement of immune function can counteract some, but not all, of the immunosuppressive effects of winter stressors. These data are consistent with the hypothesis that immune function is enhanced in short days to counteract stress-mediated immune suppression occurring during winter.

SCN efferents to peripheral tissues: implications for biological rhythms.

The suprachiasmatic nucleus (SCN) is the principal generator of circadian rhythms and is part of an entrainment system that synchronizes the animal with its environment. Here, we review the possible communication of timing information from the SCN to peripheral tissues involved in regulating fundamental physiological functions as revealed using a viral, transneuronal tract tracer, the pseudorabies virus (PRV). The sympathetic nervous system innervation of the pineal gland and the sympathetic outflow from brain to white adipose tissue were the first demonstrations of SCN-peripheral tissue connections. The inclusion of the SCN as part of these and other circuits was the result of lengthened postviral injection times compared with those used previously. Subsequently, the SCN has been found to be part of the sympathetic outflow from the brain to brown adipose tissue, thyroid gland, kidney, bladder, spleen, adrenal medulla, and perhaps the adrenal cortex. The SCN also is involved in the parasympathetic nervous system innervation of the thyroid, liver, pancreas, and submandibular gland. Individual SCN neurons appear connected to more than one autonomic circuit involving both sympathetic and parasympathetic innervation of a single tissue, or sympathetic innervation of two different peripheral tissues. Collectively, the results of these PRV studies require an expansion of the traditional roles of the SCN to include the autonomic innervation of peripheral tissues and perhaps the modulation of neuroendocrine systems traditionally thought to be controlled solely by hypothalamic stimulating/inhibiting factors.

Circadian locomotor analysis of male mice lacking the gene for neuronal nitric oxide synthase (nNOS-/-)

Nitric oxide (NO) is an endogenous gas that functions as a neurotransmitter. Because NO is very labile with a half-life of less than 5 sec, most functional studies of NO have manipulated its synthetic enzyme, NO synthase (NOS). Three isoforms of NOS have been identified: (1) in the endothelial lining of blood vessels (eNOS), (2) an inducible form found in macrophages (iNOS), and (3) in neurons (nNOS). Most pharmacological studies to date have blocked all three isoforms of NOS. Previous studies using such agents have revealed that NO might be necessary for photic entrainment of circadian rhythms; general NOS inhibitors attenuate phase shifts of free-running behavior, light-induced c-fos expression in the suprachiasmatic nucleus (SCN), and phase shifts of neural firing activity in SCN maintained in vitro. To assess the specific role of nNOS in mediating entrainment of circadian rhythms, mice with targeted deletion of the gene encoding the neuronal isoform of NOS (nNOS-/-) were used. Wild-type (WT) and nNOS-/- mice initially were entrained to a 14:10 light:dark (LD) cycle. After 3 weeks, the LD cycle was either phase advanced or phase delayed. After an additional 3 weeks, animals were held in either constant dim light or constant dark. WT and nNOS-/- animals did not differ in their ability to entrain to the LD cycle, phase shift locomotor activity, or free run in constant conditions. Animals held in constant dark were killed after light exposure during either the subjective day or subjective night to assess c-fos induction in the SCN. Light exposure during the subjective night increased c-fos expression in the SCN of both WT and nNOS-/- mice relative to animals killed after light exposure during the subjective day. Taken together, these findings suggest that NO from neurons might not be necessary for photic entrainment.

Leptin effects on immune function and energy balance are photoperiod dependent in Siberian hamsters (Phodopus sungorus).

Many adaptations have evolved in small mammals to maximize survival during winter. One such coping tactic in many species is an alteration of immune function in advance of the stressful conditions of winter. Leptin is a hormone produced by adipose tissue, and in addition to its central role in energy metabolism, leptin mediates the interactions among energy allocation, immune function, and reproduction. To examine this interaction further, exogenous leptin was administered for 2 weeks via osmotic minipumps to Siberian hamsters (Phodopus sungorus) housed in long or short days for a total of 12 weeks. Short-day hamsters displayed the expected reductions in humoral immune function, body mass, fat mass, and food intake. In Exp 1, exogenous leptin counteracted the reduction in food intake and the suppression of immune function in short days. In Exp 2, when the leptin-induced increase in food intake in short-day hamsters was prevented, leptin did not enhance immune function. In most of the measured fat pads and body mass, leptin had no effect in long days. In sum, leptin administered to short-day animals caused them to respond, in many cases, like long-day animals. Taken together, these data suggest that leptin acts indirectly to mediate energy allocation to humoral immune function. Additionally, leptin appears to act differentially, according to photoperiod, to regulate both immune and energetic parameters.

Nocturnal motor coordination deficits in neuronal nitric oxide synthase knock-out mice.

Nitric oxide is formed in the brain primarily by neurons containing neuronal nitric oxide synthase (nNOS), though some neurons may express endothelial NOS (eNOS), and inducible NOS (iNOS) only occurs in neurons following toxic stimuli. Mice with targeted disruption of nNOS (nNOS-) display distended stomachs with hypertrophied pyloric sphincters reflecting loss of nNOS in myenteric plexus neurons. nNOS- animals resist brain damage following middle cerebral artery occlusions consistent with evidence that excess release of nitric oxide mediates neurotoxicity in ischemic stroke. Neuronal NOS- mice have no grossly evident defects in locomotor activity, breeding long-term depression in the cerebellum, long-term potentiation in the hippocampus, and overall sensorimotor function. However, nNOS- animals display excessive, inappropriate sexual behavior and dramatic increases in aggression. Because the cerebellum possesses the greatest levels of nNOS neurons in the brain, it was surprising that presumed cerebellar functions such as balance and coordination were grossly normal in nNOS- mice. These previous studies were all conducted during the day (between 1400 and 1600, lights on at 0700). We now report striking, discrete abnormalities in balance and motor coordination in nNOS-mice reflected selectively at night.

Melatonin, immunity and cost of reproductive state in male European starlings.

The effects of reproductive condition and exogenous melatonin on immune function were investigated in castrated European starlings, Sturnus vulgaris. Photorefractory and photostimulated starlings exposed to long days were implanted with melatonin or with blank capsules. Photostimulated starlings with blank capsules exhibited reduced splenocyte proliferation in response to the T-cell mitogen, concanavalin A, compared with the other long-day birds. Exogenous melatonin prevented the suppression of immune function by photostimulation. Photorefractory starlings, with or without melatonin implants, exhibited enhanced immune function compared with photostimulated starlings implanted with blanks. This enhancement was not mediated by endogenous melatonin, but appeared to be related to changes in reproductive state. In addition to the traditional costs of reproduction in birds (e.g. raising of young), there may be a cost of the reproductive state of starlings (i.e. whether they are photorefractory or photostimulated). These data are, we believe, the first to indicate a direct effect of reproductive state on immune function that is independent of both photoperiod (i.e., changes in the duration of melatonin secretion) and gonadal steroids.

Photoperiod affects neuronal nitric oxide synthase and aggressive behaviour in male Siberian hamsters (Phodopus sungorus).

Many nontropical animals display physiological and behavioural changes in response to seasonal environmental cues including photoperiod (day length). Male Siberian hamsters (Phodopus sungorus) housed in short photoperiod undergo testicular regression accompanied by reduced circulating testosterone and decreased reproductive behaviour. By contrast to the majority of small mammals studied, aggressive behaviour is elevated in short-day Siberian hamsters when blood testosterone concentrations are not detectable. Because gonadal steroid hormones influence neuronal nitric oxide synthase (nNOS), and this enzyme has been implicated in aggressive behaviour, we hypothesized that nNOS expression would be decreased in short-day male Siberian hamsters and negatively correlated with the display of territorial aggression. Adult male Siberian hamsters were individually housed in either long (LD 16:8 h) or short (LD 8:16 h) photoperiods for 10 weeks. Hamsters were assigned to one of two categories by assessing testicular volume and plasma testosterone values: (i) photoperiodic responsive (i.e. regressed testes and low testosterone concentrations) or (ii) photoperiodic nonresponsive (i.e. testes size and circulating testosterone concentrations equivalent to hamsters maintained in long days). At week 10, aggression was assessed using a resident-intruder test. Latency to initial attack, frequency of attacks and duration of total attacks were recorded during a 10-min aggression trial. Brains were collected immediately after behavioural testing and stained for nNOS expression using immunohistochemistry. All short day-housed hamsters were significantly more aggressive than long-day animals, regardless of gonadal size or testosterone concentrations. Short-day animals, both reproductively responsive and nonresponsive morphs, also had significantly less nNOS-immunoreactive cells in the anterior and basolateral amygdaloid areas and paraventricular nuclei compared to long-day hamsters. Together, these results suggest that seasonal aggression in male Siberian hamsters is regulated by photoperiod, through mechanisms that are likely independent from gonadal steroid hormones.

Sympathoadrenal system differentially affects photoperiodic changes in humoral immunity of Siberian hamsters (Phodopus sungorus).

Siberian hamsters (Phodopus sungorus) rely on photoperiod as a primary cue to coordinate seasonally appropriate changes in physiology and behaviour. Among these seasonal changes is reduced immune function in short 'winter-like' days, compared to long 'summer-like' days. Previous evidence suggests that immune function is regulated, in part, by the sympathoadrenal system. The precise role of the sympathoadrenal system in regulating photoperiodic changes in immune function, however, remains unspecified. The goal of the present study was to examine the differential contributions of direct sympathetic innervation of immune target tissue, as well as adrenal medullary catecholamines, to photoperiodic changes in immune function in male Siberian hamsters. In Experiment 1, hamsters underwent either bilateral surgical removal of the adrenal medulla (ADMEDx), or sham surgeries, and were maintained in long (LD 16 : 8) or short days (LD 8 : 16). In Experiment 2, hamsters received either surgical denervation of the spleen, or sham surgeries, and were then housed in long or short days. Serum anti-KLH IgG concentrations and splenic norepinephrine (NE) content were determined in both experiments. Short-day hamsters had reduced humoral immunity compared to long-day hamsters. ADMEDx reduced immune function, but only in long-day hamsters. In contrast, splenic denervation reduced humoral immunity, but only in short-day hamsters. Splenic NE content was increased in short days and by ADMEDx. NE content was markedly reduced in denervated hamsters compared to sham-operated hamsters. Collectively, these results suggest that the sympathoadrenal system is associated with photoperiodic changes in immune function.

Novel method for localized, functional sympathetic nervous system denervation of peripheral tissue using guanethidine.

A simple technique for local chemical sympathectomy of peripheral tissues is described using guanethidine. Multiple microinjections of guanethidine were made into inguinal or epididymal white adipose tissue (IWAT and EWAT) pads or spleens of hamsters. Guanethidine virtually abolished the sympathetic innervation of both EWAT and IWAT, as measured by the absence of significant norepinephrine (NE) tissue content two weeks later and as suggested by the two-fold increase in IWAT mass characteristic of surgically induced WAT denervation. These measures were not affected in the contralateral pads given equivolumetric injections of saline. Guanethidine injections into the spleen lead to a functional sympathectomy, as indicated by significant depletions of NE content. Because guanethidine treatment did not decrease body mass, induce ptosis, or spread to closely associated adjacent tissue (contralateral EWAT pad), no chemical-induced malaise or global sympathetic denervation was suggested. Guanethidine was more effective than two other local sympathectomy treatments, injections of the sympathetic neurotoxin anti-dopamine-beta-hydroxylase saporin or surgical denervation, in decreasing IWAT NE content and increasing IWAT pad mass. Collectively, these results suggest that locally applied, chemical sympathectomy with guanethidine provides an effective, restricted method for sympathectomizing WAT, spleen and likely other peripheral tissues.

Impaired spatial working and reference memory in segmental trisomy (Ts65Dn) mice.

To evaluate the cognitive phenotype of the segmental trisomy 16 (Ts65Dn) mouse, a model of Down Syndrome (DS, trisomy 21), we assessed spatial working and reference memory using a 12-arm radial maze (RAM). Ts65Dn mice made a greater number of reference memory errors across trials compared to control mice. Both genotypes showed improvement across trials, although improvement was slower in Ts65Dn mice. Ts65Dn mice also made a greater number of working memory errors on the RAM, and in contrast to control mice, did not improve across trials, always performing at near-chance levels. These results provide evidence for both spatial working and reference memory deficits in Ts65Dn mice, characteristics of cognitive dysfunction.

Spatial memory deficits in segmental trisomic Ts65Dn mice.

Spatial memory was assessed in the segmental trisomic 16 mouse (Ts65Dn), a potential model for Down syndrome (DS), using the 12-arm radial maze (RAM). Ts65Dn mice have a portion of mouse chromosome 16 syntenic to the distal end of human chromosome 21 triplicated. On each of 8 daily trials of the RAM, Ts65Dn mice made fewer correct choices than control mice and performed at or near chance levels, indicating a deficit in spatial working memory. On trials 9 and 10, Ts65Dn mice performed as well as control mice on the initial 12 choices, but required a greater number of choices to complete the RAM. The improved performance of Ts65Dn mice on trials 9 and 10 was lost when the animals were retested after a 50-day retention period, suggesting that long-term memory is also defective. These results are not likely explained by differences in either response bias or perceptual discrimination. Ts65Dn and control mice displayed comparable levels of performance in spontaneous alternation in a T-maze, demonstrating that simple spatial memory was not impaired. In the elevated plus maze, Ts65Dn mice did not display higher anxiety levels which could affect their performance in the RAM. In fact, Ts65Dn mice visited open arms on the elevated plus maze more frequently and spent more time on open arms than did control mice. Taken together, these results provide evidence for short- and long-term spatial memory deficits in Ts65Dn mice.

Role of melatonin in mediating seasonal energetic and immunologic adaptations.

Winter is energetically demanding and stressful; thermoregulatory demands increase when food availability usually decreases. Physiological and behavioral adaptations, including termination of breeding, have evolved among nontropical animals to cope with the energy shortages during winter. Presumably, selection for the mechanisms that permit physiological and behavioral anticipation of seasonal ambient changes have led to current seasonal breeding patterns for many populations. In addition to the well-studied seasonal cycles of mating and birth, there are also significant seasonal cycles of illness and death among field populations of mammals and birds. Energetically challenging winter conditions can directly induce death via hypothermia, starvation, or shock; surviving these demanding conditions likely puts individuals under great physiological stress. The stress of coping with energetically demanding conditions may increase adrenocortical steroid levels that could indirectly cause illness and death by compromising immune function. Individuals would enjoy a survival advantage if seasonally recurring stressors could be anticipated and countered by bolstering immune function. The primary environmental cue that permits physiological anticipation of season is daily photoperiod, a cue that is mediated by melatonin. However, other environmental factors may interact with photoperiod to affect immune function and disease processes. Immune function is compromised during the winter in field studies of birds and mammals. However, laboratory studies of seasonal changes in mammalian immunity consistently report that immune function is enhanced in short day lengths. To resolve this apparent discrepancy, we hypothesize that winter stressors present in field studies counteract short-day enhancement of immune function. Prolonged melatonin treatment mimics short days, and also enhances rodent immune function. Reproductive responsiveness to melatonin appears to affect immune function. In sum, melatonin may be part of an integrative system to coordinate reproductive, immunologic, and other physiological processes to cope successfully with energetic stressors during winter.

Effects of food deprivation and metabolic fuel utilization on food hoarding by jirds (Meriones shawi).

Food hoarding plays an important role in the energetic repertoire of a variety of mammalian species. Both food hoarding and food intake have been examined in rodents using several energetic challenges including food deprivation, treatment with metabolic fuel blockers, and enhancement of fuel storage. In the present experiment, we examined food hoarding by female jirds (Meriones shawi), a desert rodent species occupying the arid steppes and desert regions of Egypt. Jirds are prodigious hoarders in the field; however, virtually nothing is known about their hoarding within controlled laboratory settings. In the present study, the effects of food deprivation as well as alterations in metabolic fuel utilization (i.e., 2-deoxy-D-glucose and isophane insulin) on food hoarding and food intake were tested in female jirds using a simulated burrow system. Jirds decreased body mass and increased food consumption following either 32 or 56-h food deprivation. Food hoarding, however, was virtually abolished after food deprivation and treatment with 2-DG. In contrast, isophane insulin treatment had no effect on food consumption or hoarding in this species. Taken together, the present results suggest that total body mass (fat), rather than short-term metabolic fuel utilization, regulates both food consumption and hoarding in female jirds. In addition, these results provide a novel set of appetitive responses to these energetic challenges in small mammals.

Castration does not inhibit aggressive behavior in adult male prairie voles (Microtus ochrogaster).

The relationship between castration and reduced male aggression is well established. However, anecdotal observations of male prairie voles (Microtus ochrogaster) suggest that castration does not reduce aggressive behavior. To investigate the role of testicular androgens on aggressive behavior, castrated or gonadally intact male prairie voles were paired in a neutral arena with a gonadally intact vole. Castration did not reduce the frequency of intermale aggression. In Experiment 2, aggressive behavior was examined further using resident-intruder, grouped aggression, and aggression against a lactating female models. Again, castration did not affect the frequency of aggression in male prairie voles. Taken together, the results of this study suggest that aggressive behavior may be independent of gonadal steroid hormones in adult male prairie voles.

Ejaculatory abnormalities in mice lacking the gene for endothelial nitric oxide synthase (eNOS-/-).

Nitric oxide (NO) has been established as a neurotransmitter in both the central and peripheral nervous systems. Three isoforms of its synthetic enzyme, NO synthase (NOS), have been identified: 1) in the endothelial lining of blood vessels (eNOS), 2) an inducible form found in macrophages (iNOS), and 3) in neurons (nNOS). Previous studies using pharmacological agents that block all three isoforms of NOS have revealed that NO mediates several aspects of reproductive physiology and behavior, including anomalies in male sexual behavior and erectile function. To determine the specific contribution of the endothelial isoform of NOS in male reproductive behavior, we studied mice missing the gene for only eNOS (eNOS-/-). Wild-type (WT) and eNOS-/- animals were placed with an estrous WT female and observed for 45 min. Both WT and eNOS-/- mice displayed equivalent motivation to mount the stimulus female. However, eNOS-/- mice exhibited striking anomalies in ejaculatory function. A higher percentage of eNOS-/- than WT mice ejaculated during the testing period (p < 0.001). This increased propensity to ejaculate was apparently due to reduced stimulation required to elicit ejaculation; eNOS-/- mice required significantly fewer mounts (p < 0.003) and intromissions (p < 0.001) to ejaculate compared to WT mice. Taken together, these results suggest that NO synthesized by eNOS may be involved in ejaculatory physiology, but not sexual motivation.