Rabbit Maternal Behavior: A Perspective from Behavioral Neuroendocrinology, Animal Production, and Psychobiology.

Rabbit maternal behavior (MB) impacts meat and fur production on the farm, survival of the species in the wild, and pet welfare. Specific characteristics of rabbit MB (i.e., three-step nest building process; single, brief, daily nursing bout) have been used as models for exploring particular themes in neuroscience, like obsessive-compulsive actions, circadian rhythms, and cognition. Particular hormonal combinations regulate nest building by acting on brain regions controlling MB in other mammals. Nonhormonal factors like type of lodging and the doe's social rank influence nursing and milk production. The concurrency of pregnancy and lactation, the display of nonselective nursing, and the rapid growth of altricial young - despite a minimal effort of maternal care - have prompted the study of mother-young affiliation, neurodevelopment, and weaning. Neurohormonal mechanisms, common to other mammals, plus additional strategies (perhaps unique to rabbits) allow the efficient, adaptive display of MB in multiple settings.

Oxytocin induces lordosis behavior in female rats through the prostaglandin E2/GnRH signaling system.

Intracerebroventricular (icv) administration of oxytocin (OT) induces robust lordosis behavior (lordosis quotient and lordosis intensity) in estrogen-primed rats. The present study explored the hypothesis that the OT-Prostaglandin E2-GnRH pathway (a pathway produced in astrocytes) is involved in the facilitation of lordosis behavior by icv infusion of OT (2 µg). In Experiment 1, we tested the involvement of the OT receptor (OTR) by infusion of the OTR antagonist, atosiban (ATO). OT-induced lordosis was significantly reduced at both 30 and 120 min by prior infusion of ATO. In Experiment 2, we studied the effects of aspirin (COX2 inhibitor) and ONO-AE3-208 (ONO; EP4 prostaglandin receptor antagonist) on OT-induced lordosis. Infusions of both compounds diminished OT-induced lordosis at both 120 and 240 min. In Experiment 3, the involvement of the GnRH-1 receptor inhibitor antide on OT-induced lordosis was evaluated. Antide significantly inhibited OT-induced lordosis at all times tested. These data indicate that the OT/PGE2/GnRH pathway is involved in the expression of OT-induced lordosis behavior, an effect that may be occurring directly in hypothalamic astrocytes.

Dataset on c-Fos expression within components of corticostriatal thalamocortical circuits during the expression of a compulsive-like behavior in the female rabbit: Brain-behavior relationships.

The dataset describes regional brain c-Fos expression and a component of maternal nest building behavior ("straw carrying") in 5 late term pregnant rabbits that had been allowed to interact with straw (a nest building material) for a discrete period (30 min), during which repetitive straw carrying behavior was initiated. Animals were sacrificed for brain c-Fos immunoreactivity 1 h after straw was placed into their cage. Regional brain c-Fos expression: Neuronal c-Fos expression is known to associate with a sustained increase in neuronal excitation above resting levels, primarily due to its induction in response to increased glutamatergic input and corresponding activation of the NMDA receptor. In practice, c-Fos expression is taken to be an indication of an increase in "neuronal activity". Importantly, there is a lag of approximately 20 to 30 min between the onset of the stimulus that caused increased excitation, and the initiation of neuronal c-Fos expression, and c-Fos has a cellular half-life of approximately 1 h. Thus, the pattern of brain c-Fos expression within a brain histological section represents a composite snapshot of "superimposed" regional activations that occurred within approximately 30 min to 2 h prior to sacrifice. Behavioral variables: Behavioral variables included in the present dataset are those that reflect the repetitive nature of straw carrying (straw carrying cycle frequency), as well as individual subcomponents of this behavior (collecting straw, interacting with the nest site), and indicators of the "rigidity" of expression of these subcomponents across all cycle repetitions (standard deviations of time spent collecting straw, time spent interacting with nest site). Exploratory Factor Analysis (EFA) with cluster rotation was applied in an exploratory manner in order to clarify correlational relationships between regional c-Fos expression and specific behavioral variables.

Protein kinase inhibitors infused intraventricularly or into the ventromedial hypothalamus block short latency facilitation of lordosis by oestradiol.

An injection of unesterified oestradiol (E2 ) facilitates receptive behaviour in E2 benzoate (EB)-primed, ovariectomised female rats when it is administered i.c.v. or systemically. The present study tested the hypothesis that inhibitors of protein kinase A (PKA), protein kinase G (PKG) or the Src/mitogen-activated protein kinase (MAPK) complex interfere with E2 facilitation of receptive behaviour. In Experiment 1, lordosis induced by i.c.v. infusion of E2 was significantly reduced by i.c.v. administration of Rp-cAMPS, a PKA inhibitor, KT5823, a PKG inhibitor, and PP2 and PD98059, Src and MAPK inhibitors, respectively, between 30 and 240 minutes after infusion. In Experiment 2, we determined whether the ventromedial hypothalamus (VMH) is one of the neural sites at which those intracellular pathways participate in lordosis behaviour induced by E2 . Administration of each of the four protein kinase inhibitors into the VMH blocked facilitation of lordosis induced by infusion of E2 also into the VMH. These data support the hypothesis that activation of several protein kinase pathways is involved in the facilitation of lordosis by E2 in EB-primed rats.

Activation of cortical and striatal regions during the expression of a naturalistic compulsive-like behavior in the rabbit.

Nest building behavior in the pregnant rabbit (Oryctolagus cuniculus) can serve as a model for compulsions in obsessive compulsive disorder (OCD). Previous work showed that the "straw carrying" phase of nest building (during which the rabbit repeatedly collects straw in its mouth, carries it into the nest box and deposits it there, and then returns to collect more) is associated with increased c-FOS expression (a marker of neuronal activity) in the orbitofrontal, anterior cingulate, and piriform cortices. In the present study, we quantified c-FOS expression in the caudate and putamen, as well as in the primary motor, somatosensory, and prefrontal cortices of: (1) pregnant rabbits given straw (PREG + STRAW); pregnant rabbits not given straw (PREG); (3) estrous rabbits given straw (ESTROUS + STRAW); and (4) estrous rabbits not given straw (ESTROUS). We found that straw carrying was associated with increased c-FOS expression in the dorsal putamen, ventral caudate, primary motor cortex, and somatosensory cortex. Additionally, a correlational analysis of PREG + STRAW animals revealed that these regions, along with the premotor and prelimbic cortices, were significantly intercorrelated with respect to c-FOS expression, suggesting their "coactivation" during repetitive straw carrying. By contrast, behavioral interactions of non-pregnant (ESTROUS) rabbits with straw (e.g., sniffing, nibbling it) were associated with a distinct pattern of c-FOS expression that included the medial and ventral putamen. c-FOS expression in PREG + STRAW rabbits is similar to patterns of regional brain activity in OCD patients exposed to obsession-provoking stimuli, as well as to those observed in healthy human mothers responding to infant-associated stimuli.

Therapeutic use of Bacillus thuringiensis in the treatment of psoroptic mange in naturally infested New Zealand rabbits.

Bacillus thuringiensis is a bacteria known for its bioinsecticidal toxins and it has been proposed as an alternative in the treatment of several parasites that infect domestic animals (helminths, ticks, mites). In this work, we evaluated the clinical efficiency of the Bacillus thuringiensis GP532 strain in the treatment of six rabbits naturally infested with the P. cuniculi mite. GP532 extract (10mg/ml) was applied by aspersion in both pinna, with a second application after seven days, and the therapeutic effect was measured in both qualitative and quantitative manner. GP532 application resulted in a decreased infestation rate, which was observed as early as 3days post-treatment. At day 14, a decrease from 4.66±0.61 to 0.50±0.10 in the left pinna and from 1.66±0.21 to 0.66±0.16 (P<0.05) in the right pinna was observed. This response was comparable to the commercial drug Ivermectin, which induced a decreased infestation rate from 4.00±0.51 to 0.16±0.10 in the left pinna and from 4.66±0.80 to 0.25±0.11 in the right pinna (P<0.05). At day 30 post-treatment, GP532 decreased the total infested area by 76.80±16.06%, whereas Ivermectin resulted in a 97.41±0.99% decrease. Neither treatment produced irritation or macroscopic lesions. Our results show that the B. thuringiensis GP532 strain has a therapeutic potential in the treatment of psoroptic mange in rabbits.

Activation of the orbitofrontal and anterior cingulate cortices during the expression of a naturalistic compulsive-like behavior in the rabbit.

We propose that maternal nest building in the female laboratory rabbit is a useful model for compulsions in obsessive-compulsive disorder (OCD). This repetitive behavior comprises collecting straw, depositing it into the nest box, and then returning to collect more straw. We reasoned that if "straw carrying" behavior is homologous to compulsive behavior, then it should be associated with activation of prefrontal regions associated with OCD, namely, the orbitofrontal and anterior cingulate cortices (OFC and ACC, respectively). In the present study, we quantified c-FOS immunoreactivity in the ACC, OFC, premotor (PM), infralimbic (IL), prelimbic (PL), and piriform (PI) cortices of: (1) pregnant female rabbits that were given straw (PREG+STRAW); (2) pregnant rabbits that were not given straw (PREG); (3) estrous rabbits that were given straw (ESTROUS+STRAW); (4) estrous rabbits that were not given straw (ESTROUS). After 1h, all females were sacrificed and processed for brain c-FOS immunoreactivity. We found that pregnant rabbits showed lower latencies to interact with the straw than estrous rabbits, and that pregnant rabbits displayed straw carrying, while estrous rabbits did not. c-FOS expression was increased in the OFC, ACC, and PI in the PREG+STRAW compared to all other groups. By contrast, c-FOS expression in all other regions was greater in PREG+STRAW compared to PREG, but not different from ESTROUS+STRAW. These results point to an important role for the OFC, ACC, and PI in initiating repetitive straw-carrying behavior, and further support the proposal that this behavior can serve as a model for compulsions in OCD.

Taenia pisiformis cysticercosis induces decreased prolificacy and increased progesterone levels in rabbits.

Reproductive alterations in hosts infected by parasites have been recognized in several phyla, especially in arthropods and mollusks, but it has been less studied in higher vertebrates, particularly in mammals. In the present study, ten eight week-old female New Zealand rabbits (Oryctolagus cuniculus) were either infected with Taenia pisiformis eggs or uninfected, and 7 weeks later they were mated. We found that serum progesterone levels were increased during pregnancy in infected does. At birth, litter size of infected does was reduced by half as compared to the control group, and, at weaning, the number of kits and the weight of litters was lower. Since serum progesterone levels have a key role in the maintenance of pregnancy and implantation, we propose that the observed prolificacy alterations in does infected with T. pisiformis infection were due to changes in the levels of circulating progesterone during pregnancy.

New dimensions in the use of rodent behavioral tests for novel drug discovery and development.

INTRODUCTION: Dimensional models of psychopathology describe mental illness in terms of natural variance along certain phenotypic dimensions that are continuous with normal. Vulnerability to psychopathology might arise when certain adaptive psychophysiological processes, conserved between humans and non-human animals, function outside of their "normal" range. Therefore, an in-depth understanding of the neurobiology and neurochemistry underlying these processes could identify possible novel drug targets. AREAS COVERED: Psychophysiological processes that might be related to anxiety disorders and depression are proposed and discussed. Those processes relevant to depressive disorders include: hedonic responsiveness, biases in the processing of stimuli, and sleep architecture. Those relevant to anxiety disorders include: startle reactivity, CO2 sensitivity, and fear generalization. Rodent behavioral tests for assessing the function of these processes and investigating their neurobiology are described. A psychophysiological process strategy for translational research is proposed, which focusses on understanding the neurobiology and neurochemistry underlying key psychophysiological processes that, when their activity deviates from normal, are associated with neuropsychiatric symptoms. This strategy emphasizes the use of analogous tests and measures in both preclinical and clinical studies, while de-emphasizing the use of preclinical animal models that attempt to replicate features of the neuropsychiatric disorder through experimental manipulations. EXPERT OPINION: Investigating the neurobiology of key psychophysiological processes in rodents should enhance our understanding of the pathophysiology of neuropsychiatric disorders. New drug development could be directed toward developing pharmacological strategies that would normalize the function of these psychophysiological processes.

Clozapine and glycinamide prevent MK-801-induced deficits in the novel object recognition (NOR) test in the domestic rabbit (Oryctolagus cuniculus).

Studies in humans indicate that acute administration of sub-anesthetic doses of ketamine, an NMDA receptor antagonist, provokes schizophrenic-like symptoms in healthy volunteers, and exacerbates existing symptoms in individuals with schizophrenia. These and other findings suggest that NMDA receptor hypofunction might participate in the pathophysiology of schizophrenia, and have prompted the development of rodent pharmacological models for this disorder based on acute or subchronic treatment with NMDA receptor antagonists, as well as the development of novel pharmacotherapies based on increasing extrasynaptic glycine concentrations. In the present study, we tested whether acute hyperlocomotory behavior and/or deficits in the novel object recognition (NOR) task, induced in male rabbits by the acute subcutaneous (s.c.) administration of MK-801 (0.025 and 0.037 mg/kg s.c., respectively), were prevented by prior administration of the atypcial antipsychotic, clozapine (0.2mg/kg, s.c.), or the glycine pro-drug glycinamide (56 mg/kg, s.c.). We found that clozapine fully prevented the MK-801-induced hyperlocomotion, and both clozapine and glycinamide prevented MK-801-induced deficits in the NOR task. The present results show that MK-801-induced hyperlocomotion and deficits in the NOR task in the domestic rabbit demonstrate predictive validity as an alternative animal model for symptoms of schizophrenia. Moreover, these results indicate that glycinamide should be investigated in pre-clinical models of neuropsychiatric disorders such as schizophrenia, obsessive compulsive disorder and anxiety disorders, where augmentation of extrasynaptic glycine concentrations may have therapeutic utility.

Hormonal and behavioral changes induced by acute and chronic experimental infestation with Psoroptes cuniculi in the domestic rabbit Oryctolagus cuniculus.

BACKGROUND: Parasitic diseases are important in animal production because they cause high economic losses. Affected animals often exhibit stereotypical behavioral alterations such as anorexia and inactivity, among others. Among the diseases that commonly affect domestic rabbits is mange, which is caused by the mite Psoroptes cuniculi. Therefore, within the context of the host-parasite relationship, it is critical to understand the mechanisms involved in the alteration of host behavior, in order to better utilize sick animal behavior as a strategy for diagnosis and treatment of disease. METHODS: Rabbits were infested placing mites in the ear conduct. We characterized changes in exploratory behavior and scent marking evoked by acute (1-9 days) and chronic (25-33 days) experimental infestation. Behavior was recorded during ten minutes while the animals were in a 120 cm × 120 cm open field arena divided into 9 squares. Serum cortisol was measured individually using radioimmunoassay kits. Locomotor activity, chinning, rearing and body weight were compared using a Friedman test, the effect of treatment (infested versus non-infested) across time was analyzed using a repeated measures ANOVA, and the Pearson test was used to determine whether chinning and ambulation scores were significantly correlated. Serum cortisol levels and food consumption were analyzed with a Kruskal-Wallis test and body temperature was analyzed with an ANOVA test. RESULTS: We observed a significant decrease in rearing behavior as early as two days post-infestation, while chinning and locomotor activity were significantly decreased four days post-infestation. Chronic infestation was associated with decreased food intake, significant weight loss, and a trend toward increased serum cortisol levels, while no changes were observed in body temperature. CONCLUSIONS: The presence of visible lesions within the ear canal is commonly used to detect mite infestation in rabbits, but this is possible only after chronic infestation. The behaviors described here may be a useful and economic tool in guiding the early diagnosis of parasitic infestation by P. cuniculi, allowing for early treatment and the application of control measures before significant weight loss occurs, thereby avoiding economic losses.

D1 and D2 dopamine receptor antagonists decrease behavioral bout duration, without altering the bout's repeated behavioral components, in a naturalistic model of repetitive and compulsive behavior.

Nest building behavior in the pregnant female rabbit (Oryctolagus cuniculus) is a model for compulsive behavior in Obsessive Compulsive Disorder (OCD). This behavior comprises a cycle of repeated, stereotyped components (collecting straw, entering nest box and depositing the straw there, returning to collect more straw), which itself is repeated 80+ times in a single bout that lasts approximately 50min. The bout, in turn, is repeated if necessary, according to the rabbit's perception of whether or not the nest is finished. We administered SCH23390 (5-100µg/kg; D1/D5 antagonist) or raclopride (0.05-1.0mg/kg; D2/D3 antagonist), subcutaneously to day 28 pregnant female rabbits, 30 or 60min before placing straw inside their home cage. At doses that minimally affected ambulatory behavior in open field (5-12.5µg/kg SCH23390, 0.5-1.0mg/kg raclopride), both antagonists dramatically reduced bout duration while not significantly affecting the initiation of straw carrying behavior, the sequential performance of the individual cycle components, maximum cycle frequency, or the total number of bouts performed. These results point to an important role for dopamine neurotransmission for the prolonged expression of a normal, repetitive and compulsive-like behavior. Moreover, the finding that dopamine receptor antagonists decrease the time spent engaged in repetitive behavior (without significantly altering the form of the repetitive behavior itself) suggests a possible explanation for why neuroleptics can be clinically effective for treating OCD.

Relevance of mating-associated stimuli, ovulation, and the progesterone receptor for the post-coital inhibition of estrous behavior in the female rabbit.

Estrous female domestic rabbits (Oryctolagus cuniculus) display scent marking ("chinning") and sexual receptivity. Mating induces ovulation, which occurs approximately 12h later, and also decreases chinning and receptivity. In the present study, we explored the participation of mating-associated stimuli, ovulation, and the progesterone receptor (PR) in mediating such behavioral effects. We found that copulatory stimuli were not necessary, and that ovulation alone was sufficient, as these behavioral changes were replicated in unmated females by intravenous administration of human chorionic gonadotropin (hCG). The post-mating administration (s.c.) of 5µg/day estradiol benzoate (EB), prevented the decline in chinning and receptivity. A lower dose of EB (1µg/day) had no effect, nor did the antiprogestin RU486 (20mg, s.c., administered 3h before mating). However, the combination of a single pre-mating administration of RU486 plus the post-mating administration of 1µg/day EB completely blocked the decline in estrous behavior. We propose that PR activation around the time of mating and a post-mating decline in ovarian estradiol secretion and/or estradiol responsiveness act in parallel to terminate estrus in this species.

Scent marking by the male domestic rabbit (Oryctolagus cuniculus) is stimulated by an object's novelty and its specific visual or tactile characteristics.

"Chinning" is a stereotyped scent marking behavior of domestic rabbits, in which the animal rubs the underside of its chin against objects in order to deposit scent gland secretions. Although the long-term maintenance of chinning requires circulating gonadal steroids, little is known about the acute regulation of this behavior. To define specific environmental stimuli that engage the chinning motor pattern, male rabbits were placed into an open field arena containing markable objects ("standard" bricks, "tall" bricks, or polished onyx spheres), observed for 30 min, returned to the home cage for 5 min, and then placed in the open field arena for another 30 min. During the 5 min interim: (1) the location of the open field or the spatial orientation of the objects within it were changed, (2) the olfactory or (3) visual characteristics of the objects were changed; or (4) no changes were made. Chinning and ambulation habituated to each type of object across the first 30 min, and bricks elicited more chinning than polished onyx spheres. In the second 30 min test, chinning was re-stimulated only when the original objects were replaced by visually different ones that had preferred characteristics. Ambulatory behavior was increased by changing the location of the open field arena, while modifying the olfactory characteristics of the objects had no effect on chinning or ambulation. These results indicate that scent marking is stimulated by object novelty and by the visual and/or tactile characteristics of the objects being marked.

Toward an understanding of the neurobiology of "just right" perceptions: nest building in the female rabbit as a possible model for compulsive behavior and the perception of task completion.

Little is known of the neural mechanisms underlying the subjective experience of task completion and the subsequent inhibition of behavior. The preparturient female rabbit displays a stereotyped "straw carrying" behavior, in which she repeatedly collects straw and carries it into a nest box (located within the home cage), resulting in a finished maternal straw nest within 3 h. Thereafter, straw carrying is inhibited for many hours, even if the original nest is removed. We tested whether the performance of straw carrying behavior or the perception of a completed nest were necessary prerequisites for the engagement of this inhibitory mechanism. On day 28 of pregnancy, we placed straw inside the home cage (at t=0 h) and recorded the female's behavior for 3 h, at which time the nest was removed from the nest box. Multiparous females began to collect straw almost immediately, whereas inexperienced females were delayed in this respect. In females that had perceived a completed nest inside the nest box during t=0-3 h, straw carrying was subsequently inhibited across t=3-24 h. In contrast, when we prevented straw from accumulating inside the nest box during the 3 h observation period (by continuously removing it through a hidden door of the nest box), straw carrying persisted across t=3-6 h. These results indicate that the perception of a completed nest is necessary to engage a mechanism that inhibits further straw carrying. However, perceiving a completed nest was not sufficient to inhibit straw carrying behavior, since females whose nest box already contained a completed nest (built by the experimenter at the beginning of the experiment) nevertheless displayed a "bout" of straw carrying lasting approximately 60 min, indicating that the performance of nest building behavior itself might also contribute to its subsequent inhibition.

Maternal and peer contact during the postnatal period participate in the normal development of maternal aggression, maternal behavior, and the behavioral response to novelty.

In the rat, social isolation during the early postnatal period disrupts the adult function of certain neuroendocrine and neurobehavioral systems. In the present study, we assessed the effects of peer and maternal contact during this period on the adult expression of aggression, maternal behavior, and the behavioral response to novelty. Female rat pups of the Wistar strain were reared across postnatal days 3-19 in one of the following conditions: (1) by their mother and with littermates (MR, control); (2) artificially reared in complete social isolation (AR); or (3) artificially reared in the presence of two same-age conspecifics (AR-Soc). As adults, all females were administered a resident-intruder aggression test, first in a non-maternal context ("territorial" aggression) and then in the presence of their pups ("maternal" aggression). Additionally, their maternal behavior and response to a novel object placed in a familiar open field arena were quantified. We found that maternal isolation impaired maternal behavior, increased maternal aggression, and increased the olfactory investigation of a novel object. The presence of peers prevented the effect of artificial rearing on aggression, but not its effects on maternal behavior or response to novelty. In the maternal aggression test, AR-Soc females engaged in significantly more sniffing of the intruder compared to the other two groups. The present results confirm and extend those of previous studies in a different strain (Sprague-Dawley), and indicate that peer-derived stimuli impact on the development of neurobehavioral systems underlying aggressive and non-aggressive social interactions, whereas the normal development of maternal behavior and response to novelty requires maternally derived stimuli.

Neuroendocrine regulation of estrous behavior in the rabbit: similarities and differences with the rat.

In this review, we compare the neuroendocrine control of estrous behavior in the rabbit, a reflex ovulator, and the rat, a more commonly studied spontaneous ovulator. Although the hormonal control of estrous behavior in both species is similar, notable differences include the absence of a stimulatory effect of progesterone (P) on sexual behavior in the rabbit and the retention of sexual behavior in a substantial proportion of female rabbits after ovariectomy. The ventrolateral component of the ventromedial hypothalamus (VMH) and an adjacent region caudal to it appear to be critical estrogen (E)-responsive regions for lordosis in the rat and rabbit, respectively. In both species the effects of E and P are largely mediated by the genomic action of their receptors (ER and PR), and in both species E similarly regulates the expression of these receptors. The prolonged, E-stimulated estrous of the rabbit is terminated after mating by unknown mechanisms, while the brief estrous of the rat is triggered by the proestrous peak of P and terminated by both the decline in P and the downregulation of hypothalamic PR. In both species, P most likely inhibits estrous behavior during pregnancy, and postpartum estrous may be triggered by a stimulatory effect of E coinciding with the withdrawal of P-mediated inhibition. Estrous behavior is inhibited in both species during lactation, most likely by the suckling-induced inhibition of gonadotropin secretion. This comparative approach can reveal neuroendocrine mechanisms underlying estrous behavior that are common to all mammals, while highlighting evolutionary adaptations unique to each species.

A murine model for neuropsychiatric disorders associated with group A beta-hemolytic streptococcal infection.

A syndrome of motoric and neuropsychiatric symptoms comprising various elements, including chorea, hyperactivity, tics, emotional lability, and obsessive-compulsive symptoms, can occur in association with group A beta-hemolytic streptococcal (GABHS) infection. We tested the hypothesis that an immune response to GABHS can result in behavioral abnormalities. Female SJL/J mice were immunized and boosted with a GABHS homogenate in Freund's adjuvant, whereas controls received Freund's adjuvant alone. When sera from GABHS-immunized mice were tested for immunoreactivity to mouse brain, a subset was found to be immunoreactive to several brain regions, including deep cerebellar nuclei (DCN), globus pallidus, and thalamus. GABHS-immunized mice having serum immunoreactivity to DCN also had increased IgG deposits in DCN and exhibited increased rearing behavior in open-field and hole-board tests compared with controls and with GABHS-immunized mice lacking serum anti-DCN antibodies. Rearing and ambulatory behavior were correlated with IgG deposits in the DCN and with serum immunoreactivity to GABHS proteins in Western blot. In addition, serum from a GABHS mouse reacted with normal mouse cerebellum in nondenaturing Western blots and immunoprecipitated C4 complement protein and alpha-2-macroglobulin. These results are consistent with the hypothesis that immune response to GABHS can result in motoric and behavioral disturbances and suggest that anti-GABHS antibodies cross-reactive with brain components may play a role in their pathophysiology.

Steroid-triggered programmed cell death of a motoneuron is autophagic and involves structural changes in mitochondria.

Neuronal death occurs during normal development and disease and can be regulated by steroid hormones. In the hawkmoth, Manduca sexta, individual accessory planta retractor (APR) motoneurons undergo a segment-specific pattern of programmed cell death (PCD) at pupation that is triggered directly and cell autonomously by the steroid hormone 20-hydroxyecdysone (20E). APRs from abdominal segment six [APR(6)s] die by 48 hours after pupal ecdysis (PE; entry into the pupal stage), whereas APR(4)s survive until adulthood. Cell culture experiments showed previously that 20E acts directly on APRs to trigger PCD, with intrinsic segmental identity determining which APRs die. The APR(6) death pathway includes caspase activation and loss of mitochondrial function. We used transmission electron microscopy to investigate the ultrastructure of APR somata before and during PCD. APR(4)s showed normal ultrastructure at all stages examined, as did APR(6)s until approximately stage PE. During APR(6) death, there was massive accumulation of autophagic bodies and vacuoles, mitochondria became ultracondensed and aggregated into compact clusters, and ribosomes aggregated in large blocks. Nuclear ultrastructure remained normal, without chromatin condensation, until the nuclear envelope fragmented late in the death process. Light microscopic immunocytochemistry showed that dying APR(6)s were TUNEL-positive, which is diagnostic of fragmented DNA. These observations indicate that the steroid-induced, caspase-dependent, cell-autonomous PCD of APR(6)s is autophagic, not apoptotic, and support an early role for mitochondrial alterations during PCD. This system permits the study of neuronal death in response to its bona fide developmental signal, the rise in a steroid hormone.