RNA-sequencing and behavioral testing reveals inherited physical inactivity co-selects for anxiogenic behavior without altering depressive-like behavior in Wistar rats.

Physical inactivity is positively associated with anxiety and depression. Considering physical inactivity, anxiety, and depression each have a genetic basis for inheritance, our lab used artificial selectively bred low-voluntary running (LVR) and wild type (WT) female Wistar rats to test if physical inactivity genes selected over multiple generations would lead to an anxiety or depressive-like phenotype. We performed next generation RNA sequencing and immunoblotting on the dentate gyrus to reveal key biological functions from heritable physical inactivity. LVR rats did not display depressive-like behavior. However, LVR rats did display anxiogenic behavior with gene networks associated with reduced neuronal development, proliferation, and function compared to WT counterparts. Additionally, immunoblotting revealed LVR deficits in neuronal development and function. To our knowledge, this is the first study to show that by selectively breeding for physical inactivity genes, anxiety-like genes were co-selected. The study also reveals molecular insights to the genetic influences that physical inactivity has on anxiety-like behavior.

Repeated neonatal isoflurane exposures in the mouse induce apoptotic degenerative changes in the brain and relatively mild long-term behavioral deficits.

Epidemiological studies suggest exposures to anesthetic agents and/or sedative drugs (AASDs) in children under three years old, or pregnant women during the third trimester, may adversely affect brain development. Evidence suggests lengthy or repeated AASD exposures are associated with increased risk of neurobehavioral deficits. Animal models have been valuable in determining the type of acute damage in the developing brain induced by AASD exposures, as well as in elucidating long-term functional consequences. Few studies examining very early exposure to AASDs suggest this may be a critical period for inducing long-term functional consequences, but the impact of repeated exposures at these ages has not yet been assessed. To address this, we exposed mouse pups to a prototypical general anesthetic, isoflurane (ISO, 1.5% for 3 hr), at three early postnatal ages (P3, P5 and P7). We quantified the acute neuroapoptotic response to a single versus repeated exposure, and found age- and brain region-specific effects. We also found that repeated early exposures to ISO induced subtle, sex-specific disruptions to activity levels, motor coordination, anxiety-related behavior and social preference. Our findings provide evidence that repeated ISO exposures may induce behavioral disturbances that are subtle in nature following early repeated exposures to a single AASD.

Marble burying as compulsive behaviors in male and female mice.

Marble burying is considered an, albeit controversial, animal model of the compulsive like behaviors of obsessive-compulsive disorder (OCD). Hallmark features of OCD patients are similarities and, more prominent, differences from anxiety disorders, e.g., the absence of sex differences and resistance to spontaneous remission. We report an experiment on marble burying by male and female C57/BL6/N mice. Animals were administered either the classic anxiolytic drug, diazepam, that targets the GABA receptor or a "pure" inhibitor of the serotonin transporter, escitalopram, that has been reported to be particularly effective in OCD. A burying paradigm that more precisely mimics the human condition was used, e.g., testing in the home environment, chronic drug exposure and acknowledging individual differences by pre-selecting for high marble burying. Results were that there were no sex differences in groups treated with drugs or in control mice. Both diazepam and escitalopram decreased numbers of marbles buried compared to vehicle-only controls in the absence of correlated changes in anxiety. Diazepam, however, was more effective than escitalopram in suppressing MB. The conclusion is that along with serotonin, GABA is involved in regulating compulsive behaviors. The marble burying paradigm may prove more useful for pharmacological drugs tests of impulsivity or attention deficit because of the involvement of serotonin and GABA in both disorders.

Cognition in female rats after blocking conversion of androgens to estrogens.

Women and non-human females have surprisingly high levels of circulating testosterone, yet the effects of androgens on non-reproductive behaviors, including cognition, of females are not well characterized. The current project used an aromatase inhibitor, letrozole, to block conversion of androgens to estrogens. Adult female rats were ovariectomized and administered either vehicle only, testosterone propionate only (400µg/kg, TP only), letrozole only (1mg/kg, Letro only), or the combination of letrozole and testosterone (TP+Letro) over 4weeks. A gonadally intact group was used for comparisons. During the last 3weeks, the animals were tested for working memory in both a spatial task (radial arm maze) and a non-spatial task (object recognition). At sacrifice, uterine weights and serum testosterone and estradiol were determined. Behavioral results were the intact animals showed better working memories on the object recognition task, but that there were no differences among the ovariectomized groups. In the radial arm maze task, groups with best to worst performance were TP only>Intact=TP+Letro>vehicle=Letro only. Highest to lowest serum titers, for testosterone, were TP+Letro>TP only>Intact=Letro only>vehicle and, for estradiol, Intact>TP only>Vehicle>Letro only=TP+Letro. Our interpretation is that testosterone enhanced spatial performance when bioavailability of both TP and E2 are high, and high testosterone can rescue spatial memory when E2 bioavailability is low.

Integrating insulin-like growth factor 1 and sex hormones into neuroprotection: Implications for diabetes.

Brain integrity and cognitive aptitude are often impaired in patients with diabetes mellitus, presumably a result of the metabolic complications inherent to the disease. However, an increasing body of evidence has demonstrated the central role of insulin-like growth factor 1 (IGF1) and its relation to sex hormones in many neuroprotective processes. Both male and female patients with diabetes display abnormal IGF1 and sex-hormone levels but the comparison of these fluctuations is seldom a topic of interest. It is interesting to note that both IGF1 and sex hormones have the ability to regulate phosphoinositide 3-kinase-Akt and mitogen-activated protein kinases-extracellular signal-related kinase signaling cascades in animal and cell culture models of neuroprotection. Additionally, there is considerable evidence demonstrating the neuroprotective coupling of IGF1 and estrogen. Androgens have also been implicated in many neuroprotective processes that operate on similar signaling cascades as the estrogen-IGF1 relation. Yet, androgens have not been directly linked to the brain IGF1 system and neuroprotection. Despite the sex-specific variations in brain integrity and hormone levels observed in diabetic patients, the IGF1-sex hormone relation in neuroprotection has yet to be fully substantiated in experimental models of diabetes. Taken together, there is a clear need for the comprehensive analysis of sex differences on brain integrity of diabetic patients and the relationship between IGF1 and sex hormones that may influence brain-health outcomes. As such, this review will briefly outline the basic relation of diabetes and IGF1 and its role in neuroprotection. We will also consider the findings on sex hormones and diabetes as a basis for separately analyzing males and females to identify possible hormone-induced brain abnormalities. Finally, we will introduce the neuroprotective interplay of IGF1 and estrogen and how androgen-derived neuroprotection operates through similar signaling cascades. Future research on both neuroprotection and diabetes should include androgens into the interplay of IGF1 and sex hormones.

Kappa Opioids, Salvinorin A and Major Depressive Disorder.

Opioids are traditionally associated with pain, analgesia and drug abuse. It is now clear, however, that the opioids are central players in mood. The implications for mood disorders, particularly clinical depression, suggest a paradigm shift from the monoamine neurotransmitters to the opioids either alone or in interaction with monoamine neurons. We have a special interest in dynorphin, the last of the major endogenous opioids to be isolated and identified. Dynorphin is derived from the Greek word for power, dynamis, which hints at the expectation that the neuropeptide held for its discoverers. Yet, dynorphin and its opioid receptor subtype, kappa, has always taken a backseat to the endogenous b-endorphin and the exogenous morphine that both bind the mu opioid receptor subtype. That may be changing as the dynorphin/ kappa system has been shown to have different, often opposite, neurophysiological and behavioral influences. This includes major depressive disorder (MDD). Here, we have undertaken a review of dynorphin/ kappa neurobiology as related to behaviors, especially MDD. Highlights include the unique features of dynorphin and kappa receptors and the special relation of a plant-based agonist of the kappa receptor salvinorin A. In addition to acting as a kappa opioid agonist, we conclude that salvinorin A has a complex pharmacologic profile, with potential additional mechanisms of action. Its unique neurophysiological effects make Salvinorina A an ideal candidate for MDD treatment research.

Comprehensive functional characterization of murine infantile Batten disease including Parkinson-like behavior and dopaminergic markers.

Infantile neuronal ceroid lipofuscinosis (INCL, Infantile Batten disease) is a neurodegenerative lysosomal storage disease caused by a deficiency in palmitoyl protein thioesterase-1 (PPT1). The PPT1-deficient mouse (Cln1(-/-)) is a useful phenocopy of human INCL. Cln1(-/-) mice display retinal dysfunction, seizures, motor deficits, and die at ~8 months of age. However, little is known about the cognitive and behavioral functions of Cln1(-/-) mice during disease progression. In the present study, younger (~1-2 months of age) Cln1(-/-) mice showed minor deficits in motor/sensorimotor functions while older (~5-6 months of age) Cln1(-/-) mice exhibited more severe impairments, including decreased locomotor activity, inferior cued water maze performance, decreased running wheel ability, and altered auditory cue conditioning. Unexpectedly, certain cognitive functions such as some learning and memory capabilities seemed intact in older Cln1(-/-) mice. Younger and older Cln1(-/-) mice presented with walking initiation defects, gait abnormalities, and slowed movements, which are analogous to some symptoms reported in INCL and parkinsonism. However, there was no evidence of alterations in dopaminergic markers in Cln1(-/-) mice. Results from this study demonstrate quantifiable changes in behavioral functions during progression of murine INCL and suggest that Parkinson-like motor/sensorimotor deficits in Cln1(-/-) mice are not mediated by dopamine deficiency.

Smoking in schizophrenic patients: A critique of the self-medication hypothesis.

A common remark among laypeople, and notably also among mental health workers, is that individuals with mental illnesses use drugs as self-medication to allay clinical symptoms and the side effects of drug treatments. Roots of the self-medication concept in psychiatry date back at least to the 1980s. Observations that rates of smokers in schizophrenic patients are multiple times the rates for regular smoking in the general population, as well as those with other disorders, proved particularly tempting for a self-medication explanation. Additional evidence came from experiments with animal models exposed to nicotine and the identification of neurobiological mechanisms suggesting self-medication with smoking is a plausible idea. More recently, results from studies comparing smoking and non-smoking schizophrenic patients have led to the questioning of the self-medication hypothesis. Closer examination of the literature points to the possibility that smoking is less beneficial on schizophrenic symptomology than generally assumed while clearly increasing the risk of cancer and other smoking-related diseases responsible for early mortality. It is a good time to examine the evidence for the self-medication concept as it relates to smoking. Our approach is to focus on data addressing direct or implied predictions of the hypothesis in schizophrenic smokers.

Differential effects of antipsychotics on lateral bias and social attention in female rats.

RATIONALE: Prior research has demonstrated that individuals with schizophrenia may exhibit lateral biases in attention and deficits in social behavior. The use of a noninvasive animal model of attentional impairments in schizophrenia and antipsychotic drugs can help elucidate the biological underpinnings of attentional processes and facilitate the study of novel therapeutics. OBJECTIVES: The purpose of this study was to compare the effects of three antipsychotic drugs on measures of lateral bias and social attention in healthy, unoperated female rats. MATERIALS AND METHODS: Female Long-Evans rats selected for a preexisting lateral bias in attention, a right behavioral orientation preference (BOP), were administered clozapine, haloperidol, sulpiride, or vehicle. Lateral bias in attention was assessed by determining which forelimb rats removed a nuisance stimulus from first. Social attention was examined by comparing the latency to remove nuisance stimuli in the presence of a social (inaccessible female rat) versus non-social (blinking clock) distractor. RESULTS: All antipsychotic drugs eliminated right lateral bias in attention, while control animals retained their initial bias. Clozapine eliminated right lateral bias more rapidly than the other drugs. Animals receiving clozapine also selectively displayed increased attention to another rat. CONCLUSIONS: The results suggest that the antipsychotic medication clozapine rapidly alters attentional bias and uniquely influences attention to a social stimulus. The right BOP paradigm is a useful animal model for comparing antipsychotic drug effects on lateralized attentional bias and attention to social stimuli.

Antidepressive effects of the κ-opioid receptor agonist salvinorin A in a rat model of anhedonia.

Salvinorin A (SalvA), the hallucinogenic derivative of the plant Salvia divinorum, is a selective κ-opioid receptor agonist that may also have antidepressant properties. Chronic mild stress (CMS) was applied to male and female Long-Evans rats to model anhedonia common in depression. The progressive loss in preference for a sucrose solution over plain water, a measure of anhedonia, and locomotor activity were monitored for 7 weeks. Because antidepressant medications often modify reproductive functions, endocrine glands and hormone-sensitive tissues were assessed at necropsy after the conclusion of the behavioral protocol. Three weeks of CMS exposure led to a decrease in sucrose preference. CMS was continued for 3 additional weeks and animals were randomly assigned to treatment with 1 mg SalvA/kg body weight or to a vehicle control group. The results indicate that SalvA reversed anhedonia whereas control animals continued to show a suppressed preference for the sucrose solution. In addition, no change in sucrose preference was observed in nonstressed rats that were exposed to the same dosage of SalvA. The results indicate that SalvA is an effective antidepressant agent when administered chronically to rats showing symptoms of depression similar to those observed in humans.

Adrenal steroids uniquely influence sexual motivation behavior in male rats.

The androgenic adrenal steroids dehydroepiandrosterone (DHEA) and 4α-androstenedione (4-A) have significant biological activity, but it is unclear if the behavioral effects are unique or only reflections of the effects of testosterone (TS). Gonadally intact male Long-Evans rats were assigned to groups to receive supplements of DHEA, 4-A, TS, corticosteroid (CORT), all at 400 µg steroid/kg of body weight, or vehicle only for 5 weeks. All males were tested in a paradigm for sexual motivation that measures time and urinary marks near an inaccessible receptive female. It was found that DHEA and 4-A supplements failed to influence time near the estrous female in the same way TS supplements did, and, indeed, 5 weeks of 4-A administration reduced the time similar to the suppressive effects of CORT after 3 weeks. Further, animals treated with DHEA or 4-A left fewer urinary marks near an estrous female than TS and control groups. These results suggest that DHEA and 4-A are not merely precursors of sex hormones, and provide support for these steroids influencing the brain and behavior in a unique fashion that is dissimilar from the effects of TS on male sexual behavior.

Behavioral consequences of NMDA antagonist-induced neuroapoptosis in the infant mouse brain.

BACKGROUND: Exposure to NMDA glutamate antagonists during the brain growth spurt period causes widespread neuroapoptosis in the rodent brain. This period in rodents occurs during the first two weeks after birth, and corresponds to the third trimester of pregnancy and several years after birth in humans. The developing human brain may be exposed to NMDA antagonists through drug-abusing mothers or through anesthesia. METHODOLOGY/PRINCIPAL FINDINGS: We evaluated the long-term neurobehavioral effects of mice exposed to a single dose of the NMDA antagonist, phencyclidine (PCP), or saline, on postnatal day 2 (P2) or P7, or on both P2 and P7. PCP treatment on P2 + P7 caused more severe cognitive impairments than either single treatment. Histological examination of acute neuroapoptosis resulting from exposure to PCP indicated that the regional pattern of degeneration induced by PCP in P2 pups was different from that in P7 pups. The extent of damage when evaluated quantitatively on P7 was greater for pups previously treated on P2 compared to pups treated only on P7. CONCLUSIONS: These findings signify that PCP induces different patterns of neuroapoptosis depending on the developmental age at the time of exposure, and that exposure at two separate developmental ages causes more severe neuropathological and neurobehavioral consequences than a single treatment.

Gender differences and the role of estrogen in cognitive enhancements with nicotine in rats.

Research has reported that nicotine can increase accuracy, response times and rates of learning with evidence of different effects on males and females. The goal of our research was to study further sex differences by examining the role played by estrogen in the effects of nicotine on learning and memory in female rats. In experiment 1, 48 male and female rats were administered 0.3 mg or 0.7 mg/kg bwt of nicotine (nic) or vehicle only (veh) and tested in a visual spatial orientation (VSO) paradigm designed to maximize the benefits of nicotine on spatial working memory. Females exposed to 0.3 mg nic performed superior to all other groups of both genders. In experiment 2, ovariectomized females (N=40) were exposed to 30 microg estradiol/kg bwt (E2), 3 mg nicotine/kg bwt, a combination of both E2 and nic, or veh, and tested as in experiment 1. The rankings of scores in the VSO task by group were E2+nic>nic alone>E2 alone>veh. The E2+nic combination group also demonstrated the highest rate of acquisition. Collectively, the findings suggest that estrogen can synergize the ability of chronic nicotine to enhance acetylcholine-hippocampal interactions underlying performance in the VSO paradigm.

Low doses of memantine disrupt memory in adult rats.

Memantine, a drug recently approved for treatment of Alzheimer's disease, has been characterized as a unique NMDA antagonist that confers protection against excitotoxic neurodegeneration without the serious side effects that other NMDA antagonists are known to cause. In the present study, we determined what dose of memantine is required to protect the adult rat brain against an NMDA receptor-mediated excitotoxic process and then tested that dose and a range of lower doses to determine whether the drug in this dose range is associated with significant side effects. Consistent with previous research, we found that memantine confers a neuroprotective effect beginning at an intraperitoneal dose of 20 mg/kg, a dose that we found, contrary to previous reports, produces locomotor disturbances severe enough to preclude testing for learning and memory effects. We then determined that, at intraperitoneal doses of 10 and 5 mg/kg, memantine disrupts both memory and locomotor behaviors. Rats treated with these doses performed at control-like levels in learning a hole-board task but were significantly impaired in demonstrating what they had learned when tested 24 h later. This impairment of memory retention was not state dependent in that it was demonstrable regardless of whether the rats were or were not exposed to memantine on the day of retention testing. We conclude that, in the adult rat, memantine behaves like other NMDA antagonists in that it is neuroprotective only at doses that produce intolerable side effects, including memory impairment.

Limits of learning enhancements with nicotine in old male rats.

Findings with young adult humans and animal models suggest that nicotine may serve both neuroprotective and cognition enhancing roles in old animals. A pair of experiments was conducted to examine drug-induced modification of the cholinergic nicotinic receptor subtype on rates of learning by young and aged rats. In experiment I males (4-7 months or 20-25 months old) were administered nicotine (0.0, 0.3 or 0.7 mg/kg injected s.c. daily) and tested in both a T-maze non-spatial discrimination paradigm and a hole board spatial task. Nicotine failed to improve acquisition by young animals on either task. Nicotine also failed to improve non-spatial learning by old animals. However, both dosages of nicotine improved performance by the old males in the spatial paradigm. In experiment II, a 5-choice serial discrimination paradigm designed to better evaluate visual attention and spatial working memory in aging was used. Groups of old male rats were administered nicotine or mecamylamine (2 or 8 mg/kg), an antagonist of the nicotinic cholinergic receptor. Results were that the 0.3 mg nicotine group learned the task fastest and achieved the highest learning asymptote. Both learning rates and final levels of performance were worst in the 8 mg mecamylamine group. However, the 2 mg mecamylamine rats were the equals of the control group and both reached a higher asymptote than the 0.7 mg nicotine group. These data suggest that healthy old animals can accrue benefits from nicotinic activation but that the benefits are complex, being limited to certain dosages and to specific cognitive skills.

Behaviorally timid rats respond differentially to conventional and atypical neuroleptics.

An inhibited temperament can be manifested as simple shyness or as social phobia and is perhaps related to the extreme social dysfunction often accompanying schizophrenia. Here, we present a methodology for selecting subjects and testing changes in social attraction in an animal model of behavioral timidity. In Experiment 1, randomly selected female rats were chronically administered either vehicle only, the conventional neuroleptic haloperidol (0.1 mg/kg) or atypical drugs sulpiride (65 mg/kg) or clozapine (18 mg/kg). The animals were tested over 3 weeks for changes in attraction to a social stimulus. Findings revealed a statistically significant decrease in social investigation in the haloperidol treated animals compared to controls but no significant differences among the other groups. Experiment 2 employed pretests to select behaviorally timid (BT) animals. Only female rats having little initial attraction to unfamiliar non-social and social stimuli were chosen to serve as subjects for the experiment using the same drug exposure regiments and behavioral measures used in experiment 1. Results with pre-selected BT animals indicated that clozapine treated animals significantly increased social investigation whereas chronic exposure to either sulpiride or haloperidol groups did not increase social investigation. Indeed, haloperidol appears to have magnified avoidance of social contact. That there were minimal differences between drug groups on a measure of non-social general activity points to the beneficial increases in investigation from clozapine being specific to social inhibition. Conclusions are that timidity may involve aspects of the serotonergic system uniquely influenced by clozapine, and the animal model of the second experiment may prove useful for studies of the biological underpinnings of behavioral timidity.

Physiology after pediatric splenic injury.

BACKGROUND: The splenic injury computed tomographic (CT) grade is used to guide nonoperative management. A study was conducted to determine whether this grade correlates with patient physiology. METHODS: Records of consecutive children with isolated spleen injuries were reviewed. Vital signs, fluids administered, urine output, and hematocrit values from the scene through 120 hours after admission were retrieved. A blinded radiologist reviewed CT scans. Statistical analyses were conducted to test for a linear relationship between injury grade and physiologic parameters. RESULTS: Eighty-two patients with isolated splenic injuries and available CT scans were located. CT injury grade correlated directly with pulse, systolic blood pressure, and diastolic blood pressure and inversely with hematocrit. No correlation was found with pulse pressure, urine output, or maximum temperature. CONCLUSION: The CT grade of splenic injury correlates directly with pulse, systolic blood pressure, and diastolic blood pressure and inversely with hematocrit. CT injury grade correlates with physiologic impact and may guide management decisions.

Chronic fluoxetine suppresses circulating estrogen and the enhanced spatial learning of estrogen-treated ovariectomized rats.

We are interested in developing animal models to evaluate cognitive processes as influenced by the interplay of steroidal hormones and drugs commonly used in psychotherapy. Two experiments with female rats were conducted to evaluate the interaction of estrogen with the serotonin specific reuptake inhibitor (SSRI) fluoxetine on spatial learning and memory and on the endocrine system. In experiment 1, estrogen (50 microg estradiol benzoate/kg body weight) was administered SC to young adult, ovariectomized (OVX) rats either alone or in combination with fluoxetine (2 mg/kg SC). After a month, the groups were compared with appropriate OVX and gonadally intact controls on trials to criterion in a hole board spatial memory task using massed training trials. Experiment 2 was a dose-response study of the influence of fluoxetine (0.5-5 mg/kg) on circulating estrogen in OVX, estrogen treated females. Results were that the OVX females administered estrogen only reached the learning criterion significantly faster than the other groups. All other groups, including the estrogen + fluoxetine animals, performed no better than the controls. Combining fluoxetine with estrogen also lowered circulating estrogen titers, with the least estrogen reductions being in the group receiving the highest dosage of fluoxetine. No differences among groups were found on measures of activity in an open field or for anxiety in a plus maze. Conclusions were that administration of estrogen improved spatial learning and memory in OVX rats, whereas concurrent fluoxetine exposure suppressed the levels of estrogen in circulation and eliminated the gains in spatial performance obtained from chronic estrogen exposure.

Cellular structure and ultrastructure of the black coral Antipathes aperta: 2. The gastrodermis and its collar cells.

The gastrodermis of the black coral Antipathes aperta is associated with eight distinct types of cells, including two types of microbasic b-mastigophores (nematocysts), spumous and vesicular mucus cells, and ganglion cells that are essentially the same as in the epidermis. Three additional types of cells are unique to the gastrodermis, and are readily distinguished from those of the epidermis by their electron-opaque inclusions. These include lipoidal cells, zymogen digestive cells, and an unusual type of epitheliomuscular collar cell. The pharyngeal region is characterized by the presence of electron-opaque nematocysts, a scattering of zymogen cells, and a large number of collar cells. The latter are distinguished in part by the presence of dense microfibrillar processes that surround the microvilli and extend into adjacent collars. This interconnection results in the formation of an extensive pharyngeal meshwork. These collar cells are additionally distinguished by the placement of the collar and flagellum adjacent to a flared cup of cytoplasm. This portion of the cell is capable of endocytosis of relatively large unicellular prey, and apparently is capable of forming digestive vesicles as well. The pharyngeal gastrodermis grades into simple lobate septal filaments toward the base of the coelenteron, where large, granular nematocysts all but replace the smaller electron-opaque types Collar cells are found here as well, but in fewer numbers compared to the zymogen cells. Ultrastructural results are compared with those of other coelenterates and discussed in terms of food and modes of nutrition.

Cellular structure and ultrastructure of the black coral Antipathes aperta: 1. Organization of the tentacular epidermis and nervous system.

The tentacular epidermis of the black coral Antipathes aperta is organized into three distinct regions, containing at least nine different types of cells. The outermost region is dominated by spirocytes along with two types of nematocytes, organized into discrete wart-like batteries. The two nematocyte types both contain microbasic b-mastigophore nematocysts. The outer boundary of the wart is marked by the presence of both spumous and vesicular mucus cells. The ciliation of the wart is contributed principally by the spirocytes. Warts are enveloped and separated from one another by an unusual neurosensory cell complex that extends from the tentacular surface to the mesogleal connective tissue foundation. Funnel-like, flagellated cells composing the complex connect with ganglion cells composing the dominant portion of the nerve net system. Branches of this complex also penetrate the central portion of the wart, making direct contact with the cnidae. The tentacular mid-region is composed of nematocytes and spirocytes in various stages of maturation, along with epitheliomuscular cell (EMC) somata. The EMC's narrow apically extend toward the tentacle surface, forming contacts with the cnidae. The basal end of the EMC expands to form the larger portion of the tentacular musculature. The inner region of tentacular epidermis is marked by a neuromuscular complex sheathed by extensions of mesoglea. The ganglion cells occur as a plexus deep within the tentacle and form polarized junctions with the EMC's, but neuromuscular synapses are not well enough defined for documentation. Polarized synapses lacking well-defined membrane thickenings characterize the interneuronal junctions. Granular cells lining the mesogleal surface appear to be responsible for mesogleal fibrillogenesis.