Chemical Lesions Research Articles

P.1.c.002 Biological peculiarities of the analgesic drug – nefopam in rats

Previous findings indicate that the brain stem descending system becomes more active in modulating spinal nociceptive processes during the development of persistent pain. The present study further identified the supraspinal sites that mediate enhanced descending modulation of behavior hyperalgesia and dorsal horn hyperexcitability (as measured by Fos-like immunoreactivity) produced by subcutaneous complete Freund's adjuvant (CFA). Selective chemical lesions were produced in the nucleus raphe magnus (NRM), the nuclei reticularis gigantocellularis (NGC), or the locus coeruleus/subcoeruleus (LC/SC). Compared to vehicle-injected animals with injection of vehicle alone, microinjection of a serotoninergic neurotoxin 5,7-dihydroxytryptamine into the NRM significantly increased thermal hyperalgesia and Fos protein expression in lumbar spinal cord after hindpaw inflammation. In contrast, the selective bilateral destruction of the NGC with a soma-selective excitotoxic neurotoxin, ibotenic acid, led to an attenuation of hyperalgesia and a reduction of inflammation-induced spinal Fos expression. Furthermore, if the NGC lesion was extended to involve the NRM, the behavioral hyperalgesia and CFA-induced Fos expression were similar to that in vehicle-injected rats. Bilateral LC/SC lesions were produced by microinjections of a noradrenergic neurotoxin, DSP-4. There was a significant increase in inflammation-induced spinal Fos expression, especially in the ipsilateral superficial dorsal horn following LC/SC lesions. These results demonstrated that multiple specific brain stem sites are involved in descending modulation of inflammatory hyperalgesia. Both NRM and LC/SC descending pathways are major sources of enhanced inhibitory modulation in inflamed animals. The persistent hyperalgesia and neuronal hyperexcitability may be mediated in part by a descending pain facilitatory system involving NGC. Thus, the intensity of perceived pain and hyperalgesia is fine-tuned by descending pathways. The imbalance of these modulating systems may be one mechanism underlying variability in acute and chronic pain conditions.

Inhibition of corneal neovascularization after alkali burn: comparison of different doses of bevacizumab in monotherapy or associated with dexamethasone

To compare the effects of different doses of bevacizumab with both saline and dexamethasone on inflammatory angiogenesis in the rat cornea induced by small chemical lesions. Corneal chemical cauterization was performed on 24 rats. Animals were divided randomly into six groups and received a daily subconjunctival injection for 7 days of: balanced salt solution 0.1 mL or dexamethasone phosphate 4 mg/day or bevacizumab 2.5 mg/day, 3.75 mg/day, 5.0 mg/day or bevacizumab 5.0 mg/day + dexamethasone phosphate 4 mg/day. Clinical examination under slitlamp was performed daily for 7 days to evaluate corneal opacity and vessel size evolution. Computer assisted quantitative image analysis was used to measure the total corneal area covered by neovascularization. At final examination, the dexamethasone, bevacizumab 5.0 mg/day and dexamethasone + bevacizumab groups showed a significant lowering in corneal opacity score as compared with control (P = 0.024, P = 0.006 and P = 0.013, respectively). Also, a significant reduction on new vessels size score was observed. Surface of corneal neovascularization was significantly reduced in dexamethasone, bevacizumab 5.0 mg/day and dexamethasone + bevacizumab groups compared with control (P =0.045, P = 0.047 and P = 0.044, respectively). Our study demonstrates the ability of a 5.0 mg/day bevacizumab subconjunctival injection, in monotherapy or associated with dexamethasone, to cause a short-term involution of corneal neovascularization after corneal alkali burn. Combination of both of these treatments may have advantages to monotherapy approaches.

Regulation of subthalamic neuron activity by endocannabinoids

High levels of anandamide are located in the basal ganglia. The subthalamic nucleus (STN) is considered to be an important modulator of basal ganglia output. The present study aims at characterizing the modulation of the electrical activity of STN neurons by exogenous anandamide or endocannabinoids. Single-unit extracellular recordings in anesthetized rats and patch-clamp techniques in rat brain slices containing the STN were performed. Immunohistochemical assays were used. In vivo, anandamide administration produced two opposite effects (inhibition or stimulation) on STN neuron firing rates, depending of the precise location of the neuron within the nucleus. These effects were enhanced by prior inhibition of fatty acid amide hydrolase with URB597, but not by the inhibitor of carrier-mediated anandamide transport AM404. Rimonabant, a specific CB(1) receptor antagonist, also produced inhibition or stimulation of STN neuron activity when administered alone or after anandamide. These effects seem to be mediated by indirect mechanisms since: (1) STN neuron activity is not modified by the cannabinoid agonist Delta(9)-tetrahydrocannabinol (Delta(9)-THC) in vitro; (2) no depolarization-induced suppression of inhibition phenomena were observed; and (3) CB(1) receptor immunolabeling was not detected in the STN, but was abundant in areas which project efferents to this nucleus. Moreover, chemical lesion of the globus pallidus abolished the stimulatory effect of anandamide and microinfusion of anandamide into the prefrontal cortex led to inhibition of STN neuron activity. The present results show that endocannabinoids exert a tonic control on STN activity via receptors located outside the nucleus. These findings may contribute to enhance our understanding of the role of the endocannabinoid system in motor control.

Role of adrenoceptors in the regulation of dopamine/DARPP‐32 signaling in neostriatal neurons

Studies in animal models of Parkinson's disease have revealed that degeneration of noradrenaline neurons is involved in the motor deficits. Several types of adrenoceptors are highly expressed in neostriatal neurons. However, the selective actions of these receptors on striatal signaling pathways have not been characterized. In this study, we investigated the role of adrenoceptors in the regulation of dopamine/dopamine- and cAMP-regulated phosphoprotein of M(r) 32 kDa (DARPP-32) signaling by analyzing DARPP-32 phosphorylation at Thr34 [protein kinase A (PKA)-site] in mouse neostriatal slices. Activation of beta(1)-adrenoceptors induced a rapid and transient increase in DARPP-32 phosphorylation. Activation of alpha(2)-adrenoceptors also induced a rapid and transient increase in DARPP-32 phosphorylation, which subsequently decreased below basal levels. In addition, activation of alpha(2)-adrenoceptors attenuated, and blockade of alpha(2)-adrenoceptors enhanced dopamine D(1) and adenosine A(2A) receptor/DARPP-32 signaling. Chemical lesioning of noradrenergic neurons mimicked the effects of alpha(2)-adrenoceptor blockade. Under conditions of alpha(2)-adrenoceptor blockade, the dopamine D(2) receptor-induced decrease in DARPP-32 phosphorylation was attenuated. Our data demonstrate that beta(1)- and alpha(2)-adrenoceptors regulate DARPP-32 phosphorylation in neostriatal neurons. G(i) activation by alpha(2)-adrenoceptors antagonizes G(s)/PKA signaling mediated by D(1) and A(2A) receptors in striatonigral and striatopallidal neurons, respectively, and thereby enhances D(2) receptor/G(i) signaling in striatopallidal neurons. alpha(2)-Adrenoceptors may therefore be a therapeutic target for the treatment of Parkinson's disease.

Rapid Degeneration and Regeneration of the Zebrafish Olfactory Epithelium after Triton X-100 Application

The effects of Triton X-100 on the olfactory epithelium (OE) of adult zebrafish were examined to study neuronal turnover in this model system. Fish were killed at various time points after detergent application and stained with hematoxylin and eosin to examine olfactory structures, immunocytochemistry to examine cell types, or DiI to examine connections to the olfactory bulb. A significant decrease in epithelial thickness of treated sides was observed 1-day posttreatment. Epithelium thickness recovered by 5 days. The most significant reduction in the OE following Triton X-100 treatment corresponded to the region of supporting cells and mature olfactory sensory neurons. Labeling for all neurons with anti-Hu and for the 3 sensory neuron subtypes of the zebrafish OE (ciliated, microvillous, and crypt neurons) diminished 1 day after lesion and returned by 5 days posttreatment. Retrograde labeling from the olfactory bulb showed that the majority of mature olfactory sensory neurons disappeared in 1 day and reappeared by 5 days after treatment. Anti-proliferating cell nuclear antigen was used to show mitotic activity, and after chemical lesion, there was an increase in proliferation in specific regions of the OE. Thus, chemical ablation causes temporary reduction with swift regeneration of the OE occurring within a week.

Activation of neurons in the hypothalamic dorsomedial nucleus via hypothalamic projections of the nucleus of the solitary tract following refeeding of fasted rats

We report that satiation evokes neuronal activity in the ventral subdivision of the hypothalamic dorsomedial nucleus (DMH) as indicated by increased c-fos expression in response to refeeding in fasted rats. The absence of significant Fos activation following food presentation without consumption suggests that satiation but not craving for food elicits the activation of ventral DMH neurons. The distribution pattern of the prolactin-releasing peptide (PrRP)-immunoreactive (ir) network showed remarkable correlations with the distribution of activated neurons within the DMH. The PrRP-ir fibers and terminals were immunolabeled with tyrosine hydroxylase, suggesting their origin in lower brainstem instead of local, hypothalamic PrRP cells. PrRP-ir fibers arising from neurons of the nucleus of the solitary tract could be followed to the hypothalamus. Unilateral transections of these fibers at pontine and caudal hypothalamic levels resulted in a disappearance of the dense PrRP-ir network in the ventral DMH while PrRP immunoreactivity was increased in transected fibers caudal to the knife cuts as well as in perikarya of the nucleus of the solitary tract ipsilateral to the transections. In accord with these changes, the number of Fos-expressing neurons following refeeding declined in the ipsilateral but remained high in the contralateral DMH. However, the Fos response in the ventral DMH was not attenuated following chemical lesion (neonatal monosodium glutamate treatment) of the hypothalamic arcuate nucleus, another possible source of DMH inputs. These findings suggest that PrRP projections from the nucleus of the solitary tract contribute to the activation of ventral DMH neurons during refeeding, possibly by transferring information on cholecystokinin-mediated satiation.

Two opposite effects of Δ9-tetrahydrocannabinol on subthalamic nucleus neuron activity: Involvement of GABAergic and glutamatergic neurotransmission

Activation of CB1 cannabinoid receptors in the basal ganglia interferes with movement regulation. The aim of this study was to characterize the effect of Delta(9)-tetrahydrocannabinol (Delta(9)-THC) on neurons in the subthalamic nucleus (STN) and to elucidate the mechanisms involved in this effect using single-unit extracellular recordings in anesthetized rats. Administration of Delta(9)-THC (0.25-2 mg/kg, i.v.) stimulated (by 107% +/- 32%) neurons mainly recorded in the ventromedial portion of the caudal STN, whereas it inhibited (by 65% +/- 4%) neurons recorded in the dorsolateral portion of the rostral STN. The CB1 receptor antagonist rimonabant (1 mg/kg, i.v.) completely reverted these effects. The excitatory effect of Delta(9)-THC on STN neurons was not observed after antagonism of GABA(A) receptors by bicuculline administration (10 ng, icv.) or after chemical lesion of the globus pallidus with ibotenic acid. The inhibitory effect was abolished when excitatory amino acid receptors were blocked by kynurenic acid (0.5 mumol, icv.). These results indicate that CB1 receptor activation modulates STN neuron activity by indirect mechanisms involving glutamatergic and GABAergic neurotransmission.

Modulation of excessive neuronal activity by fibroblasts: Potential use in treatment of Parkinson's disease

A number of neurological disorders are marked by increased or aberrant frequency of neuronal discharge in specific parts of the brain. Administration of drugs such as antiepileptic compounds results in the depression of neuronal activity in the whole brain, with the potential for serious side-effects. In the search for additional therapies to reduce the unphysiological electrical activity of over-active brain foci, we have examined the effect of fibroblasts transplanted to areas responsible for motor dysfunction in hemi-parkinsonian rats, since bursting synchronous discharges in internal segment of globus pallidus (GPi) are thought to be partially responsible for the movement disorders of PD. Fibroblasts express gap junctions and ion channels, and so, when transplanted to brain tissue, can potentially modulate excessive electrical activity. Neonatal cortical neurons were cultured on multi-electrode arrays, and their electrical activity was evaluated before and after fibroblast seeding. Unilateral 6-hydroxydopamine (6-OHDA) lesion was carried out in Fischer rats. Lesioned or control rats were transplanted with either syngeneic dermal fibroblasts, microfine glass beads, ibotenic acid, or physiological saline, in the entopeduncular nucleus (EP). Apomorphine-induced rotational behavior and L-dopa-induced dyskinetic movements were evaluated before transplantation (baseline) and 2, 4, 8, 12, and 24 weeks following transplantation. Following behavioral experiments, rats were perfused with 4% formaldehyde in PBS for immunohistochemical study of the brain. We demonstrate in vitro that the introduction of fibroblasts into a network of neurons does not interfere with overall functional measures of activity, while moderately altering the characteristics of synchronous neuronal discharge. In rats with unilateral 6-hydroxydopamine lesions of the nigro-striatal dopaminergic pathway, apomorphine-induced rotations were reduced by more than 60% following ipsilateral transplantation of fibroblasts to the EP. L-Dopa-induced dyskinesia was also significantly reduced. Transplantation of inert microspheres, or chemical lesion of the same area with ibotenic acid, did not produce beneficial effects on parkinsonian symptomatology. Fibroblast transplantation could be an alternative treatment strategy for the parkinsonian patient.

Role of Locus coeruleus noradrenergic neurons in cardiorespiratory and thermal control during hypoxia

We investigated the specific role of LC noradrenergic (NA) neurons in the cardiorespiratory and thermal responses to hypoxia. Aiming this, LC NA neurons were selectively destroyed by microinjecting 6-OHDA bilaterally in the LC of male rats. Pulmonary ventilation (VE, plethysmograph), core body temperature (Tc, dataloggers), mean arterial pressure (MAP) and heart rate (fH) were measured during normoxia followed by 7% O(2) in air. Tyrosine hydroxylase (TH) immunoreactivity was performed to verify the chemical lesions effectiveness. Hypoxia caused increase in VE, decrease in Tc and did not affect fH in both sham-operated and 6-OHDA-lesioned groups. In addition, hypoxia decreased MAP in the sham-operated group and did not affect MAP in the 6-OHDA-lesioned group. LC lesion did not affect VE, MAP, fH and Tc. Thus, these results indicate that LC noradrenergic neurons do not play a role in cardiorespiratory control and thermoregulation under basal and hypoxic conditions.

Ichthyosiform Irritant Contact Dermatitis Caused by Antiseptics for Personal Hygiene

Background: Antiseptics are commonly used for personal hygiene in Taiwan. We have previously reported a series of ichthyosiform irritant contact dermatitis induced by cetrimide-containing antiseptics. Objectives: To report a new series of antiseptics-induced ichthyosiform irritant contact dermatitis and bring attention to the potential irritation effects of antiseptics. Methods: We reviewed the clinicopathologic features of cases of cetrimide dermatitis, Savlon dermatitis or ichthyosiform irritant contact dermatitis diagnosed in our department (October 1993-August 2008). Cases with characteristic ichthyosiform, scarlet to dusky red or chemical burn-like skin lesions, primarily affecting the flexures and/or genital areas, and relevant exposure histories were included. Results: There were 35 patients (14 men and 21 women), aged from 3 to 79 years (mean 45 years) Most patients were seen in summer with a recognizable underlying pruritic dermatosis, and had used commercial products containing cetrimide. The exposure time was from 4 days to 2 months (mean 3.4 weeks). The great majority (80%) had moderate or severe skin lesions characterized by scarlet to dusky red or brownish lesions with a collodion membrane-like surface, and showed a preferential involvement of the flexural and genital areas, especially the groin and axilla where the lesions often manifested centrifugal rings. In no case was antiseptics-related dermatitis suspected by the clinicians or the patients. Skin biopsy was performed in 5 patients; all revealed changes consistent with cetrimide ichthyosiform irritant contact dermatitis. Conclusions: Our study showed that self-application of cetrimide-containing antiseptics is still an important cause of irritant contact dermatitis in southern Taiwan. This dermatitis should be differentiated from ichthyosis, tinea circinata, erythema annulare centrifugum, and nutritional deficiency diseases. Recognition of the clinical characteristics can prompt clinicians to inquire about exposure to antiseptics and facilitate correct diagnosis.

Descending glutamatergic pathways of PFC are involved in acute and chronic action of methylphenidate

Progressive augmentation of behavioral response following repeated psychostimulant administrations is known as behavioral sensitization, and is an indicator of a drug's liability for abuse. It is known that methylphenidate (MPD) (also known as Ritalin), a drug used to treat attention-deficit hyperactivity disorder (ADHD), induces sensitization in animals following repeated injections. It was recently reported that bilateral electric (non-specific) lesion of prefrontal cortex (PFC) prevented MPD elicited behavioral sensitization. Since PFC sends glutamatergic afferents to both ventral tegmental area (VTA) and nucleus accumbens (NAc), sites that are involved in induction and expression of behavioral sensitization respectively and glutamate from PFC is known to modulate dopamine cell activity in VTA and NAc, this study investigated the role of descending glutamate from PFC in MPD elicited behavioral sensitization. Locomotor activity of three groups of rats–control, sham operated and group with specific chemical lesion of glutamate neurons of PFC–was recorded using an open-field assay. On experimental day (ED) 1, the locomotor activity was recorded post a saline injection. The sham and lesion groups underwent respective surgeries on ED 2, and were allowed to recover for 5 days (from ED 3 to ED 7). The post-surgery baseline was recorded on ED 8 following a saline injection. On ED's 9 through 14, 2.5 mg/kg MPD was given, followed by a 4-day washout period (ED 15 -18). All three groups received a rechallenge injection of 2.5 mg/kg on ED 19 and their locomotor activity on various days was analyzed. It was found that ibotenic acid lesion modulated the acute and chronic effects of MPD and hence suggests that PFC glutamatergic afferents are involved in the acute effect of MPD as well as in its chronic effects such as behavioral sensitization to MPD.

Evidence for Developmentally Regulated Local Translation of Odorant Receptor mRNAs in the Axons of Olfactory Sensory Neurons

Odorant receptor mRNAs are transported within axons of olfactory sensory neurons that project into the olfactory bulb. Odorant receptor proteins have been identified along the distal part of these axons, which raises the possibility of their local synthesis in axons. We took advantage of the anatomical separation between the olfactory mucosa (which contains the sensory neuron cell bodies) and the bulb (which contains sensory axons but no sensory neuron cell bodies) to address this issue using a quantitative biochemical approach. Combining a method that separates polysome-associated mRNAs from untranslated mRNAs with a reverse transcription-quantitative PCR approach, we demonstrate that significant amounts of odorant receptor mRNAs are associated with polysomes in the sensory axons of the adult mouse bulb. We thus provide the first evidence for local synthesis of odorant receptor proteins in these axons. Interestingly, the rate of odorant receptor mRNA translation in axons is significantly greater during periods when the proportion of immature axons is higher (i.e., at postnatal day 4 or on regeneration after chemical lesion in adults). In contrast, the olfactory marker protein mRNA, which is restricted to mature axons, is translated at a low and constant level. Overall, we demonstrate that translation levels of odorant receptor mRNAs in axons are developmentally regulated, and positively correlated to the stage of axonal growth into the bulb. Given the established function of odorant receptors in the axonal wiring of sensory projections, we propose that this regulated axonal translation may play a role in the development and maintenance of the glomerular array.

Reversal of acute streptozotocin-induced glycemic changes by chemical ventro-medial hypothalamic lesions.

This report describes the effect of chemical gold thioglucose (GTG) lesions of the ventromedial hypothalamic (VMH) nuclei in male Sprague-Dawley rats previously made diabetic by injection of streptozotocin (50 mg/kg, i.v.). Injection of streptozotocin resulted in hyperglycemia (blood glucose greater than 200 mg%) within one week; treatment of these rats with intracerebroventricular (ICV) injections of GTG (0.025 M) + Insulin (1 microU) reversed these glycemic changes within six weeks. On the basis of these initial experiments, it has been shown that GTG lesions of the VMH may reverse glycemic changes in acute streptozotocin-induced diabetes.

Mechanisms of Base Selection by Human Single-stranded Selective Monofunctional Uracil-DNA Glycosylase

hSMUG1 (human single-stranded selective monofunctional uracil-DNA glyscosylase) is one of three glycosylases encoded within a small region of human chromosome 12. Those three glycosylases, UNG (uracil-DNA glycosylase), TDG (thymine-DNA glyscosylase), and hSMUG1, have in common the capacity to remove uracil from DNA. However, these glycosylases also repair other lesions and have distinct substrate preferences, indicating that they have potentially redundant but not overlapping physiological roles. The mechanisms by which these glycosylases locate and selectively remove target lesions are not well understood. In addition to uracil, hSMUG1 has been shown to remove some oxidized pyrimidines, suggesting a role in the repair of DNA oxidation damage. In this paper, we describe experiments in which a series of oligonucleotides containing purine and pyrimidine analogs have been used to probe mechanisms by which hSMUG1 distinguishes potential substrates. Our results indicate that the preference of hSMUG1 for mispaired uracil over uracil paired with adenine is best explained by the reduced stability of a duplex containing a mispair, consistent with previous reports with Escherichia coli mispaired uracil-DNA glycosylase. We have also extended the substrate range of hSMUG1 to include 5-carboxyuracil, the last in the series of damage products from thymine methyl group oxidation. The properties used by hSMUG1 to select damaged pyrimidines include the size and free energy of solvation of the 5-substituent but not electronic inductive properties. The observed distinct mechanisms of base selection demonstrated for members of the uracil glycosylase family help explain how considerable diversity in chemical lesion repair can be achieved.

The anterior cingulate cortex is involved in retrieval of long-term/long-lasting but not short-term memory for step-through inhibitory avoidance in rats

It is known that the anterior cingulate cortex (ACC) is involved in the formation of contextual fear memory. It has been shown that the ACC is important for the retrieval of long-term contextual fear memory. In order to further examine the role of the ACC in fear memory, we investigated the effects of chemical lesion to or reversible inactivation of the ACC on the retrieval of long-term and short-term step-through inhibitory avoidance (IA) memory. Chemical lesion to the ACC by quinolinic acid severely impaired the retrieval of 15-day and 29-day memories for one-trial step-through IA. Pre-retrieval inactivation of the ACC by locally infusing muscimol, a selective GABA A receptor agonist, produced a severe deficit in 7-day, 4-day and 1-day IA memories, with no effect on 2-h and 6-h memories. Thus, the ACC is required for the retrieval of long-term/long-lasting IA memory, but is dispensable for short-term one.

Effects of chemical lesion of Nucleus Raphe Obscurus on the thermoregulatory and ventilatory responses to hypoxia

The role of Nucleus Raphe Obscurus (ROb) on the hypoxic ventilatory response (HVR) and anapyrexia is not yet understood. Our objective was to study the effects of chemical lesion of ROb on ventilatory and thermoregulatory responses to hypoxia. Seven days before the experiments, male Wistar rats received a microinjection of ibotenic acid (0.2 ug/0.1 uL) or vehicle (PBS) into the ROb. The Fluoro‐Jade (FJ) histochemistry was used for staining the degenerative neurons (FJ+) in the microinjection site and surroundings, the latter named peri‐ROb group. Pulmonary ventilation (Ve, plethismography) and body temperature (data loggers) were measured followed by 1 h of hypoxic exposure (7% O2). FJ+ROb lesions decreased significantly HVR in the initial stage (from 10 to 20min) of hypoxia exposure (1136±65 mL.kg‐1.min‐1) compared to FJ+peri‐ROb (1315±63 mL.kg‐1.min‐1) and vehicle group (1326±81 mL.kg‐1.min‐1).The decrease in HVR was mediated by respiratory frequency (RF) while tidal volume (VT) did not change. There were no differences in hypoxia‐induced anapyrexia between the three groups. The results suggest that ROb neurons exert an excitatory modulation (acting via RF) playing a role in the early stages of HVR. In the present study, there are no evidences supporting the involvement of ROb in thermoregulatory responses elicited by hypoxia. Financial Support: FAPESP

Hippocampal lesions impair spatial memory performance, but not song—A developmental study of independent memory systems in the zebra finch

Songbirds demonstrate song- and spatial-learning, forms of memory that appear distinct in formal characteristics and fitting the descriptions and criteria of procedural and episodic-like memory function, respectively. As in other vertebrates, the neural pathways underlying these forms of memory may also be dissociable, and include the corresponding song circuit and hippocampus (HP). Whether (or not) these two memory systems interact is unknown. Interestingly, the HP distinguishes itself as a site of immediate early gene expression in response to song and as a site of estrogen synthesis, a steroid involved in song learning. Thus, an interaction between these memory systems and their anatomical substrates appears reasonable to hypothesize, particularly during development. To test this idea, juvenile male or female zebra finches received chemical lesions of the HP at various points during song learning, as did adults. Song structure, singing behavior, song preference, and spatial memory were tested in adulthood. Although lesions of the HP severely compromised HP-dependent spatial memory function across all ages and in both sexes, we were unable to detect any effects of HP lesions on song learning, singing, or song structure in males. Interestingly, females lesioned as adults, but not as juveniles, did lose the characteristic preference for their father's song. Since compromise of the neural circuits that subserve episodic-like memory does very little (if anything) to affect procedural-like (song learning) memory, we conclude that these memory systems and their anatomical substrates are well dissociated in the developing male zebra finch.

Neural Regulation of Ejaculation

Delineation of the underlying neurophysiological mechanisms of ejaculatory behavior is crucial for the treatment of male sexual dysfunction, including premature ejaculation. Recent studies provide compelling evidence that a population of lumbar spinothalamic (LSt) cells may play a role in the regulation of the ejaculatory response. Subsequent to ejaculation, LSt cells exhibit markers of activation that are not only highly correlated with ejaculatory behavior, but are also absent following the expression of other components of sexual behavior, such as mounts or intromissions. Similarly, targeted chemical lesion of LSt cells using substance P-saporin abolishes ejaculatory behavior explicitly. Early evidence suggests that pharmacological manipulation of LSt cells may offer additional evidence of crucial LSt cell involvement in the generation of ejaculation. This review is intended to summarize what has currently been revealed regarding the role of LSt cells in the regulation and generation of ejaculatory behavior, and also to discuss the direction of future behavioral investigations. Information presented in this discussion was derived from analysis of numerous recent articles detailing the delineation of anatomical and physiological correlates of sexual behavior, as well as numerous literature searches using the National Library of Medicine PubMed Services. A great deal of the work that has led to the implication of LSt cells in ejaculatory behavior is reviewed in the present article, including clinical data, as well as anatomical, physiological, and behavioral examinations. The rationale for ongoing pharmacological studies is also discussed. LSt cells appear to play a vital role in the generation and regulation of ejaculatory behavior. Additional elucidation of this "ejaculation generator" could prove invaluable for the future treatment of male sexual dysfunction. Studies are currently in progress to further reveal the precise function of these cells and mechanisms of action through which they operate.

Notch signaling and Hes labeling in the normal and drug-damaged organ of Corti

During the development of the inner ear, the Notch cell signaling pathway is responsible for the specification of the pro-sensory domain and influences cell fate decisions. It is assumed that Notch signaling ends during maturity and cannot be reinitiated to alter the fate of new or existing cells in the organ of Corti. This is in contrast to non-mammalian species which reinitiate Delta1–Notch1 signaling in response to trauma in the auditory epithelium, resulting in hair cell regeneration through transdifferentiation and/or mitosis. We report immunohistochemical data and Western protein analysis showing that in the aminoglycoside-damaged guinea pig organ of Corti, there is an increase in proteins involved in Notch activation occurring within 24 h of a chemical hair cell lesion. The signaling response is characterized by the increased presence of Jagged1 ligand in pillar and Deiters cells, Notch1 signal in surviving supporting cell nuclei, and the absence of Jagged2 and Delta-like1. The pro-sensory bHLH protein Atoh1 was absent at all time points following an ototoxic lesion, while the repressor bHLH transcription factors Hes1 and Hes5 were detected in surviving supporting cell nuclei in the former inner and outer hair cell areas, respectively. Notch pathway proteins peaked at 2 weeks, decreased at 1 month, and nearly disappeared by 2 months. These results indicate that the mammalian auditory epithelium retains the ability to regulate Notch signaling and Notch-dependent Hes activity in response to cellular trauma and that the signaling is transient. Additionally, since Hes activity antagonizes the transcription of pro-sensory Atoh1, the presence of Hes after a lesion may prohibit the occurrence of transdifferentiation in the surviving supporting cells.

Effect of central serotonin depletion on 5‐HT receptor‐mediated vasomotor responses in the middle meningeal artery of anaesthetized rats

1 It has been hypothesized that craniovascular 5-HT receptors mediating dilatation of cranial vessels undergo sensitization on decreased serotonergic transmission in migraine. This study analysed the effect of chemical lesion of the 5-HT system in the brain with 5,7-dihydroxytryptamine (5,7-DHT) on 5-HT receptor-mediated dilator responses to 5-carboxamidotryptamine (5-CT) in the middle meningeal artery of anaesthetized rats. 5-CT has recently been shown to elicit dilator responses in this cranial vessel via 5-HT(7) receptors and, to a much lesser extent, 5-HT(1B/1D) receptors. 2 Pretreatment with 5,7-DHT produced a drastic and selective decrease of 5-HT levels in the brain (78 +/- 6% and 94 +/- 2% in dorsal raphe and hypothalamic paraventricular nuclei, respectively) compared with controls (1% ascorbic acid). 3 Topical application of 5-CT (1-1000 microm) to exposed dura mater encephali produced concentration-dependent decreases in diastolic blood pressure and dilator responses in the middle meningeal artery that were similar in vehicle- and 5,7-DHT-pretreaed animals. 4 Hypotensive and meningeal dilator responses to 5-CT were unaltered by the 5-HT(1B/1D) receptor antagonist, GR-127935 (1 mg kg(-1), i.v.), but were strongly inhibited by the 5-HT(7) receptor antagonist, SB-269970 (1 mg kg(-1), i.v.), with similar efficacy, in both groups of animals. Treatment with GR-127935 + SB-269970 (1 mg kg(-1), i.v. each), produced a stronger inhibitory effect than individual treatments on hypotensive but not on meningeal responses to 5-CT. Meningeal 5-HT(7) receptor-mediated responses (i.e. in GR-127935-pretreated animals) were unchanged by 5,7-DHT pretreatment. 5 Results suggest that the sensitivity of craniovascular 5-HT(7) receptors mediating dilatation is unaffected by a decrease of 5-HT levels in the brain. A neuronal involvement of 5-HT in migraine seems more likely, therefore.