Early microglia activation in a mouse model of chronic glaucoma

Changes in microglial cell activation and distribution are associated with neuronal decline in the central nervous system (CNS), particularly under pathological conditions. Activated microglia converge on the initial site of axonal degeneration in human glaucoma, yet their part in its pathophysiology remains unresolved. To begin with, it is unknown whether microglia activation precedes or is a late consequence of retinal ganglion cell (RGC) neurodegeneration. Here we address this critical element in DBA/2J (D2) mice, an established model of chronic inherited glaucoma, using as a control the congenic substrain DBA/2J Gpnmb(+/SjJ) (D2G), which is not affected by glaucoma. We analyzed the spatial distribution and timecourse of microglial changes in the retina, as well as within the proximal optic nerve prior to and throughout ages when neurodegeneration has been reported. Exclusively in D2 mice, we detected early microglia clustering in the inner central retina and unmyelinated optic nerve regions, with microglia activation peaking by 3 months of age. Between 5 and 8 months of age, activated microglia persisted and concentrated in the optic disc, but also localized to the retinal periphery. Collectively, our findings suggest microglia activation is an early alteration in the retina and optic nerve in D2 glaucoma, potentially contributing to disease onset or progression. Ultimately, detection of microglial activation may have value in early disease diagnosis, while modulation of microglial responses may alter disease progression.     

重组人白细胞介素-2玻璃体注射慢性高眼压大鼠神经节细胞层神经元的凋亡

BACKGROUND: The majority of studies addressing neuroprotection have focused on drugs, proteins and cytokines. A recent study reports that recombinant human interleukin-2 (rhIL-2) achieves good therapeutic effects, but the underlying mechanisms remain poorly understood. OBJECTIVE: To observe the neuroprotection of recombinant human interleukin-2 (rhIL-2) by intravitreous injection on the retinal ganglion cells (RGCs) following chronic elevated IOP model in Wistar rats. DESIGN, TIME AND SETTING: A randomized, controlled, animal experiment was performed at the Center Laboratory, Second Xiangya Hospital, Central South University, China, from July 2008 to March 2009. MATERIALS: LY294006 and AG490 were provided by Invitrogen, USA. Rabbit anti-AKT1 polyclonal antibody and rabbit anti-STAT3 polyclonal antibody were purchased from American Basic Gene Associate Bioscience, USA. METHODS: Chronic elevated IOP model of Wistar rats were established by cauterizing the limbal episcleral veins to block the reflux of aqueous humor by 532nm viridis-lite diode laser in right eyes. Intravitreous injection of 2µl PBS, rhIL-2 or rhIL-2 LY294002, AG490 mixed liquor for eyes were administered 1 week after chronic elevated IOP model established. Ocular tissue was immunostained to investigate ED-1 expressions. Rats were sacrificed after intravitreous injection 3 days, 14 days, 21 days to identify RGC, and the 3% fluorogold was used to the epithalamus 7 days before sacrifice. Western blot analysis Akt1 and STAT3 protein in the retina after intravitreous injection 3h,12h, 96h. MAIN OUTCOME MEASURES: ED-1 expression of activated microglia/macrophages was determined by immunofluorescence. The RGCs levels were detected using fluorogold staining. The efficacy of the intraocular rhIL-2 and rhIL-2, LY294002, AG490 mixed liquor injections in inhibiting PI3K/Akt、Jak/Stat3 signal transductions were confirmed by Western blot analysis. RESULTS: Numerous ED-1 positive macrophages were seen in the vitreous and along the inner retinal surface in rhIL-2 treatment group. The number of RGCs was much higher than that in control groups and PBS group (P<0.01). The numbers of RGCs in rhIL-2 and LY294002, AG490 mixed liquor group was as much lower than that in rhIL-2 group (P<0.01). Western blotting analysis showed very little Akt1 and STAT3 protein in the retina after 3h intravitreous injections of rhIL-2 in rats, started to increase rapidly at 12 reperfusion,96h decreased slightly. LY294002 and AG490 prevented the Akt1 and STAT3 protein expression in the retina. CONCLUTION: rhIL-2 has neuroprotective effection after intravitreous injection. It can decrease or avoid RGCs layer damage in chronic glaucoma model of rats. Intravitreal injection of rhIL-2 activates macrophages, which derived factors might enhance the RGCs survival. After intravitreal injection of rhIL-2 PI3K/Akt、Jak/Stat3 signal transductions may play an important role following a chronic elevated IOP model, and LY294002, AG490 can block this effect.     

Increased thalamic inhibition in the absence seizure prone DBA/2J mouse

Genetic background plays a significant role in producing variable seizure outcomes in patients and animal models. The neurobiological mechanism underlying this heterogeneity is not clear. Here we compare GABAergic synaptic properties within the thalamocortical circuit of two commonly used inbred mice strains, the C57B/6 and spike-and-wave discharge (SWD) prone DBA/2J. Differences between the strains occur in amplitude, kinetics, and frequency of miniature inhibitory postsynaptic currents (mIPSCs) in a region-specific manner. The biggest difference in synaptic inhibition was seen in the thalamus where DBA/2J mice showed a doubling of mIPSC frequency when compared to C57B/6. EEG analysis revealed higher power in the 6-12 Hz band during non-rapid eye movement sleep in DBA/2J mice. Increased susceptibility of the DBA2/J strain to develop SWDs and increase in the 6-12 Hz EEG power may be due to a hypersynchronous ventrobasal thalamus as a consequence of increased GABAergic input.     

Persistence of intact retinal ganglion cell terminals after axonal transport loss in the DBA/2J mouse model of glaucoma

Axonal transport defects are an early pathology occurring within the retinofugal projection of the DBA/2J mouse model of glaucoma. Retinal ganglion cell (RGC) axons and terminals are detectable after transport is affected, yet little is known about the condition of these structures. We examined the ultrastructure of the glaucomatous superior colliculus (SC) with three‐dimensional serial block‐face scanning electron microscopy to determine the distribution and morphology of retinal terminals in aged mice exhibiting varying levels of axonal transport integrity. After initial axonal transport failure, retinal terminal densities did not vary compared with either transport‐intact or control tissue. Although retinal terminals lacked overt signs of neurodegeneration, transport‐intact areas of glaucomatous SC exhibited larger retinal terminals and associated mitochondria. This likely indicates increased oxidative capacity and may be a compensatory response to the stressors that this projection is experiencing. Areas devoid of transported tracer label showed reduced mitochondrial volumes as well as decreased active zone number and surface area, suggesting that oxidative capacity and synapse strength are reduced as disease progresses but before degeneration of the synapse. Mitochondrial volume was a strong predictor of bouton size independent of pathology. These findings indicate that RGC axons retain connectivity after losing function early in the disease process, creating an important therapeutic opportunity for protection or restoration of vision in glaucoma. J. Comp. Neurol. 524:3503–3517, 2016. © 2016 Wiley Periodicals, Inc.     

Evidence supporting a role of serotonin in modulation of sudden death induced by seizures in DBA/2 mice

PURPOSE: Sudden unexpected death in epilepsy (SUDEP) is a serious concern for epilepsy patients. DBA/2 mice are proposed as a SUDEP model, because these mice exhibit respiratory arrest (RA) after audiogenic seizures (AGSs), and RA is also implicated in human SUDEP. Respiratory mechanisms are modulated, in part, by serotonin. Therefore we evaluated the effects of serotoninergic agents on RA incidence in DBA/2 mice. METHODS: DBA/2 mice (75%) exhibited AGS and RA, and approximately 99% of animals could be resuscitated. The mice exhibiting RA were given a selective serotonin reuptake inhibitor, fluoxetine, 24 h after the initial AGS, and RA susceptibility was evaluated 30 min later. Ten percent of DBA/2 mice exhibited tonic hindlimb extension (TE) without RA, and a serotonin antagonist (cyproheptadine) was administered to these mice. RESULTS: Fluoxetine (15-25 mg/kg, i.p.) significantly reduced the incidence of RA in DBA/2 mice after AGSs, and this effect was reversible by 72 h. Only the 25-mg/kg dose reduced AGS severity. In mice exhibiting TE without RA, the incidence of RA was significantly increased 30 min after cyproheptadine (1-2 mg/kg i.p.). Most of these mice exhibited AGSs without RA again by 72 h. CONCLUSIONS: These findings indicate that fluoxetine reduced RA in DBA/2 mice at doses that did not reduce seizure severity. Because DBA/2 mice are a proposed model for human SUDEP, these data support evaluation of fluoxetine for SUDEP prevention in the patient population most susceptible to SUDEP. The data raise concern about the use of serotonin antagonists in this patient population.     

Retinal ganglion cell death during optic nerve regeneration in the frog Hyla moorei

In the frog Hyla moorei we have estimated there to be between approximately 450,000 and 750,000 cells in the retinal ganglion cell layer. Optic axon counts and retrograde transport of horseradish peroxidase (HRP) indicated that 72-76% of these were ganglion cells. Cells of this type were distributed as a temporally situated area centralis within a horizontal visual streak. Cell and optic axon counts showed that there was an approximately 40% loss of ganglion cells during optic nerve regeneration. Ganglion cells appeared chromatolysed by 6-8 days after an extracranial nerve crush but there was no indication of cell death until 15 days. By this stage anterograde transport of HRP indicated that axons had reached the chiasma. Death was first seen in the area centralis, extended along the streak, and finally was observed in the periphery by 65 days; cell counts demonstrated that at this time the wave of death was almost complete. We have previously shown by electrophysiological visual mapping (Humphrey and Beazley, '82) and confirmed in this study that visuotectal projections were retinotopically organized during regeneration. Multiunit receptive fields were initially large but progressively refined starting in nasal field (temporal retina) to restore a normal projection. The similar sequences whereby the visuotectal projection became refined and death took place in the retinal ganglion cell layer suggested that death may be related to a process of organization within the regenerating projection. In normal animals primary visual pathways revealed by anterograde transport of HRP were essentially similar to those of Rana pipiens and R. esculenta. Regenerating axons generally remained within optic pathways. Exceptions were a retinoretinal projection which was not completely withdrawn even after 1,028 days and a direct projection to the ipsilateral tectum via an inappropriate part of the optic tract.     

Characterization of cortical spindles in DBA/2 and C57BL/6 inbred mice

Continuous twenty-four hour EEG recordings were conducted on freely-moving DBA/2 and C57BL/6 inbred mice. No brief spindle episodes (BSEs: 6-7 cps, 1-5 sec duration, high amplitude spindle bursts) were seen in the waking EEG of C57BL/6 mice. BSEs were a conspicuous element of the EEG during active waking (AW) and quiet waking (QW) in DBA/2 mice. BSEs occurred at a 10X faster rate in QW than in AW and had a longer duration. Sleep spindle bursts resembling BSEs were seen in both C57BL/6 and DBA/2 mice, and occasionally were observed to follow a K-complex. Rostropontine, but not midpontine, brainstem transection released spindles in both strains. Pentobarbital produced spindles in both strains. The waveforms of the waves comprising BSEs, sleep spindles, transection-induced spindles and barbiturate spindles were quite similar, though differing in frequencies and amplitude. Genetic factors may be critical for the lack of BSEs during AW and QW in C57BL/6 mice and for the occurrence of BSEs during AW in DBA/2 mice. In contrast, most other rodents whow a third pattern: BSEs only during QW. Since C57BL/6 mice can generate spindles under some circumstances, the absence of spindles during waking reflects some alteration in the mechanisms that control the initiation of BSEs rather than a lack of the circuits required to generate a BSE. These mechanisms are distinct from those processes of arousal that produce the background EEG desynchronization of waking. Following both rostropontine and midpontine transection, the background EEG is desynchronized, yet after rostropontine, but not midpontine transection, BSEs occur freely, at a rate over 200 per hour.     

Characterization of multiple bistratified retinal ganglion cells in a purkinje cell protein 2‐Cre transgenic mouse line

Retinal ganglion cells are categorized into multiple classes, including multiple types of bistratified ganglion cells (BGCs). The recent use of transgenic mouse lines with specific type(s) of ganglion cells that are labeled by fluorescent markers has facilitated the morphological and physiological studies of BGCs, particularly the directional‐selective BGCs. The most important benefit from using transgenic animals is the capability to perform in vivo gene manipulation. In particular, the Cre/LoxP recombination system has become a powerful tool, allowing gene deletion, overexpression, and ectopic expression in a cell type‐specific and temporally controlled fashion. The key to this tool is the availability of Cre mouse lines with cell or tissue type‐specific expression of Cre recombinase. In this study we characterized the Cre‐positive retinal ganglion cells in a PCP2 (Purkinje cell protein 2)‐cre mouse line. We found that all of the Cre‐positive retinal ganglion cells were BGCs. Based on morphological criteria, we determined that they can be grouped into five types. The On‐ and Off‐dendrites of three of these types stratified outside of the cholinergic bands and differed from directional selective ganglion cells (DSGCs) morphologically. These cells were negative for Brn‐3b and positive for both calretinin and CART retina markers. The remaining two types were identified as putative On‐Off and On‐DSGCs. This Cre mouse line could be useful for further studies of the molecular and functional properties of BGCs in mice. J. Comp. Neurol. 521:2165–2180, 2013. © 2012 Wiley Periodicals, Inc.     

Genetic analysis of cerebellar folial pattern in crosses of C57BL/6J and DBA/2J inbred mice

Variation in the cerebellar folial pattern of mice is influenced by genetic elements [Inouye, M. and Oda, S.,J. Comp. Neurol., 190 (1980) 357–362]. In crosses of C57BL/6J and DBA/2J inbred mice, the presence or absence of a specific fissure, the intraculminate fissure, is largely determined by a single genetic locus (Cfp-1), which is located on distal Chromosome 4 [Neumann et al.,Brain Res., 524 (1990) 85–89]. In the present study, the mid-sagittal cerebellar folial pattern has been examined in crosses of C57BL/6J and DBA/2J mice and in BXD recombinant inbred strains. At least three loci, includingCfp-1, are involved in variation in vermian pattern formation. Genetic variation in thyroid hormone function may be involved in the inheritance of folial pattern. A locus (Cfp-2) that appears to be partially responsible for this negative genetic correlation in mice may be linked toAfp on Chromosome 5. This hypothesis was suggested by the negative correlation between neonatal serum T₄ level and the number of folia in rats given neonatal injections of thyroxine or propylthiouracil [Lauder, J.M. et al.,Brain Res., 76 (1974) 33–40].     

Retinal ganglion cell and nonneuronal cell responses to a microcrush lesion of adult rat optic nerve

Injury of the optic nerve has served as an important model for the study of cell death and axon regeneration in the CNS. Analysis of axon sprouting and regeneration after injury by anatomical tracing are aided by lesion models that produce a well-defined injury site. We report here the characterization of a microcrush lesion of the optic nerve made with 10-0 sutures to completely transect RGC axons. Following microcrush lesion, 62% of RGCs remained alive 1 week later, and 28% of RGCs, at 2 weeks. Optic nerve sections stained by hematoxylin-based methods showed a thin line of intensely stained cells that invaded the lesion site at 24 h after microcrush lesion. The lesion site became increasingly disorganized by 2 weeks after injury, and both macrophages and blood vessels invaded the lesion site. The microcrush lesion was immunoreactive for chondroitin sulfate proteoglycans (CSPG), and an adjacent GFAP-negative zone developed early after the lesion, disappearing by 1 week. Luxol fast blue staining showed a myelin-free zone at the lesion site, and myelin remained distal to the lesion at 8 weeks. To study the axonal response to microcrush lesion, anterograde tracing was used. Within 6 h after injury all RGC axons retracted back from the site of lesion. By 1 week after injury, axons regrew toward the lesion, but most stopped abruptly at the injury scar. The few axons that were able to cross the injury site did not extend further in the optic nerve white matter by 8 weeks postlesion. Our observations suggest that both the CSPG-positive scar and the myelin-derived growth inhibitory proteins contribute to the failure of RGC regeneration after injury.     

Critical neuroprotective roles of heme oxygenase‐1 induction against axonal injury‐induced retinal ganglion cell death

Although axonal damage induces significant retinal ganglion cell (RGC) death, small numbers of RGCs are able to survive up to 7 days after optic nerve crush (NC) injury. To develop new treatments, we set out to identify patterns of change in the gene expression of axonal damage‐resistant RGCs. To compensate for the low density of RGCs in the retina, we performed retrograde labeling of these cells with 4Di‐10ASP in adult mice and 7 days after NC purified the RGCs with fluorescence‐activated cell sorting. Gene expression in the cells was determined with a microarray, and the expression of Ho‐1 was determined with quantitative PCR (qPCR). Changes in protein expression were assessed with immunohistochemistry and immunoblotting. Additionally, the density of Fluoro‐gold‐labeled RGCs was counted in retinas from mice pretreated with CoPP, a potent HO‐1 inducer. The microarray and qPCR analyses showed increased expression of Ho‐1 in the post‐NC RGCs. Immunohistochemistry also showed that HO‐1‐positive cells were present in the ganglion cell layer (GCL), and cell counting showed that the proportion of HO‐1‐positive cells in the GCL rose significantly after NC. Seven days after NC, the number of RGCs in the CoPP‐treated mice was significantly higher than in the control mice. Combined pretreatment with SnPP, an HO‐1 inhibitor, suppressed the neuroprotective effect of CoPP. These results reflect changes in HO‐1 activity to RGCs that are a key part of RGC survival. Upregulation of HO‐1 signaling may therefore be a novel therapeutic strategy for glaucoma. © 2014 Wiley Periodicals, Inc.     

Serotonin transporter, 5‐HT1A receptor,and behavior in DBA/2J mice in comparison with four inbred mouse strains

Prepulse inhibition (PPI), the reduction in acoustic startle produced when it is preceded by a weak prepulse stimulus, is impaired in schizophrenic patients. The DBA/2J mouse strain displayed deficient PPI and is therefore suggested as an experimental animal model for the loss of sensorimotor gating in schizophrenia. Brain serotonin (5‐HT) has been implicated in the pathophysiology of several psychiatric disorders, including major depressive disorder and schizophrenia. In the present study, behavior, 5‐HT transporter (5‐HTT) mRNA level, 5‐HT_1A receptor mRNA level, and 5‐HT_1A receptor density in the brain regions were studied in DBA/2J mice in comparison with four inbred mouse strains (CBA/Lac, C57BL/6, BALB/c, and ICR). A decrease in 5‐HTT mRNA level in the midbrain and a reduced density of 5‐HT_1A receptors in the frontal cortex without significant changes in 5‐HT_1A receptor mRNA level in DBA/2J mice were found. It was shown that, along with decreased PPI, DBA/2J mice demonstrated considerably reduced immobility in the tail suspension test and in the forced swim test. No significant interstrain differences in intermale aggression, or in light‐dark box and elevated plus‐maze tests, were found. The results suggested the involvement of decreased 5‐HTT gene expression and 5‐HT_1A receptor density in genetically defined PPI deficiency and showed a lack of any association between PPI deficiency and predisposition to aggressive, anxiety, and depressive‐like behaviors. © 2009 Wiley‐Liss, Inc.     

Retinal ganglion cell response to nerve growth factor in the regenerating and intact visual system of the goldfishCarassius auratus

Light and electron microscopic observations indicated that 1 μg NGF injected into the goldfish eye at the time of optic nerve crush initiated significant retinal ganglion cell body, nuclear and nucleolar hypertrophy when compared to controls at 7 days post-axotomy (7 DPA). In addition, the ultrastructural morphometric values for NGF-treated retinal ganglion cells at 7 DPA were not significantly different from the normal hypertrophy found in 14 DPA controls. Therefore, it appears that NGF treatment caused an acceleration of the normal cell body response to axotomy by about a week. These responses were found to be specific for the NGF molecule and dose-dependent over a 100–1000 ng NGF concentration range.In contrast to the lesioned state, NGF treatment had no significant influence on intact, non-lesioned retinal ganglion cell morphology as measured by morphometric analysis. These results strongly indicate that the responsiveness of the goldfish retinal ganglion cells to NGF is initiated by the axotomy.Eyes treated with NGF in the same manner that elicited the dramatic retinal ganglion cell morphological changes at 7 DPA could also be shown to significantly increase neurite outgrowth from retinal explants cultured at the same post-operative period (i.e. 7 DPA).     

Aberrant synaptic input to retinal ganglion cells varies with morphology in a mouse model of retinal degeneration

Retinal degeneration describes a group of disorders which lead to progressive photoreceptor cell death, resulting in blindness. As this occurs, retinal ganglion cells (RGCs) begin to develop oscillatory physiological activity. Here we studied the morphological and physiological properties of RGCs in rd1 mice, aged 30–60 days, to determine how this aberrant activity correlates with morphology. Patch‐clamp recordings of excitatory and inhibitory currents were performed, then dendritic structures were visualized by infusion of fluorescent dye. Only RGCs with oscillatory activity were selected for further analysis. Oscillatory frequency and power were calculated using power spectral density analysis of recorded currents. Dendritic arbor stratification, total length, and area were measured from confocal microscope image stacks. These measurements were used to sort RGCs by cluster analysis using Ward's Method. This resulted in a total of 10 clusters, with monostratified and bistratified cells having five clusters each. Both populations exhibited correlations between arbor stratification and aberrant inhibitory input, while excitatory input did not vary with arbor distribution. These findings illustrate the relationship between aberrant activity and RGC morphology at early stages of retinal degeneration. J. Comp. Neurol. 522:4085–4099, 2014. © 2014 Wiley Periodicals, Inc.     

Beta and gamma range EEG power-spectrum correlation with spiking discharges in DBA/2J mice absence model: role of GABA receptors

PURPOSE: To describe the correlations between spiking pattern and EEG power spectrum frequency in DBA/2J mice, a model for murine absence seizures, after gamma-aminobutyric acid (GABA)(B) modulation. METHODS: The animals were first tested with the GABA(B) agonist l-baclofen followed by the GABA(B) antagonist SCH 50911. Moreover, digital EEGs recorded under experimental conditions were processed at baseline and 10 and 20 min after l-baclofen injection. This procedure was followed by injection of the GABA(B) antagonist SCH50911 and by an additional EEG evaluation at 10 and 20 min from drug administration. The power spectra analysis of signals was obtained for delta (0.5-3 Hz), theta (3.5-7.5 Hz), alpha (8-12 Hz), beta (13-20 Hz), and gamma (21-50 Hz) frequencies. RESULTS: The spiking pattern and power spectrum of beta activity was increased by 80%), whereas gamma power increased (correlation, 0.92; p < 0.001). The remaining frequency bands were unaffected. CONCLUSIONS: This study confirms the potential of GABA(B) antagonists in contrasting seizure absence in rodent models and suggests the application of drugs with a similar mechanism in humans. In addition, because GABA(B) antagonists not only contrast seizure in rodent models of absence but also improve "cognitive" performance, it could be hypothesized that gamma increase, correlated with optimized cortical binding during coherent percepts, may produce potential cognition-enhancing effects.     

Audiogenic seizure susceptibility of C57BL/6 ↔ DBA/2 allophenic mice

Allophenic mice composed of cells from a strain (DBA/2) susceptible to sound-induced seizures and cells from a strain (C57BL/6) resistant to them were produced by embryo aggregation techniques. Twenty-eight allophenic mice were tested for audiogenic seizure susceptibility. The results were compared with the genotypic composition of the coat melanocytes. For those animals with a predominance of one genotype or the other in the coat, their seizure phenotype was the same as that of the strain most represented in the coat. In contrast, those animals with major contributions ofboth genotypes in the coat demonstrated the entire spectrum of susceptibility phenotypes. Such results are likely to be a manifestation of a relatively small target tissue for the genes influencing the development of audiogenic seizure susceptibility.     

Retinal ganglion cell loss produced by intraocular kainic acid in cats: variation with somal size and eccentricity

Eight eyes of adult cats were injected with different doses of kainic acid (KA) and examined following survival times of either 5 or 12 days. At a survival time of 12 days, a dose of 76 nmol produced an 18% lossof ganglion cells in the center of the area centralis (AC), 70% loss at a location 2 mm from the AC, and 95% loss at a location 6 mm from the AC. Larger doses (240, 760 and 2400 nmol) produced losses comparable to that observed for 76 nmol. For example, 2400 nmol produced a 35% loss in the AC, 81% at 2 mm, and 88% at 6 mm. At a survival time of 5 days, doses of 240 and 760 nmol produced a loss of ganglion cells comparable to that seen at 12 days. In one eye, a large dose of KA (7600 nmol) produced total loss of ganglion cells at a survival time of 5 days. By comparing loss of cells in restricted somal diameter ranges at different retinal eccentricities, it was possible to distinguish two significant correlations that were largely independent of survival time and dose: (1) at 2 mm, loss of cells with somal diameters larger than 21 μm significantly exceeded loss of cells with smaller somata. In particular, alpha cells were totally eliminated in 6 of the 8 KA-treated eyes. (2) The mean loss of ganglion cells with somal diameters less than 21 μm was significantly greater at 2 mm and 6 mm than in the AC. Together, these results show that loss of ganglion cells produced by KA varies somal size and retinal eccentricity.     

Omega-3 fatty acid deficiency augments amphetamine-induced behavioral sensitization in adult DBA/2J mice: relationship with ventral striatum dopamine concentrations

The principal polyunsaturated fatty acid acids found in brain, arachidonic acid (AA) and docosahexaenoic acid (DHA), preferentially accumulate in synaptic membranes. Although neurochemical studies have found that dietary-induced deficits in rat brain DHA composition significantly alter mesocorticolimbic dopamine (DA) neurotransmission, its impact on DA-mediated behavior remains poorly understood. In the present study, we determined the effects of dietary-induced deficits in brain DHA composition on amphetamine (AMPH)-induced locomotor activity and sensitization in DBA/2J mice, an inbred strain previously found to be hyporesponsive to AMPH, as well as monoamine concentrations in the PFC and ventral striatum following the AMPH challenge. Chronic dietary omega-3 fatty acid deficiency significantly decreased PFC (-25%) and ventral striatum (-20%) DHA composition, increased PFC (+7%) and ventral striatum (+6%) AA composition, and increased the AA:DHA ratio in PFC (+30%) and ventral striatum (+24%). The development and expression of AMPH-induced sensitization was significantly increased in DHA-deficient mice, whereas novelty- and acute AMPH-induced locomotor activity were not altered. DHA-deficient mice exhibited significantly greater ventral striatum, but not PFC, DA and DA metabolite concentrations following the AMPH challenge, whereas serotonin and noradrenalin concentrations were not altered. Ventral striatum AA composition and the AA:DHA ratio were both positively correlated with DA concentrations, and both ventral striatum AA composition and DA concentrations were positively correlated with locomotor activity during the preceding AMPH challenge. These results demonstrate that dietary-induced brain DHA deficiency, and associated elevation in the AA:DHA ratio, augment AMPH-induced sensitization in DBA/2J mice, and that this augmented response is associated with selective alterations in the mesolimbic DA pathway.     

Effect of Lycium barbarum Polysaccharides on the expression of endothelin-1 and its receptors in an ocular hypertension model of rat glaucoma

Lycium barbarum, a traditional Chinese anti-aging herb, has been shown to protect retinal ganglion cells (RGCs) in a rat chronic ocular hypertension (COH) model. Here, we investigated the expression of endothelin-1 (ET-1), a strong vasoconstrictor, and its receptors, ETA and ETB, in the COH model and assessed the effects of Lycium barbarum on the ET-1 axis. Elevated intraocular pressure (IOP) was induced in the right eye of SD rats using argon laser photocoagulation. (1) The expression of ET-1, ETA and ETB in normal and COH retinas was studied. (2) Some COH rats were fed daily with Lycium barbarum Polysaccharides (LBP) using 1 mg/kg or phosphate-buffered saline (PBS) for 3 weeks (started 1 week before photocoagulation). The effects of LBP on the expression of ET-1 and its receptors, ETA and ETB, in COH retina were evaluated. A semi-quantitative analysis of staining intensity was used to evaluate the expression levels of ET-1, ETA and ETB in retinal vasculature. We found that (1) Under COH condition, the immunoreactivity of ET-1 was increased in retina associated with an increase of ETB receptor immunoreactivity and a decrease of ETA receptor immunoreactivity. (2) After feeding COH rats with LBP, the expression of ET-1 was decreased with an increase of ETA expression and a decrease of ETB expression in the retina, especially in RGCs. (3) By comparing the staining intensity in the vasculature of COH retina in LBP-fed group with PBS-fed group, there was a decrease in the expression of ET-1 and ETA and an increase in ETB. In summary, ET-1 expression was up-regulated in the retina in COH model. LBP could decrease the expression of ET-1 and modulate the expression of its receptors, ETA and ETB, under the condition of COH. The neuroprotective effect of LBP on RGCs might be related to its ability to regulate the ET-1-mediated biological effects on RGCs and retinal vasculature.