targeted-toxins

Treece BR, Speth RC, Ritter RC, Burns GA (2003) Altered CCK binding in the dorsal vagal complex following cytotoxic lesion of the nodose ganglion. Neuroscience 2003 Abstracts 830.5. Society for Neuroscience, New Orleans, LA.

Summary: CCK reduces meal size by activating a specific subpopulation (30%-38%) of vagal afferent neurons. MK-801, attenuates reduction of food intake by CCK and increases meal size. We hypothesized that vagal afferents that are sensitive to excitatory amino acids might express CCK receptors and thereby mediate effects of both CCK and MK-801. Therefore we examined I125CCK-8 binding in the dorsal vagal complex, following unilateral nodosectomy or cytotoxic lesion of nodose cell bodies. To destroy vagal afferent cell bodies and their central projections we made unilateral intranodose injections of NMDA/kainic acid, which has been shown to destroy about 40% of vagal afferents. We also made intranodose injections of CCK-saporin (CCK-SAP), a novel cytotoxin, which appears to destroy vagal afferents when the ribosomal toxin, saporin, is selectively internalized along with the CCK/receptor complex. In some rats we completely eliminated vagal afferents on one side via nodosectomy. We found that this produced significant ipsilateral reduction in CCK binding in the nucleus of the solitary tract (NTS). Intranodose injection of NMDA/kainate or CCK-SAP also resulted in significantly reduced CCK binding in the NTS, ipsilateral to the injected nodose. The reduction of NTS CCK binding following nodosectomy, NMDA/kainate or CCK-SAP injection did not differ significantly. Interestingly, CCK binding appeared to be reduced in the vagal dorsal motor nucleus, ipsilateral to nodose removal, but was increased ipsilateral to CCK-SAP injection. We conclude that CCK-SAP destroys a subpopulation of vagal afferents that express CCK receptors and that intranodose NMDA/kainate destroys a partially overlapping afferent subpopulation that also expresses CCK receptors. Finally, our data suggest that expression of CCK binding in the dorsal motor nucleus may be controlled by feedback from CCK sensitive vagal afferents.

Related Products: CCK-SAP (Cat. #IT-31)

Kolasa K, Harrell LE (2003) Combined lesions of central cholinergic and noradrenergic denervation in the rat using 192 IgG-saporin and DSP-4 as an animal model of Alzheimer’s disease. Neuroscience 2003 Abstracts 842.6. Society for Neuroscience, New Orleans, LA.

Summary: To better model the consequences of persistent cholinergic hypofunction observed in Alzheimer’s disease, medial septum (MS) lesions were made by using specific cholinotoxin 192-IgG saporin. In this study the effect of simultaneous hippocampal cholinergic denervation, induced by intraseptal injection of 192-IgG saporin, and central noradrenergic denervation, induced by systemic injection of DSP-4 (N-[2-chloroethyl]-N-ethyl-2-bromobenzylamine) was examined in the rat dorsal hippocampus. DSP-4, an adrenergic neurotoxin selective for locus coeruleus innervated brain regions, induced a decrease in norepinephrine (NE) concentration in hippocampus. MS lesions resulted not only in selective cholinergic denervation of hippocampus (CD; superior cervical ganglion removed to prevent ingrowth of peripheral NE fibers), but also ingrowth of NE fibers into the hippocampus (HI; superior cervical ganglion left intact). MS lesions also resulted in a significant loss of choline-acetyltransferase activity in HI and CD groups, and an increase in NE in the HI group. In the HI group, but not in CD or control groups, visualization of hippocampus revealed a dense NE innervation with fine NE fibers with varicosities. Combination of MS lesion and DSP-4 treatment resulted in a reduction of NE concentration in HI group, with concomitant decrease in visualization of NE fibers. Those that remained were thick with sparse varicosities, possibly derived from peripheral sympathetic ingrowth. Elevated NE concentration and NE fiber number following specific cholinergic lesions might reflect compensatory sprouting of both central and peripheral adrenergic fibers into the hippocampus. Thus, noradrenergic sprouting in response to cholinergic denervation of hippocampus might be a valuable model for studying mechanisms as well as the consequences of neuronal plasticity in the mature CNS.

Related Products: 192-IgG-SAP (Cat. #IT-01)

Hummer TA, Johnson AD, Givens B (2003) The effects of ethanol and cholinergic lesions on anxiety in mice. Neuroscience 2003 Abstracts 859.16. Society for Neuroscience, New Orleans, LA.

Summary: Ethanol’s anxiolytic properties in mice are well-documented as measured with an elevated plus-maze. The effect of IP ethanol injections and cholinergic lesions on anxiety was investigated. Animals were injected with 0.5 µg of the cholinergic immunotoxin anti-murine-p75-saporin or control saline into the right lateral ventricle. After recovery (14-17 d), mice were given an IP 1.6 g/kg ethanol (10% v/v) or vehicle injection. After 60 min, animals were placed on the elevated plus-maze for a 10 min session and then perfused. Mice displayed increased open arm entries and time following ethanol. Saporin did not effect measures of anxiety nor interact with the effects of ethanol. Choline acetyltransferase staining was performed to confirm the extent of the cholinergic lesioning, and ethanol assays confirmed the presence or absence of ethanol in the blood stream. The research demonstrates that, as measured in the plus-maze, the basal forebrain cholinergic system has no significant involvement in anxiety nor in ethanol’s anxiolytic effects.

Related Products: mu p75-SAP (Cat. #IT-16)

Carstens EE, Iodi-Carstens M, Simons CT (2003) Reduced scratching in rats receiving intracisternal substance-saporin to ablate cervical superficial dorsal horn neurons that express NK-1 receptors. Neuroscience 2003 Abstracts 908.2. Society for Neuroscience, New Orleans, LA.

Summary: While glutamate and the neuropeptide substance P (SP) are involved in the spinal neurotransmission of nociceptive signals, little is known about transmitters involved in itch. We investigated a role for SP in itch by determining if scratching behavior is affected by selective neurotoxic destruction of cervical superficial dorsal neurons that express NK-1 receptors for SP. Sprague-Dawley rats received intracisternal microinjection of SP conjugated to saporin (SP-SAP; 2.27 μM/ 20 μl). Controls received saporin (SAP) only. At least 2 wk post-surgery, rats were tested for dose-related hindlimb scratching directed toward the site of intradermal microinjection of serotonin (5-HT; 50, 100 or 200 μg/10 μl) or saline (control) into the nape of the neck, with at least 1 wk between sessions for each dose. After the intradermal injection, rats were videotaped for 44 min. The numbers and durations of individual scratching bouts were counted and averaged for each dose. Rats receiving SAP exhibited a dose-related increase in scratching bouts similar to naïve rats. Rats receiving SP-SAP exhibited significantly reduced scratching (to ~38%) at all 5-HT doses. Individual bout durations (~2 s) did not vary significantly between groups or by dose of 5-HT. After behavioral testing, rats were perfused with fixative and caudal medullary and cervical spinal cord sections processed immunohistochemically for NK-1 receptors. Tissue from SAP-treated rats exhibited a normal distribution of pronounced NK-1 immunoreactivity in superficial layers of the dorsal horn at caudal medullary through C5 levels, while in SP-SAP-treated rats there was a complete absence of NK-1 immunoreactivity at these levels. These results indicate that SP plays an important role in neurotransmission from itch-signaling primary afferent fibers to second-order neurons in the superficial dorsal horn.

Related Products: SP-SAP (Cat. #IT-07)

Sarter MF, Martinez V, Bruno JP (2003) Lateralization of the attentional functions mediated via cortical cholinergic inputs. Neuroscience 2003 Abstracts 921.20. Society for Neuroscience, New Orleans, LA.

Summary: The role of basal forebrain (BF) corticopetal cholinergic projections in mediating attentional processing has been well established. For example, bilateral cortical cholinergic deafferentation produces robust impairments in attention. Neuropsychological and neuroimaging studies have postulated that attentional functions and capacities are mediated primarily via a lateralized, right-hemispheric network. The present study tested the general hypothesis that the attentional functions of cortical cholinergic inputs likewise are lateralized and thus that right-hemisphere cortical cholinergic deafferentation yields more severe attentional impairments. Rats were trained to perform an operant sustained attention task. Upon reaching criterion performance, unilateral cortical cholinergic deafferentation was produced by infusions of 192-IgG saporin into either the left or right BF. Compared with the performance of sham-operated animals and animals with left-hemispheric lesions, right-hemispheric cortical cholinergic deafferentation resulted in a persistent and selective decrease in the detection of signals (hits), mirroring the more potent but similarly selective effects of bilateral lesions. In contrast, left cortical cholinergic deafferentation did not affect hits but decreased the number of rejections in non-signal trials. These data extend previous studies suggesting that the integrity of the right cortical cholinergic input system is necessary for signal detection (Bushnell et al. 1998). Furthermore, the present data substantiate the assumption that the detection of signals and the rejection of non-signals are based on fundamentally different cognitive operations, and that the cholinergic mediation of these two operations is lateralized.

Related Products: 192-IgG-SAP (Cat. #IT-01)

Eichenbaum HB, Ross R, Raji A, McGaughy JA (2003) Noradrenergic, but not cholinergic, deafferentation of the infralimbic/prelimbic cortex impairs attentional set-shifting. Neuroscience 2003 Abstracts 940.7. Society for Neuroscience, New Orleans, LA.

Summary: Damage to the prefrontal cortex in humans produces deficits in the ability to shift attention to a previously irrelevant stimulus dimension (extra-dimensional shift; EDS) while sparing reversal and intra-dimensional shifts (a novel discrimination without a change in the relevant dimension; IDS). Data from human subjects has also shown the administration of noradrenergic agonists and antagonists disrupts EDS suggesting a role for coeruleal-cortical norepinephrine (NE). Usher and colleagues have proposed that high, tonic levels of NE may maximize behavioral flexibility to allow an optimal responsiveness to changes in the environment e.g. the predictability of reinforcement (Usher et al., Science 283, 1999). Based on these data, a loss of NE would be predicted to impair attentional set-shifting. In a rodent model of attentional set-shifting developed by Brown and colleagues, excitotoxic lesions of the infralimbic/prelimbic cortex (IL/PL) produced impairments in EDS but not IDS or reversal learning (J. Neurosci. 20, 2000). This study confirmed the importance of IL/PL to EDS, but did not address the role of NE in this type of cognition. In the current study, rats were infused with anti-dopamine beta-hydroxylase-saporin (NE-SAP)in IL/PL (0.01 µ g/ µl; 0.5 µl/hemisphere) to produce noradrenergic deafferentation, 192 IgG saporin (ACH-SAP; 0.01 µ g/ µl; 0.5 µl/hemisphere)to produce cholinergic deafferentation, or vehicle then tested in an attentional set-shifting task. NE-SAP rats were impaired in EDS but not in IDS or reversal learning. In contrast, ACH-SAP rats showed no impairment in any aspect of the task. The effect of DBH-SAP lesions on EDS support the hypothesis that NE, but not ACH, is critical to the adaptation of behavior to changes in reinforcement contingencies. The lack of effect of these lesions on reversal learning suggest the robustness of this effect may vary with the extent of behavioral adaptation required.

Related Products: 192-IgG-SAP (Cat. #IT-01)

Dudkin KN, Chueva IV, Makarov FN, Beach TG, Roher AE (2003) Impairments of working memory processes on a model of Alzheimer’s disease in monkeys. Neuroscience 2003 Abstracts 626.8. Society for Neuroscience, New Orleans, LA.

Summary: We have investigated the characteristics of visual working memory in a delayed-discrimination task in a model of Alzheimer’s disease (AD) in rhesus monkeys. Three animals received unilateral stereotaxic intracerebroventricular injection of the nucleus basalis of Meynert and three monkeys received sterile saline injections and thus served as controls. The lesioning agent consisted of a ribosomal toxin, saporin, conjugated to monoclonal antibodies against (the nbM lesion) the p75 neurotrophin receptor (p75NTR), which is expressed almost exclusively on cholinergic neurons of the nbM. The rationale for the model is the same as for a rabbit model of AD (Roher et al, Ann. NY Acad Sci. 2000). The monkeys were trained to discriminate stimuli with different types of visual information (spatial frequency gratings, color, spatial choice, spatial relationships between components of objects). The data obtained demonstrate that the nbM lesioning agent had a weak effect on visual differentiation without delay (long-term memory), but significantly decreased the duration of information storage (by a factor of 2 – 3) in working memory later two months after injection. These changes depended on temporal stage after injection and stimulus properties, and were accompanied by increase of motor reaction time and of refusal of task decision. In monkeys that were sham injected, there were no alterations in working memory characteristics. The results suggest that considerable worsening of the working memory characteristics for monkeys after lesion of the nbM reflects the formation of an AD model in these monkeys. The principles of functional organization of working memory and role of pathology of the cortical mechanisms in an impairment of memory characteristics are discussed.

Related Products: ME20.4-SAP (Cat. #IT-15)

Kim I, Wilson RE, Wellman CL (2003) Cholinergic deafferentation increases expression of the glur-1 subunit of the AMPA receptor in frontal cortex of young adult but not aging rats. Neuroscience 2003 Abstracts 633.9. Society for Neuroscience, New Orleans, LA.

Summary: Previously, we demonstrated that plasticity of frontal cortex is altered in aging rats: cholinergic lesions of the nucleus basalis magnocellularis (NBM) produce larger declines in dendritic morphology in frontal cortex of aged rats compared to young adults. In addition, these lesions result in upregulation of dendritic spines in frontal cortex of young adult but not aging rats. To begin to identify possible mechanisms underlying age-related differences in plasticity after NBM lesion, we assessed immunohistochemical labeling of the AMPA receptor subunit GluR1 in young adult and aging rats after either sham or 192 IgG saporin lesions of the NBM. Young adult (N=17), middle-aged (N=16), and aged rats (N=13) received unilateral sham or 192 IgG saporin lesions of the NBM. Two weeks later, brains were processed for immunohistochemical labeling of the GluR1 subunit of the AMPA receptor. An unbiased stereological technique was used to estimate density of labeled neurons in layer II-III of frontal cortex. Cells were identified as neurons based on standard morphological criteria and counted. Using a computerized image analysis system interfaced with a microscope, the average optical density of the white matter below frontal cortex was determined; neurons with optical densities at least one standard deviation above this mean were identified as intensely labeled. In young adult rats, lesions produced a 55% increase in the density of intensely GluR1-immunopositive neurons in frontal cortex. On the other hand, lesions had no effect on counts of GluR1-immunoreactive neurons in middle-aged and aged rats. This age-related difference in lesion-induced expression of AMPA receptor subunit protein could underlie the age-related differences in dendritic plasticity after NBM lesions.

Related Products: 192-IgG-SAP (Cat. #IT-01)

Janssen WG, Andrews G, Tomey MI, Baxter MG, Morrison JH (2003) Combined bilateral perforant path lesions with lesions of the cholinergic system: an ultrastructural immunogold analysis of nmdar1 representation within the dentate gyrus. Neuroscience 2003 Abstracts 676.26. Society for Neuroscience, New Orleans, LA.

Summary: Alzheimer’s disease is characterized by deterioration of cholinergic input to the hippocampus, as well as degeneration of input from the entorhinal cortex to the dentate gyrus(DG). Studies have demonstrated an upregulation of the NMDA receptor subunit, NR1, following unilateral ablatement of the perforant path(pp). We hypothesized that cholinergic innervation might be essential for DG plasticity following pp ablation. Our study was designed to investigate the synaptic distribution of NR1 following combined 192 IgG-Saporin lesions of the medial septum/vertical diagonal band(MS/VDB) and bilateral(bilat) pp knife cut ablation. Animals received bilat-pp lesions 2-3 weeks days post MS/VDB and were sacrificed 17 days following pp lesion. Four groups of rats were tested: 1)MS-VDB with sham bilat-pp; 2)sham MS-VDB with bilat-pp; 3)MS-VDB with bilat-pp; 4)sham MS-VDB with sham bilat-pp. Using postembedding immunogold electron microscopy and SynBin, a program designed for quantification and compartmentalization of immunogold particles at the synaptic level, we investigated these effects in the outer molecular layer of the DG in a pilot study with 2 animals/group. Initial results suggest that the synaptic pools of NR1 within post-synaptic compartments were not affected with single MS/VDB, but that a long term synaptic down regulation of NR1 follows bilat pp lesion that is not affected by the additional removal of cholinergic input. While these combined lesions do not alter the pattern of synaptic NR1 receptor distribution following pp lesions, these data has important implications for lesion-induced hippocampal plasticity as well as structural and functional recovery.

Related Products: 192-IgG-SAP (Cat. #IT-01)

Yoder RM, Pang KCH (2003) Medial septal and entorhinal cortical involvement in hippocampal theta rhythm. Neuroscience 2003 Abstracts 719.19. Society for Neuroscience, New Orleans, LA.

Summary: Hippocampal theta rhythm (HPCΘ) may be involved in various phenomena, including attention and the acquisition of sensory information. Two projections to HPC, the medial septum-diagonal band of Broca (MSDB) and entorhinal cortex (EC), are involved in the activation or synchronization of HPCΘ. MSDB contains excitatory (cholinergic) and inhibitory (GABAergic) hippocampal projections via the fimbria/fornix. EC contains excitatory (glutamatergic) hippocampal projections via the perforant path (PP). MSDB GABAergic, MSDB cholinergic, or bilateral PP lesions eliminate HPCΘ during urethane anesthesia (HPCΘ-II). In unanesthetized recordings, each of these lesions reduced but did not eliminate HPCΘ during locomotion (HPCΘ-I); MSDB cholinergic and EC lesions caused similar reductions in HPCΘ, and MSDB GABAergic lesions produced a greater amplitude reduction. In an attempt to determine whether interactions exist between MSDB projections and EC, we examined the effects of MSDB GABAergic or cholinergic lesions combined with PP lesions on HPCΘ-I. MSDB GABAergic and cholinergic lesions were produced by intraseptal injection of kainic acid and 192 IgG-saporin, respectively. Bilateral PP lesions were produced by passing cathodal current through an electrode located in the medial PP. HPCΘ amplitude was calculated as the square root of power at peak frequency (Fourier analysis) within the HPCΘ range. The combination of MSDB GABAergic and PP lesions eliminated HPCΘ-I. The combination of MSDB cholinergic and PP lesions did not reduce HPCΘ-I amplitude further than MSDB cholinergic or PP lesions alone. These results suggest the inhibitory (MSDB GABAergic) and excitatory (MSDB cholinergic or EC glutamatergic) projections interact to support HPCΘ-I. Furthermore, MSDB cholinergic and EC glutamatergic projections may be redundant for HPCΘ-I.

Related Products: 192-IgG-SAP (Cat. #IT-01)