Developmental changes in cellular prion protein in primate visual cortex

Cellular prion protein (PrP(c)) is a cell surface glycoprotein highly expressed in neurons, and a protease-resistant conformer of the protein accumulates in the brain parenchyma in prion diseases. In human prion diseases, visual cortex and visual function can be affected. We examined both the levels and the localization of PrP(c) in developing visual cortex of the common marmoset. Western blot analysis showed that PrP(c) increased from the day of birth through adulthood, and this increase correlated with the progression of synapse formation. Immunohistochemistry showed that PrP(c) was present in fiber tracts of the neonate, and this immunoreactivity was lost with maturation. Within the neuropil, the laminar distribution of PrP(c) changed with age. In the neonate, PrP(c) immunoreactivity was strongest in layer 1, where the earliest synapses form. At the end of the first postnatal week, layer 4C, as identified by its strong cytochrome oxidase activity, was noticeably lighter in terms of PrP(c) immunoreactivity than the adjacent layers. The contrast between the strong immunoreactivity in both supragranular and infragranular layers and weak immunoreactivity in layer 4C increased with age. Layers 2/3 and 5 contained more intense PrP(c) immunoreactivity; these layers receive thalamic input from the koniocellular division of the LGN, and these layers of the LGN also had strong PrP(c) immunoreactivity. Together, these results provide evidence for PrP(c) localization in an identified functional pathway and may shed some light on prion disease pathogenesis.     

MPTP treatment of common marmosets impairs proteasomal enzyme activity and decreases expression of structural and regulatory elements of the 26S proteasome

Dysfunction of the ubiquitin‐proteasome system occurs in the substantia nigra (SN) in Parkinson's disease (PD). However, it is unknown whether this is a primary cause or a secondary consequence of other components of the pathogenic process. We have investigated in nonhuman primates whether initiating cell death through mitochondrial complex I inhibition using 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine hydrochloride (MPTP) altered proteasomal activity or the proteasomal components in the SN. Chymotrypsin‐like, trypsin‐like and peptidylglutamyl‐peptide hydrolase (PGPH) activating of 20S proteasome were decreased in SN homogenates of MPTP‐treated marmosets compared to naïve animals. Western blotting revealed a marked decrease in the expression of 20S‐α subunits, but no change in 20S‐β subunits in the SN of MPTP‐treated marmoset compared to naïve animals. There was a marked decrease in the expression of the proteasome activator 700 (PA700) and proteasome activator 28 (PA28) regulatory complexes. The 20S‐α4 subunit immunoreactivity was decreased in the nucleus of colocalized tyrosine hydroxylase (TH)‐positive cells of MPTP‐treated animals compared to naïve animals but no difference in the intensity of 20S‐β1i subunit staining. Immunoreactivity for PA700‐Rpt5 and PA28‐α subunits within surviving TH‐positive cells of MPTP‐treated marmoset was reduced compared to naïve controls. Overall, the changes in proteasomal function and structure occurring follow MPTP‐induced destruction of the SN in common marmosets were very similar to those found in PD. This suggests that altered proteasomal function in PD could be a consequence of other pathogenic processes occurring in SN as opposed to initiating cell death as previously suggested.     

GDNF reverses priming for dyskinesia in MPTP-treated, L-DOPA-primed common marmosets

Parkinson's disease (PD) is associated with a progressive loss of dopamine neurons in the substantia nigra and degeneration of dopaminergic terminals in the striatum. Although L-DOPA treatment provides the most effective symptomatic relief for PD it does not prevent the progression of the disease, and its long-term use is associated with the onset of dyskinesia. In rodent and primate studies, glial cell line-derived neurotrophic factor (GDNF) may prevent 6-OHDA- or MPTP-induced nigral degeneration and so may be beneficial in the treatment of PD. In this study, we investigate the effects of GDNF on the expression of dyskinesia in L-DOPA-primed MPTP-treated common marmosets, exhibiting dyskinesia. GDNF or saline was administered by two intraventricular injections, 4 weeks apart, to MPTP-treated, L-DOPA-treated common marmosets primed to exhibit dyskinesia. Prior to GDNF or saline administration, all animals displayed marked dyskinesia when treated with L-DOPA. GDNF administration produced a significant improvement in motor disability and, following the second injection of GDNF, a significant improvement in the locomotor activity was observed. Following the administration of L-DOPA there was a greater reversal of disability and a reduction in the intensity of L-DOPA-induced dyskinesia in GDNF-treated animals compared to saline-treated controls. However, there was no significant difference in L-DOPA's ability to increase locomotor activity between GDNF-treated and saline-treated animals. GDNF treatment caused a significant increase in the number of tyrosine hydroxylase-positive neurons in the substantia nigra, but no change in [(3)H]mazindol binding to dopamine terminals was found in the striatum of GDNF-treated animals compared to saline-treated controls. In GDNF-treated animals a small but significant reduction in enkephalin mRNA was observed in the caudate nucleus but not in the putamen or the nucleus accumbens. Substance P mRNA expression was equally reduced in the caudate nucleus and the putamen of the GDNF-treated animals but not in the nucleus accumbens. Intraventricular administration of GDNF improved MPTP-induced disability and reversed dopamine cell loss in the substantia nigra. GDNF also diminished L-DOPA-induced dyskinesia, which may relate to its ability to partly restore nigral dopaminergic transmission or to modify the activity of striatal output pathways.     

Avoiding hepatic metastasis naturally: Lessons from the cotton top tamarin (Saguinus oedipus)

Much has been written about hepatic metastasis and animal models abound. In terms of the human experience, progress in treating this final common pathway, a terminal event of many human malignancies has been relatively slow. The current thinking is that primary prevention is best served by early detection of cancer and eradication of early stage cancers by screening. Some cancers spread early in their course and the role of screening may be limited. Until relatively recently there has not been a pathfinder model that makes the evasion of this unfortunate event a reality. This review discusses such an animal model and attempts to relate it to human disease in terms of intervention. Concrete proposals are also offered on how scientists may be able to intervene to prevent this deadly progression of the cancer process.     

Establishment of lethal inhalational infection with Francisella tularensis (tularaemia) in the common marmoset (Callithrix jacchus)

Susceptibility and lethality studies of inhalational tularaemia were undertaken using the common marmoset ( Callithrix jacchus ) to determine its suitability as a non-human primate model. Pairs of marmosets were exposed to varying challenge doses of Francisella tularensis by the airborne route and monitored for up to 14 days postchallenge (p.c.). Lethal infection was achieved following a retained dose of less than 10 bacterial colony-forming units (CFU). However, precise LD₅₀ determination was not possible. The model was characterized using a target challenge dose of approximately 100 CFU. Increased core body temperature was the first indicator of disease, at approximately 2.5 days p.c. Overt clinical signs were first observed 12–18 h after the temperature increase. Significantly decreased activity was observed after approximately 3 days. All animals succumbed to infection between 4.5 and 7 days p.c. At postmortem examination, gross pathology was evident in the liver, spleen and lungs of all animals and high bacterial numbers were detected in all the organs assessed. Bacteraemia was demonstrated in all animals postmortem. Histopathological observations included severe suppurative bronchopneumonia, severe multifocal pyogranulomatous hepatitis, splenitis and lymphadenitis. Tularaemia disease progression in the common marmoset therefore appears to be consistent with the disease seen in humans and other animal models. The common marmoset may therefore be considered a suitable model for further studies of inhalational tularaemia.     

Head-cocking as a form of exploration in the common marmoset and its development

Head-cocking of 15 infant marmosets (Callithrix jacchus) was scored from Day 1 to 60 of postnatal life, the growth period with overproduction of interneuronal synapses. Head-cocking was scored during four 30 min intervals daily, including angle of head-cocking and objects being fixated. Mean age of onset of head-cocking was Day 13 (+/-1.3) and frequency increased to a fixed rate by Day 24-29, at the time of maturation of the foveal representation in layer 6 of the visual cortex, thus lending further support to the importance of head-cocking to visual processing. The most common distance of objects fixated during head-cocking was up to .5 m. Angle of head-cocking increased with age, and some asymmetry of direction was noted. Fewer head-cocking events occurred in the morning than in the afternoon. We also scored anogenital licking of offspring. Head-cocking occurred at higher levels in marmosets receiving more anogenital licking. As this was associated positively with increased exploration, head-cocking may be regarded as an exploratory behavior.     

Unravelling the subcortical and retinal circuitry of the primate inferior pulvinar

The primate visual brain possesses a myriad of pathways, whereby visual information originating at the retina is transmitted to multiple subcortical areas in parallel, before being relayed onto the visual cortex. The dominant retinogeniculostriate pathway has been an area of extensive study, and Vivien Casagrande's work in examining the once overlooked koniocellular pathway of the lateral geniculate nucleus has generated interest in how alternate subcortical pathways can contribute to visual perception. Another subcortical visual relay center is the inferior pulvinar (PI), which has four subdivisions and numerous connections with other subcortical and cortical areas and is directly recipient of retinal afferents. The complexity of subcortical connections associated with the PI subdivisions has led to differing results from various groups. A particular challenge in determining the exact connectivity pattern has been in accurately targeting the subdivisions of the PI with neural tracers. Therefore, in the present study, we used a magnetic resonance imaging (MRI)-guided stereotaxic injection system to inject bidirectional tracers in the separate subdivisions of the PI, the superior layers of the superior colliculus, the retina, and the lateral geniculate nucleus. Our results have determined for the first time that the medial inferior pulvinar (PIm) is innervated by widefield retinal ganglion cells (RGCs), and this pathway is not a collateral branch of the geniculate and collicular projecting RGCs. Furthermore, our tracing data shows no evidence of collicular terminations in the PIm, which are confined to the centromedial and posterior PI.     

Neural coding of temporal information in auditory thalamus and cortex

How the brain processes temporal information embedded in sounds is a core question in auditory research. This article synthesizes recent studies from our laboratory regarding neural representations of time-varying signals in auditory cortex and thalamus in awake marmoset monkeys. Findings from these studies show that 1) the primary auditory cortex (A1) uses a temporal representation to encode slowly varying acoustic signals and a firing rate-based representation to encode rapidly changing acoustic signals, 2) the dual temporal-rate representations in A1 represent a progressive transformation from the auditory thalamus, 3) firing rate-based representations in the form of monotonic rate-code are also found to encode slow temporal repetitions in the range of acoustic flutter in A1 and more prevalently in the cortical fields rostral to A1 in the core region of marmoset auditory cortex, suggesting further temporal-to-rate transformations in higher cortical areas. These findings indicate that the auditory cortex forms internal representations of temporal characteristics of sounds that are no longer faithful replicas of their acoustic structures. We suggest that such transformations are necessary for the auditory cortex to perform a wide range of functions including sound segmentation, object processing and multi-sensory integration.     

Terfenadine t-butyl hydroxylation catalyzed by human and marmoset cytochrome P450 3A and 4F enzymes in livers and small intestines

1.?Roles of human cytochrome P450 (P450) 3A4 in oxidation of an antihistaminic drug terfenadine have been previously investigated in association with terfenadine–ketoconazole interaction. Several antihistamine drugs have been recently identified as substrates for multiple P450 enzymes. In this study, overall roles of P450 3A4, 2J2, and 4F12 enzymes in terfenadine t-butyl hydroxylation were investigated in small intestines and livers from humans, marmosets, and/or cynomolgus monkeys.

2.?Human liver microsomes and liver and small intestine microsomes from marmosets and cynomolgus monkeys effectively mediated terfenadine t-butyl hydroxylation. Ketoconazole and N-hydroxy-N′-(4-butyl-2-methylphenyl)-formamidine (a P450 4A/F inhibitor) almost completely and moderately inhibited these activities, respectively, in human liver microsomes; however, these chemicals did not show substantially suppression in marmoset liver. Anti-human P450 3A and 4F antibodies showed the roughly supportive inhibitory effects.

3.?Recombinant P450 3A4/90 and 4F12 showed high terfenadine t-butyl hydroxylation activities with substrate inhibition constants of 84–144?μM (under 26–76?μM of K_m values), in similar manners to liver and intestine microsomes.

4.?These results suggest that human and marmoset P450 3A4/90 and 4F12 in livers or small intestines played important roles in terfenadine t-butyl hydroxylation. Marmosets could be a model for humans during first pass extraction of terfenadine and related substrates.     

Effects of aging and rifampicin pretreatment on the pharmacokinetics of human cytochrome P450 probes caffeine,warfarin, omeprazole,metoprolol and midazolam in common marmosets genotyped for cytochrome P450 2C19

1. The pharmacokinetics were investigated for human cytochrome P450 probes after single intravenous and oral administrations of 0.20 and 1.0?mg/kg, respectively, of caffeine, warfarin, omeprazole, metoprolol and midazolam to aged (10–14?years old, n?=?4) or rifampicin-treated/young (3?years old, n?=?3) male common marmosets all genotyped as heterozygous for a cytochrome P450 2C19 variant.

2. Slopes of the plasma concentration–time curves after intravenous administration of warfarin and midazolam were slightly, but significantly (two-way analysis of variance), decreased in aged marmosets compared with young marmosets. The mean hepatic clearances determined by in silico fitting for individual pharmacokinetic models of warfarin and midazolam in the aged group were, respectively, 23% and 56% smaller than those for the young group.

3. Significantly enhanced plasma clearances of caffeine, warfarin, omeprazole and midazolam were evident in young marmosets pretreated with rifampicin (25?mg/kg daily for 4?days). Two- to three-fold increases in hepatic intrinsic clearance values were observed in the individual pharmacokinetic models.

4. The in vivo dispositions of multiple simultaneously administered drugs in old, young and P450-enzyme-induced marmosets were elucidated. The results suggest that common marmosets could be experimental models for aged, induced or polymorphic P450 enzymes in P450-dependent drug metabolism studies.