Gary E. Baker

Neuroprotection of midbrain dopaminergic cells in MPTP-treated mice after near-infrared light treatment

This study explores whether near‐infrared (NIr) light treatment neuroprotects dopaminergic cells in the substantia nigra pars compacta (SNc) and the zona incerta‐hypothalamus (ZI‐Hyp) from degeneration in 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine (MPTP)‐treated mice. BALB/c albino mice were divided into four groups: 1) Saline, 2) Saline‐NIr, 3) MPTP, 4) MPTP‐NIr. The injections were intraperitoneal and they were followed immediately by NIr light treatment (or not). Two doses of MPTP, mild (50 mg/kg) and strong (100 mg/kg), were used. Mice were perfused transcardially with aldehyde fixative 6 days after their MPTP treatment. Brains were processed for tyrosine hydroxylase (TH) immunochemistry. The number of TH+ cells was estimated using the optical fractionator method. Our major finding was that in the SNc there were significantly more dopaminergic cells in the MPTP‐NIr compared to the MPTP group (35%–45%). By contrast, in the ZI‐Hyp there was no significant difference in the numbers of cells in these two groups. In addition, our results indicated that survival in the two regions after MPTP insult was dose‐dependent. In the stronger MPTP regime, the magnitude of loss was similar in the two regions (≈60%), while in the milder regime cell loss was greater in the SNc (45%) than ZI‐Hyp (≈30%). In summary, our results indicate that NIr light treatment offers neuroprotection against MPTP toxicity for dopaminergic cells in the SNc, but not in the ZI‐Hyp. J. Comp. Neurol. 518:25–40, 2010.

An investigation of the role of auditory cortex in sound localization using muscimol-releasing Elvax.

Lesion studies suggest that primary auditory cortex (A1) is required for accurate sound localization by carnivores and primates. In order to elucidate further its role in spatial hearing, we examined the behavioural consequences of reversibly inactivating ferret A1 over long periods, using Elvax implants releasing the GABAA receptor agonist muscimol. Sub‐dural polymer placements were shown to deliver relatively constant levels of muscimol to underlying cortex for >5 months. The measured diffusion of muscimol beneath and around the implant was limited to 1 mm. Cortical silencing was assessed electrophysiologically in both auditory and visual cortices. This exhibited rapid onset and was reversed within a few hours of implant removal. Inactivation of cortical neurons extended to all layers for implants lasting up to 6 weeks and throughout at least layers I–IV for longer placements, whereas thalamic activity in layer IV appeared to be unaffected. Blockade of cortical neurons in the deeper layers was restricted to ≤ 500 µm from the edge of the implant, but was usually more widespread in the superficial layers. In contrast, drug‐free Elvax implants had little discernible effect on the responses of the underlying cortical neurons. Bilateral implants of muscimol–Elvax over A1 produced significant deficits in the localization of brief sounds in horizontal space and particularly a reduced ability to discriminate between anterior and posterior sound sources. The performance of these ferrets gradually improved over the period in which the Elvax was in place and attained that of control animals following its removal. Although similar in nature, these deficits were less pronounced than those caused by cortical lesions and suggest a specific role for A1 in resolving the spatial ambiguities inherent in auditory localization cues.

Prechiasmatic Reordering of Fibre Diameter Classes in the Retinofugal Pathway of Ferrets.

This study examines the distribution of fibre diameter classes at various sites along the retinofugal pathway of adult ferrets. Light microscopic observations were made on semi‐thin sections, and regional fibre diameter spectra were constructed from diameter measurements taken from electron micrographs of thin sections of the intraorbital optic nerve (2.5 mm from the optic disc), the intracranial optic nerve (1 mm rostral to the fusion of the nerves), and the optic tract (just caudal to the optic chiasm).

The Course of Fibre Diameter Classes Through the Chiasmatic Region in the Ferret

The present study has examined the transition of fibre order from the optic nerve through the optic chiasm and into the optic tract in the ferrets retinofugal pathway. Semi‐thin sections through the chiasmatic region were examined in normal and in monocularly enucleated ferrets in order to display the segregation of the different axon diameter classes as the fibres pass from the optic nerve into the optic tract, and to determine, for each diameter class, where the crossed and uncrossed components become separated in the chiasmatic region.

Photobiomodulation enhances nigral dopaminergic cell survival in a chronic MPTP mouse model of Parkinson’s disease

We have shown previously that photobiomodulation or near-infrared light (NIr) treatment protects dopaminergic cells of the substantia nigra pars compacta (SNc) in an acute MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) model of Parkinsons disease (PD). In this study, we tested the protective and rescue action of NIr treatment in a chronic MPTP model, developed to resemble more closely the slow progressive degeneration in PD patients. We examined three regions of dopaminergic cells, the SNc, periaqueductal grey matter (PaG) and zona incerta-hypothalamus (ZI-Hyp). BALB/c mice had MPTP or saline injections over five weeks, followed by a three-week survival. NIr treatment was applied either at the same time as (simultaneous series) or after (post-treatment series) the MPTP insult. There were four groups within each series; Saline, Saline-NIr, MPTP and MPTP-NIr. Brains were processed for tyrosine hydroxylase (TH) immunochemistry and cell number was analysed using the optical fractionator method. In the SNc, there was a significant reduction (≈ 45%) in TH(+) cell number in the MPTP groups compared to the saline controls of both series. In the MPTP-NIr groups of both series, TH(+) cell number was significantly higher (≈ 25%) than in the MPTP groups, but lower than in the saline controls (≈ 20%). By contrast in the PaG and ZI-Hyp, there were no significant differences in TH(+) cell number between the MPTP an MPTP-NIr groups of either series. In summary, exposure to NIr either at the same time or well after chronic MPTP insult saved many SNc dopaminergic cells from degeneration.

Development and organization of ocular dominance bands in primary visual cortex of the sable ferret

Thalamocortical afferents in the visual cortex of the adult sable ferret are segregated into eye‐specific ocular dominance bands. The development of ocular dominance bands was studied by transneuronal labeling of the visual cortices of ferret kits between the ages of postnatal day 28 (P28) and P81 after intravitreous injections of either tritiated proline or wheat germ agglutinin‐horseradish peroxidase. Laminar specificity was evident in the youngest animals studied and was similar to that in the adult by P50. In P28 and P30 ferret kits, no modulation reminiscent of ocular dominance bands was detectable in the pattern of labeling along layer IV. By P37 a slight fluctuation in the density of labeling in layer IV was evident in serial reconstructions. By P50, the amplitude of modulation had increased considerably but the pattern of ocular dominance bands did not yet appear mature. The pattern and degree of modulation of the ocular dominance bands resembled that in adult animals by P63. Flat mounts of cortex and serial reconstructions of layer IV revealed an unusual arrangement of inputs serving the two eyes in the region rostral to the periodic ocular dominance bands. In this region, inputs serving the contralateral eye were commonly fused along a mediolateral axis, rostral to which were large and sometimes fused patches of ipsilateral input. J. Comp. Neurol. 407:151–165, 1999.

Maturational gradients in the retina of the ferret

In the present set of studies, we have examined the site for the initiation of retinal maturation in the ferret. A variety of maturational features across the developing inner and outer retina were examined by using standard immunohistochemical, carbocyanine dye labelling, and Nissl‐staining techniques, including 1) two indices of early differentiation of the first‐born retinal ganglion cells, the presence of β‐tubulin and of neuron‐specific enolase; 2) the receding distribution of chondroitin sulfate proteoglycans within the inner retina; 3) the distribution of the first ganglion cells to grow axons along the optic nerve; 4) the emergence of the inner plexiform layer; 5) the emergence of the outer plexiform layer and 6) the onset of synaptophysin immunoreactivity within it; 7) the differentiation of calbindin‐immunoreactive horizontal cells; and 8) the cessation of proliferative activity at the ventricular surface. Although we were able to define distinct maturational gradients that are associated with many of these features of inner and outer retinal development (each considered in detail in this report), with dorsal retina maturing before ventral retina, and with peripheral retina maturing last, none showed a clear initiation in the region of the developing area centralis. Rather, maturation began in the peripapillary retina dorsal to the optic nerve head, which is consistent with previous studies on the topography of ganglion cell genesis in the ferret. These results make clear that the order of retinal maturation and the formation of the area centralis are not linked, at least not in the ferret.

Spatial-frequency tuning and geniculocortical projections in the visual cortex (areas 17 and 18) of the pigmented ferret

We have examined the spatial‐frequency selectivity of neurons in areas 17 and 18 of the adult pigmented ferret, by measuring how the amplitude of response depends on the spatial‐frequency of moving sinusoidal gratings of optimal orientation and fixed contrast. Neurons in area 17 of the ferret respond optimally to low spatial frequencies [average 0.25 cycles per degree (c/deg)], much lower than the optima for cat area 17. The tuning curves are of the same form as those found in cat and monkey: unimodal with bandwidths in the range 0.8–3.5 octaves. Neurons in area 18 of the ferret respond optimally to even lower spatial frequencies (average 0.087 c/deg) than area 17 neurons, and the distributions of optimal spatial frequency for areas 17 and 18 hardly overlap. In both cortical areas, the bandwidth of the tuning curves is inversely correlated with optimal spatial frequency. This marked difference in tuning between the two cortical areas is probably attributable to differential geniculo‐cortical projections. Small injections of fluorescent latex microspheres or horseradish peroxidase (HRP) were made into area 17 or area 18 in order to investigate the populations of geniculate neurons projecting to the two cortical areas. After injections into area 17, labelled neurons are found predominantly in the geniculate A layers, with a few neurons labelled in the C layers. Conversely, after an area 18 injection, similar numbers of labelled neurons are found in the C layers as in the A layers. Soma‐size analysis of the neurons in the A‐layers suggests the existence of two populations of relay neurons, which project differentially to areas 17 and 18. The different geniculate inputs and the different spatial‐frequency tuning in areas 17 and 18 may imply that the two cortical areas process visual information more in parallel than in series.

The re-establishment of the representation of the dorso-ventral retinal axis in the chiasmatic region of the ferret.

This study has examined the representation of the dorso-ventral retinal axis in the optic nerve and tract of the ferret, as well as the associated fiber transformations which take place within the chiasmatic region. In one series of experiments, dorsal or ventral retinal lesions were made to induce fiber degeneration along the pathway, from which semi-thin sections were then stained for degenerating myelin. In a second series, implants of the carbocyanine dye, DiI, were made into the caudo-medial or rostro-lateral optic tract in order to label retrogradely the axons as they course through the chiasmatic region. Additional observations were made from the optic pathways of ferrets that had been similarly lesioned or implanted, but employing either a reduced-silver technique to reveal the degenerating axons or horseradish peroxidase as the retrograde label. The axons arising from the dorsal and ventral retina course in the dorsal and ventral parts of the optic nerve posterior to the eye, but as they continue along the nerve they disperse producing a highly impoverished retinotopy in the prechiasmatic portion of the nerve. As they course through the chiasmatic region, however, they become segregated again: dorsal fibers cross the midline relatively caudally while ventral fibers cross further rostrally, although there is overlap between them. Nearer the threshold of the optic tract, the fibers from dorsal and ventral retina undergo a further and more striking segregation, placing the dorsal fibers caudo-medially and the ventral fibers rostro-laterally within the tract. This re-emergence of retinotopic order implicates a fiber-substrate interaction as being responsible for the axonal reordering, and suggests that fiber pre-ordering in the tract contributes to the formation of the orderly projection of the dorso-ventral retinal axis upon central visual targets.

Topography of fibre organisation in the corticofugal pathways of rats

The organisation of the long descending corticofugal pathways is poorly understood. We have examined these pathways to determine the fibre relationships along the extent of their course through the internal capsule, cerebral peduncle, longitudinal pontine fasciculus, pyramid, pyramidal decussation, and dorsal column of the spinal cord. Different cytoarchitectonic regions (e.g., lateral agranular and granular) of the rats neocortex were injected with the axonal tracer biotinylated dextran. In other experiments, each animal had different‐coloured fluorescent tracers (Fluoro Ruby and dextran‐fluorescein) injected into separate cortical areas. Our results show that in the anterior and posterior limbs of the internal capsule, axons arising from spatially separate sites in rat neocortex occupy distinct regions of the cross‐sectional area of the pathway. More caudally, within the cerebral peduncle and the longitudinal pontine fasciculus, axons from more distant cortical areas remain largely separate, but those from adjacent cortical areas begin to overlap. By the medullary pyramid, the pyramidal decussation, and the dorsal column of the spinal cord, the representations of all the cortical regions injected overlap completely; in these structures, the axons arising from each cortical area are widely intermingled. Thus, along the rostral‐to‐caudal course of the corticofugal pathways, there is a change in the organisation of axons. At rostral levels, the order corresponds roughly to the spatial distribution of the cells of origin, but more caudally, this changes to an arrangement of axons that has no readily apparent order. A similar change has been observed along the course of the retinofugal pathway, where a decrease of spatial order in the fibre distribution has been associated with a reordering of axons according to their temporal sequence of outgrowth.J.Comp. Neurol. 381:143‐157, 1997.