Search

Intraocular injection of the Bartha strain of pseudorabies virus (PRV Bartha) results in transsynaptic infection of the hypothalamic suprachiasmatic nucleus (SCN), a retinorecipient circadian oscillator. PRV Bartha infection of a limited number of retinorecipient structures, including the SCN, was initially interpreted as the differential infection of a subpopulation of rat retinal ganglion cells, followed by replication and anterograde transport via the optic nerve. A recent report that used a recombinant strain of PRV Bartha (PRV152) expressing enhanced green fluorescent protein demonstrated that SCN infection actually results from retrograde transneuronal transport of the virus via the autonomic innervation of the eye in the golden hamster. In the present study using the rat, the pattern of infection after intravitreal inoculation with PRV152 was examined to determine if infection of the rat SCN is also restricted to retrograde transsynaptic transport. It was observed that infection in preganglionic autonomic nuclei (i.e., Edinger-Westphal nucleus, superior salivatory nucleus, and intermediolateral nucleus) precedes infection in the SCN. Sympathetic superior cervical ganglionectomy did not abolish label in the SCN after intraocular infection, nor did lesions of parasympathetic preganglionic neurons in the Edinger-Westphal nucleus. However, combined Edinger-Westphal nucleus ablation and superior cervical ganglionectomy eliminated infection of the SCN. This observation allowed a detailed examination of the SCN contribution to descending autonomic circuits afferent to the eye. The results indicate that in the rat, as in the hamster, SCN infection after intraocular PRV152 inoculation is by retrograde transsynaptic transport via autonomic pathways to the eye. J. Comp. Neurol. 471:298–313, 2004. © 2004 Wiley-Liss, Inc.

Melanopsin is a novel opsin synthesized in a small subset of retinal ganglion cells. Ganglion cells expressing melanopsin are capable of depolarizing in response to light in the absence of rod or cone input and are thus intrinsically light sensitive. Melanopsin ganglion cells convey information regarding general levels of environmental illumination to the suprachiasmatic nucleus, the intergeniculate leaflet, and the pretectum. Typically, retinal ganglion cells communicate information to central visual structures by receiving input from retinal photoreceptors via bipolar and amacrine cells. Because melanopsin ganglion cells do not require synaptic input to generate light-induced signals, these cells need not receive synapses from other neurons in the retina. In this study, we examined the ultrastructure of melanopsin ganglion cells in the mouse retina to determine the type (if any) of synaptic input these cells receive. Melanopsin immunoreaction product was associated primarily with the plasma membrane of (1) perikarya in the ganglion cell layer, (2) dendritic processes in the inner plexiform layer (IPL), and (3) axons in the optic fiber layer. Melanopsin-immunoreactive dendrites in the inner (ON) region of the IPL were postsynaptic to bipolar and amacrine terminals, whereas melanopsin dendrites stratifying in the outer (OFF) region of the IPL received only amacrine terminals. These observations suggested that rod and/or cone signals may be capable of modifying the intrinsic light response in melanopsin-expressing retinal ganglion cells.

The suprachiasmatic nucleus (SCN), a circadian oscillator, receives glutamatergic afferents from the retina and serotonergic (5-HT) afferents from the median raphe. 5-HT1B and 5-HT7 receptor agonists inhibit the effects of light on SCN circadian activity. Electron microscopic (EM) immunocytochemical procedures were used to determine the subcellular localization of 5-HT1B and 5-HT7 receptors in the SCN. 5-HT1B receptor immunostaining was associated with the plasma membrane of thin unmyelinated axons, preterminal axons, and terminals of optic and nonoptic origin. 5-HT1B receptor immunostaining in terminals was almost never observed at the synaptic active zone. To a much lesser extent, 5-HT1B immunoreaction product was noted in dendrites and somata of SCN neurons. 5-HT7 receptor immunoreactivity in γ-aminobutyric acid (GABA), vasoactive intestinal polypeptide (VIP), and vasopressin (VP) neuronal elements in the SCN was examined by using double-label procedures. 5-HT7 receptor immunoreaction product was often observed in GABA-, VIP-, and VP-immunoreactive dendrites as postsynaptic receptors and in axonal terminals as presynaptic receptors. 5-HT7 receptor immunoreactivity in terminals and dendrites was often associated with the plasma membrane but very seldom at the active zone. In GABA-, VIP-, and VP-immunoreactive perikarya, 5-HT7 receptor immunoreaction product was distributed throughout the cytoplasm often in association with the endoplasmic reticulum and the Golgi complex. The distribution of 5-HT1B receptors in presynaptic afferent terminals and postsynaptic SCN processes, as well as the distribution of 5-HT7 receptors in both pre- and postsynaptic GABA, VIP, and VP SCN processes, suggests that serotonin plays a significant role in the regulation of circadian rhythms by modulating SCN synaptic activity. J. Comp. Neurol. 432:371–388, 2001. © 2001 Wiley-Liss, Inc.

The hypothalamic suprachiasmatic nucleus (SCN) is the primary mammalian circadian clock that regulates rhythmic physiology and behavior. The SCN is composed of a diverse set of neurons arranged in a tight intrinsic network. In the rat, vasoactive intestinal peptide (VIP)- and gastrin-releasing peptide (GRP)-containing neurons are the dominant cell phenotypes of the ventral SCN, and these cells receive photic information from the retina and the intergeniculate leaflet. Neurons expressing vasopressin (VP) are concentrated in the dorsal and medial aspects of the SCN. Although the VIP/GRP and VP cell groups are concentrated in different regions of the SCN, the separation of these cell groups is not absolute. The inhibitory neurotransmitter gamma-aminobutyric acid (GABA) is expressed in most SCN neurons irrespective of their location or peptidergic phenotype. In the present study, immunoperoxidase labeling, immunofluorescence confocal microscopy, and ultrastructural immunocytochemistry were used to examine the spatial distribution of several markers associated with SCN GABAergic neurons. Glutamate decarboxylase, a marker of GABA synthesis, and vesicular GABA transporter were more prominently observed in the ventral SCN. KCC2, a K(+)/Cl(-) cotransporter, was highly expressed in the ventral SCN in association with VIP- and GRP-producing neurons, whereas VP neurons in the dorsal SCN were devoid of KCC2. On the other hand, GABA(B) receptors were observed predominantly in VPergic neurons dorsally, whereas, in the ventral SCN, GABA(B) receptors were associated almost exclusively with retinal afferent fibers and terminals. The differential expression of GABAergic markers within the SCN suggests that GABA may play dissimilar roles in different SCN neuronal phenotypes.

The afferent connections of the hypothalamic suprachiasmatic nucleus (SCN) of the golden hamster were examined using horseradish peroxidase (HRP) as the retrograde tracer molecule. Unilateral iontophoretic deposition of HRP into the SCN labeled ganglion cells bilaterally in the retinae. The labeled ganglion cells all had large somata and were randomly distributed across the retina. A similar number were labeled in each retina, which contrasted with the findings from injections into the optic chiasm and lateral geniculate body. Chiasm and geniculate injections both labeled three classes of ganglion cell (small, large, and giant) predominantly in the contralateral retinae. Telencephalic afferent projections to the SCN included the ventral subicular cortex and the septum. Notable diencephalic afferents included the dorsal lamina of the internal division of the ventral lateral geniculate nucleus (vLGN); the ipsilateral input was twice that of the contralateral projection. The same region of the vLGN was also noted to be reciprocally connected to the contralateral vLGN. The thalamic paraventricular nucleus was also heavily labeled but only ipsilaterally. Of functional significance, the SCN was discovered to innervate its contralateral homologue. Other less numerous afferents in the midbrain included the dorsal and median raphe nuclei and the dorsal nucleus of the lateral lemniscus. The afferent projections to the SCN determined in this study are discussed in regard to the known physiological role of the SCN as part of the circadian clock system.

The central projections of the retinal ganglion cells of the golden hamster were examined using horseradish peroxidase (HRP) as the anterograde tracer molecule. Following monocular injections of HRP into the vitreous, retinofugal fibers were histochemically demonstrated using the chromagen tetramethylbenzidine. This procedure, being more sensitive than the 3H-amino acid radioautographic technique, provided a clear demonstration of previously controversial retinal projections, clearer definition of established projections, and the discovery of new retinal pathways. An inferior accessory optic system was shown to be unequivocally present in this species and to consist of both crossed and uncrossed components. A direct retinal projection to the suprachiasmatic nucleus (SCN) of the hypothalamus was confirmed in this study. But the distribution of terminals as seen by this procedure was substantially different than previously reported; both rostrocaudal and mediolateral asymmetries in the distribution of label between the ipsilateral and contralateral SCN were observed. Substantial differences in the retinal projection to the SCN in the hamster and the rat were also noted. It is suggested that these differences may reflect the different effects photic input has on the neuro-endocrine-gonadal axis in these two species. Finally, labeled retinal axons were followed leaving the optic tract and coursing anteriorly through the plexiform layer of the piriform cortex; other labeled fibers were seen to enter the septal region. The physiological significance of these previously undescribed retinal projections is not known.