Teresa L. Steininger

Sleep–waking discharge patterns of ventrolateral preoptic/anterior hypothalamic neurons in rats

Numerous lesion, stimulation and recording studies in experimental animals demonstrate the importance of neurons within the preoptic/anterior hypothalamic area (POA) in the regulation of sleep induction and sleep maintenance. Recently, a discrete cluster of cells in the ventrolateral POA (vlPOA) of rats was found to exhibit elevated c-fos gene expression during sleep, indicating that these neurons are strongly activated during nonREM and/or REM sleep stages. We examined neuronal discharge during wakefulness and sleep throughout the dorsal to ventral extent of the lateral POA in rats, using chronic microwire technique. We found that neurons with elevated discharge rates during sleep, compared to waking, were localized to the vlPOA. As a group, vlPOA neurons displayed elevated discharge rates during both nonREM and REM sleep. Discharge of vlPOA neurons reflected the depth of sleep, i.e., discharge rates increased significantly from light to deep nonREM sleep. During recovery sleep following 12-14 h of sleep deprivation, vlPOA neurons displayed increased sleep-related discharge, compared to baseline sleep. Neurons in the vlPOA displaying increased neuronal discharge during sleep were located in the same area where neurons exhibit increased c-fos gene expression during sleep. Such neurons are likely components of a rostral hypothalamic mechanism that regulates sleep onset and sleep maintenance.

Subregional organization of preoptic area/anterior hypothalamic projections to arousal-related monoaminergic cell groups.

Pathways mediating the generation and/or maintenance of sleep reside within the preoptic/anterior hypothalamus (POAH). Reproduction, water balance, thermoregulation, and neuroendocrine functions are also associated with POAH, but it is not fully understood whether sleep is consolidated with these behavioral and physiological functions, or whether sleep‐related circuitry is segregated from other POAH regions. Recent studies indicate that sleep mechanisms may be localized to the ventrolateral preoptic area (VLPO) and that this region sends inhibitory projections to waking/arousal‐related neurons in the histaminergic tuberomammillary nucleus (TM), the noradrenergic locus coeruleus (LC), and the serotonergic dorsal raphe (DR). The present study is a quantitative investigation of preoptic area efferents to these monoaminergic groups. The results demonstrate that biotinylated dextran injections in the VLPO region reveal a robust innervation of TM that was as much as five times greater than innervation derived from other POAH subregions. The innervation of TM originated almost exclusively from injection sites in the region of galanin neurons. VLPO projections to the LC were moderately dense and were greater than in other POAH regions except for equivalent input from the medial preoptic area. Projections to the dorsal raphe were equivalent to LC innervation and were generally two to three times greater from VLPO than from other POAH regions, except for projections from the lateral preoptic region, which were similar in magnitude. The rostral and caudal levels projected more to the TM, whereas the midrostral region of VLPO strongly innervated the LC core. These findings, with recent studies demonstrating medial and lateral extensions of the sleep‐related VLPO neuronal group, indicate that descending arousal state control may be mediated by this specific galaninergic/γ‐aminobutyric acid (GABA)ergic cell group. J. Comp. Neurol. 429:638–653, 2001.

High-affinity nerve growth factor receptor (Trk) immunoreactivity is localized in cholinergic neurons of the basal forebrain and striatum in the adult rat brain

Trk-immunoreactivity was observed in basal forebrain and striatal cholinergic neurons, whereas low-affinity NGF receptor immunoreactivity was observed in basal forebrain but not striatal cholinergic neurons. Since NGF exerts trophic actions on both basal forebrain and striatal cholinergic populations, the presence of Trk in these neurons lends strong support for an essential role of Trk in NGF-responsive neurons, but suggests that the low affinity receptor is not necessary for NGF actions in the striatum.

Serotonergic dorsal raphe nucleus projections to the cholinergic and noncholinergic neurons of the pedunculopontine tegmental region: a light and electron microscopic anterograde tracing and immunohistochemical study.

The serotonergic dorsal raphe nucleus is considered an important modulator of state‐dependent neural activity via projections to cholinergic neurons of the pedunculopontine tegmental nucleus (PPT). Light and electron microscopic analysis of anterogradely transported biotinylated dextran, combined with choline acetyltransferase (ChAT) immunohistochemistry, were employed to describe the synaptic organization of mesopontine projections from the dorsal raphe to the PPT. In a separate set of experiments, we utilized immunohistochemistry for the serotonin transporter (SERT), combined with ChAT immunohistochemistry at the light and electron microscopic levels, to determine whether PPT neurons receive serotonergic innervation. The results of these studies indicate that: (1) anterogradely labeled and SERT‐immunoreactive axons and presumptive boutons invest the PPT at the light microscopic level; (2) at the ultrastructural level, dorsal raphe terminals in the PPT pars compacta synapse mainly with dendrites and axosomatic contacts were not observed; (3) approximately 12% of dorsal raphe terminals synapse with ChAT‐immunoreactive dendrites; and (4) at least 2‐4% of the total synaptic input to ChAT‐immunoreactive dendrites is of dorsal raphe and/or serotonergic origin. This serotonergic dorsal raphe innervation may modulate cholinergic PPT neurons during alterations in behavioral state. The role of these projections in the initiation of rapid eye movement (REM) sleep and the ponto‐geniculo‐occipital waves that precede and accompany REM sleep is discussed. J. Comp. Neurol. 382:302‐322, 1997.

Chapter 2: Ascending cholinergic pathways: functional organization and implications for disease models

Publisher Summary Acetylcholine (ACh) is known to exert profound modulatory effects on information processing in the brain. For example, experimental blockade of cholinergic transmission can lead to defects in memory and cognitive function, and loss of cerebral cholinergic innervation occurs in Alzheimers disease. This latter cholinergic deficit contributes, at least in part, to the dementia seen in Alzheimers patients. The development of antibodies to choline acetyltrarfsferase (ChAT), the synthesizing enzyme for ACh, allowed for specific visualization of cholinergic neurons and immunohistochemical studies have revealed two major ascending cholinergic systems that are likely to mediate the behavioral effects of ACh: the magnocellular basal forebrain and mesopontine tegmental cholinergic groups. More recently, the availability of molecular probes to visualize neuronal populations expressing ChAT mRNA have confirmed the identity of these cell groups, as well as other major cholinergic neuronal populations in the nervous system. This chapter describes the current level of understanding concerning the functional organization of the ascending cholinergic pathways cited above, as well as addressing some issues relating to the identification of trophic mechanisms that support the viability of the basal forebrain system. This latter topic bears special relevance to cholinergic dysfunction in disease and is considered in more detail in elsewhere in this volume. While the present focus is on ascending cholinergic pathways, it is important to bear in mind that these pathways represent part of a larger group of so-called diffuse ascending systems, including dopaminergic, noradrenergic, serotonergic and histaminergic transmitter systems. All of these pathways exert important modulatory effects on information processing in the nervous system and are likely to interact significantly with one another.

Localization of immunoreactive epidermal growth factor receptor in neonatal and adult rat hippocampus.

The regional and developmental expression of epidermal growth factor (EGF) receptor in rat hippocampus was investigated utilizing immunocytochemical techniques at the light and electron microscopic levels. EGF receptor immunoreactivity in adult hippocampus was compared to that found at postnatal day 7 (P7). While the receptor was observed in P7 hippocampus, immunostaining was more prominent in the adult hippocampus, especially in the pyramidal CA2 field. Ultrastructural analysis of this region revealed that the receptor was localized to the cell bodies of both P7 and adult neurons rather than the axons or dendrites. The expression of EGF receptor in selected regions of the adult brain was verified by Western blotting. These results demonstrate the presence of EGF receptor in rat hippocampus as early as P7, localize the receptor to the pyramidal cell body, and establish the hippocampal formation, particularly CA2, as a major site of EGF receptor expression in rat brain.

Differences in the retinohypothalamic tract in albino Lewis versus brown Norway rat strains.

Differences in sleep-wake patterns in response to light-dark stimulation have been observed between albino Lewis and pigmented Brown Norway strains of rats, which may be associated with albinism. Since several anatomical differences have been demonstrated in the visual pathways of albino and pigmented mammals, the present study was undertaken to determine whether additional differences in visual pathways of these rat strains exist that might account for their behavioral differences. Using anterograde tracing techniques and image analysis, we have investigated the retinal projections of Lewis and Brown Norway rats. Our results demonstrate that the distribution of retinal terminals in the hypothalamic suprachiasmatic nucleus extends over a greater area in Lewis compared to Brown Norway rats. This zone of termination corresponds to a cytoarchitectonically definable ventrolateral subdivision of the suprachiasmatic nucleus (SCN), which is also greater in Lewis than in Brown Norway rats. These results may have implications for behaviors related to the SCN.