Mary Behan

Intrinsic and efferent connections of the endopiriform nucleus in rat.

The endopiriform nucleus is a large group of multipolar cells located deep to the piriform cortex. The function of this nucleus is unknown, but studies with animal models suggest that it plays an important role in temporal lobe epileptogenesis. To address questions concerning mechanisms of epileptogenesis and to gain insights into its normal function, efferent axons from the endopiriform nucleus were labeled by anterograde transport from small extracellular injections of Phaseolus vulgaris leucoagglutinin.

Sex steroid hormones and the neural control of breathing

We review evidence that sex steroid hormones including estrogen, progesterone and testosterone are involved in the central neural control of breathing. Sex hormones may exert their effects on respiratory motoneurons via neuromodulators, in particular, the serotonergic system. Recent studies have shown that levels of serotonin (5HT) in the hypoglossal and phrenic nuclei are greater in female than in male rats. Serotonin-dependent plasticity in hypoglossal and phrenic motor output also differs in male and female rats. Changing levels of gonadal hormones throughout the estrus cycle coincide with changing levels of 5HT in respiratory motor nuclei, and gonadectomy in male rats results in a decrease in 5HT-dependent plasticity in respiratory motor output. We speculate that sex steroid hormones are critically involved in adaptations in the neural control of breathing throughout life, and that decreasing levels of these hormones with increasing age may have a negative influence on the respiratory control system in response to challenge.

A new subdivision of anterior piriform cortex and associated deep nucleus with novel features of interest for olfaction and epilepsy.

The anterior part of the piriform cortex (the APC) has been the focus of cortical‐level studies of olfactory coding and associative processes and has attracted considerable attention as a result of a unique capacity to initiate generalized tonic‐clonic seizures. Based on analysis of cytoarchitecture, connections, and immunocytochemical markers, a new subdivision of the APC and an associated deep nucleus are distinguished in the rat. As a result of its ventrorostral location in the APC, the new subdivision is termed the APCVR. The deep nucleus is termed the pre‐endopiriform nucleus (pEn) based on location and certain parallels to the endopiriform nucleus. The APCVR has unique features of interest for normal function: immunostaining suggests that it receives input from tufted cells in the olfactory bulb in addition to mitral cells, and it provides a heavy, rather selective projection from the piriform cortex to the ventrolateral orbital cortex (VLO), a prefrontal area where chemosensory, visual, and spatial information converges. The APCVR also has di‐ and tri‐synaptic projections to the VLO via the pEn and the submedial thalamic nucleus. The pEn is of particular interest from a pathological standpoint because it corresponds in location to the physiologically defined “deep piriform cortex” (“area tempestas”) from which convulsants initiate temporal lobe seizures, and blockade reduces ischemic damage to the hippocampus. Immunostaining revealed novel features of the pEn and APCVR that could alter excitability, including a near‐absence of γ‐aminobutyric acid (GABA)ergic “cartridge” endings on axon initial segments, few cholecystokinin (CCK)‐positive basket cells, and very low γ‐aminobutyric acid transporter‐1 (GAT1)‐like immunoreactivity. Normal functions of the APCVR‐pEn may require a shaping of neuronal activity by inhibitory processes in a fashion that renders this region susceptible to pathological behavior. J. Comp. Neurol. 434:289–307, 2001.

Sex steroidal hormones and respiratory control.

There is a growing public awareness that sex hormones can have an impact on a variety of physiological processes. Yet, despite almost a century of research, we still do not have a clear picture as to the effects of sex hormones on the regulation of breathing. Considerable data has accumulated showing that estrogen, progesterone and testosterone can influence respiratory function in animals and humans. Several disorders of breathing such as obstructive sleep apnea (OSA) and sudden infant death syndrome (SIDS) show clear sex differences in their prevalence, lending weight to the importance of sex hormones in respiratory control. This review focuses on questions such as: how early do sex hormones influence breathing? Which is the most effective? Where do sex hormones exert their effects? What mechanisms are involved? Are there age-associated changes? A clearer understanding of how sex hormones influence the control of breathing could enable sex- and age-specific therapeutic interventions for diseases of the respiratory control system.

Long term facilitation of respiratory motor output decreases with age in male rats

1 Long term facilitation (LTF) is a serotonin‐dependent augmentation of respiratory motor output (phrenic and hypoglossal) following episodic hypoxia. Since ageing influences respiratory control mechanisms and serotonergic function, we tested the hypothesis that LTF decreases with age in male rats. 2 Young (3‐4 month) and aged (13 month) male Sprague‐Dawley rats were anaesthetized with urethane, vagotomized, paralysed and pump ventilated. Integrated phrenic and hypoglossal (XII) nerve activities were measured before (baseline), during and for 60 min after three 5 min episodes of isocapnic hypoxia (Pa,O2 35‐45 mmHg) separated by 5 min of hyperoxia (Pa,O2 > 150 mmHg). 3 In young rats, LTF was observed as an augmentation in peak integrated phrenic (n = 8) and XII (n = 7) amplitudes following episodic hypoxia (56 ± 14 and 73 ± 16 % (means ±s.e.m.) at 60 min post‐hypoxia, respectively; both P < 0.05). In aged rats, LTF was significantly increased compared to baseline in phrenic (25 ± 8 % at 60 min, P < 0.05), but not in XII (4 ± 7 %, P > 0.05) motor output. LTF was significantly greater in young than in aged rats in both motor outputs (P < 0.05). 4 Decreased phrenic and XII LTF suggests that serotonergic modulation of respiratory motor output decreases in ageing male rats. We speculate that decreased serotonergic modulation may contribute to age‐related breathing disorders.

Age and gender effects on serotonin-dependent plasticity in respiratory motor control

We review recent evidence indicating that serotonin-dependent plasticity in respiratory control is influenced by age and gender. Specifically, respiratory long-term-facilitation following intermittent hypoxia decreases with age in male rats, but increases in female rats. We speculate about a possible relationship between age and gender effects on serotonin-dependent plasticity in upper airway motoneuron pools and the prevalence of obstructive sleep apnea.

The precise origin of the tectospinal pathway in three common laboratory animals: A study using the horse-radish peroxidase method

The horseradish peroxidase tracing method has been used to study the cells of origin of the tectospinal projections in the opossum, the tree shrew, and the cat. The present data show that only those collicular neurons which occupy the deep (ventral to the stratum opticum) tectal laminae send axons to the cervical spinal cord. In particular, layer IV contains the greatest number of spinal projecting neurons. Our results also reveal that while only the large sized collicular neurons project upon the cervical spinal cord in the opossum and the tree shrew, neurons comprising several different size categories do so in the cat. We thus suggest that several different descending channels exist over which the superior colliculus can influence the neck musculature in the cat.

Comparison of hypocretin/orexin and melanin-concentrating hormone neurons and axonal projections in the embryonic and postnatal rat brain

Hypocretin/orexin (H/O) and melanin-concentrating hormone (MCH) are peptide neuromodulators found in separate populations of neurons located within the lateral and perifornical hypothalamic regions. H/O has been linked to sleep-wakefulness regulation and to the sleep disorder narcolepsy, and both systems have been implicated in energy homeostasis, including the regulation of food intake. In the present study we compared the development of H/O and MCH-expressing neuronal populations with in situ hybridization and immunohistochemistry on adjacent sections in the embryonic and postnatal rat brain. We found that MCH mRNA and protein were present in developing neurons of the hypothalamus by embryonic day 16 (E16), whereas H/O mRNA and protein were not detected until E18. We also identified previously undescribed populations of MCH-immunoreactive cells in the lateral septum, paraventricular hypothalamic nucleus, lateral zona incerta, and ventral lateral geniculate nucleus that may play a specific role in the development of these regions. MCH immunoreactive axonal processes were also evident earlier than H/O stained fibers and at the time H/O immunoreactive processes were first identified in the hypothalamus at E20, extensive MCH axonal fiber systems were already present in many brain regions. Interestingly, however, the density of axonal fibers immunoreactive for H/O in the locus coeruleus reached peak levels at the same developmental age (P21) as MCH immunoreactive axons in the diagonal band of Broca (DBB). The peak of axon density coincided with the developmental stage at which adult patterns of feeding and sleep-waking activity become established. The present results demonstrate developmental differences and similarities between the MCH and H/O systems that may relate to their respective roles in feeding and sleep regulation.

Sex hormone receptors are expressed in identified respiratory motoneurons in male and female rats

Sex hormones including estrogen, progesterone and testosterone can influence breathing. However, it is not clear whether such hormones exert their effects directly on respiratory motoneurons. We used immunocytochemistry to demonstrate that estrogen receptor alpha, estrogen receptor beta and androgen receptor are localized in respiratory motor neurons. Motoneurons in the hypoglossal (XII) and the phrenic nuclei were retrogradely labeled from the tongue and the diaphragm respectively. Double-label fluorescence immunocytochemistry was used to show that sex hormone receptors are present in respiratory motoneurons of both male and female rats. These data suggest that in male and female rats, sex hormones can modulate the output of respiratory motoneurons directly.

Age-related changes in serotonin in the hypoglossal nucleus of rat: implications for sleep-disordered breathing.

We are attempting to determine the neuronal factors that influence upper airway patency during sleep in the elderly. Serotonin has a facilitatory effect on hypoglossal motoneurons that innervate the tongue, and manipulations of the serotonergic system alter airway patency. We hypothesized that age-associated changes in serotonergic input to the hypoglossal nucleus might be a factor in the increased susceptibility to sleep-disordered breathing in the elderly. We used light microscopic immunocytochemistry to study the distribution of serotonin in the hypoglossal nucleus in young and old rats. Rats > 18 months had fewer serotonin immunoreactive axons and boutons in the hypoglossal nucleus than rats < 6 months. These data suggest that normal aging may result in a change in the availability of serotonin that results in decreased facilitation of hypoglossal motoneurons.