David E. Scott

Prophylactic Treatment of Pediatric Migraine

Background.—Migraine occurs in 3% to 5% of young children and up to 18% of adolescents. Management requires a tailored regimen of pharmacological and behavioral measures that consider the headache burden and disability. Patients with frequent or disabling attacks (or both) may warrant preventive agents.

Increased Expression of Nitric Oxide Synthase in Hypothalamic Neuronal Regeneration

This investigation deals with the histochemical and scanning electron microscopic (SEM) correlates that depict regeneration of the neurohypophyseal system that may be nitric oxide dependent following hypophysectomy in the rodent hypothalamus. NOS histochemistry and correlative SEM were employed to establish the rates of regrowth and appearance of NOS-positive supraoptic (SON) and paraventricular (PVN) neurites and their cell bodies following hypophysectomy. NOS activity increased significantly in SON and PVN neuronal perikarya and regenerating axons by 2 weeks. NOS-positive neurites were observed to regrow into the adjacent median eminence and insinuate into the lumen of the third cerebral ventricle. By 4 weeks posthypophysectomy, NOS staining of SON and PVN neurons and their regrown neurites had returned to normal control levels. Despite this fact, large complexes of apparent magnocellular neurites remained upon the floor of the third cerebral ventricle as observed with SEM. These observations support the hypothesis that NO may play a fundamental role in the process of regeneration, plasticity, and retargeting of SON and PVN axons following injury.

Treatment of paediatric headache

Headaches are very common during childhood and become increasingly frequent during adolescence. The diagnosis of primary headache disorders (e.g., migraine and tension-type headache) rests principally on clinical criteria as set forth by the International Headache Society. Treatment options include acute or episodic measures, prophylactic agents and non-pharmacological or behavioural interventions. From review of available evidence, the most efficacious acute treatments of paediatric migraine include the non-steroidal anti-inflammatory agent ibuprofen at 7.5 – 10 mg/kg/dose or nasal sumatriptan at doses of 5 or 20 mg. For those patients with headaches that occur with sufficient frequency and severity to warrant daily prophylaxis, controlled data are limited. Agents which are likely to be beneficial include amitriptyline, flunarizine (not available in the US) and cyproheptadine. Clinical experience with the anti-epileptic agents topiramate and valproate suggests an expanding role for the prevention of paediatric migraine in the future.

Neural regeneration and neuronal migration following injury. I. The endocrine hypothalamus and neurohypophyseal system

Central to this investigation are several basic hypotheses that are designed to test the role of nitric oxide (NO) in the complex process of central regeneration and plasticity in a well established model system of the mammalian brain. We have employed histochemical techniques at the light and ultrastructural level coupled with correlative scanning electron microscopy, immunoelectron microscopy, and in situ hybridization in order to determine the functional significance of the increased expression of nitric oxide synthase (NOS) in neurons of the supraoptic (SON) and paraventricular (PVN) nuclei which accompanies regeneration of their axotomized neurites following hypophysectomy. The aim of this investigation was to determine the potential role and temporal up-regulation of NOS in this basic regenerative process and to establish the ultrastructural and neuroanatomical correlates during critical periods of regeneration and regrowth of SON and PVN axons following hypophysectomy in the endocrine hypothalamus of the rat. Our data support the hypothesis that NO may serve as a second messenger molecule that may act in some fashion to govern not only the process of central regeneration and regrowth of magnocellular (SON/PVN) axons into the median eminence, neural stem, and neural lobe (the neurohypophyseal system) but may also influence the regeneration of neurites into new neuroanatomical domains such as the adjacent lumen of the third cerebral ventricle. We have demonstrated a distinct temporal relationship between injury (axotomy) of SON/PVN axons and the establishment of new neurovascular zones following hypophysectomy with the up-regulation of NOS. This up-regulation appears to correlate well with successful regeneration in the mammalian neurohypophyseal system. We have also successfully inhibited axonal regeneration with the use of nitroarginine, a competitive antagonist of NO. NOS up-regulation attendant to regeneration of SON and PVN axons may have inestimable clinical implications, particularly with respect to closed head injury and cerebral contusion that involves the mechanical shearing of the infundibular stalk. In addition, this investigation has reaffirmed that large numbers of bona fide neurons migrate and emerge upon the floor of the adjacent third cerebral ventricle shortly following hypophysectomy (within 2 weeks). The origin and mechanisms of neuronal migration and plasticity following hypophysectomy are the subject of interpretation and discussion in this investigation.

Vasoactive intestinal polypeptide in sacral primary sensory pathways in the cat

Unmyelinated sensory axons in the sacral spinal cord may play a role in bladder reflexes under certain pathological conditions. Previous data suggested vasoactive intestinal polypeptide (VIP) might be contained exclusively in sensory C‐fibers, some of which innervate the bladder. This study was undertaken to describe the morphology of these VIP fibers in the sacral cord of the cat. VIP immunoreactivity was confined to unmyelinated axons observed at several levels of the sensory pathway including the dorsal root ganglia, dorsal roots, Lissauers tract, and the lateral collateral pathway. A combination of light and electron microscopic observations showed VIP‐immunoreactive fibers with labeled varicosities and synaptic terminals in laminae I, IIo, V, VII, and X. VIP‐immunolabeled varicosities had a mean diameter of 1.6 μm (range = 0.11–7.4 μm, S.D. = 1.01, n = 311) with a small percentage (8%) being relatively large (3–7.4 μm). VIP varicosities contained a mixture of small clear vesicles (CLV) and large dense core vesicles (LDV). Although most varicosities contained a moderate number of LDVs (14.86 LDVs/μm2), some varicosities contained a large number of LDVs, whereas others contained very few. Varicosities that possessed synaptic specializations were classed as terminals and were divided into three morphological classes. Two of these resembled Grays Type I terminal, whereas a third was similar to the Grays Type II terminal. There was no consistent relationship between vesicle content of the terminal and the type of synaptic contact it possessed. This study shows that in the sacral spinal cord of the cat, VIP terminals originate only from C‐fibers, terminate primarily in laminae I and V, and exhibit a variety of morphologies consistent with heterogeneous origins and functions of the lower urinary tract. J. Comp. Neurol. 407:381–394, 1999.

Transplantation of the Mammalian Pineal Gland: Studies of Survival, Revascularization, Reinnervation, and Recovery of Function

The survival, revascularization, reinnervation, and recovery of function of transplanted rat pineal glands were studied following grafting into four different locations in pinealectomized rats. Pineal grafts were well vascularized by fenestrated capillaries. Pinealocytes in the grafts maintained high-metabolic activity. More nerve fibers and terminals were observed in the grafts within the anterior chamber of the eye than in the third cerebral ventricle and the pineal region (in situ transplantation). No fibers or terminals were found in grafts placed beneath the renal capsule. Nighttime serum melatonin levels increased significantly in pinealectomized rats with transplants into either the third cerebral ventricle or the anterior eye chamber. This increase might reflect graft reinnervation. Yet day-night differences in serum melatonin were observed only in host rats receiving transplants in the anterior eye chamber. In conclusion, pinealocytes survived transplantation into different locations and exhibited ultrastructural features indicative of active secretory processes; however, day-night differences in serum melatonin are only restored following transplants into the anterior eye chamber. Reinnervation of the grafts by the host superior cervical ganglion is necessary for this restoration.

Correlative scanning-immunoelectromicroscopic analysis of neuropeptide localization and neuronal plasticity in the endocrine hypothalamus

Thirty-two Sprague-Dawley rats were divided into four groups, eight rats per group. Animals were hypophysectomized with removal of both the pars distalis and the neural lobe of the neurohypophysis. Groups of eight rats were euthanized 1, 2, 4 and 8 weeks following hypophysectomy and prepared for routine scanning electron microscopy (SEM) and correlative immunoelectron microscopy employing antisera against arginine vasopressin (AVP). Eight normal rats served as controls. In experimental rats that survived one to eight weeks posthypophysectomy, remarkable neuroanatomical alterations were notable in the median eminence and adjacent third cerebral ventricular lumen. In contrast to normal control rats, large numbers of neurites were observed with SEM to insinuate from the lateral recess into the cerebral ventricular lumen and as early as one week following hypophysectomy they overgrew the apical surfaces of ependymal cells that constitute the lining of the cerebral ventricle. Immunoelectron microscopy revealed that a significant proportion of these neurites were magnocellular in origin in that they harbored AVP-positive neurosecretory vesicles. In addition to large numbers of invading magnocellular neurites, neuronal perikayria with apparent axosomatic synapses were observed to emerge upon the thick feltwork of invading axons, the latter of which appeared to freely terminate within the ventricular lumen. AVP-positive axon profiles were, in addition, seen to terminate upon the basal lamina of portal perivascular spaces in the zona externa of the median eminence. These data are consistent with the idea that following hypophysectomy (to include high stalk section of the neurohypophyseal system), that there is rapid, and dynamic sprouting and regrowth of AVP-positive axons into the adjacent third cerebral ventricular lumen and to the contact zone of the median eminence as well. This phenomenon may represent a compensatory physiological response to injury of the neurohypophyseal system characterized by a highly plastic neuroanatomical reorganization of magnocellular elements which appear to utilize the CSF of the third cerebral ventricle as a functional terminus for the neurocisternal secretion of AVP which ultimately enters the systemic circulation.

POST-TRAUMATIC MIGRATION AND EMERGENCE OF A NOVEL CELL LINE UPON THE EPENDYMAL SURFACE OF THE THIRD CEREBRAL VENTRICLE IN THE ADULT MAMMALIAN BRAIN

This investigation describes the migration and emergence of significant numbers of what appear to be neuron‐like cells upon the surface of the median eminence of the adult rodent neurohypophyseal system of the endocrine hypothalamus following the trauma of hypophysectomy. These cells appear to migrate through the neuropil of the underlying median eminence and emerge in large numbers upon the surface of the third cerebral ventricle within 7 days following hypophysectomy (axotomy) of supraoptic (SON) and paraventricular neurites (PVN) of the adult neurohypophyseal system. Previous investigations have demonstrated regeneration of the neural stem and neural lobe in a variety of mammalian species (Adams et al., J Comp Neurol, 1969;135:121–144; Beck et al., Neuroendocrinology, 1969;5:161–182; Scott et al., Exp Neurol, 1995;131–1:23–39; Scott and Hansen, Vir Med 1997;124:249–261). It also has been demonstrated that the process of regeneration is invariably accompanied by the up‐regulation of nitric oxide synthase (NOS), the enzyme that catalyzes arginine to nitric oxide (NO) and that both neurohypophyseal regeneration, as well as migration and emergence of neuron‐like cells upon the surface of the adjacent third cerebral ventricle, is associated with the up‐regulation of NOS and increased expression of NO. It also has been amply demonstrated that this entire process of neurohypophyseal regeneration and cell migration is completely inhibited by the introduction of the antagonist of nitric oxide, namely, nitroarginine (Scott et al., Exp Neurol, 1995;131–1:23–39; Scott and Hansen, Vir Med, 1997;124:249–261). The emergence and migratory dynamics of this novel cell line upon the floor of the rodent third cerebral ventricle are discussed with respect to the role of the ubiquitous free radical NO and the implications and potential clinical applications of neuronal migration following trauma in the human central nervous system (CNS). Anat Rec 256:233–241, 1999.

Transplantation of the pineal gland in the mammalian third cerebral ventricle

Fine structural correlates and functional parameters were measured in pinealectomized rats following grafting of the pineal gland into the third cerebral ventricle. Pinealectomy caused a significant decrease in serum melatonin concentration of animals compared to that in normal controls. No significant difference was observed in the serum melatonin concentration between pinealectomized rats and those receiving sham transplantation with fragments of occipital cortex. By 6 weeks nearly 50% of pinealectomized rats receiving pineal transplants demonstrated a significant increase in the serum melatonin concentration in contrast to that of pinealectomized rats and pinealectomized animals receiving sham transplants. Pinealocytes survived and flourished following transplantation from the epithalamic region to the third cerebral ventricle of the hypothalamus in host rats. These cells were found to be arranged individually or in clusters surrounding fenestrated capillaries of the graft. Moreover, these pinealocytes demonstrated ultrastructural features indicative of an active secretory process, including dense-core and clear vesicles as well as vacuoles containing flocculent material. Additional characteristics distinctive of normal control pinealocytes were observed in surviving cells of grafts, such as synaptic ribbons, synaptic ribbon fields, and myeloid bodies. Bundles of unmyelinated axons and apparent adrenergic nerve endings were observed with transmission electron microscopy and immunocystochemistry using antisera against tyrosine hydroxylase (TH). Nerve fibers and terminals were found within perivascular spaces surrounding fenestrated capillaries of viable grafts. These reported observations suggest that a significant population of transplanted pinealocytes recover functional activity (e.g., heightened melatonin secretion) following stereotaxic grafting into the third cerebral ventricles of pinealectomized animals. This apparent recovery of function may be linked directly to reinnervation of the gland by nerve fibers that appear to arise from the underlying median eminence.

In Situ Fixation of the Neonatal Brain and Spinal Cord

A delay in the autopsy can result in significant tissue autolysis, especially in the central nervous system. We have developed a rapid technique of in situ fixation that preserves central nervous system tissues until the formal autopsy can be performed. Through the lateral margin of the anterior fontanelle, Zambonis solution is injected percutaneously into the lateral ventricles and allowed to exit via an intrathecal spinal needle. The choice of fixative allows a wide array of postmortem studies to be done.