Biomedical subjects

J R O'Kusky

Publications and source records attributed to J R O'Kusky.

At least 19 recordsLinked to original sources

PubMed2000

Insulin-like growth factor-I promotes neurogenesis and synaptogenesis in the hippocampal dentate gyrus during postnatal development.

The in vivo actions of insulin-like growth factor-I (IGF-I) on the growth and development of the hippocampal dentate gyrus were investigated in transgenic mice that overexpress IGF-I postnatally in the brain and in normal nontransgenic littermate controls. Stereological analyses of the dentate gyrus were performed by light and electron microscopy on days 7, 14, 21, 28, 35, and 130 to determine postnatal changes in the numerical density and total number of neurons and synapses. The volumes of both the granule cell layer and the molecular layer of the dentate gyrus were significantly increased by 27-69% in transgenic mice after day 7, with the greatest relative increases occurring by day 35. Although the numerical density of neurons in the granule cell layer did not differ significantly between transgenic and control mice at any age studied, the total number of neurons was significantly greater in transgenic mice by 29-61% beginning on day 14. The total number of synapses in the molecular layer was significantly increased by 42-105% in transgenic mice from day 14 to day 130. A transient increase in the synapse-to-neuron ratio was found in transgenic mice at postnatal days 28 and 35 but not at day 130. This finding indicates a disproportionate increase in synaptogenesis, exceeding that expected for the observed increase in neuron number. Our results demonstrate that IGF-I overexpression produces persistent increases in the total number of neurons and synapses in the dentate gyrus, indicating that IGF-I promotes both neurogenesis and synaptogenesis in the developing hippocampus in vivo.

Analysis of Variance↗

PubMed1999

Increased insulin-like growth factor-I (IGF-I) expression during early postnatal development differentially increases neuron number and growth in medullary nuclei of the mouse.

Morphometric analyses of the medulla were performed in transgenic mice that overexpress insulin-like growth factor-I (IGF-I) postnatally and in non-transgenic littermates. The total volume of the medulla was increased in transgenic mice at all postnatal ages studied: 14 days (18%), 21 days (23%), 28 days (23%), and 35 days (27%). By 35 days of age, the volumes of individual medullary nuclei were also increased: nucleus of the tractus solitarius (NTS, 59%), dorsal motor nucleus of the vagus (DMV, 84%), hypoglossal nucleus (HN, 29%) and facial nucleus (FN, 21%). Neuron number in transgenic mice was significantly greater in NTS (50%) and DMV (53%), but not in the HN or the FN. Motor neurons in DMV, HN and FN of transgenic mice exhibited increases in mean profile areas of the soma and decreased numerical densities, suggesting increases in neuritic outgrowth. These results point to IGF-I actions in promoting neuron survival and growth, and suggest that IGF-I has differential effects on distinct neuron populations, possibly depending upon its time of expression.

Animals↗

PubMed1999

Neuronal degeneration in the basal ganglia and loss of pallido-subthalamic synapses in mice with targeted disruption of the Huntington's disease gene.

Huntington's disease (HD) is a progressive neurodegenerative disorder associated with CAG repeat expansion within a novel gene (IT15). We have previously created a targeted disruption in exon 5 of Hdh (Hdhex5), the murine homologue of the HD gene. Homozygotes for the Hdhex5 mutation exhibit embryolethality before embryonic day 8.5, while heterozygotes survive to adulthood and display increased motor activity and cognitive deficits. Detailed morphometric and stereological analyses of the basal ganglia in adult heterozygous mice were performed by light and electron microscopy. Morphometric analyses demonstrated a significant loss of neurons from both the globus pallidus (29%) and the subthalamic nucleus (51%), with a normal complement of neurons in the caudate-putamen and substantia nigra. The ultrastructural appearance of sporadic degenerating neurons in these regions indicated apoptosis. The highest frequency of apoptotic neurons was observed in the globus pallidus and subthalamic nucleus. Stereological analyses in the subthalamic nucleus revealed a significant decrease in the numerical density of symmetric synapses (43%), suggesting a relatively selective loss of inhibitory pallido-subthalamic afferents. Immunohistochemistry using antibodies against enkephalin and substance-P was unremarkable in heterozygotes, indicating a normal complement of enkephalin-immunoreactive striatopallidal afferents and substance-P-immunoreactive striatopeduncular and striatonigral afferents in these animals. These findings show that loss of an intact huntingtin protein is associated with significant morphological alterations in the basal ganglia of adult mice, indicating an important role for this protein during development of the central nervous system.

Afferent Pathways↗

PubMed1998

Postnatal changes in the numerical density and total number of asymmetric and symmetric synapses in the hypoglossal nucleus of the rat.

Morphometric analyses were performed to investigate the progressive and regressive phases of synaptogenesis in the hypoglossal nucleus of the rat during normal postnatal development. The total volume of the hypoglossal nucleus and both the numerical density (NV, contacts per mm3) and total number of synapses were measured at 5-day intervals from birth to postnatal day 30 and in young adults. Values of NV were calculated separately for asymmetric and symmetric synapses as well as for axospinous, axodendritic and axosomatic contacts. The volume of the hypoglossal nucleus increased significantly from birth to postnatal day 30 (414%) with no further increase in the adult. The NV of all synapses increased significantly from birth to day 20 (131%), followed by a significant decreases in adults (45%). The total number of synapses increased significantly from birth to day 20 (843%), followed by a significant decrease in adults (30%). Similar developmental changes in density and total number were observed for asymmetric and symmetric synapses. The magnitude of synapse elimination, occurring after day 20, was approximately 30% for both morphological types. During postnatal development the vast majority of synapses in the hypoglossal nucleus were found to form axodendritic contacts (85-95%). Synapse elimination was observed for axospinous, axodendritic and axosomatic contacts. These findings indicate that the progressive and regressive phases of synaptogenesis occur earlier in the hypoglossal nucleus than in the cerebral cortex of the rat, suggesting a caudal-to-rostral gradient. Synapse elimination was not restricted on the basis of morphological type or postsynaptic target site.

Age Factors↗

PubMed1997

Toward understanding the molecular pathology of Huntington's disease.

Huntington's Disease (HD) is caused by expansion of a CAG trinucleotide beyond 35 repeats within the coding region of a novel gene. Recently, new insights into the relationship between CAG expansion in the HD gene and pathological mechanisms have emerged. Survival analysis of a large cohort of affected and at-risk individuals with CAG sizes between 39 and 50 repeats have yielded probability curves of developing HD symptoms and dying of HD by a certain age. Animals transgenic for the first exon of huntingtin with large CAG repeats lengths have been reported to have a complex neurological phenotype that bears interesting similarities and differences to HD. The repertoire of huntingtin-interacting proteins continues to expand with the identification of HIP1, a protein whose yeast homologues have known functions in regulating events associated with the cytoskeleton. The ability of huntingtin to interact with two of its four known protein partners appears to be influenced by CAG length. Caspase 3 (apopain), a key cysteine protease known to play a seminal role in neural apoptosis, has also been demonstrated to specifically cleave huntingtin in a CAG length-dependent manner. Many of these features are combined in a model suggesting mechanisms by which the pathogenesis of HD may be initiated. The development of appropriate in vitro and animal models for HD will allow the validity of these models to be tested.

Age of Onset↗

PubMed1996

Synaptogenesis in hemimegalencephaly: the numerical density of asymmetric and symmetric synapses in the cerebral cortex.

Specimens of cerebral cortex were prepared for electron microscopy from cortical resections performed for the treatment of intractable seizures in four cases of hemimegalencephaly (HME). Morphometric analyses were performed to determine mean cortical thickness, the numerical density of synapses (NV, contacts per mm3) and the number of synapses in a column of cortex beneath 1 mm2 of pial surface. The NV were calculated separately for asymmetric and symmetric synapses as well as for axospinous, axodendritic and axosomatic contacts. Four cases of Rasmussen's encephalitis served as controls, with tissue being sampled from a region distant to the site of the inflammatory lesion without obvious necrosis. The NV of synapses did not differ significantly between HME cases and controls. The proportions or asymmetric and symmetric synapses were similar in both groups, as were the proportions of axospinous, axodendritic and axosomatic contacts. However, there was a significant increase in mean cortical thickness in HME cases (130%, P < 0.05). Consequently, there was a significant increase in the total number of synapses in a column of cortex (126%, P < 0.05). In HME the cerebral cortex is characterized by synaptic dysgenesis. Although synaptic density per unit volume of tissue appears relatively normal, the increased thickness and volume of the cerebral cortex provides for an increase in the total number of synapses in a given cytoarchitectonic area.

Brain Diseases↗

PubMed1995

Targeted disruption of the Huntington's disease gene results in embryonic lethality and behavioral and morphological changes in heterozygotes.

Huntington's disease (HD) is an incurable neuropsychiatric disease associated with CAG repeat expansion within a widely expressed gene that causes selective neuronal death. To understand its normal function, we have created a targeted disruption in exon 5 of Hdh (Hdhex5), the murine homolog of the HD gene. Homozygotes die before embryonic day 8.5, initiate gastrulation, but do not proceed to the formation of somites or to organogenesis. Mice heterozygous for the Hdhex5 mutation display increased motor activity and cognitive deficits. Neuropathological assessment of two heterozygous mice shows significant neuronal loss in the subthalamic nucleus. These studies show that the HD gene is essential for postimplantation development and that it may play an important role in normal functioning of the basal ganglia.

Animals↗

PubMed1995

Sudden infant death syndrome: increased number of synapses in the hypoglossal nucleus.

The medulla was sampled from nine cases of sudden infant death syndrome (SIDS) and from six age-matched control cases without neurological disease. Morphometric analyses were performed on serial Nissl sections through the hypoglossal nucleus on the left side of the medulla. The total volume of the nucleus and both the numerical density (Nv, cells per mm3) and total number of neurons were measured. Tissue from the remaining hypoglossal nucleus was prepared for electron microscopy using the ethanolic phosphotungstic acid method to stain synaptic contacts. Stereological analyses were performed to determine the Nv and total number of synapses. Total volume of the hypoglossal nucleus was significantly greater (36%) in SIDS cases than in controls. The Nv of neurons was significantly less than in controls (28%), although the total number of neurons did not differ significantly. The mean profile area of motor neuron cell bodies was significantly greater (30%) in SIDS cases, with no differences in the mean profile areas for interneurons or glia. The Nv of synapses did not differ significantly between SIDS cases and controls, although the total number of synapses was greater (61%) in SIDS. These abnormalities in growth indicate a greater volume of neuropil in a hypoglossal nucleus containing a normal complement of neurons. The greater number of synapses in SIDS cases is consistent with a failure to eliminate normally extraneous synapses during early development.

Brain↗

PubMed1995

Sudden infant death syndrome: postnatal changes in the volumes of the pons, medulla and cervical spinal cord.

The brainstem and cervical spinal cord were sampled from 45 cases of sudden infant death syndrome (SIDS), from 17 control cases without neurological disease, and from three negative control cases with abnormal growth of the central nervous system (36-98 postconceptional weeks). Morphometric analyses were performed on serial Nissl sections to determine the total volumes of the pons, nucleus pontis, medulla and cervical spinal cord. Normal development was characterized by a linear increase in the volumes of these regions during the first postnatal year. Regression analysis revealed that in SIDS cases the rates of increase in the volumes of the pons and nucleus pontis were significantly greater than in controls (56% and 83%, respectively), while growth rates did not differ significantly for the medulla and cervical spinal cord. By direct comparison, there was a significant increase in the mean volumes of the pons (33%), nucleus pontis (38%) and medulla (19%) in SIDS cases when compared to controls. No evidence of excessive edema or gliosis was noted in the brainstem by light and electron microscopy to account for the increased volumes. Subtle morphological abnormalities in brainstem neurons from SIDS cases, including an increased size of Nissl bodies in the cytoplasm of large motor neurons and the presence of paranucleolar coiled bodies, were consistent with an increased synthesis and transport of ribosomal RNA, an increased synthesis of cellular proteins and neuronal hypertrophy.

Apnea↗

PubMed1994

Sudden infant death syndrome: increased synaptic density in the central reticular nucleus of the medulla.

The medulla was sampled from nine cases of sudden infant death syndrome (SIDS) and from six age-matched control cases without neurological disease. Morphometric analyses were performed on serial Nissl sections through the left half of each medulla to determine the total volume of the hemimedulla and the numerical density of neurons (Nv, neurons per mm3) in the central reticular nucleus. Blocks of tissue from the right half were prepared for electron microscopy using the ethanolic phosphotungstic acid (EPTA) method to stain synaptic contacts. Stereological analyses were performed to determine the Nv of synapses in the central reticular nucleus. Total volume of the hemimedulla did not differ significantly between SIDS cases and controls. However, in the central reticular nucleus of SIDS cases, the Nv of neurons was significantly less than in controls (30%), while the mean profile area of reticular neurons was significantly greater (39%). Synaptogenesis in control cases was characterized by a gradual decrease in the Nv of synapses from approximately 150 million at 40 postconceptional weeks to 110 million at 84 weeks. In SIDS cases the Nv of synapses was significantly greater (38%). These results demonstrate a disorder of synaptogenesis in the central reticular nucleus of SIDS cases. In view of previous reports of a persistence of dendritic spines on reticular neurons in SIDS, the results are consistent with a failure to eliminate normally extraneous synapses during early development.

Female↗

PubMed1992

Sudden infant death syndrome: postnatal changes in the numerical density and total number of neurons in the hypoglossal nucleus.

Tissue specimens from the medulla were sampled from 28 sudden infant death syndrome (SIDS) victims and from 15 control cases without neurological disease (36-95 postconceptional weeks). Morphometric analyses were performed on serial Nissl sections through the hypoglossal nucleus. The total volume of the hypoglossal nucleus, the numerical density (Nv, cells per mm3) and the total number of motor neurons, interneurons and glia were determined. Normal development was characterized by a linear increase in the volume of the hypoglossal nucleus during the first postnatal year. While the Nv of neurons decreased, the total number of neurons remained relatively constant at approximately 7,600 motor neurons and 3,100 interneurons. In SIDS cases the rate of increase in the volume of the hypoglossal nucleus was significantly greater than in controls (79%). The Nv of neurons was less than in controls (25-30%), although the total number of motor neurons and interneurons did not differ significantly. In SIDS cases the mean profile area of motor neuron cell bodies was significantly greater than in controls (29%), while the mean profile areas of interneurons and glia did not differ. These abnormalities in growth indicate a greater volume of neuropil in a hypoglossal nucleus containing a normal complement of neurons. The disproportionately rapid increase in volume of neuropil in the hypoglossal nucleus of SIDS cases may result from an increased arborization of dendrites on the motor neurons.

Autopsy↗

PubMed1991

Cytomegalovirus infection of the developing brain alters catecholamine and indoleamine metabolism.

Tissue concentrations of noradrenaline (NA), serotonin (5-HT), dopamine (DA) and selected metabolites were measured in the spinal cord, cerebellum, cerebral cortex and caudate-putamen of developing mice following intraventricular inoculation with murine cytomegalovirus (MCMV) on postnatal day 10. MCMV-infected animals exhibited transient signs of neurological impairment, including apparent hypertonicity of hindlimb extensors and abnormal gait, beginning on days 14-16 and continuing for 3-5 days. At the onset of neurological impairment, tissue concentrations of NA were significantly reduced in the spinal cord (20%), cerebellum (32%) and cerebral cortex (40%) of infected animals. Levels of 5-HT were significantly increased in the caudate-putamen (50%), while 5-hydroxyindoleacetic acid (5-HIAA) was increased in both the spinal cord (94%) and caudate-putamen (65%). The ratio of 5-HIAA/5-HT, which is frequently used as an estimate of turnover of 5-HT, was significantly increased in the spinal cord (90%) at the onset of neurological impairment. In the caudate-putamen of MCMV-infected animals, there were significant increases in the tissue levels of DA (37%), homovanillic acid (HVA, 41%) and 3,4-dihydroxyphenylacetic acid (DOPAC, 34%). All neurochemical parameters were normal in the MCMV-infected animals by postnatal day 70, approximately 50 days after the resolution of neurological signs. These results indicate transient alterations in monoamine metabolism in the developing nervous system during the pathogenesis of cytomegalovirus-induced movement and postural disorders.

Animals↗

PubMed1989

In situ hybridization analysis for herpes simplex virus nucleic acids in Alzheimer disease.

Histological sections of brain from patients showing evidence of advanced pathology of Alzheimer disease (AD) were examined for the presence of herpes simplex type-1 (HSV-1) nucleic acids by a sensitive in-situ hybridization technique. Samples from neurologically normal patients were examined in parallel. Sensitivity of the assay was verified by the detection of HSV-1 nucleic acids in neurons of trigeminal ganglia taken from cases of AD and normal controls. This indicated that the hybridization reaction was sufficiently sensitive to detect latent HSV-1 infections. Positive hybridization in the brain was only detected in a confirmed case of herpes simplex virus encephalitis. These results appear to confirm previous reports that HSV-1 infection is not directly involved in the pathology associated with AD.

Aged↗

PubMed1989

Methylmercury-induced movement and postural disorders in developing rat: high-affinity uptake of choline, glutamate, and gamma-aminobutyric acid in the cerebral cortex and caudate-putamen.

Subcutaneous administration of methylmercuric chloride to neonatal rats resulted in movement and postural disorders during the fourth postnatal week. Sodium-dependent high-affinity uptake of radiolabeled choline, glutamate, and gamma-aminobutyric acid (GABA) was measured in homogenates of cerebral cortex and caudate-putamen. There was a significant decrease in the uptake of [3H]choline in the cerebral cortex, but not in the caudate-putamen, at the onset of neurological impairment (73-75%) and at one subclinical stage of toxicity (58-64%). No significant differences in [3H]glutamate uptake were detected in either region. The uptake of [3H]GABA in the presence of 1 mM beta-alanine, which was employed to inhibit the glial uptake process, was reduced significantly in both the cerebral cortex and caudate-putamen at the onset of neurological impairment (50-62%) and at one subclinical stage (40-51%). This decrease in [3H]GABA uptake is consistent with the results of previous studies using this animal model, which demonstrated a preferential degeneration of GABAergic neurons in the cerebral cortex and caudate-putamen of methylmercury-treated animals. Because the high-affinity uptake of choline is the rate-limiting step for acetylcholine synthesis by cholinergic neurons, the decrease in [3H]choline uptake may reflect an abnormal development of cholinergic innervation of the cerebral cortex.

Animals↗

PubMed1989

Increased uric acid in the developing brain and spinal cord following cytomegalovirus infection.

Tissue concentrations of uric acid were determined in the spinal cord, cerebellum, caudate-putamen, and cerebral cortex of developing mice following intraventricular inoculation with murine cytomegalovirus (MCMV) on postnatal day 10. Transient signs of neurological impairment were observed in MCMV-infected animals beginning on days 13-16 and continuing until days 19-21. At the onset of neurological impairment, uric acid concentrations in tissues from infected animals were 17-60-fold greater than in control animals. On postnatal day 70, 60 days after inoculation and 40 days after resolution of neurological signs, uric acid levels were still two- to threefold greater in infected animals. Histological examination revealed signs of focal ischemia in the cerebral and cerebellar cortices of MCMV-infected mice only at the onset of neurological impairment, with ischemic cell changes in some pyramidal neurons of the cerebral cortex. These results indicate that uric acid may be a sensitive marker of persistent vascular pathology resulting from cytomegalovirus infection of the developing nervous system.

Aging↗

PubMed1988

Increased cytochrome oxidase activity of mesencephalic neurons in developing rats displaying methylmercury-induced movement and postural disorders.

Subcutaneous administration of the neurotoxin methylmercuric chloride to developing rats produced movement and postural disorders during the 4th postnatal week. Cytochrome oxidase histochemistry revealed an increase in the oxidative metabolic activity of small neurons within the magnocellular red nucleus (RMC) and the interrubral mesencephalon. A concurrent suppression of cytochrome oxidase activity in the large neurons and neuropil of RMC was apparent relative to controls. Decortication on postnatal day 3 did not alter the course of motor impairment or the cytochrome oxidase histopathology, suggesting that the role of neocortex in the pathogenesis of methylmercury-induced movement and postural disorders is minimal.

Animals↗

PubMed1988

Methylmercury-induced movement and postural disorders in developing rat: loss of somatostatin-immunoreactive interneurons in the striatum.

Tissue concentrations of the neuropeptide somatostatin and the specific activities of glutamic acid decarboxylase (GAD) were measured in several regions of the central nervous system in young rats, following chronic postnatal administration of methylmercuric chloride. By the beginning of the fourth postnatal week, these animals exhibited clinical signs of a mixed spastic/dyskinetic syndrome with visual deficits. At the onset of neurological impairment, a significant decrease in GAD activity was detected in the occipital cortex (48-49%) and striatum (45-50%) when compared to either normal or weight-matched controls. At one subclinical stage of toxicity, decreased GAD activity was detected only in the occipital cortex (29-30%). Tissue levels of somatostatin did not change significantly in the occipital cortex of methylmercury-treated animals at any stage of the experiment. However, somatostatin levels in the striatum were significantly reduced at the onset of neurological impairment (55-57%) and at one subclinical stage of toxicity (49-54%). Immunohistochemistry for somatostatin- and neuropeptide Y-immunoreactive neurons confirmed a marked loss of cells in the dorsolateral region of the striatum with atrophy of the surviving neurons. In the cerebral cortex of methylmercury-treated animals the morphology and distribution of somatostatin-positive neurons appeared normal. In view of the reported co-localization of GAD and somatostatin in some non-pyramidal neurons of the cerebral cortex, these results indicate that methylmercury-induced lesions of the developing cerebral cortex involve a subpopulation of GABAergic neurons which are not co-localized with somatostatin. In the striatum, where GAD and somatostatin are not co-localized within the same neurons, methylmercury-induced lesions involve both GABAergic and somatostatin-positive neurons.

Aging↗

PubMed1988

Methylmercury-induced movement and postural disorders in developing rat: regional analysis of brain catecholamines and indoleamines.

Subcutaneous administration of methylmercury (MeHg) to rats during early postnatal development resulted in movement and postural disorders by day 22-24. Tissue concentrations of norepinephrine (NE), serotonin (5-HT), dopamine (DA) and selected metabolites were measured in the cerebral cortex, spinal cord and caudate-putamen at the onset of neurological impairment and at two subclinical stages of toxicity. In the cerebral cortex there was a significant increase in tissue concentrations of 5-HT (54-81%) and 5-hydroxyindoleacetic acid (HIAA, 133-178%) at the onset of neurological impairment. Similar increases were detected in the spinal cord for 5-HT (19-43%) and HIAA (98-123%) as well as an increase in the concentration of NE (42-51%). In the caudate-putamen there were significant increases in the concentrations of NE (98-116%), HIAA (108-124%) and DA (28-29%) with a significant decrease in the concentration of 3,4-dihydroxyphenylacetic acid (DOPAC, 20-27%); however, tissue levels of homovanillic acid (HVA) did not change significantly. Many of these changes were detected at subclinical stages of MeHg toxicity. The ratio of HIAA/5-HT, which is frequently used as an estimate of turnover for 5-HT, was significantly increased in all 3 tissues at the onset of neurological impairment (38-94%) and at one subclinical stage (47-114%). The ratio of (DOPAC + HVA)/DA was significantly decreased in caudate-putamen at all 3 stages of toxicity (18-40%). These changes indicate altered metabolism in aromatic amine systems in the developing central nervous system during the pathogenesis of MeHg-induced movement and postural disorder.

Aging↗