The NIH MRI study of normal brain development (Objective-2): newborns, infants, toddlers, and preschoolers.

The Magn. Reson. Imaging (MRI) study of normal brain development currently conducted by the Brain Development Cooperative Group represents the most extensive MRI study of brain and behavioral development from birth through young adulthood ever conducted. This multi-center project, sponsored by four Institutes of the National Institutes of Health, uses a combined longitudinal and cross-sectional design to characterize normal, healthy brain and behavioral development. Children, ages newborn through 18-plus years of age, receive comprehensive behavioral, neurological and multimodal MRI evaluations via Objective-2 (birth through 4-years 5-months of age) and Objective-1 (4-years 6-months through 18 years of age and older). This report presents methods (e.g., neurobehavioral assessment, brain scan) and representative preliminary results (e.g., growth, behavior, brain development) for children from newborn through 4-years 5-months of age. To date, 75 participants from birth through 4-years 5-months have been successfully brain scanned during natural sleep (i.e., without sedation); most with multiple longitudinal scans (i.e., 45 children completing at least three scans, 22 completing four or more scans). Results from this younger age range will increase our knowledge and understanding of healthy brain and neurobehavioral development throughout an important, dynamic, and rapid growth period within the human life span; determine developmental associations among measures of brain, other physical characteristics, and behavior; and facilitate the development of automated, quantitative MR image analyses for neonates, infants and young children. The correlated brain MRI and neurobehavioral database will be released for use by the research and clinical communities at a future date.

A prospective, longitudinal diffusion tensor imaging study of brain injury in newborns.

OBJECTIVE: To establish the magnitude and time course of the changes in water diffusion coefficient (D(av)) following newborn infant brain injury. METHODS: Ten newborn infants at high risk for perinatal brain injury were recruited from the neonatal intensive care unit. Conventional and diffusion tensor MRI was performed on three occasions during the first week of life. Regions of injury were determined by evaluating conventional MR images (T1, T2, fluid-attenuated inversion recovery) at 1 week after injury. D(av) values were determined for these regions for all three scans. RESULTS: D(av) values were decreased in most infants 1 day after injury, but injury was not evident or underestimated in 4 of 10 infants despite the presence of injury on conventional imaging at 1 week. By the third day, D(av) values were decreased in injured areas in all infants, reaching a nadir of approximately 35% less than normal values. By the seventh day after injury, D(av) values were returning to normal (pseudonormalization). CONCLUSIONS: MR diffusion images (for which contrast is determined by changes in D(av)) obtained on the first day after injury do not necessarily show the full extent of ultimate injury in newborn infants. Images obtained between the second and fourth days of life reliably indicate the extent of injury. By the seventh day, diffusion MR is less sensitive to perinatal brain injury than conventional MR because of transient pseudonormalization of D(av). Overall, diffusion MR may not be suitable as a gold standard for detection of brain injury during the first day after injury in newborn infants.

Normal brain maturation during childhood: developmental trends characterized with diffusion-tensor MR imaging.

PURPOSE: To characterize the maturational changes in water diffusion within central gray matter nuclei and central white matter pathways of the human brain by using diffusion-tensor magnetic resonance (MR) imaging. MATERIALS AND METHODS: Retrospective analysis of normal MR examination findings in 153 subjects (age range, 1 day to 11 years) referred for clinical neuroimaging was performed. All studies included diffusion tensor-encoded echo-planar MR imaging. Isotropic diffusion coefficient (D) and diffusion anisotropy (A(sigma)) were measured in the corpus callosum, internal capsule, caudate nucleus, lentiform nucleus, and thalamus. RESULTS: exhibited biexponential decay with age in gray and white matter regions, except for monoexponential decay in the genu of the corpus callosum. There was a steep nonlinear increase of A(sigma) in white matter tracts that paralleled the time course of the decline in D. In basal ganglia, only a small linear increase in A(sigma) was observed in patients. A(sigma) changes in the thalamus were intermediate between basal ganglia and white matter structures. CONCLUSION: Changes in magnitude and anisotropy of water diffusion follow stereotypical time courses during brain development that can be empirically described with multiexponential regression models, which suggests that quantitative scalar parameters derived from diffusion-tensor MR imaging may provide clinically useful developmental milestones for brain maturity.

Human fetal and neonatal movement patterns: Gender differences and fetal-to-neonatal continuity.

Longitudinal quantification of leg movements per minute for human subjects during both fetal and neonatal periods was accomplished from videotapings conducted antenatally (ultrasonography 30, 34, and 37 weeks gestational age) and postnatally (birth and 6 weeks of age). Fetal/neonatal subjects displayed decreasing numbers of leg movements per minute during antenatal development (30 to 37 weeks), followed by increasing numbers of leg movements per minute during postnatal development (birth to 6 weeks of age). Male subjects displayed greater numbers of leg movements per minute than female subjects during both antenatal and postnatal development. Fetal-to-neonatal continuity for numbers of leg movements per minute was found for comparisons between fetal (37 weeks gestational age) and neonatal (during sleep states at birth) measures, and females displayed a stronger and different movement continuity pattern than males. These results indicate a differential time course for neurobehavioral development of male and female fetuses/neonates, and the findings have implications for the clinical assessment of fetal neurobehavioral development and well-being.

BDNF protects against spatial memory deficits following neonatal hypoxia-ischemia.

Hypoxic-ischemic (H-I) brain injury in the human perinatal period often leads to significant long-term neurobehavioral dysfunction in the cognitive and sensory-motor domains. Using a neonatal H-I injury model (unilateral carotid ligation followed by hypoxia) in postnatal day seven rats, previous studies have shown that neurotrophins, such as brain-derived neurotrophic factor (BDNF), can be protective against neural tissue loss. The present study explored potential relationships between neural protective and behavioral protective strategies in this neonatal H-I model by determining if neonatal H-I was associated with behavioral spatial learning and memory deficits and whether the neurotrophin BDNF was protective against both brain injury and spatial learning/memory dysfunction. Postnatal day seven rats received vehicle or BDNF pretreatments (intracerebroventricular injections) followed by H-I or sham treatments and then tested for spatial learning and memory on the simple place task in the Morris water maze from postnatal days 20 to 30, and their brains were histologically analyzed at 4 weeks following treatments. H-I rats with vehicle pretreatment displayed significant tissue loss in the hippocampus (including CA1 neurons), cortex, and striatum, as well as severe spatial memory deficits (e.g., short probe times). BDNF pretreatment resulted in significant protection against both H-I-induced brain tissue losses and spatial memory impairments. These findings indicate that unilateral H-I brain injury in a neonatal rodent model is associated with cognitive deficits, and that BDNF pretreatment is protective against both brain injury and spatial memory impairment.

Normal brain in human newborns: apparent diffusion coefficient and diffusion anisotropy measured by using diffusion tensor MR imaging.

PURPOSE: To establish quantitative standards for the directionally averaged water apparent diffusion coefficient (D) and quantitative diffusion anisotropy (A sigma) of normal brains in newborns by using diffusion tensor magnetic resonance (MR) imaging. MATERIALS AND METHODS: Diffusion tensor MR imaging was performed during the first 36 hours of life in 22 newborns (gestational age range, 31-41 weeks). Values of D and A sigma were measured in regions of interest chosen in the cortical gray matter, centrum semiovale, caudate nuclei, lentiform nuclei, thalami, internal capsules, and cerebellar hemispheres. RESULTS: The D values in the gray and white matter in newborns are considerably higher than those in adults. There is a striking correlation between gestational age and D, with D decreasing as gestational age increases. The A sigma values in the white matter in newborns are lower than those in adults. Values of A sigma show statistically significant correlations with gestational age only in the white matter of the centrum semiovale, in which A sigma values increase sharply near term. CONCLUSION: The D values primarily reflect overall brain water content. The A sigma values are more sensitive to tissue microstructure (e.g., white matter packing and myelination). The D and A sigma images reveal information and not apparent on T1- and T2-weighted images.

Higher neonatal cerebral blood flow correlates with worse childhood neurologic outcome.

Cerebral blood flow (CBF) in newborn infants is often below levels necessary to sustain brain viability in adults. Controversy exists regarding the effects of such low CBF on subsequent neurologic function. We determined the current childhood neurologic status and IQ in 26 subjects who had measurements of CBF performed with PET in the neonatal period between 1983 and 1989 as part of a study of hypoxic-ischemic encephalopathy. Follow-up information at ages 4 to 12 years was obtained on all 26 subjects. Ten subjects had died. All 16 survivors underwent clinical neurologic evaluation, and 14 also underwent intelligence testing. Eight had abnormal clinical neurologic evaluations; eight were normal. The mean neonatal CBF in those with abnormal childhood neurologic outcome was significantly higher than in those with normal childhood neurologic outcome (35.64 +/- 11.80 versus 18.26 +/- 8.62 mL 100 g(-1) min(-1), t = 3.36, p = 0.005). A significant negative correlation between neonatal CBF and childhood IQ was demonstrated (Spearman rank correlation r = -0.675, p = 0.008). Higher CBF was associated with lower IQ. The higher CBF in subjects with worse neurologic and intellectual outcome may reflect greater loss of cerebrovascular autoregulation or other vascular regulatory mechanisms due to more severe brain damage.

Carboxyfullerenes as neuroprotective agents.

Two regioisomers with C3 or D3 symmetry of water-soluble carboxylic acid C60 derivatives, containing three malonic acid groups per molecule, were synthesized and found to be equipotent free radical scavengers in solution as assessed by EPR analysis. Both compounds also inhibited the excitotoxic death of cultured cortical neurons induced by exposure to N-methyl-D-aspartate (NMDA), alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), or oxygen-glucose deprivation, but the C3 regioisomer was more effective than the D3 regioisomer, possibly reflecting its polar nature and attendant greater ability to enter lipid membranes. At 100 microM, the C3 derivative fully blocked even rapidly triggered, NMDA receptor-mediated toxicity, a form of toxicity with limited sensitivity to all other classes of free radical scavengers we have tested. The C3 derivative also reduced apoptotic neuronal death induced by either serum deprivation or exposure to Abeta1-42 protein. Furthermore, continuous infusion of the C3 derivative in a transgenic mouse carrying the human mutant (G93A) superoxide dismutase gene responsible for a form of familial amyotrophic lateral sclerosis, delayed both death and functional deterioration. These data suggest that polar carboxylic acid C60 derivatives may have attractive therapeutic properties in several acute or chronic neurodegenerative diseases.

Neonatal hippocampal damage in rats: long-term spatial memory deficits and associations with magnitude of hippocampal damage.

This study investigated the effects of neonatal hippocampal ablation on the development of spatial learning and memory abilities in rats. Newborn rats sustained bilateral electrolytic lesions of the hippocampus or were sham-operated on postnatal day 1 (PN1). At PN20-25, PN50-55, or PN90-95, separate groups of rats were tested in a Morris water maze on a visible "cue" condition (visible platform in a fixed location of the maze), a spatial "place" condition (submerged platform in a fixed location), or a no-contingency "random" condition (submerged platform in a random location). Rats were tested for 6 consecutive days, with 12 acquisition trials and 1 retention (probe) trial per day. During acquisition trials, the rat's latency to escape the maze was recorded. During retention trials (last trial for each day, no escape platform available), the total time the rat spent in the probe quadrant was recorded. Data from rats with hippocampal lesions tested as infants (PN20-25) or as adults (PN50-55 and PN90-95) converged across measures to reveal that 1) spatial (place) memory deficits were evident throughout developmental testing, suggesting that the deficits in spatial memory were long-lasting, if not permanent, and 2) behavioral performance measures under the spatial (place) condition were significantly correlated with total volume of hippocampal tissue damage, and with volume of damage to the right and anterior hippocampal regions. These results support the hypothesis that hippocampal integrity is important for the normal development of spatial learning and memory functions, and show that other brain structures do not assume hippocampal-spatial memory functions when the hippocampus is damaged during the neonatal period (even when testing is not begun until adulthood). Thus, neonatal hippocampal damage in rats may serve as a rodent model for assessing treatment strategies (e.g., pharmacological) relevant to human perinatal brain injury and developmental disabilities within the learning and memory realm.

Modulation of lateral hypothalamic activity by olfactory bulb and sciatic stimulation.

Lateral hypothalamic area (LHA) single unit activity (extracellular) was studied in response to electrical stimulation of the olfactory bulb (OB) or sciatic nerve in adult albino rats (n = 39) anesthetized with dialurethane. Olfactory stimulation resulted in a greater proportion of LHA units showing inhibitory rather than excitatory responses, while sciatic nerve stimulation resulted in similar proportions of units showing inhibitory and excitatory responses. Of the 76 LHA units tested with both OB and sciatic nerve stimulation, 36% responded to both stimulation sites, 18% responded only to OB stimulation, 26% responded only to sciatic nerve stimulation, and 20% were unresponsive to either stimulation. The locations of responsive units were diffuse throughout the LHA sampled. The response characteristics of LHA neurons to external sensory stimulation are consistent with the anatomy and putative integrative functions of this brain region.

Locus coeruleus stimulation modulates olfactory bulb evoked potentials.

Field-evoked potentials from the main olfactory bulb in response to stimulation of the olfactory nerve and lateral olfactory tract were measured without and with conditioning stimulation of the locus coeruleus noradrenergic system. The locus coeruleus conditioning stimulus suppressed or inhibited the late components of the olfactory bulb potential evoked by orthodromic olfactory nerve stimulation; this inhibitory effect was suppressed by the microinjection of the alpha-adrenergic blocker prazosin into the olfactory bulb. Results indicate that noradrenergic fibers projecting from the locus coeruleus exert modulatory influences on neuronal networks underlying orthodromic evoked responses in the main olfactory bulb.

Tactile-visual acquisition and reversal learning deficits in rats with prefrontal cortical lesions.

Rats with prefrontal cortex (PFC) lesions or sham operations were tested for acquisition and reversal learning with tactile-visual stimuli. PFC rats performed extremely poorly during acquisition and also differed from control rats in reversal learning. Both higher-order processing and subtle motoric dysfunctions may have combined to account for the marked effects of these cortex lesions.

Large and small medial frontal cortex lesions and spatial performance of the rat.

Rats with sham operations, or small or large medial frontal cortex lesions were compared for the acquisition of a position habit and for learning a series of 4 position habit reversals. No differences were found during acquisition of the position habit. On the reversals, the rats with the larger lesions were markedly impaired, while less severe deficits or control-like performance characterized the rats with the smaller ablations. Analyses of the error scores showed that the rats with the larger lesions made more perseverative errors than the other groups and performed more sporadically even after breaking a previous position habit. These results confirm the hypothesis that a strong relationship exists between medial frontal cortex lesion features (depth and length) and performance. The differences found in acquisition vs. reversal learning, and the nature of the errors observed, also suggest that this part of the brain functions as more than simply a spatial analyzer.

Effects of medial frontal cortex lesions on DRL performance in rats.

Rats with medial frontal cortical lesions (MFC) and control animals with sham operations were tested on an ascending series of 8 DRL schedules of reinforcement ranging from 5 to 70 seconds. Rats with MFC lesions did not differ from the control group in performance when response rate, responses per reinforcement and efficiency were analyzed. These results indicate that deficits in "response inhibition" often displayed by rats with MFC lesions may be task-specific.

Behavioral effects of large and small lesions of the rat medial frontal cortex.

Rats with either sham operations, small medial frontal cortex lesions (SMF), or large medial frontal cortex lesions (LMF) were tested in the open field, for spatial alternation and for 8-arm maze learning. The behaviors of sham-operated and SMF rats were similar on the spatial learning tasks, while the LMF group performed poorly in relation to these two groups. In contrast, the two lesion groups differed from the sham group, but not from each other, on locomotor activity in the open field. The differences in performance between the SMF and LMF groups on the spatial learning measures suggest that medial frontal cortex lesion size and locus may be important factors underlying the variable results of previous studies evaluating spatially-oriented behaviors of rats with varying degrees of medial frontal cortex damage.

Behavioral effects of severe and moderate early malnutrition.

Rats with a history of prenatal and early postnatal undernutrition (6 or 8% casein diets) were "nutritionally rehabilitated" at weaning, and were compared to well-fed animals (25% casein) at maturity. The severely-malnourished (6%) animals were hyperactive in the open field and when tested in a stabilimeter. They also appeared to be highly active during the early trials in 8-arm radial maze testing where they made more arm entries and re-entry errors than the well-fed rats. In terms of trials to criterion, however, their scores on the radial maze and on a spatial alternation task fell within normal limits. The moderately-malnourished (8%) rats tended to perform at control levels on the learning measures, but these rats were not as active as the 6% rats on the measures of activity. Brain size and weight differences among the three groups of rats also are presented and discussed.

Severe early malnutrition and DRL performance in the rat.

Rats with a history of severe early malnutrition (6% casein) were compared to well-fed control animals on an ascending series of DRL values ranging from 5 to 60 seconds. The 6% rats who were dietarily-rehabilitated at weaning did not differ from control animals in efficiency, responses per reinforcement or response rate. In contrast, rats chronically exposed to 6% diets performed so poorly during training with continuous reinforcement that they did not advance to even the first DRL (5-sec) condition. These findings show that severely-undernourished rats can perform within normal limits on even high DRL values, provided they are well trained and that they have adequate nutritional rehabilitation.

Brain damage and neuroplasticity: mechanisms of recovery or development?

Brain damage may be followed by a number of dynamic events including reactive synaptogenesis, rerouting of axons to unusual locations and altered axon retraction processes. In the present theoretical review, the relationship between these morphological changes and behavioral recovery of function is examined from two perspectives. First, an examination of the research literature reveals that the association between these reorganizational events and recovery of function is inconsistent, and it is proposed that in most cases a causal relationship between neural reorganization and behavioral recovery remains speculative at best. It is further noted that aberrant neural circuitry has been associated with neurological dysfunction in many studies. Second, evolutionary considerations suggest that there is little reason to believe that neural reorganizational events emerged to 'heal' damaged brains. Both experimental and evolutionary orientations support the idea that neuronal circuitry changes in response to injury can be better understood as developmental growth processes that are triggered or potentiated in response to cell loss, rather than as recovery or healing processes. The contribution of 'growth' to behavioral recovery of function may be inconsistent because these growth processes are occurring against the backdrop of a damaged brain and may make connections different from those ordinarily seen. Further, they must be considered in conjunction with phenomena such as diaschisis and compensation which may also influence behavioral changes following neural injury.

Ontogeny of electrical activity of main olfactory bulb in freely-moving normal and malnourished rats.

Electroencephalographic activity (EEG) was recorded from the main olfactory bulb (MOB) in freely-moving, normally-nourished, (NP, normal-protein diet) and malnourished (LP, low-protein diet) rats from 4 days of age through adulthood. MOB EEG was analyzed for dominant frequency components using power spectral techniques. For NP rats, a single dominant frequency component (induced wave) was present in the MOB EEG at 4-6 days of age. From 10 days of age through adulthood, the MOB EEG contained two dominant frequency components (induced and intrinsic waves). Both the induced wave and intrinsic waves increased in center-frequency to reach maturity at approximately 30 days of age. Rats reared on low-protein diets (8% casein, prenatal and postnatal) displayed relatively permanent retardation in the development of induced wave center-frequencies and a delay in the development of the intrinsic wave center-frequencies. These results closely parallel the morphological development of the MOB in normally-nourished and malnourished rats.

Postnatal development of sensory influences on neurons in the ventromedial hypothalamic nucleus of the rat.

Extracellular unitary records were obtained from neurons in the ventromedial hypothalamic nucleus (VMH) of very young (1-25 days of postnatal age) and adult rats. Spontaneous unitary activity and evoked responses to both external (somatic, gustatory, and olfactory) and internal sensory (systemic administration of hypertonic saline and glucose solutions) stimulation were determined in order to assess the functional development of VMH neurons and their afferents. The basic electrophysiological characteristics of VMH neurons were established prenatally. From the date of birth, many VMH neurons had: spontaneous action potential generation; evoked responses to external or internal sensory stimulation; and convergent sensory inputs. In contrast, the major developmental change in the neurophysiological properties of VMH neurons was the diminution with increasing age of the convergence of external and internal sensory influences. This developmental 'fine-tuning' of a complex functional feature of VMH neurons is important because the maturation of convergence coincides with a 'critical period' of VMH ontogenesis demonstrated in behavioral and experimental brain damage reports.