Early EEG power predicts MRI injury in infants with hypoxic-ischemic encephalopathy.

OBJECTIVE: Early identification of infants with hypoxic-ischemic encephalopathy who have adverse outcomes despite neuroprotection with therapeutic hypothermia (TH) is urgently needed. Recent studies have found limited value of amplitude integrated EEG (aEEG) for predicting short-term outcomes in this population. Other quantitative electroencephalography (EEG) variables reflecting EEG amplitude, such as EEG power, could provide early stratification of a high-risk cohort in this population. The aim of the study was to evaluate and compare early EEG power and aEEG as predictors of magnetic resonance imaging (MRI) injury in neonatal hypoxic-ischemic encephalopathy. STUDY DESIGN: We conducted a retrospective cohort analysis of 78 encephalopathic infants treated with TH between January 2009 and April 2013. About 56 infants had no/mild injury on MRI (group A), whereas 22 had moderate/severe MRI injury (group B). Total EEG power (TEP) and aEEG were obtained soon after initiation of hypothermia and then compared for their ability to predict future MRI injury. RESULTS: TEP, calculated at a mean age of 8.9 h, was significantly higher in infants in group A as compared to group B (71.6±64.8 vs 26.9±65.3, P=0.02). Odds ratios for predicting moderate-severe MRI injury for TEP<10 µV2, TEP<20 µV2, burst Suppression or worse aEEG pattern were 55 (confidence interval (CI) 6.4 to 471), 12.5 (CI 3.8 to 40.7) and 6.7 (CI 2.0 to 19.8), respectively. CONCLUSION: Early TEP is a reliable predictor of moderate-severe MRI injury in encephalopathic infants undergoing TH and may enable early stratification of infants who may benefit from adjuvant therapeutic interventions.

Intrinsic functional architecture in the anaesthetized monkey brain.

The traditional approach to studying brain function is to measure physiological responses to controlled sensory, motor and cognitive paradigms. However, most of the brain's energy consumption is devoted to ongoing metabolic activity not clearly associated with any particular stimulus or behaviour. Functional magnetic resonance imaging studies in humans aimed at understanding this ongoing activity have shown that spontaneous fluctuations of the blood-oxygen-level-dependent signal occur continuously in the resting state. In humans, these fluctuations are temporally coherent within widely distributed cortical systems that recapitulate the functional architecture of responses evoked by experimentally administered tasks. Here, we show that the same phenomenon is present in anaesthetized monkeys even at anaesthetic levels known to induce profound loss of consciousness. We specifically demonstrate coherent spontaneous fluctuations within three well known systems (oculomotor, somatomotor and visual) and the 'default' system, a set of brain regions thought by some to support uniquely human capabilities. Our results indicate that coherent system fluctuations probably reflect an evolutionarily conserved aspect of brain functional organization that transcends levels of consciousness.

Differences in the expression of GABA(A) receptors between functionally innervated and non-innervated granule neurons in neonatal rat cerebellar cultures.

We had earlier found that granule neurons in cultures of small explants of neonatal rat cerebellar cortex could be placed in two groups: cells in one group showed spontaneous synaptic activity and also had a large response to applications of 1 microM gamma-aminobutyric acid (GABA) while cells in the other lacked spontaneous activity and also showed much lower sensitivity to GABA [25]. For convenience, the more responsive cells will be termed A-type neurons, while the less responsive cells will be termed B-type. We have undertaken a comparison of the responses mediated by activation of GABA A receptors for the two types of neurons. A-type neurons have a larger maximal response to GABA (about 10 times that for B-type neurons), suggesting that they express more functional GABA(A) receptors. The concentration of GABA producing half-maximal activation of A-type neurons is somewhat less (12 mu M) than that for B-type neurons (41 microM), while the Hill coefficients are similar. Responses of both types of cell desensitize to prolonged applications of GABA. At a given concentration of GABA the responses of A-type neurons desensitize more rapidly than the responses of B-type neurons, indicating that the physiological properties of the receptors differ. Responses of A-type neurons are also potentiated to a significantly lesser extent by either chlordiazepoxide or alphaxalone than are the responses of B-type neurons, indicating that the pharmacological properties of the receptors differ. These data indicate that A-type and B-type granule neurons in our cultures express GABA(A) receptors which differ in number, physiological properties and pharmacological responsiveness. We have also confirmed the observation that almost all A-type neurons also show spontaneous synaptic currents, while almost no B-type neurons do.

Neonatal rat cerebellar granule and Purkinje neurons in culture express different GABAA receptors.

We have established a culture system for microexplants of rat cerebellar cortical tissue in which cells develop morphologically, express type-A receptors for the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) and form GABAergic synaptic connections. Criteria of cell size and shape allow reliable identification of granule and Purkinje neurons, criteria confirmed by studies of the binding of antibodies to calbindin D28K and GABA. Both granule and Purkinje neurons express GABAA receptors, but granule neurons fall into two classes in terms of their sensitivity. Granule neurons which do not show spontaneous synaptic currents are relatively insensitive to GABA, while granule neurons with synaptic currents are much more sensitive. The responses of Purkinje neurons to application of 1 microM GABA are relatively insensitive to Zn2+ ion (10 microM), and are potentiated by chlordiazepoxide (100 microM) and La3+ ions (100 microM). Responses of innervated granule neurons, on the other hand, are blocked more strongly by Zn2+ ions, are less affected by chlordiazepoxide and are equally potentiated by La3+ ions. Hence these cultures provide a source of identifiable, functionally innervated cells which express distinct types of GABAA receptors.

How quickly can GABAA receptors open?

We have examined GABAA receptor activation by making rapid applications of GABA to outside-out patches excised from cultured postnatal rat cerebellar neurons. The rate of development of current increases with increasing GABA concentration from a low to a high concentration asymptote. The low concentration asymptote is about 10 s-1 for patches taken from granule cells and 4 s-1 for patches from Purkinje cells. The high concentration asymptote is about 6000 s-1 for patches taken from either granule cells or Purkinje cells. The high concentration asymptote gives an estimate of the fastest rate at which these channels can open and indicates that agonist binding steps are not rate limiting. The concentration dependence of the development of current indicates that more than one GABA molecule is bound to most receptors with open channels and that the final binding step is of low affinity (about 500 microM). A comparison with GABA-mediated postsynaptic currents suggests that the properties of the GABAA receptor play a major role in determining the shape of inhibitory synaptic responses and that the cleft concentration of GABA reaches at least 500 microM.

Immunocytochemical localization of GABA in the cochlear nucleus of the guinea pig.

The immunocytochemical distribution of gamma-aminobutyric acid (GABA) was determined in the cochlear nucleus of the guinea pig using affinity-purified antibodies made against GABA conjugated to bovine serum albumin. Light microscopic immunocytochemistry shows immunoreactive puncta, which appear to be GABA-positive presynaptic terminals, distributed throughout the cochlear nucleus. In the ventral cochlear nucleus, these puncta are often found around unlabeled neuronal cell bodies. While occasional labeled small cells are found in the ventral cochlear nucleus, most GABA-immunoreactive cell bodies are present in the superficial layers of the dorsal cochlear nucleus. Based on size and shape, immunoreactive cells in the dorsal cochlear nucleus are divided into 3 classes: medium round cells with diameters averaging 16 microns, small round cells with average diameters of 9 microns and small flattened cells with major and minor diameters averaging 11 and 6 microns, respectively. Labeled fusiform and granule cells are not seen. A similar distribution of label was seen using antibodies against glutamic acid decarboxylase. Electron microscopic immunocytochemistry of the anteroventral cochlear nucleus shows GABA immunoreactive boutons containing oval/pleomorphic synaptic vesicles on cell bodies and dendrites. Other major classes of terminals, including those with small round, large round and flattened synaptic vesicles are unlabeled.

GABA visualized by immunocytochemistry in the guinea pig cochlea in axons and endings of efferent neurons.

Antiserum raised against GABA coupled with glutaraldehyde to bovine serum albumin was applied to the guinea pig cochlea. Immunoreactivity was visualized as horseradish peroxidase reaction product in surface preparations of the organ of Corti using immunocytochemical techniques. Bright-field, differential interference contrast and video-enhanced contrast light microscopy were used. GABA-like immunoreactivity was found in axons and endings of efferent neurons in all turns of the cochlear spiral, but predominantly in the third turn and first half of the fourth turn. In these apical turns, immunoreactivity was seen in the efferent components: inner spiral bundle, tunnel spiral bundle, tunnel-crossing fibers, large nerve endings synapsing on outer hair cell bases, nerve endings high up on outer hair cells, nerve endings or varicosities close to outer hair cells, and outer spiral fibers. Some immunoreactive large nerve endings at outer hair cells were found in the apical half of the fourth turn. This study shows that axons and endings of efferent neurons in the organ of Corti of guinea pig contain GABA-like immunoreactivity with a distribution similar to that of GAD-like immunoreactivity as shown in a previous study. In both studies, many efferent nerve axons and endings were unstained, even in regions of maximal density of immunoreactivity in the apical turns. The evidence indicates that a subpopulation of efferent neurons projecting to the organ of Corti is GABAergic and very likely different from the lateral and the medial olivocochlear efferent systems.