Electroencephalograms of patients with initial signs of depression: Independent component analysis

Independent component analysis (ICA) of 19-channel background EEG was performed in 111 patients with the early signs of depressive disorders and in 526 healthy subjects. The power spectra of the independent components were compared in the depressive patients and in healthy subjects at the eyes closed and eyes opened states. Statistically significant differences between the groups were detected in three frequency bands: θ (4–7.5 Hz), α (7.5–14 Hz), and β (14–20 Hz). Increased θ and α activities in parietal and occipital derivations of depressive patients may have been caused by a reduced cortical activity in the projection of these derivation. Diffuse enhancement of the β activity may be correlated with anxiety symptoms that are pronounced in the clinical picture of depressive disorders at early stages of the disease. ICA used to compare quantitative EEG parameters in different groups of patients and in healthy persons makes it possible to localize the differences more accurately than the traditional analysis of EEG spectra.     

Cytokines and Absence Seizures in a Genetic Rat Model

We investigated the role of two cytokines, IL-1β and TNF-α, in the development of absence seizures using a genetic model of absence epilepsy in WAG/Rij rats. We administered these cytokines to animals systemically and measured the number of spike-wave discharges (SWDs) in the EEG. We also coadministered IL-1β with the GABA reuptake inhibitor tiagabine and measured the levels of IL-1β and TNF-α in the brain and blood plasma of 2-, 4-, and 6-month-old WAG/Rij rats and animals that served as a non-epileptic control (ACI). We found that IL-1β induced a significant increase in SWDs 2-5 h after administration, while TNF-α enhanced SWDs much later. Both cytokines enhanced passive behavior; body temperature was elevated only after TNF-α. The action of tiagabine was potentiated by earlier IL-1β injection, even when IL-1β was no longer active. Young WAG/Rij rats showed higher levels of TNF-α in blood serum than young ACI rats; the effects in the brain tended to be opposite. The marked differences in timing of the increase in SWDs suggest different time scales for the action of both cytokines tested. It is proposed that the results found after TNF-α are due to the de novo synthesis of IL-1β. TNF-α may possess neuroprotective effects. IL-1β might increase GABA-ergic neurotransmission. The changes in the efficacy of antiepileptic drugs related to changes in the cytokine systems may have some clinical relevance.     

Reflection of individual typological features in the structure of spatial interaction of EEG waves of various frequency ranges

The role of EEG β-, α-, and θ-activity in the formation of the structure of the spatial interaction of cortical formations and interhemispheric balance (IB), evaluated on the basis of the matrix of conditional probabilities, were studied in healthy test subjects occupied with oil refining. Preliminarity, the groups of test subjects with high (I), average (II), and low (III) adaptivity were distinguished. Individuals with a positive IB prevailed in groups I and II and among the test subjects with the length of service less than 20 years, and those with negative IB were in the majority in group III and among the test subjects with the length of service over 20 years. The structure of spatial interaction of EEG waves of test subjects with high and average adaptivity, as well as those with positive IB and length of service below 20 years, was determined mostly by α-activity. The spatial pattern of EEG of test subjects with low adaptivity, as well as those with negative IB and length of service above 20 years, was determined by the interaction of bioelectrical activity mainly in the α- and β-ranges.     

Electroencephalographic characteristics of cognitive-specific alerting attention in verbal learning: III. Localized characteristics of EEG spatial synchronization

Electroencephalograms (EEGs) were recorded in 19 standard derivations in 88 healthy subjects (students) in the state of rest with eyes open and during memorization (learning) of verbal bilingual semantic pairs (the Latin and Russian languages) and retrieval of information from memory (control). The estimates of EEG coherence in these states were compared for the following frequency bands: θ (4–7 Hz), α₁ (7–10 Hz), α₂ (10–13 Hz), β₁ (13–18 Hz), β₂ (18–30 Hz), and γ (30–40 Hz). Compared to the state of rest, the decrease in coherence in the pairs of derivations from the frontal and central cortical areas in all EEG frequency bands was the most pronounced for memorization, and the increase in coherence in the interhemispheric derivation pairs of the parietal-occipital region in most of the frequency bands was the most pronounced for retrieval. In addition, in the pairs formed by derivations from the parietal-occipital region with derivations from the frontal and central regions, retrieval is also characterized by an increase in coherence in the β₂ and γ bands along with its decrease in the low-frequency ranges. The dynamics of EEG coherence, when comparing the states of memorization and retrieval, is more statistically significant in the interhemispheric and cross-hemispheric pairs of derivations than in the intrahemispheric pairs. The revealed topographic specificity of the dynamics of EEG coherence owing to the change of state is considered in terms of the notion on cognitive-specific forms of sustained goal-directed mental attention.     

Background cerebral electrical activity in healthy mental aging

EEG power mapping in the frequency bands from Δ to γ₂ was used to study the changes in the background cerebral activity during “successful” cognitive aging, with the cognitive ability preserved, in subjects engaged in complex occupational activities. The sample consisted of an older age group (OAG) of 32 subjects (14 men and 18 women with a mean age of 65.1 ± 1.18 years) and a younger age group (YAG) of 33 subjects (18 men and 15 women with a mean age of 22.1 ± 0.38 years). The mean power of the slow (Δ, θ, and α₂) rhythms decrease with age, and that of the fast (β and γ) rhythms increase with age. The heterogeneity of the power parameters recorded at different sites was decreased in the OAG compared to the YAG. The centro-lateral power gradient was smoothed in the frequency bands from Δ- to β₂-, and both the centro-lateral and fronto-parietal power gradients and interhemispheric differences, in the α- and β₁ bands in the OAG. The results suggest that the observed age-related changes are prerequisites for the involvement of compensatory mechanisms, which may be related to both mobilization of larger resources ensuring cognitive activity and reorganization of cortical networks in the areas prone to age-related physiological changes.     

Laminin chains in the basement membranes of human thymus

Summary In recent studies, the α2 chain of laminin (Ln) has been suggested to be the only laminin α chain expressed in mouse and human thymus. We have now used chain-specific monoclonal antibodies and indirect immunofluorescence microscopy to study the expression of laminin chains in samples of foetal and 6-year-old human thymus. The subepithelial basement membrane of the capsule of foetal 16- to 18-week thymus presented a bright immunoreactivity for Ln α1, α3, β1, β3 and γ1 chains but not for α2 chain, suggesting the expression of laminins-1 and-5. Most cortical and medullary epithelial cells, including Hassall's corpuscles, however, lacked laminin immunoreactivity. Immunoreactivity for Ln β2 chain was only seen in basal laminae of larger blood vessels. In thymic specimens from 6-year-old children, immunoreactivity for the laminin α1, α3, β1, β3 and γ1 chains was invariably found in subepithelial basement membrane of the capsule and that for laminin α2 chain was now also distinct but more heterogeneous. Furthermore, the thymic subepithelial basement membrane of the capsule at all stages showed immunore-activity for collagen type VII, forming the anchoring fibres in epithelial basement membranes. The subcapsular thymic epithelium also showed immunoreactivity for the BP 230 antigen and β₄ integrin subunit, both components of hemidesmosomes. The present results show that the thymic subepithelial basement membrane of the capsule presents properties which are commonly seen in stratified and combined epithelia, and are compatible with suggestions of the antigenic similarity of thymic epithelial cells and keratinocytes.     

Establishment of a monoclonal antibody directed against Gb3Cer/CD77: a useful immunochemical reagent for a differentiation marker in Burkitt's Iymphoma and germinal centre B cells

A new monoclonal antibody (TU-1) directed against the Galα1-4Galβ1-4Glc residue of the Gb3Cer/CD77 antigen was prepared by the hybridoma technique following immunization of mice with an emulsion composed of monophosphoryl lipid A, trehalose dimycolate, and Gb3Cer isolated from porcine erythrocytes. TU-1 showed reactivity towards Gb3Cer and lyso-Gb3Cer (Galα1-4Galβ1-4Glcβ1-1′Sph), although the reactivity towards lyso-Gb3Cer was about 10-fold lower than that to Gb3Cer. But it did not react with other structurally-related glycolipids, such as LacCer (Galβ1-4Glcβ1-1′Cer), Gg3Cer, Gg4Cer, Gb4Cer (GalNAcβ1-3Galα1-4Galβ1-4Glcβ1-1′Cer), galactosylparagloboside (Galα1-3Galβ1-4GlcNAcβ1-3Galβ1-4Glcβ1-1′Cer), sulfatide (HSO3-3Galβ1-1′Cer), other gangliosides (GM3, GM2, GM1a, GD1a and GT1b), or P1 antigen (Galα1-4Galβ1-4GlcNAcβ1-3Galβ1-4Glcβ1-1′Cer) among neutral glycolipids prepared from P1 phenotype red blood cells. Furthermore, TU-1 reacted with viable lymphoma cells, such as human Burkitt lymphoma cell line, Daudi, and Epstein-Barr virus (EBV)-transformed B cells by the immunofluorescence method, and also with germinal centre B cells in human tonsil and vessel endothelial cells in human thymus histochemically. These results indicate that TU-1 is a monoclonal antibody directed against Gb3Cer/CD77 antigen and can be utilized as a diagnostic reagent for Burkitt's lymphoma and also for detection of the blood group Pk antigen in glycolipid extracts of erythrocytes. Abbreviations: ATL, adult T-cell leukaemia; BSA, bovine serum albumin; Cer, ceramide; DPPC, L-α-dipalmitoylphosphatidylcholine; EBV, Epstein-Barr virus; FCS, fetal calf serum; GalCer, Galβ1-1′Cer; GlcCer, Glcβ1-1′Cer; LacCer, Galβ1-4Glcβ1-1′Cer; Gb3Cer, Galα1-4Galβ1-4Glcβ1-1′Cer; Iyso-Gb3Cer, Galα1-4Galβ1-4Glc1-1′Sph; Gb4Cer, GalNAcβ1-3Galα1-4Galβ1-4Glc1-1′Cer; galactosylparagloboside, Galα1-3Galβ1-4GlcNAcβ1-3Galβ1-4Glcβ1-1′Cer; Gg3Cer, GalNAcβ1-4Galβ1-4Glcβ1-1′Cer; Gg4Cer, Galβ1-3GalNAcβ1-4Galβ1-4Glcβ1-1′Cer; GM3, Neu5Acα2-3Galβ1-4Glcβ1-1′Cer; GM2, GalNAcβ1-4(Neu5Acα2-3) Galβ1-4Glcβ1-1′Cer; GM1a, Galβ1-3GalNAcβ1-4(Neu5Acα2-3)Galβ1-4Glcβ1-1′Cer; GD1a, Neu5Acα2-3Galβ1-3GalNAcβ1-4(Neu5Acα2-3)Galβ1-4Glcβ1-1′Cer; GD1b, Galβ1-3GalNAcβ1-4(Neu5Acα2-8Neu5Acα2-3)Galβ1-4Glcβ1-1′Cer; GT1b, Neu5Acα2-3Galβ1-3GalNAcβ1-4(Neu5Acα2-8Neu5Acα2-3) Galβ1-4Glcβ1-1′Cer; HRP, horseradish peroxidase; LDH, lactate dehydrogenase; MAb, monoclonal antibody; MPL, monophosphoryl lipid A; P1 antigen, Galα1-4Galβ1-4GlcNAcβ1-3Galβ1-4Glcβ1-1′Cer; PVP, polyvinylpyrolidone; Sph, sphingosine; sulfatide, HSO3-Galβ1-1′Cer; TDM, trehalose dimycolate; TLC, thin-layer chromatography This revised version was published online in November 2006 with corrections to the Cover Date.     

Effects of unstructured video exposure on EEG power in situations of forced attention and rest

Group 1 (N = 30) and Group 2 (N = 22) of healthy volunteers participated in the experiment. In both groups, the EEG was recorded in the state of rest with the eyes closed (REC); at rest with eyes open (REO); and during passively watching TV channel noises (white noise). EEG was also recorded in the state of forced visual attention: when counting colored symbols appearing on the screen (group 1) and when searching for an image of a real object in the noise (group 2). The averaged values of EEG power in each derivation were calculated for every subject and for each state in the Δ, θ, α₁, α₂, β₁, β₂, and γ frequency bands. The results demonstrate that the exposure to unstructured noninformative video noise may lead to significant changes in the EEG power in various frequency bands, with the most prominent changes taking place in the α₂ band. These changes are topically wide, reflecting systemic changes in the corresponding brain mechanisms, but are much less intense compared to the difference between the states of rest with the eyes open and closed.     

Locus of Psychological Control and Characteristics of the EEG Frequency Components

In a group including 65 adults of both sexes, we examined correlations of the internal/external personality type (locus of psychological control diagnosed by the Rotter questionnaire) with parameters of the EEG frequency components (rhythms). Multichannel recording of ongoing EEG was carried out in the resting state; leads were located according to the 10-20 system. Despite natural high interindividual variability, the subgroup of internals was, in general, characterized by higher spectral powers (SPs) of the δ, θ, α, and β1 rhythms and a lower SPs of β2 and γ oscillations recorded in the resting state with the eyes closed. In internals, the modal frequencies of practically all EEG ranges were, on average, somewhat lower. In this subgroup, reaction of EEG activation related to opening of the eyes was stronger, while the interhemisphere asymmetry was weaker.     

Importance of signaling via the IFN-α/β receptor on host cells for the realization of the therapeutic benefits of cyclophosphamide for mice bearing a large MOPC-315 tumor

Here we show that low-dose cyclophosphamide (CY), that depends for its therapeutic effectiveness on the immunopotentiating activity of the drug for T cell-mediated tumor-eradicating immunity, is curative for ~80% of wild-type (WT) mice bearing a large s.c. MOPC-315 tumor, but only for ~10% of IFN-α/βR^−/− mice bearing a large s.c. MOPC-315 tumor. Histopathological examination of the s.c. tumors of such mice on day 4 after the chemotherapy revealed that the low dose of CY led to accumulation of T lymphocytes in both the WT and the IFN-α/βR^−/− mice. However, in the CY treated tumor bearing WT mice the T lymphocytes were present throughout the tumor mass and in direct contact with tumor cells, but in the CY treated tumor bearing IFN-α/βR^−/− mice most of the T lymphocytes remained in blood vessels. In addition to being important for CY-induced transendothelial migration of T lymphocytes into the tumor mass, we show here that signaling via the IFN-α/βR is also important for CY-induced control of metastatic tumor progression in the spleen and liver of the tumor bearing mice. Finally, CY cured tumor bearing WT mice were resistant to a subsequent challenge with MOPC-315 tumor cells, but the few CY cured tumor bearing IFN-α/βR^−/− mice were not. Thus, signaling via the IFN-α/βR on host cells in MOPC-315 tumor bearers is important for CY-induced: (a) transendothelial migration of T lymphocytes into the tumor mass and the eradication of the primary tumor, (b) control of metastatic tumor progression, and (c) resistance to a subsequent tumor challenge. This work was supported by Research Grant 03-19 from the American Cancer Society-Illinois Division.     

EEG changes upon unblocking of the acoustic and visual channels

Modality-specific and supramodal EEG components that are related to the reorientation of involuntary anticipatory attention from the internal to the external type were studied by comparing the unblocking of the visual and acoustic channels. EEG recording was performed while subjects were at rest with their eyes open, with their eyes closed, or with their eyes closed while using noise-protective earplugs. The means of both the EEG power in each of the derivations and EEG coherence in each of the derivation pairs were calculated for every subject and for each of the states. The estimations have been done for the Δ, θ, α₁, α₂, β₁, β₂, and γ frequency bands. The obtained data proved the hypothesis on the manifestation of both modality-specific and supramodal components in the brain mechanisms of involuntary anticipatory attention. These results are of interest for discussion on the degree of similarity and distinction between the systemic supports of the brain mechanisms of visual and auditory attention.     

Peculiarities of EEG in Subjects with Coronary-Prone (Type A) Behavior

In a group including 72 adults of both sexes, we studied correlations between the estimates of the so-called coronary-prone personality type (type A) diagnosed using the Jenkins questionnaire and the spectral powers (SPs) of the frequency components (rhythms) of background EEGs recorded in the resting state (leads C3 and C4 according to the 10-20 system). Despite natural high interindividual variability, estimates that characterized the subject as belonging to the behavioral type A corresponded, on average, to relatively low SPs of the δ, θ, and α EEG components, intermediate values of the β₁ rhythm SP and coefficient of reactivity of the α rhythm, and higher SPs of the high-frequency (β₂ and γ) rhythms. Estimates characterizing type B personality corresponded to significantly higher δ-rhythm SPs, intermediate SPs of the θ and α rhythms, and smaller SPs of the β and γ rhythms. The interhemisphere asymmetry coefficient for the α rhythm was usually negative in type-A individuals and positive in the cases of types B and AB. The peculiarities observed are probably determined, to a certain extent, by the fact that both the characteristics of the behavioral types of the personality and the amplitude parameters of EEG rhythms depend significantly on inherited (in particular neurochemical) factors. Such peculiarities of the neurodynamic constitution of the individual are determined, to a considerable extent, by the specificity of organization and functioning of a few neurotransmitter (in particular aminergic) and neurohumoral systems.     

A comparative electrophysiological study of regulatory components of working memory in adults and seven- to eight-year-old children: An analysis of coherence of EEG rhythms

Coherence at the frequency of θ, α, and β EEG rhythms was analyzed in 14 adults and 23 children of 7–8 years old while they performed cognitive tasks requiring an involvement of working memory (WM). We used the pair matching paradigm in which subjects had to match a pair of stimuli shown in succession in the central visual field. The pairs of verbal and visuo-spatial stimuli were mixed together and presented in a pseudo random order. Each pair was preceded by a warning signal that did not specify a modality of upcoming stimuli. We analyzed EEG segments recorded (i) in the rest condition, (ii) prior to the first (reference) stimulus (maintenance of nonspecific voluntary attention), and (iii) prior to the second (test) stimulus (retention of information in WM). In the present study we focused on the regulatory functional components of WM, and therefore, the stimulus modality has not been taken into account. In adults, maintaining nonspecific voluntary attention was accompanied by an increase of the strength of θ-related functional coupling between medial areas of the frontal cortex and temporal cortical zones and by a strengthening of local β-related functional connectivity in the fronto-central areas of the cortex. In children, no such increase was found for θ rhythm; for β rhythm the increase was limited to several short-range functional links. In adults, the retention of information in WM was accompanied by the growth in α coherence in distant fronto-parietal links, predominantly in the right hemisphere, while in children information retention was accompanied by the growth of θ coherence in the inferio-temporal and parietal cortical regions. The results of the study point to a relative immaturity of the mechanisms of executive control of WM in children of 7–8 years old.     

Interrelations between Peculiarities of the Motivational Sphere of a Personality and Characteristics of the Current EEG

In a group of 70 adult subjects of both sexes, we estimated the levels of the need for achievement and its motivation (interpreted as generalized characteristics of the state of the motivational sphere of a personality and diagnosed using questionnaires of Orlov and Mehrabian) and examined the correlations of these estimates with spectral powers (SPs) of the frequency components (rhythms) of the current background EEG (С3 and С4 leads according to the 10–20 system). Despite the fact that naturally high interindividual variability exists, high estimates of the need for achievement correlated with medium values of the SPs of most EEG frequency components (δ, θ, α, and β₁ rhythms), low SPs of the β₂ and γ rhythms, and higher values of the coefficient of reactivity (CR) of the α rhythm. A high level of the need for achievement corresponded to higher SPs of the θ and γ rhythms and high CR of the α rhythm, a medium SP of this EEG component, and lower SPs of the δ and β rhythms. The coefficient of interhemisphere asymmetry of the α rhythm was the highest at low values of the studied psychological indices. Interrelations that we found in our study can be explained as follows: both the characteristics of the need for achievement and motivation typical of a certain personality and the amplitude parameters of the EEG rhythms observed in this individual depend significantly on genetically determined factors (in particular, on neurochemical ones). These peculiarities of neurodynamic constitution of the individual are, to a considerable extent, due to the specificity of organization and functioning of a few neurotransmitter (aminergic, in particular) and neurohumoral systems. Neirofiziologiya/Neurophysiology, Vol. 41, No. 1, pp. 61–69, January–February, 2009.     

Correlation between level of aggressiveness of personality and characteristics of EEG frequency components

In a group of 80 adults of both sexes, we studied the correlation between the characteristics of aggressiveness of the individual (diagnosed using the Buss-Durkee questionnaire) and parameters of the frequency components (rhythms) of EEG recorded in the resting state from leads C3 and C4 according to the 10–20 system. Despite the natural high interindividual variability, the higher spectral powers of nearly all EEG frequency components (δ-, θ-, α-, and β rhythms) and coefficient of reactivity of the α rhythm corresponded in general to lower indices on the scales of the Buss-Durkee questionnaire and smaller values of the index of aggressiveness and index of hostility. These correlations probably reflect a significant dependence of both characteristics of aggressiveness of personality and amplitude parameters of the EEG rhythms on hereditary factors. Both these aspects of the neurodynamic constitution of the individual are to a significant extent determined by the specificity of organization and functioning of a few neurotransmitter (in particular, aminergic) and neurohumoral systems. Neirofiziologiya/Neurophysiology, Vol. 38, Nos. 5/6, pp. 448–457, September–December, 2006.     

Reflection of the emotional experience, strength, intensity of drive and probability of its satisfaction in interhemispheric distribution of frequency-amplitude parameters of EEG

The character of interhemispheric distribution of frequency-amplitude parameters of EEG as a function of the strength of emotional experience, the intensity of drive, and the expected probability of its satisfaction were determined in human test subjects. The emotional state was aroused by an imaginary reproduction of emotionally colored events. The modified test by D. Price and co-authors was used where the subjects estimated the intensity of drive (the degree of undesirability of imaginary event) and the expected probability of its occurrence by the length of line segment. In this study, the simulation of anger was used. The highest correlation coefficients were observed between the expectation and power in the α-EEG-range in the left frontal region. The strength of emotional experience negatively correlated with the α-, θ-, and Δ-ranges. Statistically significant correlation was revealed between the intensity of drive and power of Δ- and θ-waves. The power of β-rhythm correlated with the strength of emotion as well as with the expectation and drive indices.     

Modulation of High-Frequency EEG Rhythms under Conditions of the Activation Reaction: Dependence on Psychological Characteristics of Personality

In a group of 118 adults of both sexes, we studied modulation of high-frequency EEG rhythms (β and γ components) under conditions of EEG activation related to opening of the eyes. Typical changes in the α rhythm (significant depression in the overwhelming majority of the tested subjects) was accompanied by different (in direction and intensity) shifts of the spectral powers (SPs) of the β and γ components in different subjects. The power of the β1 subcomponent under conditions of the activation reaction increased in 6 persons and dropped in 109 persons; changes in this index in two hemispheres were opposite in their directions in 3 subjects. Thus, changes in the β1 SP upon opening of the eyes were nearly parallel to those of the α rhythm but less intense. The subgroups differentiated according to the pattern of reactivity of the β2 subcomponent included 23, 85, and 10 subjects; for the γ rhythm, the corresponding numbers were 31, 72, and 8. In other words, the patterns of reactivity of β2 and γ oscillations were rather similar to each other but differed significantly from the pattern for the β1 rhythm. Values of the differential coefficients of reactivity (DCRs) of the β1, β2, and γ rhythms demonstrated significant correlations that were especially close for the β2 and γ activity. Within the entire examined group, we found significant negative correlations of the DCR values for β and γ activities with the estimates of some psychological characteristics of the subjects (levels of neuroticism, psychoticism, plasticity, self-control of behavior, and some others) measured using the RTS (Strelau), EPQ (Eysenck), OST (Rusalov), and 16PF (Kettel) questionnaires. The dispersion analysis demonstrated that the above-mentioned subgroups of the subjects (with increase and decrease in the SPs of the β and γ rhythms in both hemispheres and opposite changes of these indices in the right and left hemisphere) showed significant or close to significant specific intergroup differences of a few mean values of the psychological characteristics estimated according to the above questionnaires. Possible mechanisms of modulation of high-frequency EEG components under conditions of the reaction of EEG activation and relations of such changes to the psychological characteristics of the personality are discussed.     

Co-expression of α′ and β subunits of β-conglycinin in rice seeds and its effect on the accumulation behavior of the expressed proteins

A transgenic rice that produces both the α′ and β subunits of β-conglycinin has been developed through the crossing of two types of transgenic rice. Although the accumulation level of the α′ subunit in the α′β-transgenic rice was slightly lower than that in the transgenic rice producing only the α′ subunit, the accumulation level of the β subunit in the α′β-transgenic rice was about 60% higher than that in the transgenic rice producing only the β subunit. Results from sequential extraction and gel-filtration experiments indicated that part of the β subunit formed heterotrimers with the α′ subunit in a similar manner as in soybean seeds and that the heterotrimers interacted with glutelin via cysteine residues. These results imply that the accumulation level of the β subunit in the α′β-transgenic rice increases by an indirect interaction with glutelin. Immunoelectron microscopy revealed that the α′ and β subunits are localized in a low electron-dense region of protein body-II (PB-II) and that α′ homotrimers in the α′β-transgenic rice seeds seem to accumulate outside of this low electron-dense region.