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PubMed2006

Cytochrome oxidase staining reveals functionally important activity bands in the olfactory epithelium of newborn rat.

We used cytochrome oxidase (CytOx) staining intensity, which is correlated with neuronal functional activity, to evaluate maturity and functionality of newborn rat olfactory epithelium (OE) and olfactory receptor neurons (ORNs). Nasal olfactory tissue of neonatal rats was stained with CytOx and analyzed qualitatively and quantitatively. Results revealed that newborn OE shows six differentially stained horizontal bands. Bands run parallel to the OE surface and were categorized as very light, medium or darkly stained. A narrow and pale Band 1 overlapped with horizontal basal cells. Next, a wide and lightly stained Band 2 was observed that coincides with the globose basal cell layer and immature ORNs, deep in OE. Next apically, a medium-staining Band 3 overlapped with ORN perikarya. Closer to the surface, a medium to light Band 4 was discerned where dendrites of mature ORNs normally occur. This band was interrupted with lighter areas due to the presence of supporting cells nuclei. Next, a superficial but dark Band 5 occurred, populated by the apical portions of ORN dendrites and their ciliated knobs and by supporting cell apices; mitochondria in apices of supporting cells contribute predominantly to dense staining of this Band 5. Apical to Band 5, a thin and fairly light Band 6 was observed which overlaps with the mucus layer that contains part of the ORN knobs, their cilia and supporting cell microvilli. Along the length of ORN dendrites, apical segments just below the ORN knobs, and wide basal segments showed a darker staining than the middle segments implying "microzones" of higher neural activity within the most apical and basal regions of dendrites. Our findings agree with ultrastructural studies showing a presence of mitochondria in knobs, basal portions of ORN dendrites and in OE supporting cell apices, suggesting that apical regions of both olfactory and supporting cells near the surfaces are metabolically most active, in odorant detection, signal processing, and detoxification, the latter for supporting cells.

Animals↗
PubMed1992

Cytochrome oxidase activity in vagal and glossopharyngeal visceral sensory neurons of the rat: effect of peripheral axotomy.

Cytochrome oxidase (CO) activity, an endogenous metabolic marker, was examined in visceral sensory neurons of the rat nodose and petrosal ganglia by using enzyme histochemistry. In the normal nodose and petrosal ganglia, nerve cells showed various degrees of staining intensity. The population of darkly stained neurons in the nodose ganglion was higher than in the petrosal ganglion. Axotomy of the peripheral axons of these bipolar sensory neurons was used to study potential changes in ganglionic cellular metabolism associated with loss of afferent inputs and/or injury. Peripheral axotomy had a significant effect on CO activity in the nodose ganglion. By 3 days after axotomy, darkly stained neurons decreased in number and lightly stained neurons, which were not observed in the normal ganglion, appeared in the nodose ganglion. At 7 days after axotomy, the average population of these lightly stained neurons increased to 29% in the nodose ganglion. Subsequently, the population decreased so that at 14 days and 21 days, 19% and 7% respectively of neurons were stained lightly. Even at 28 days after axotomy, the lightly stained neurons were still observed. In the petrosal ganglion, no remarkable change was observed at any stage after axotomy. These results suggest that metabolic activity decreases in some nodose neurons after peripheral nerve section.

Animals↗
PubMed1995

Cardiolipin modulates the secondary structure of the presequence peptide of cytochrome oxidase subunit IV: a 2D 1H-NMR study.

The secondary structure of the presequence of cytochrome oxidase subunit IV (p25) was studied by circular dichroism and 2D nuclear magnetic resonance in micelles of dodecylphosphocholine (DPC) and mixed micelles of DPC and mitochondrial cardiolipin (CL). In both systems, alpha-helix formation was observed. The alpha-helix stretches from the N- to the C-terminus with a break at the proline residue at position 13. Upon introduction of CL in the DPC micellar system, an increased stability of the helix was observed around proline13 and in the C-terminal half. This observation, together with reported results on specific interactions between CL and p25, led to the proposal of a two-state equilibrium of the alpha-helical conformation of p25, modulated by CL.

Amino Acid Sequence↗
PubMed1987

Changes of cytochrome oxidase activity in the rat subcortical visual centers after unilateral eye enucleation.

Histochemical densitometric measurements of cytochrome oxidase (CO) activity in the dorsal and ventral lateral geniculate nuclei (LGNd and LGNv) and the superficial gray stratum of the superior colliculus (SC) were done between 2 and 12 weeks after unilateral enucleation in the adult albino rat. This deafferentation resulted in a transient, slight but statistically significant decrease in CO activity in the contralateral LGNd and LGNv. Whereas in the contralateral SC, a marked reduction in CO activity was observed, and this persisted over the postoperative periods studied. The regional difference in the recovery pattern of CO activity were discussed from the aspect of synaptic reorganization.

Animals↗
PubMed1989

Cytochrome oxidase activity in the rat retina following unilateral thalamic lesion.

The present study has investigated the cytochrome oxidase (CO) activity in retinas of normal rats and rats which received central lesions at birth or young adult stage. The results show that a thalamic lesion which injured the retinal ganglion cell axons in young adult rats led to severe loss of CO activity in the ganglion, inner and outer plexiform layers in the retina contralateral to the lesion as compared to those of normal rats. In contrast, distinct CO-reactive bands and cells were clearly observed in corresponding laminae in retinas in which almost the entire population of retinal ganglion cells was eliminated by a neonatal thalamic lesion. These results indicate that retinal ganglion cells contribute significantly to the CO activity observed in the inner retinal laminae under normal but not under abnormal conditions, and suggest that considerable changes in the activity of the remaining neurons and possibly reorganization of neural circuitry within the retina in rats which received neonatal lesions has taken place, as revealed by CO histochemistry.

Animals↗
PubMed1990

Quantitative light- and electron-microscopic analysis of cytochrome-oxidase distribution in neurons of the lateral geniculate nucleus of the adult monkey.

The distribution of cytochrome oxidase (CO) in the lateral geniculate nucleus (LGN) of normal adult macaque monkeys was analyzed in quantitative light- and electron-microscope (EM) studies. Both reactive and nonreactive neurons were found throughout laminae 1-6. Darkly reactive neurons were more numerous in laminae 1, 2, and 6. Within each lamina, there was a positive correlation between cell size and level of CO activity in neurons. The cell density was greater in laminae 1, 4, and 6, which received input from the contralateral eye, than in corresponding laminae representing the ipsilateral eye (2, 3, and 5). The cell density in each of the magnocellular laminae was less than that in each of the parvicellular laminae. Three types of neurons could be distinguished within magnocellular laminae at the EM level. Types I and II were interpreted to be relay neurons, while the small size of type III qualified them to be interneurons. Two populations were found within parvicellular laminae, corresponding to relay neurons and interneurons, respectively. EM quantitative analyses revealed that the darkly reactive neurons contained a high proportion of moderate to darkly reactive mitochondria, which comprised 44.2%, 42.5%, and 39.2% of the mitochondrial population within neurons of laminae 1, 2, and 6 respectively. In contrast, a relatively low proportion of reactive mitochondria were found in neurons within laminae 3 (22.8%), 4 (26.4%), and 5 (32.4%). Moreover, magnocellular laminae contained a higher density of synapses on cell bodies than parvicellular laminae. All laminae, except layer 5, contained predominantly symmetric synapses on cell bodies. However, the proportion of symmetrical synapses in the magnocellular laminae (65.6%) was greater than that in the parvicellular laminae (59%). Our quantitative light and EM results indicate that the level of metabolic activity in neurons of the monkey LGN is higher in magnocellular, ON-center, and contralateral than in parvicellular, OFF-center, and ipsilateral visual pathways, respectively. The metabolic activity of neurons is likely to reflect the chronic level of spontaneous and synaptically evoked discharges of these cells.

Animals↗
PubMed2003

Molecular characterization of the mitochondrial cytochrome oxidase I gene of Oestridae species causing obligate myiasis.

A 688-bp region of the mitochondrial cytochrome oxidase I gene was sequenced from larvae of 18 species of Oestridae causing obligate myiasis. Larvae belonged to the four Oestridae subfamilies (Cuterebrinae, Gasterophilinae, Hypodermatinae and Oestrinae), which are commonly found throughout the world. Analysis of both nucleotide and amino acid data was performed. Nucleotide sequences included 385 conserved sites and 303 variable sites; mean nucleotide variation between all species was 18.1% and variation within each subfamily ranged from 5.3% to 13.34%. Intraspecific pairwise divergences ranged from 0.14% to 1.59%, and interspecific variation ranged from 0.7% to 27%. Of the 229 amino acids, 76 were variable (60 of which were phylogenetically informative), with some highly conserved residues identified within each subfamily. Phylogenetic analysis showed a strong divergence among the four subfamilies, concordant with classical taxonomy based on morphological and biological features. This study provides the first molecular data set for myiasis-causing Oestridae species, providing an essential database for the molecular identification of these parasites and the assessment of phylogenetic relationships within family Oestridae.

Animals↗
PubMed2004

Projections from primary visual cortex to cytochrome oxidase thin stripes and interstripes of macaque visual area 2.

It has been controversial whether the cytochrome oxidase (CO)-dense blobs in primate primary visual cortex (V1) and CO-dense thin stripes in visual area 2 (V2) are parts of a cortical color-processing stream that is segregated from other functional streams. One of the key pieces of evidence for the segregated color stream is the previous report of specific connections between blobs and thin stripes, which is parallel to the connections between interblobs and interstripes. To study the degree of the segregation between the proposed different streams, in the current study, anatomical tracers were injected into different V2 compartments with the functional guidance of optical imaging. The spatial relationship between each labeled cell and the CO blobs in V1 were analyzed quantitatively. After tracer injections in the color-preferring modules in CO thin stripes, equal amounts of labeled cells were found in the blobs and interblobs. However, the density of the labeled cells was more than twice as high in the blobs as that in the interblobs, and most of the clusters of labeled cells partially overlapped with the blobs. Tracer injections in the interstripes labeled cells predominantly in the interblobs. Our results suggest that both the blobs and interblobs project to the thin stripes and call into question the proposition that different CO compartments in V1 and V2 are connected in parallel to form highly segregated functional streams.

Animals↗
PubMed2003

Postnatal changes in cytochrome oxidase expressions in brain stem nuclei of rats: implications for sensitive periods.

Previously, we reported that cytochrome oxidase (CO) activity in the rat pre-Bötzinger complex (PBC) exhibited a plateau on postnatal days (P) 3-4 and a prominent decrease on P12 (Liu and Wong-Riley, J Appl Physiol 92: 923-934, 2002). These changes were correlated with a concomitant reduction in the expression of glutamate and N-methyl-d-aspartate receptor subunit 1 and an increase in GABA, GABAB, glycine receptor, and glutamate receptor 2. To determine whether changes were limited to the PBC, the present study aimed at examining the expression of CO in a number of brain stem nuclei, with or without known respiratory functions from P0 to P21 in rats: the ventrolateral subnucleus of the solitary tract nucleus, nucleus ambiguus, hypoglossal nucleus, nucleus raphe obscurus, dorsal motor nucleus of the vagus nerve, medial accessory olivary nucleus, spinal nucleus of the trigeminal nerve, and medial vestibular nucleus (MVe). Results indicated that, in all of the brain stem nuclei examined, CO activity exhibited a general increase with age from P0 to P21, with MVe having the slowest rise. Notably, in all of the nuclei examined except for MVe, there was a plateau or decrease at P3-P4 and a prominent rise-fall-rise pattern at P11-P13, similar to that observed in the PBC. In addition, there was a fall-rise-fall pattern at P15-P17 in these nuclei, instead of a plateau pattern in the PBC. Our data suggest that the two postnatal periods with reduced CO activity, P3-P4 and especially P12, may represent common sensitive periods for most of the brain stem nuclei with known or suspected respiratory control functions.

Aging↗
PubMed1989

Marked and variable inhibition by chemical fixation of cytochrome oxidase and succinate dehydrogenase in single motoneurons.

The effect of tissue fixation on succinate dehydrogenase and cytochrome oxidase activity in single motoneurons of the rat was demonstrated using a computer image processing system. Inhibition of enzyme activity by chemical fixation was variable, with some motoneurons being affected more than others. It was concluded that quantification of enzymatic activity in chemically fixed tissue provides an imprecise estimate of enzyme activities found in fresh-frozen tissues.

Animals↗
PubMed1976

Detection of macrophages in atherosclerotic lesions with cytochrome oxidase.

The anaerobic metabolism of the arterial wall allows macrophages to be demonstrated therein by the cytochrome oxidase histochemical method. Monocytes (macrophages) in human fatty streaks (WHO grade I) or fibrofatty (WHO grade II) human atherosclerotic lesions are normally confined to subendothelial regions. Lesions complicated by ulceration, mural thrombosis or intimal haemorrhage (WHO grade III) showed numerous monocytes (macrophages) around newly-formed capillaries in focal areas of organization. By contrast with grades I and II atherosclerotic lesions, macrophages in lipid granulomas induced by subcutaneous injection of cholesterol oleate are more numerous and distributed throughout the lesion. The slow or absent resorption of lipid from atheromatous lesions may in part result from the paucity of macrophages therein.

Adult↗
PubMed1990

Observation of the Fe4+ = O stretching Raman band for cytochrome oxidase compound B at ambient temperature.

Resonance Raman and visible absorption spectra were simultaneously observed for cytochrome oxidase reaction intermediates at 5 degrees C by using the artificial cardiovascular system (Ogura, T., Yoshikawa, S., and Kitagawa, T. (1989) Biochemistry 28, 8022-8027) and a device for Raman/absorption simultaneous measurements (Ogura, T., and Kitagawa, T. (1988) Rev. Sci. Instrum. 59, 1316-1320). The Fe4+ = O stretching (nu FeO) Raman band was observed at 788 cm-1 for compound B for the first time. This band showed the 16O/18O isotopic frequency shift (delta nu FeO) by 40 cm-1, in agreement with that for horseradish peroxidase compound II (nu FeO = 787 cm-1 and delta nu FeO = 34 cm-1). In the time region when the FeII-O2 stretching band for compound A and the nu FeO band for compound B were coexistent, a Raman band assignable to the Fe3+-O-O-Cu2+ linkage was not recognized.

Animals↗
PubMed1979

Studies of the orientation of the mitochondrial redox carriers. III. Orientation of the gx and gy axes of the hemes of cytochrome oxidase with respect to the plane of the membrane in oriented membrane multilayers.

The EPR absorption properties of the hemes of cytochrome oxidase and their liganded derivatives were examined in oriented multilayers from isolated oxidase, mitochondrial membranes and membrane fragments of a bacterium, Paracoccus denitrificans. The hemes of the oxidase in all the systems investigated were oriented normal to the plane of the multilayers. The directions of the g signals corresponding to the gx and gy axes of the g tensor were found to be different in low-spin ferric heme in fully oxidized oxidase and in half-reduced liganded oxidase. It is suggested that this different orientation of gx and gy in fully oxidized oxidase and half-reduced liganded oxidase arises because the respective EPR signals belong to two different hemes, those of cytochrome a and a3.

Animals↗
PubMed2001

The mitochondrial cytochrome oxidase II gene in Bacillus stick insects: ancestry of hybrids, androgenesis, and phylogenetic relationships.

Sequencing of a cytochrome oxidase II (COII) gene fragment in Bacillus taxa provided evidence that the bisexual B. rossius is the maternal ancestor of the hybridogenetic B. rossius-grandii strains and revealed the same ancestry for both parthenogenetic hybrids: the diploid B. whitei (B. rossius/grandii grandii) and the triploid B. lynceorum (B. rossius/grandii grandii/atticus). Present data clearly demonstrate that all Bacillus unisexuals arose through asymmetrical hybridization events and realized a paraphyletic derivation from the B. rossius redtenbacheri subspecies. The invention of B. rossius mitochondrial DNA haplotypes in specimens with B. grandii grandii nuclear genomes revealed the occurrence of androgenesis in nature. Natural androgens represent a peculiar escape from hybridity and can help maintain the hybridogenetic system through the production of the fathering taxon via hybrid females. Results from the COII gene support the phyletic relationships among taxa suggested by previous taxonomical approaches, but also indicate a departure of B. grandii subspecies from the established taxonomy. Assuming the existence of a molecular clock, the evaluated substitution rate brings the splitting between B. rossius and B. grandii/B. atticus back to 22.79 +/- 2.65 myr before present, while the origin of hybrids appears to be much more recent (1.06 +/- 0.53 myr).

Androgens↗
PubMed1994

PET1402, a nuclear gene required for proteolytic processing of cytochrome oxidase subunit 2 in yeast.

The nuclear mutation pet ts1402 prevents proteolytic processing of the precursor of cytochrome oxidase subunit 2 (cox2) in Saccharomyces cerevisiae. The structural gene PET1402 was isolated by genetic complementation of the temperature-sensitive mutation. DNA sequence analysis identified a 1206-bp open reading frame, which is located 215 bp upstream of the PET122 gene. The DNA sequence of PET1402 predicts a hydrophobic, integral membrane protein with four transmembrane segments and a typical mitochondrial targeting sequence. Weak sequence similarity was found to two bacterial proteins of unknown function. Haploid cells containing a null allelle of PET1402 are respiratory deficient.

Alleles↗
PubMed1993

X-ray absorption spectroscopy of oriented cytochrome oxidase.

The polarized X-ray absorption spectra of the copper, iron and zinc sites of mitochondrial cytochrome oxidase in oriented membrane multilayers have been examined. The copper X-ray absorption edge spectra indicate the presence of a tetragonal copper, which we assign as CuB, oriented with the long axis approximately orthogonal to the membrane normal. We have also detected the presence of a relatively long (2.6 A) Cu-S or Cu-Cl interaction, which we assign to a copper-thioether (probably Met210) coordination at the CuA site, with the bond oriented along the membrane normal. The coordination of the zinc, the iron and the CuB heme a3 binuclear site are discussed.

Animals↗
PubMed1985

Heme-copper relationship of cytochrome oxidase in rat brain in situ.

The role of copper aa3 in relation to heme aa3 of cytochrome oxidase in electron transfer and oxygen utilization is poorly understood in vitro. In an attempt to study this in situ, we have simultaneously monitored the steady state redox changes of heme aa3 and copper aa3 in an isolated perfused rat head model (skull intact, muscle removed). By means of reflectance spectrophotometry the redox reactions of heme aa3 and copper aa3 were continuously monitored using 605-625 nm and 815-920 nm wavelength pairs, respectively. The reaction kinetics of these components in response to transient perfusion interruption in energized and de-energized preparations were then examined. We found that in response to perfusion interruption, soon after full reduction, the copper signal begins to change toward oxidation despite continuation of anoxic insult and progressive reduction of heme aa3. This phenomenon disappeared by pretreatment of the preparation with 2,4-dinitrophenol. A schematic sequence of electron transport in situ is proposed which emphasizes an active role for Cua in this sequence.

Animals↗
PubMed1993

Localization of cytochrome oxidase (COX) activity and COX mRNA in the perirhinal and superior temporal sulci of the monkey brain.

Cytochrome oxidase (COX) activity and COX II mRNA expression were localized in the perirhinal and superior temporal sulci of the rhesus monkey brain. In both regions, a laminar distribution of COX activity and COX II mRNA was observed. COX activity was intense in layers I and IV and were localized to the neuropil. In contrast, COX II mRNA was localized to neuronal cell bodies. In the prorhinal region, highest levels of COX II mRNA was detected in cell bodies of layers II and IV, and in the perirhinal region, in cell bodies of layers III and V-VI. In the superior temporal sulcus, COX II mRNA was detected in cell bodies of layers III and V-VI. Thus, COX II mRNA and COX activity are uniquely localized in the cortical layers and to those neurons that support cortico-cortical connections.

Animals↗