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Spindle activity in children during cardiac surgery and hypothermic cardiopulmonary bypass.

Hypothermia has marked effects on the electrical activity of the brain, which has been shown in animals as well as in humans. The aim of this study was to investigate EEG spindle activity in children during cardiac surgery and hypothermic cardiopulmonary bypass. The authors obtained intraoperative 21-channel EEG recordings in 36 children (mean age, 22 months; range, 6 days to 69 months) with congenital heart disease. Bipolar EEG derivations were analyzed visually for rhythmic spindle activity based on morphology, frequency, duration, and amplitude. Linear regression analysis for duration, frequency, and amplitude versus rectal temperature was performed in each individual. Spindle activity was observed in 17 children (16 children < 12 months of age). Progressive slowing of spindle frequency with decreasing rectal temperature was found (mean decrease, 0.54 +/- 0.31 Hz/ degrees C). Spindle duration increased on average by 0.69 +/- 0.39 second/ degrees C during cooling procedures. Spindle amplitude did not show any correlation to changes in rectal temperature. The current study demonstrates spindle activity during hypothermic cardiopulmonary bypass with temperature-dependent spindle modifications of frequency and duration. Although the temperature-dependent changes in this study confirm temperature coefficients of other EEG studies, the reasons for the clear age relationship and the "nature" of these spindles remain unknown.

Body Temperature↗

Immunohistochemical analysis of mucinous tubular and spindle cell carcinoma and papillary renal cell carcinoma of the kidney: significant immunophenotypic overlap warrants diagnostic caution.

Mucinous tubular and spindle cell carcinoma, a rare, recently described distinctive subtype of renal cell carcinoma, may have some morphologic similarities to the more common papillary renal cell carcinoma, particularly the basophilic (type 1) tumors with prominent solid growth pattern. Tumor circumscription, compact tubular architecture, focal papillations, mucin production and foam cells (features seen in both papillary renal cell carcinoma and mucinous tubular and spindle cell carcinoma), as well as spindle cell morphology, have resulted in some cases sent to us in consultation with a question of possible sarcomatoid papillary renal cell carcinoma. In this study, tissue microarrays with triplicate samples each from 27 mucinous tubular and spindle cell carcinomas and 20 papillary renal cell carcinomas were created to simulate experience in renal biopsy specimens. From immunohistochemistry (IHC) data, published in the contemporary literature, a panel consisting of alpha-methylacyl-CoA racemase (AMACR), cytokeratin 7 (CK7), epithelial membrane antigen (EMA), renal cell carcinoma marker (RCC Ma), CD10, high molecular weight cytokeratin (HMWK), and c-kit was designed to test its utility in differential diagnosis. The immunoreactivity in mucinous tubular and spindle cell carcinoma was AMACR 93%, CK7 81%, EMA 95%, RCC Ma 7%, CD10 15%, HMWK 15%, and c-kit 5% and in papillary renal cell carcinoma was AMACR 95%, CK7 65%, EMA 88%, RCC Ma 25%, CD10 80%, HWMK 15%, and c-kit 18%. This largest study to date on IHC of mucinous tubular and spindle cell carcinoma dispels the specificity of AMACR for papillary renal cell carcinoma among the RCC subtypes. The histogenesis of mucinous tubular and spindle cell carcinoma from the distal nephron continues to be debatable, as our study showed the expression of the proximal convoluted tubule-related marker AMACR among these tumors. Thus, in tumors with predominant compact tubular growth and focal papillary architectures, careful attention to the presence of a low-grade spindle cell population may be helpful in the distinction of mucinous tubular and spindle cell carcinoma, as the key immunohistochemical stains for papillary renal cell carcinoma are also expressed in this subtype of renal cell carcinoma.

Adenocarcinoma, Mucinous↗

Spindle cell carcinoma of the breast: a case report and an immunohistochemical study including p53 and Ki-67 expression.

A rare case of spindle cell carcinoma (SpCC) of the breast occurring in a 51-year-old Japanese woman is reported. A firm and well-circumscribed tumor, measuring 9 x 8.5 x 8.5 cm, was located on the upper lateral region of the right breast. Microscopically, the tumor consisted of sheets of both malignant spindle cells and poorly differentiated ductal carcinoma containing squamoid islands with gradual transition to the spindle cell component. The immunocytochemical expression of epithelial markers was recognized in the spindle cells, as well as in the carcinomatous cells. Moreover, the spindle cell component expressed vimentin, alpha-smooth muscle actin and S-100 protein. Ultrastructurally, in addition to the features of adenocarcinoma, squamous or myoepithelial differentiation was confirmed in the spindle cell component. These findings thus suggest an epithelial origin with squamous differentiation and myoepithelial participation in the genesis of SpCC. In a comparative study, the expression of p53 protein and Ki-67 as a proliferation marker in each component of this tumor was also investigated. The mean p53 labeling index (LI) in both the carcinomatous and spindle cell area was similar, however the mean MIB-1 LI in the spindle cell area was significantly higher than that in the carcinomatous area. The results indicate that p53 overexpression is involved in the tumorigenesis of both components in the SpCC, and the spindle cell component shows a higher degree of proliferative activity than the carcinomatous component.

Biomarkers↗

Spindle microtubules: thermodynamics of in vivo assembly and role in chromosome movement.

In this paper I have presented results of experiments in which spindle microtubules were depolymerized by hydrostatic pressure, in order to examine the Inoué dynamic equilibrium concept of spindle assembly and the possible role of microtubule depolymerization-polymerization in the movement of chromosomes. Using a newly developed optical hydrostatic pressure chamber, I investigated with polarization microscopy the quantitative effects of pressure on the polymerization of spindle microtubules and, with phase contrast microscopy, the relationship of pressure-induced spindle microtubule depolymerization to chromosome movement in living cells. From results of earlier experiments, principally those of Inoué et al. with low temperature and colchicine as microtubule-depolymerizing agents, and from results of my own research, I have concluded that: (1) spindle fiber microtubules are sensitive to depolymerization by pressure (3000-7000 psi), spindle microtubules do exist in a labile equilibrium with a pool of subunits, and the Inoué simple equilibrium model does predict changes in spindle microtubule assembly at metaphase induced by pressure; (2) the stability of microtubules depends on the number of "attached ends;" (3) the longest interpolar microtubules and the longest chromosomal fiber microtubules regulate the spindle interpolar length and the chromosome-to-pole positions; (4) chromosome velocity is independent of the number of spindle microtubules, as well as of the drag force of the chromosomes; (5) the chromosomal fiber microtubules transmit the forces between the poles and between the chromosomes and the poles; and (6) polymerization of microtubules does produce pushing forces and, if controlled microtubule depolymerization does not actually produce pulling forces, at least it governs the velocity of chromosome-to-pole movement.

Animals↗

Some factors involved in the thalamic control of spontaneous barbiturate spindles.

1. The origin of thalamic and cortical spontaneous spindles was studied in cats anaesthetized with sodium pentobarbital.2. Complete removal of all cortical grey matter left the thalamic rhythmic spindle activity unchanged.3. Removal of the entire thalamus or pronounced oedema in the thalamus abolished completely the spindle activity in the corresponding hemisphere.4. In a neuronally isolated cortical area, a fast, low voltage background activity appeared, interrupted by occasional irregular rapid potential changes (sharp waves) of high voltage. Regular spindle rhythms were seldom observed unless excited by a depolarizing drug. Spontaneous spindles did not invade the isolated cortex via a bridge of intact cortical tissue.5. With increasingly larger destruction of the thalamus in a rostro-caudal direction, the activity in the post-cruciate cortex did not change until the anterior third of the thalamus was encroached upon. A transverse section in front of the thalamus nearly eliminated the cortical spindles.6. Complete removal of the mid line and intralaminar nuclei left the spontaneous rhythmic activity of the lateral thalamic nuclei and of the frontal cortex principally unchanged.7. Removal of the laterally located thalamic nuclei, including the n. ventralis posterolateralis (VPL), abolished virtually all spontaneous spindle activity of the frontal cortex, including the post-cruciate area.8. Local cortical cooling reduced the amplitude but not the frequency of the cortical spindles.9. Cooling of the whole brain reduced both the amplitude and the frequency of the spindles. At low temperatures, all spindle activity in the cortex disappeared, and occasional sharp waves occurred, as with de-afferentation.10. It is concluded that the rhythm of the cortical spontaneous barbiturate spindles is generated exclusively by thalamic neurones. The electromotive force of the corticographic waves, however, has a cortical origin.

Animals↗

Cytologic findings of spindle cell ductal carcinoma in situ of the breast: a case report.

BACKGROUND: Ductal carcinoma in situ (DCIS), spindle cell type, is a rare and recently recognized entity in the breast. While the histologic features have been described in detail, no cytologic descriptions exist. We describe the cytologic and corresponding histologic findings in a case of spindle cell DCIS. CASE: A 35-year-old woman underwent fine needle aspiration (FNA) of a 1.5-cm, palpable mass. The aspirate showed predominantly clusters and a smaller population of single cells in a necrotic background. The cells were mostly spindled, The cells were mostly spindled, with a minor population of epithelioid cells. They had a high nuclear/cytoplasmic ratio, with hyperchromatic and pleomorphic nuclei that contained prominent nucleoli, altogether diagnostic of malignancy. Given the biphasic cell composition, a diagnosis of metaplastic carcinoma was favored and excision recommended; it showed spindle cell DCIS. CONCLUSION: The cytologic features of spindle cell DCIS and metaplastic spindle cell carcinoma are similar and almost indistinguishable. Despite its rarity, spindle cell DCIS should be considered in the differential diagnosis of biphasic and spindle cell lesions in the breast.

Adult↗

Frontal spindle activity that appears in conjunction with nontraumatic diffuse encephalopathy.

This peculiar 11-14 Hz spindle activity appears predominantly in the frontal area, and was observed in eight patients with impaired consciousness caused by nontraumatic diffuse encephalopathy. Characteristic of this frontal spindle activity is its transience and accordance with changes in the arousal level of the patient. When the degree of impaired consciousness in the patient was minimal and clinically not very apparent, this spindle activity appeared during light drowsiness. In lethargic patients, it was observed when the patient's level of consciousness rose (e.g. immediately after opening and closing the eyes). These frontal spindles disappeared at the onset of Stage 2 sleep, when normal physiologic spindle waves that are dominant in the vertex area appeared. A paroxysmal discharge was sometimes recorded in association with the frontal spindle activity and it disappeared at about the same time as these spindles. The prognosis was satisfactory for all patients in whom frontal spindle activity was observed; its correlation to spindle coma is also studied.

Adolescent↗

Development of sleep spindles in young children and adolescents.

The development of sleep spindles was studied quantitatively in 32 healthy subjects between the ages of 4 and 24 years. The peak frequency distribution of the spindles showed a bimodal pattern with 11.0 to 12.75 Hz in the frontal area and 12.5 to 14.5 Hz in the centroparietal area. The two types of spindle activity showed different courses of maturation. The peak frequency of the centroparietal spindles gradually increased linearly with age, whereas the frontal spindles abruptly increased in frequency during early adolescence. Regarding the power spectra, while centroparietal spindles showed little change in power from 4 to 24 years of age, frontal spindles decreased remarkably in power and became stable at about 13 years of age. The two types of spindles and the difference in their development may suggest the existence of different generators or a topographical difference during maturation in the thalamocortical network. The frontal spindle activity could be a good indicator to evaluate CNS maturation in young children and adolescents.

Adolescent↗

Pattern-specific associative long-term potentiation induced by a sleep spindle-related spike train.

Spindles are non-rapid eye movement (non-REM) sleep EEG rhythms (7-14 Hz) that occur independently or in association with slow oscillations (0.6-0.8 Hz). Despite their proposed function in learning and memory, their role in synaptic plasticity is essentially unknown. We studied the ability of a neuronal firing pattern underlying spindles in vivo to induce synaptic plasticity in neocortical pyramidal cells in vitro. A spindle stimulation pattern (SSP) was extracted from a slow oscillation upstate that was recorded in a cat anesthetized with ketamine-xylazine, which is known to induce a sleep-like state. To mimic the recurrence of spindles grouped by the slow oscillation, the SSP was repeated every 1.5 s (0.6 Hz). Whole-cell patch-clamp recordings were obtained from layer V pyramidal cells of rat somatosensory cortex with infrared videomicroscopy, and composite EPSPs were evoked within layers II-III. Trains of EPSPs and action potentials simultaneously triggered by the SSP induced an NMDA receptor-dependent short-term potentiation (STP) and an L-type Ca2+ channel-dependent long-term potentiation (LTP). The number of spindle sequences affected the amount of STP-LTP. In contrast, spindle trains of EPSPs alone led to long-term depression. LTP was not consistently induced by a regular firing pattern, a mirrored SSP, or a randomized SSP; however, a synthetic spindle pattern consisting of repetitive spike bursts at 10 Hz reliably induced STP-LTP. Our results show that spindle-associated spike discharges are efficient in modifying excitatory neocortical synapses according to a Hebbian rule. This is in support of a role for sleep spindles in memory consolidation.

Action Potentials↗

[Periodic discharges of sleep spindles with placebo and zopiclone].

According to our experiences (parietal) sleep spindles often show a periodical appearance. Successive sleep spindles in series have a distance of about four seconds. In 95% the duration of such series of sleep spindles is not longer than 40 s, so that there appears no more than ten successive periodical sleep spindles. In the present study ten subjects aged between 57 and 77 years were given Placebo and 7.5 mg Zopiclone. Under the effect of verum we found a doubling of sleep spindle series. However, the distance between the periodical spindles and the length of the spindle series remained unchanged. The variance in the ability to generate sleep spindle series was very high across subjects. One subject for example produced only one sleep spindle series in the Placebo-night (three under verum) and another subject on the other hand 95 (under verum 130). The results of the present study well corresponding to the results of a former investigation comparing Placebo, Pentobarbital, Methaqualon, Carbromal, Flunitrazepam, Triazolam and Lormetazepam.

Aged↗

The fine structure of human extraocular muscle spindles and their potential proprioceptive capacity.

Twenty seven muscle spindles from six extraocular muscles removed following ocular enucleation from patients aged 58, 76 and 74 years were examined throughout all or most of their length by means of light and electron microscopy using serial transverse sections. Five others were prepared in longitudinal section. Twelve spindles of the superior rectus muscle from three sheep orbits were studied in a similar manner to provide a comparison. The human spindles contained a total of 90 (42%) nuclear chain and 5 (2%) nuclear bag fibres with the usual complement of sensory endings, and 120 (56%) fibres were anomalous with continuous, unattenuated myofibrils throughout their length, a constant width and peripherally placed nuclei. Eight anomalous fibres received sensory terminals similar in form to those of chain and bag fibres. Most (26) spindles contained at least one chain and one anomalous fibre. The periaxial space was limited or absent and the inner capsule was often segmented and in contact with the outer capsule. Abrupt termination of some chain fibres including several with one pole missing, together with evidence of fibre fragmentation and other structural anomalies, were indicative of degeneration. Eight further encapsulated fibre groups were identified as false spindles containing only anomalous fibres; associated nerves failed to terminate in the encapsulations. Sheep spindle content was of regular form, all spindles containing several chain and at least one bag fibre enclosed by an inner capsule and surrounded by a substantial periaxial space equatorially. The human extraocular muscle spindles have lost, either by aging or phylogenetically, the privilege of contractile chambers isolated by a fluid periaxial space from extrafusal fibre activity and sensory terminals are subject to the direct mechanical influences of anomalous intrafusal fibres. These, and the other departures from normal structure described, must jeopardize monitoring of muscle activity in the manner normally attributed to spindles and their capacity to provide useful proprioceptive information is questionable.

Aged↗

Neocortical spindling during wakefulness in the rat.

Neocortical spindling that frequently occurs in rats during wakefulness was studied to evaluate the hypotheses that spindle bursts are either the electrophysiological manifestation of a short-lasting sleep episode that briefly interrupts wakefulness (due to an urge to sleep) or a short decrease of the vigilance level. In order to evaluate sleep need, the latency to the onset of natural sleep, the percentual composition of the sleep-wakefulness cycles, and the durations and intervals of desynchronized sleep episodes were determined in six male Wistar rats weighing 250-350 g and having chronically implanted electrodes for frontal electrocorticogram and cervical electromyogram. These animals were selected on the basis of spindling manifestation during wakefulness. The occurrence of spindling during a period of repeated painful tail-pinching was subsequently measured to determine the vigilance level in the same animals. Two rats were also studied during forced immobilization for the same purpose. Sleep parameters were found to be normal in all rats studied, thus excluding the hypothesis that spindling in wakefulness is a manifestation of a high sleep need. Spindling also occurred in both situations requiring a high level of vigilance (frequent tail-pinching and forced immobilization). Natural sleep cycles never started with this type of spindling, which is not related to the typical synchronization patterns of synchronized sleep. The frequency of the potentials that make up spindles in wakefulness were systematically 1 to 2 Hz lower than those of synchronized sleep in all animals studied. The possibility that spindling during wakefulness may be associated to brief episodes of distraction is discussed.

Animals↗

Spindle positioning during the asymmetric first cell division of Caenorhabditis elegans embryos.

Cell division during development in many cases generates daughter cells that differ not only in fate, but also in size. We investigate the mechanisms that ensure proper spindle positioning during such asymmetric divisions using the one-cell stage Caenorhabditis elegans embryo as a model system. We utilized a UV laser microbeam as an in vivo microtubule-severing device to probe the forces driving spindle positioning. Our results indicate that extra-spindle pulling forces acting on the spindle poles dictate spindle position along the anterior-posterior embryonic axis. Importantly, forces acting on the posterior spindle pole appear more extensive than those acting on the anterior one, thus explaining the overall posterior spindle displacement that leads to the asymmetric division of the wild-type one-cell stage embryo. In separate work, we analysed a locus called zyg-8, which plays a key role in ensuring proper spindle positioning. Our data show that zyg-8 is required to promote microtubule growth and/or stability during anaphase. We identified the molecular nature of the zyg-8 locus in the course of a large-scale RNAi-based functional genomics screen. ZYG-8 harbours two notable protein domains: a Ca2+/calmodulin-dependent kinase domain, and a domain related to doublecortin, a human microtubule-associated protein involved in neuronal migration.

Animals↗

A comparative study of muscle spindles in slow and fast neonatal muscles of normal and dystrophic mice.

Muscle spindles from the slow-twitch soleus and the fast-twitch extensor digitorum longus (EDL) muscles of genetically dystrophic mice of the dy2J/dy2J strain were compared with age-matched normal animals at neonatal ages of 1-3 weeks according to histochemical, quantitative, and ultrastructural parameters. Intrafusal fibers in both the soleus and EDL exhibited similar regional differences in myosin ATPase activity, and conformed to those noted previously in various adult species. In distal polar regions, all nuclear bag fibers resembled extrafusal fibers of the type 1 variety, whereas in capsular zones they could be divided into two subtypes. Nuclear chain fibers possessed a staining pattern similar to type 2 extrafusal fibers, and in contrast to the bag fibers they exhibited no regional variations. These features were consistently observed in both the normal and dystrophic muscles at all ages. Spindles varied only slightly in their number and distribution in the two types of muscle, and their location followed the neurovascular branching pattern in each. Irrespective of age or genotype, spindles in the soleus were more homogeneously dispersed, but those in the EDL were concentrated along the dorsal aspect of the muscle. No significant differences were noted in the total number of spindles between normal and dystrophic muscles. In addition, no dramatic differences were observed in the muscle spindle index for soleus and EDL. The first obvious disease-related changes were noted in extrafusal fibers of the soleus of 3-week-old mice, and spindles were often located close to these areas of fiber degeneration. Despite alterations in the surrounding tissue, however, spindles appeared morphologically unaltered in dystrophy. These observations indicate that intrafusal fibers of spindles in neonatal mice appear enzymatically and histologically unaffected in incipient stages of progressive muscular dystrophy.

Adenosine Triphosphatases↗

Kinetochore microtubules in crane-fly spermatocytes: untreated, 2 degrees C-treated, and 6 degrees C-grown spindles.

We investigated the involvement of kinetochore microtubules (kMTs) in mediating chromosome-to-pole connections in crane-fly (Nephrotoma suturalis and Nephrotoma ferruginea) spermatocytes. Two experimental treatments were used to yield spindles with reduced numbers of nonkinetochore microtubules (nkMTs). Short-term (10-15 min) exposure of spermatocytes to 2 degrees C caused depolymerization of the majority of nkMTs, resulting in a kMT:(kMT + nkMT) ratio of 0.76. Long-term (24h) exposure to 2 degrees C followed by recovery at 6 degrees C resulted in a kMT:(kMT + nkMT) ratio of 0.55, the spindle having more nkMTs than a 2 degrees C-treated spindle but fewer than an untreated spindle, in which the kMT:(kMT + nkMT) ratio was 0.27. The numbers and lengths of kMTs in 6 degrees C-grown spindles were similar to those in untreated cells, suggesting that the overall inhibition of MT assembly at 6 degrees C apparently did not affect the mechanism by which kMTs are formed. We observed most kMTs of early anaphase spindles to be long (greater than 3 microns), and many extended to the polar regions of the spindle. Thus, the crane-fly spindle appears not to be as atypical as it was previously suggested to be.

Animals↗

Reactivation of isolated mitotic apparatus: metaphase versus anaphase spindles.

Mitotic spindles isolated from sea urchin eggs can be reactivated to undergo mitotic processes in vitro. Spindles incubated in reactivation media containing sea urchin tubulin and nucleotides undergo pole-pole elongation similar to that observed in living cells during anaphase-B. The in vitro behavior of spindles isolated during metaphase and anaphase are compared. Both metaphase and anaphase spindles undergo pole-pole elongation with similar rates, but only in the presence of added tubulin. In contrast, metaphase but not anaphase spindles increase chromosome-pole distance in the presence of exogenous tubulin, suggesting that in vitro, tubulin can be incorporated at the kinetochores of metaphase but not anaphase chromosomes. The rate of spindle elongation, ultimate length achieved, and the increase in chromosome-pole distance for isolated metaphase spindles is related to the concentration of available tubulin. Pole-pole elongation and chromosome-pole elongation does not require added adenosine triphosphate (ATP). Guanosine triphosphate (GTP) will support all activities observed. Thus, the force generation mechanism for anaphase-B in isolated sea urchin spindles is independent of added ATP, but dependent on the availability of tubulin. These results support the hypothesis that the mechanism of force generation for anaphase-B is linked to the incorporation of tubulin into the mitotic apparatus. (If, in addition, a microtubule-dependent motor-protein(s) is acting to generate force, it does not appear to be dependent on ATP as the exclusive energy source.

Adenosine Triphosphate↗

Location and completeness of reinnervation by two types of neurons at a single target: the feline muscle spindle.

Muscle spindles from the tenuissimus muscle of the cat were examined microscopically to assess the precision and completeness of reinnervation of intrafusal muscle fibers by efferent and afferent neurons. Positions of motor and sensory nerve terminals were charted relative to the cross-sectional area enclosed by the outer capsule of the spindle. Profiles of nerve endings were measured for normally innervated and reinnervated spindles. The tenuissimus was deprived of innervation by freezing its nerve, sometimes in conjunction with either spinal ganglion removal or ventral rhizotomy. Sensory and motor terminals occupied separate locales along the length of normal muscle spindles. Nerve terminals of efferent and afferent neurons were located in appropriate positions along the length of spindles when axons of both types of neurons regrew together and when either category of axon regenerated alone. Precise reinnervation of muscle spindles occurred in spite of a diminished diameter of intrafusal fibers. Repopulation of the spindle with motor endings was less complete than that by sensory endings, based on the proportion and size of the regenerated terminals. We conclude that under optimal conditions for axonal regrowth, efferent and afferent neurons reinnervate their respective regions along intrafusal muscle fibers but motor lags sensory reinnervation within the spindle. The mechanism by which positional specificity happens during reinnervation of intrafusal fibers requires neither an interaction between terminals of the two types of neurons nor target cells of normal bulk.

Animals↗

Histological and histochemical comparisons of muscle spindles in three hind limb muscles of the guinea pig.

Guinea pig soleus, medial gastrocnemius and vastus lateralis muscles were compared for spindle density and distribution, number of intrafusal fibers per spindle and histochemical appearance of the axial bundle. A total of 326 spindles was used in the comparisons. Spindle density was over four times greater in the soleus than in either the medial gastrocnemius or vastus lateralis. In the soleus the spindles were distributed at random, but in the other two muscles no spindles were found in those fascicles in which fast-twitch glycolytic extrafusal fibers predominated. The average number of intrafusal fibers per spindle varied by less than 5% between the three kinds of muscles. About 80% of all spindles located had four intrafusal fibers, two of the nuclear bag type and two of the nuclear chain type. The histochemical appearance of the axial bundle was the same in each kind of muscle. Based on intensities of the myofibrillar adenosine triphosphatase reaction product at polar regions nuclear bag fibers were separable into two histochemical groups; nuclear chain fibers were of only one histochemical type.

Adenosine Triphosphatases↗