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ENG FR. Please try again. Send It. OTM Series Scouts. Need a Quote? Filopodia were identified as long, thin structures without enlarged heads generally twice as long as the average spine length, head: neck diameter ratio less than 1. The rest of the protrusions were classified as spines.
Spines or filopodia in the second view were considered different if they were more than 0. Tests for differences between populations were performed using two-tailed Student t tests with n being the number of animals. Use of Mann—Whitney U test also confirmed all the conclusions. To study the effect of general anesthetics on synapse development, we measured the formation and elimination rates of dendritic spines in the primary somatosensory cortex of 1-month-old mice with or without K-X anesthesia fig.
In awake control mice that had received K-X the day before imaging but none during the imaging session, we found that the rates of newly formed and eliminated dendritic spines were 1.
K-X anesthesia for 1 h or 4 h had no significant effect on the formation and elimination of dendritic spines compared with the controls 1 h K-X: 0. Together, these results suggest that exposure to K-X for 1—4 h has no significant effect on the dynamics or density of dendritic spines. Administration of ketamine-xylazine rapidly increased the formation of dendritic filopodia but not spines over hours.
A and B , In vivo time-lapse imaging of the same dendritic segments over 4 h in the primary somatosensory cortex of 1-month-old animals that received no anesthesia A or ketamine-xylazine K-X anesthesia B. Most dendritic spines on the same dendritic branches remained stable over 4 h whereas filopodia asterisks underwent rapid turnover. C and D , Percentage of newly formed C and eliminated D dendritic spines over 1 and 4 h.
Administration of K-X did not alter spine dynamics. E and F , Percentage of newly formed E and eliminated F dendritic filopodia over 1 and 4 h. K-X anesthesia led to a rapid increase of filopodial formation but had no effect on filopodial elimination. Each filled circle represents a single animal. Dendrites in the developing cortex contain not only dendritic spines but also filopodia, which are long, thin protrusions lacking a bulbous head.
Notably, the rate of filopodial formation over 1 h was significantly higher in mice with K-X anesthesia K-X anesthesia for 4 h further increased the formation of filopodia We also found that the effect of K-X on filopodial formation was dose-dependent: a lower dose 2. Thus, although exposure to K-X has no significant effect on spine dynamics or density, it causes a significant increase in the formation rate of dendritic filopodia.
To determine whether K-X has long-lasting effects on dendritic spines and filopodia, we imaged the same dendritic branches 8 h after the animals recovered from 4 h K-X anesthesia fig. The animals appeared awake and reacted to visual and auditory stimuli between the 4 h and 12 h time points.
We found no significant difference in spine formation 3. Moreover, the formation These results suggest that 4 h K-X anesthesia has no significant long-lasting effect on spine and filopodial dynamics. Ketamine-xylazine has no long-lasting effects on the formation and elimination rates of dendritic spines and filopodia. A , Animals were under ketamine-xylazine K-X anesthesia for the first 4 h and recovered for the next 8 h.
B and C , Percentage of newly formed B and eliminated C dendritic spines over 12 h. D and E , Percentage of newly formed D and eliminated E dendritic filopodia over 12 h. There was no significant difference in spine or filopodial formation and elimination over 12 h between animals with and without K-X anesthesia. Previous studies have suggested that filopodia serve as the precursors of spines. For filopodia formed within the 4 h K-X anesthesia, most of them This elimination rate was comparable to that of filopodia formed without anesthesia A small fraction of filopodia persisted over the next 8 h in mice with Notably, 9.
This percentage of transformation from filopodia to spines was not significantly different from that of nonanesthetized animals 6. Because more filopodia were formed during the 4 h K-X anesthesia, there were approximately 0.
Most newly formed filopodia and spines do not persist. A , The percentage of new filopodia formed over the first 4 h that were eliminated, that persisted as filopodia, or that were transformed to spines over the next 8 h. There was no significant difference between filopodia formed with and without ketamine-xylazine K-X.
B , Percentage of new spines persisting for 8 h. Fewer than half of the new spines formed within the first 4 h persisted for the next 8 h. There was no significant difference between spines formed with and without K-X.
C , Percentage of new spines persisting for 1 month. It is important to note that new spines transformed from filopodia are largely unstable. In fact, more than half of the new spines that were transformed from filopodia within 4 h were eliminated in the next 8 h, regardless of whether they were formed while the mice were awake or anesthetized fig. Furthermore, when the fate of new spines formed over hours to days was followed over a period of 1 month, only 4.
These results are consistent with previous findings showing that most new spines are eliminated over subsequent weeks to months. Previous studies have shown that ketamine is an antagonist of NMDA receptors and produces unconsciousness with analgesia. We observed that MK injection caused a high rate of filopodial formation Over 12 h, the formation rate of filopodia was comparable between MK and saline-injected control animals The elimination rates of filopodia over 4 h Together, these results suggest that the transient effect of 4 h K-X anesthesia on filopodial dynamics is likely mediated by NMDA receptor blockade.
Systemic administration of MK mimics ketamine-xylazine induced filopodial formation. A , Percentage of newly formed dendritic filopodia over 4 and 12 h. Animals were injected with MK after the first imaging session and reimaged 4 and 12 h later. MK injection caused a rapid increase of filopodial formation over 4 but not 12 h. B , Percentage of eliminated dendritic filopodia over 4 and 12 h.
MK had no significant effects on filopodial elimination. Percentages were calculated as the number of filopodia formed or eliminated divided by the number of preexisting filopodia. To further investigate the effect of anesthetics on synapse development, we examined the dynamics of dendritic spines and filopodia after the animals were exposed to isoflurane anesthesia fig.
Similar to K-X, we found that isoflurane had no significant effect on the formation 0. Interestingly, the rate of filopodia elimination was significantly lower in isoflurane-anesthetized mice than in nonanesthetized control mice The rate of filopodial formation over 4 h in isoflurane-anesthetized mice was also lower than that in control mice, although not statistically significant After the animals woke up for 8 h, there was no difference in filopodial elimination between animals with and without isoflurane Administration of isoflurane affects the dynamics of dendritic filopodia but not spines.
A , In vivo time-lapse imaging of the same dendritic segments over 4 h in 1-month-old, isoflurane-anesthetized animals. Most dendritic spines remained stable over 4 h whereas filopodia asterisks underwent rapid turnover. B and C , Percentage of newly formed B and eliminated C dendritic spines over 4 h.
Administration of isoflurane did not alter spine formation and elimination during this time period. D and E, Percentage of newly formed D and eliminated E dendritic filopodia over 4 h. Isoflurane anesthesia decreased the elimination of filopodia but had no significant effect on the formation of filopodia over 4 h. There is increasing evidence that anesthetics induce changes in the developing brain.
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