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Differences in the hippocampal frequency of creatine inclusions between the acute and latent phases of pilocarpine model defined using synchrotron radiation-based FTIR microspectroscopy.

Kutorasinska J, Setkowicz Z, Janeczko K, Sandt C, Dumas P, Chwiej J - Anal Bioanal Chem (2013)

Bottom Line: The results obtained for the period when stabilization of animal behavior and EEG occurs were afterwards compared with ones obtained for the acute phase of pilocarpine-induced SE and for naive controls.Moreover, for the hilus of the DG and CA3 area, the number of creatine deposits was higher in the latent than in the acute phase after pilocarpine injection.In light of the obtained results, an anomaly in the hippocampal accumulation of creatine is the long-term effect of pilocarpine-evoked seizures, and the intensity of this phenomenon may increase with time passing from the primary injury.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Physics and Applied Computer Science, AGH University of Science and Technology, 30 Mickiewicza Av., 30-059, Krakow, Poland. Justyna.Kutorasinska@fis.agh.edu.pl

ABSTRACT
Temporal lobe epilepsy (TLE) is the most common type of epilepsy in adults. Of the animal models developed to investigate the pathogenesis of TLE, the one with pilocarpine-induced seizures is most often used. After pilocarpine administration in animals, three distinct periods--acute, latent, and chronic--can be distinguished according to their behavior. The present paper is the continuation of our previous study which has shown an increased occurrence of creatine inclusions in rat hippocampal formations from the acute phase of pilocarpine-induced status epilepticus (SE) and positive correlation between their quantity and the total time of seizure activity within the observation period. In this paper, we tried to verify if anomalies in hippocampal creatine accumulation were the temporary or permanent effect of pilocarpine-evoked seizures. To realize this purpose, male Wistar rats in the latent phase (3 days after pilocarpine administration) were examined. The results obtained for the period when stabilization of animal behavior and EEG occurs were afterwards compared with ones obtained for the acute phase of pilocarpine-induced SE and for naive controls. To investigate the frequency of creatine inclusions within the hippocampal formation as well as in its selected areas (sectors 1-3 of Ammon's horn (CA1-CA3), dentate gyrus (DG), and hilus of DG) and cellular layers (pyramidal, molecular, multiform, and granular cell layers), synchrotron radiation-based Fourier-transform infrared microspectroscopy was used. The applied technique, being a combination of light microscopy and infrared spectroscopy, allowed us to localize microscopic details in the analyzed samples and provided information concerning their chemical composition. Moreover, the use of a synchrotron source of IR radiation allowed us to carry out the research at the diffraction-limited spatial resolution which, because of the typical size of creatine inclusions (from a few to dozens of micrometers), was necessary for our study. The comparison of epileptic animals in the latent phase with controls showed statistically significant increase in the number of creatine inclusions for most of the analyzed hippocampal regions, all examined cellular layers, as well as the whole hippocampal formation. Moreover, for the hilus of the DG and CA3 area, the number of creatine deposits was higher in the latent than in the acute phase after pilocarpine injection. In light of the obtained results, an anomaly in the hippocampal accumulation of creatine is the long-term effect of pilocarpine-evoked seizures, and the intensity of this phenomenon may increase with time passing from the primary injury.

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The mean and median values of creatine inclusion quantities in particular cellular layers of hippocampal formations. Statistically significant differences (p value < 0.1) between SE72H and N groups were marked with *, between SE6H and N groups with **, and between SE72H and SE6H groups with #. The differences significant with Bonferroni-corrected p value were additionally marked in red
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Fig5: The mean and median values of creatine inclusion quantities in particular cellular layers of hippocampal formations. Statistically significant differences (p value < 0.1) between SE72H and N groups were marked with *, between SE6H and N groups with **, and between SE72H and SE6H groups with #. The differences significant with Bonferroni-corrected p value were additionally marked in red

Mentions: In Tables 1 and 2, the results of quantitative analysis carried out for the three examined animal populations were presented as the mean, median, minimal, and maximal values of the number of creatine deposits in analyzed hippocampal areas and cellular layers. Additionally, the data were shown graphically as the box plots in Figs. 4 and 5.Table 1


Differences in the hippocampal frequency of creatine inclusions between the acute and latent phases of pilocarpine model defined using synchrotron radiation-based FTIR microspectroscopy.

Kutorasinska J, Setkowicz Z, Janeczko K, Sandt C, Dumas P, Chwiej J - Anal Bioanal Chem (2013)

The mean and median values of creatine inclusion quantities in particular cellular layers of hippocampal formations. Statistically significant differences (p value < 0.1) between SE72H and N groups were marked with *, between SE6H and N groups with **, and between SE72H and SE6H groups with #. The differences significant with Bonferroni-corrected p value were additionally marked in red
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3756859&req=5

Fig5: The mean and median values of creatine inclusion quantities in particular cellular layers of hippocampal formations. Statistically significant differences (p value < 0.1) between SE72H and N groups were marked with *, between SE6H and N groups with **, and between SE72H and SE6H groups with #. The differences significant with Bonferroni-corrected p value were additionally marked in red
Mentions: In Tables 1 and 2, the results of quantitative analysis carried out for the three examined animal populations were presented as the mean, median, minimal, and maximal values of the number of creatine deposits in analyzed hippocampal areas and cellular layers. Additionally, the data were shown graphically as the box plots in Figs. 4 and 5.Table 1

Bottom Line: The results obtained for the period when stabilization of animal behavior and EEG occurs were afterwards compared with ones obtained for the acute phase of pilocarpine-induced SE and for naive controls.Moreover, for the hilus of the DG and CA3 area, the number of creatine deposits was higher in the latent than in the acute phase after pilocarpine injection.In light of the obtained results, an anomaly in the hippocampal accumulation of creatine is the long-term effect of pilocarpine-evoked seizures, and the intensity of this phenomenon may increase with time passing from the primary injury.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Physics and Applied Computer Science, AGH University of Science and Technology, 30 Mickiewicza Av., 30-059, Krakow, Poland. Justyna.Kutorasinska@fis.agh.edu.pl

ABSTRACT
Temporal lobe epilepsy (TLE) is the most common type of epilepsy in adults. Of the animal models developed to investigate the pathogenesis of TLE, the one with pilocarpine-induced seizures is most often used. After pilocarpine administration in animals, three distinct periods--acute, latent, and chronic--can be distinguished according to their behavior. The present paper is the continuation of our previous study which has shown an increased occurrence of creatine inclusions in rat hippocampal formations from the acute phase of pilocarpine-induced status epilepticus (SE) and positive correlation between their quantity and the total time of seizure activity within the observation period. In this paper, we tried to verify if anomalies in hippocampal creatine accumulation were the temporary or permanent effect of pilocarpine-evoked seizures. To realize this purpose, male Wistar rats in the latent phase (3 days after pilocarpine administration) were examined. The results obtained for the period when stabilization of animal behavior and EEG occurs were afterwards compared with ones obtained for the acute phase of pilocarpine-induced SE and for naive controls. To investigate the frequency of creatine inclusions within the hippocampal formation as well as in its selected areas (sectors 1-3 of Ammon's horn (CA1-CA3), dentate gyrus (DG), and hilus of DG) and cellular layers (pyramidal, molecular, multiform, and granular cell layers), synchrotron radiation-based Fourier-transform infrared microspectroscopy was used. The applied technique, being a combination of light microscopy and infrared spectroscopy, allowed us to localize microscopic details in the analyzed samples and provided information concerning their chemical composition. Moreover, the use of a synchrotron source of IR radiation allowed us to carry out the research at the diffraction-limited spatial resolution which, because of the typical size of creatine inclusions (from a few to dozens of micrometers), was necessary for our study. The comparison of epileptic animals in the latent phase with controls showed statistically significant increase in the number of creatine inclusions for most of the analyzed hippocampal regions, all examined cellular layers, as well as the whole hippocampal formation. Moreover, for the hilus of the DG and CA3 area, the number of creatine deposits was higher in the latent than in the acute phase after pilocarpine injection. In light of the obtained results, an anomaly in the hippocampal accumulation of creatine is the long-term effect of pilocarpine-evoked seizures, and the intensity of this phenomenon may increase with time passing from the primary injury.

Show MeSH
Related in: MedlinePlus