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1

The role of interleukin 6 in stress response and anxiety-driven behavior in mice

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Aniszewska A., Chlodzinska N., Turlejski K., Djavadian R.
Acta Neurobiologiae Experimentalis
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2013
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tom 73
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nr 1
Interleukin 6 (IL-6) plays an important role in stress response and glucocorticoid action on the brain. It has been also shown that IL-6 plays a significant role in physiological and pathological brain development and is a crucial factor in the effects of prenatal immune challenge on physiological and behavioral abnormalities in adult offspring. We examined involvement of IL-6 in stressinduced changes in behavior and brain mechanisms in the adult mice. The PhenoRack system was used to non-invasively monitor mice home cage activity. Behavior of wild type mice C57BL/6J and IL-6 -/- knockout (IL-6 KO) mice were observed for 3 days using the PhenoRack system, which enables non-invasive monitoring of the mice home-cage activity (distance travelled, speeds and duration of movement, freezing). Mice were also subjected to standard behavioral tests. The open field test was used to establish the balance between exploratory behavior and anxiety evoked by the unknown, potentially dangerous situation. We measured the distance travelled in the open part of the arena, as well as time spent in it. We observed the sex-dependent effect of IL-6 on exploratory behavior. In all tested parameters IL-6 deficient females showed less anxiety than the wild type females. There was no difference in behavior in the open field between wild type and IL-6 deficient males. After finishing behavioral tests, the animals were killed with an overdose of pentobarbital and their brains were perfused transcardially with saline (0.9% NaCl) followed by 4% paraformaldehyde in 0.1 M phosphate buffer. The brains were cut on a cryostat into 40 µm sections and collected into 10 parallel series. Two of these series were stained with antibodies against glucocorticoid (GR) and mineralocorticoid (MR) receptors. One series from each brain was Nissl-stained for delineation of borders of the brain structures and evaluation of the number of cells in some of them. The number of hippocampal neurons and GRimmunopositive neurons in an area was estimated using the StereoInvestigator system (MicroBrightField Inc). Due to the crucial role of IL-6 in development of the hippocampus we evaluated the total number of cells in the CA1 field. We found that IL-6- deficient mice had significantly lower number of cells in the CA1 field and that almost all cells, unlike in wild-type mice, expressed the glucocorticoid receptor. Our preliminary results demonstrated that IL-6 is implicated in stress. Supported by the Polish National Science Center grant No 1577
2

Lipopolysaccharide injected to pregnant mice affects behavior of their offspring in adulthood

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Chlodzinska N., Gajerska M., Bartkowska K., Turlejski K., Djavadian R. L.
Acta Neurobiologiae Experimentalis
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2011
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tom 71
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nr 4
We studied consequences of maternal immune response on the course of pregnancy and the behavior of adult offspring. Mice in late gestation (day 16-17) were injected with lipopolysaccharide (LPS). Treatment of pregnant mice with high doses of LPS resulted in fetal resorption or stillbirths. Pregnant mice treated with low doses (100 or 300 ^g/kg) of LPS gave birth to normal numbers of pups. However, behavior of the offspring was altered. Adult offspring of dams injected at a dose of 300 ^g/kg of LPS traveled longer distances in the open field and spent more time in the central part of the arena, than mice in the control group. Female mice of this group spent more time in open arms of the elevated plus maze, in comparison to female control mice. Results of the Morris water maze test showed impairment of spatial learning and memory in male offspring born to LPS-injected dams. Furthermore, in the nest building test adult mice born from LPS challenged pregnancies constructed worse quality nests, which points to the presence of hippocampal dysfunction. These findings indicate that maternal bacterial infections during pregnancy may alter offspring behavior in adult life.
3

The role of interleukin 6 in behavior of aged mice

88%
Aniszewska A., Chlodzinska N., Winnicka M., Turlejski K., Djavadian R. L.
Acta Neurobiologiae Experimentalis
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2013
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tom 73
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nr Suppl.1
Interleukin 6 (IL-6) is a cytokine playing an important pleiotropic role in the immune system. IL-6 is also involved in stress response, etiology of the age-related diseases and plays a role of mediator between the central nervous system and the immune system. To study effects of IL-6 on behavior during aging we examined aged (13 to 15 months) IL-6 deficient and wild type (WT) mice. Behavior was tested using the open field test, elevated plus maze test and registration of spontaneous activity in the individual home cages for 72 hours. These registrations showed that IL-6 deficient animals were less active than WT mice. The difference was more distinct during the dark phase. Interestingly, in the open field IL-6 deficient mice displayed higher locomotor activity than control WT mice and spent more time in the central part of the arena. In the elevated plus maze IL-6 deficient mice spent more time exploring open arms than WT mice. We conclude that IL-6 deficient aging animals show lower level of anxiety than WT control animals. After tests mice were perfused and brains were cut into 40 μm sections. Brain sections were immunohistochemically labeled for IL-6 and its receptor (IL-6R), also known as CD126. We found that cells immunopositive for both IL-6 and CD126 were present in the hippocampus and other brain structures. Supported by the National Science Center grant No 1577.
4

Delineation of brain structures in the Monodelphis domestica opossum brain

76%
Chlodzinska N., Majka P., Banasik T., Djavadian R. L., Weglarz W. P., Wojcik D., Turlejski K.
Acta Neurobiologiae Experimentalis
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2013
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tom 73
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nr Suppl.1
The Monodelphis opossum became an important laboratory animal and is often used in biomedical research. However, data on the brain anatomy are scarce and there is no reliable brain anatomy reference. The aim of this study is to present neuroanatomical delineation of basic brain structures. Data which served for construction of the 3-dimensional atlas were magnetic resonance images (MRI) and stained brain sections. MRI was obtained 48 h after perfusion of the animal with 4% paraformaldehyde and gadotheridol contrast (ProHance 20:1 v:v). The second MRI was performed 30 days after perfusion of the same animal. Both MRIs were aquired using Bruker Biospin system with voxel reolution of 50 µm3. For Nissland myelin staining, coronal brain sections were cut in cryostat at 40 µm thickness. To minimize tissue deformation, sections were transferred from the cutting blade to slides using the Tape-Transfer System. Then brain sections stained either with Nissl or for myelin were imaged with a high resolution scanner and were transformed to three-dimensional form. By superimposing all three-dimensional data, several brain structures were delineated, e.g., the olfactory bulb, cerebral cortex, hippocampus, white matter and other. Supported by grant from the Polish Ministry of Regional Development POIG.02.03.00-00-003/09.
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