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Neurotrophins belonging to the class of growth factors and including nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and neurotrophin-4/5 (NT-4/5) are widely recognized as essential factors in the developing central nervous system (CNS). Neurotrophins are synthesized as precursor forms (proneurotrophins). Mature forms of neurotrophins exert their effect by binding to specific tyrosine kinases receptors (TrkA, TrkB and TrkC) as well as via the p75 receptor, a member of the tumor necrosis factor receptor superfamily while proneurotrophins interact with the receptor p75 or co-receptor complex of p75 and sortilin, that is a Vps10p domain-containing transmembrane protein. Expression of neurotrophins corresponds with the onset of neurogenesis in developing mammalian species. BDNF is low in early embryonic stages of development, while NT-3 highly expresses in the developing CNS. Expression of neurotrophins receptors mainly overlaps at early development. Data concerning early distribution of neurotrophins and their receptors in the nervous system and results in mice with targeted disruptions of neurotrophin or receptor genes show that neurotrophins and their receptors play distinct roles in control and regulation of the most crucial developmental processes such as proliferation, migration, differentiation, survival, apoptosis and synaptic plasticity.
Neurotrophins are important regulators of neuronal function in the developing and adult brain. We studied expression of TrkB receptors in the postnatally (P) developing brain of the opossum (Monodelphis domestica). The Western blot analysis showed presence of the full-length catalytical isoform of TrkB and three truncated kinase-lacking isoforms in the opossum brain. Expression of the fulllength TrkB receptor was present in the newborn opossum, whereas truncated forms of TrkB receptors were almost undetectable at this period. The level of the full-length TrkB protein gradually increased with a developing opossum brain, reaching maximum at P12–20. The highest levels of expression of the full-length TrkB correspond to the time of cortical layers generation. The level of truncated TrkB rapidly increased at P20 and started to dominate since P35 (when opossums open eyes). Immunohistochemical staining for TrkB receptors showed that the majority of labeled neurons were placed in the olfactory bulb, cerebral cortex, hippocampus, thalamus, hypothalamus, cerebellum and various brainstem structures. Interestingly, TrkB receptors were predominantly expressed in neurons. Lack of TrkB receptors in glial cells, especially astrocytes and oligodendrocytes, provides the evidence that TrkB receptors can play functionally different role in marsupials than in eutherians. Supported by the NSC grant No 2011/01/B/NZ4/01575.
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.
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.
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.
This study describes the topography, borders and divisions of the globus pallidus in the Brazilian short-tailed opossum (Monodelphis domestica) and distribution of the three calcium binding proteins, parvalbumin (PV), calbindin D-28k (CB) and calretinin (CR) in that nucleus. The globus pallidus of the opossum consists of medial and lateral parts that are visible with Nissl or Timm’s staining and also in PV and CR immunostained sections. Neurons of the globus pallidus expressing these proteins were classified into three types on the basis of size and shape of their soma and dendritic tree. Type 1 neurons had medium-sized fusiform soma with dendrites sprouting from the opposite poles. Neurons of the type 2 had medium-to-large, multipolar soma with scarce, thin dendrites. Cell bodies of type 3 neurons were small and either ovoid or round. Immunostaining showed that the most numerous were neurons expressing PV that belonged to all three types. Density of the PV-immunopositive fibers and puncta correlated with the density of the PV-labeled neurons. Labeling for CB resulted mainly in the light staining of neuropil in both parts of the nucleus, while the CB-expressing cells (mainly of the type 2) were scarce and placed only along the border of the globus pallidus and putamen. Staining for calretinin resulted in labeling almost exclusively the immunoreactive puncta and fibers that were distributed with medium-to-high density throughout the nucleus. Close to the border of globus pallidus with the putamen these fibers (probably dendrites) were long, thin and varicous, while more medially bundles of thick, short and smooth fibers predominated. Single CR-ir neurons (all of the type 3) were scattered through the globus pallidus. Colocalization of two calcium binding proteins in one neuron was never observed. The CB-ir puncta (probably terminals of axons projecting to the nucleus) frequently formed basket-like structures around the PV-ir neurons. Therefore, the globus pallidus in the opossum, much as that in the rat, consists of a heterogeneous population of neurons, probably playing diversified functions.
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