Course of macroscopic anatomy in Magdeburg under pandemic conditions.

Introduction and objectives: The Covid-19 pandemic has created major challenges for university teaching. At the beginning of the summer semester 2020, teaching at the Medical Faculty in Magdeburg was almost completely online. Also the course in macroscopic anatomy had to be replaced by virtual e-learning offers. Methods: Videos and photo presentations of the preparation steps and structures to be displayed were made available online. The reactions of the students showed very quickly that the three-dimensionality, the independent preparation and the haptics of the object to be studied make up a large part of this subject. Results and conclusions: Virtual e-learning offerings are a useful supplement to, but not a substitute for, active dissecting on body donors. By changing the course offerings in compliance with hygiene and distance rules, we were able to offer a classroom course again during the semester, which was expressly welcomed by the students.

Predator odor induced defensive behavior in wild and laboratory rats: A comparative study.

Laboratory rats are frequently used as animal models in research. Since the 1920s rats are bred and reared in laboratories which affects anatomy, physiology, and behavior responses. In the present study we exposed laboratory and wild rats to predator odor and comparatively analyzed their behavioral and physiological responses. In detail, Warsaw Wild Captive Pisula Stryjek (WWCPS) rats and Lister Hooded (LH) rats were exposed to the predator odor 2,3,5-trimethyl-3-thiazoline (TMT), their behavior was videotaped and blood samples were collected for subsequent serum corticosterone analysis. In both rat stocks, exposure to TMT induced avoidance behavior and increased freezing behavior. Notably, the increase in freezing was based on an increase number of freezing events in LH rats whereas WWCPS rats prolonged the mean duration of the single freezing events. Interestingly, TMT exposure lead to a serum corticosterone increase in WWCPS rats but not in LH rats. Furthermore, WWCPS rats generally expressed decreased but faster locomotor activity, as well as more grooming behavior than LH rats. Taken together, these data indicate differences in behavioral and physiological defensive responses to predator odors in the two rat stocks.

Altered synaptic phospholipid signaling in PRG-1 deficient mice induces exploratory behavior and motor hyperactivity resembling psychiatric disorders.

Plasticity related gene 1 (PRG-1) is a neuron specific membrane protein located at the postsynaptic density of glutamatergic synapses. PRG-1 modulates signaling pathways of phosphorylated lipid substrates such as lysophosphatidic acid (LPA). Deletion of PRG-1 increases presynaptic glutamate release probability leading to neuronal over-excitation. However, due to its cortical expression, PRG-1 deficiency leading to increased glutamatergic transmission is supposed to also affect motor pathways. We therefore analyzed the effects of PRG-1 function on exploratory and motor behavior using homozygous PRG-1 knockout (PRG-1-/-) mice and PRG-1/LPA2-receptor double knockout (PRG-1-/-/LPA2-/-) mice in two open field settings of different size and assessing motor behavior in the Rota Rod test. PRG-1-/- mice displayed significantly longer path lengths and higher running speed in both open field conditions. In addition, PRG-1-/- mice spent significantly longer time in the larger open field and displayed rearing and self-grooming behavior. Furthermore PRG-1-/- mice displayed stereotypical behavior resembling phenotypes of psychiatric disorders in the smaller sized open field arena. Altogether, this behavior is similar to the stereotypical behavior observed in animal models for psychiatric disease of autistic spectrum disorders which reflects a disrupted balance between glutamatergic and GABAergic synapses. These differences indicate an altered excitation/inhibition balance in neuronal circuits in PRG-1-/- mice as recently shown in the somatosensory cortex [38]. In contrast, PRG-1-/-/LPA2-/- did not show significant changes in behavior in the open field suggesting that these specific alterations were abolished when the LPA2-receptor was lacking. Our findings indicate that PRG-1 deficiency led to over-excitability caused by an altered LPA/LPA2-R signaling inducing a behavioral phenotype typically observed in animal models for psychiatric disorders.

Neuregulin-1 mutant mice indicate motor and sensory deficits, indeed few references for schizophrenia endophenotype model.

Neuregulins (Nrg) are a gene family that binds to tyrosine kinase receptors of the ErbB family. The protein of Nrg1 is to be involved in heart formation, migration of neurons, axonal pathfinding and synaptic function. A relation between Nrg1 and schizophrenia is assumed. Chronic impairment in schizophrenia is characterized by different positive and negative symptoms. Detectable markers of this disease in human and in animal models are activity, social behavior and sensory processing. In this study we compared heterozygous Nrg1 mutant mice in behavior and quantification of dopaminergic and serotoninergic neurons with wild type-like littermates. In the Nrg1 mutant mice the epidermal growth factor-like domain is replaced by the neomycin resistance gene. We found significant differences in locomotor and pain perception behavior. No differences were found in specific schizophrenia social interaction and prepulse inhibition behavior. The number of dopaminergic and serotoninergic neurons did not differ in the investigated regions ventral tegmental area, substantia nigra, periaqueductal grey and raphe nuclei. In conclusion, this analyzed Nrg1 mutant mice model did not serve as a complete schizophrenia model. Particular aspects of schizophrenia disease in locomotor and sensory behavior deficits could represent in this Nrg1 mutant mice. Beside several different models could Nrg1 deficiency represent an endophenotype of schizophrenia disease.

A Jacob/Nsmf Gene Knockout Results in Hippocampal Dysplasia and Impaired BDNF Signaling in Dendritogenesis.

Jacob, the protein encoded by the Nsmf gene, is involved in synapto-nuclear signaling and docks an N-Methyl-D-Aspartate receptor (NMDAR)-derived signalosome to nuclear target sites like the transcription factor cAMP-response-element-binding protein (CREB). Several reports indicate that mutations in NSMF are related to Kallmann syndrome (KS), a neurodevelopmental disorder characterized by idiopathic hypogonadotropic hypogonadism (IHH) associated with anosmia or hyposmia. It has also been reported that a protein knockdown results in migration deficits of Gonadotropin-releasing hormone (GnRH) positive neurons from the olfactory bulb to the hypothalamus during early neuronal development. Here we show that mice that are constitutively deficient for the Nsmf gene do not present phenotypic characteristics related to KS. Instead, these mice exhibit hippocampal dysplasia with a reduced number of synapses and simplification of dendrites, reduced hippocampal long-term potentiation (LTP) at CA1 synapses and deficits in hippocampus-dependent learning. Brain-derived neurotrophic factor (BDNF) activation of CREB-activated gene expression plays a documented role in hippocampal CA1 synapse and dendrite formation. We found that BDNF induces the nuclear translocation of Jacob in an NMDAR-dependent manner in early development, which results in increased phosphorylation of CREB and enhanced CREB-dependent Bdnf gene transcription. Nsmf knockout (ko) mice show reduced hippocampal Bdnf mRNA and protein levels as well as reduced pCREB levels during dendritogenesis. Moreover, BDNF application can rescue the morphological deficits in hippocampal pyramidal neurons devoid of Jacob. Taken together, the data suggest that the absence of Jacob in early development interrupts a positive feedback loop between BDNF signaling, subsequent nuclear import of Jacob, activation of CREB and enhanced Bdnf gene transcription, ultimately leading to hippocampal dysplasia.

Plasticity-Related Gene 1 Affects Mouse Barrel Cortex Function via Strengthening of Glutamatergic Thalamocortical Transmission.

Plasticity-related gene-1 (PRG-1) is a brain-specific protein that modulates glutamatergic synaptic transmission. Here we investigated the functional role of PRG-1 in adolescent and adult mouse barrel cortex both in vitro and in vivo. Compared with wild-type (WT) animals, PRG-1-deficient (KO) mice showed specific behavioral deficits in tests assessing sensorimotor integration and whisker-based sensory discrimination as shown in the beam balance/walking test and sandpaper tactile discrimination test, respectively. At P25-31, spontaneous network activity in the barrel cortex in vivo was higher in KO mice compared with WT littermates, but not at P16-19. At P16-19, sensory evoked cortical responses in vivo elicited by single whisker stimulation were comparable in KO and WT mice. In contrast, at P25-31 evoked responses were smaller in amplitude and longer in duration in WT animals, whereas KO mice revealed no such developmental changes. In thalamocortical slices from KO mice, spontaneous activity was increased already at P16-19, and glutamatergic thalamocortical inputs to Layer 4 spiny stellate neurons were potentiated. We conclude that genetic ablation of PRG-1 modulates already at P16-19 spontaneous and evoked excitability of the barrel cortex, including enhancement of thalamocortical glutamatergic inputs to Layer 4, which distorts sensory processing in adulthood.

NT-3 protein levels are enhanced in the hippocampus of PRG1-deficient mice but remain unchanged in PRG1/LPA2 double mutants.

The plasticity-related gene 1 (PRG1) modulates bioactive lipids at the postsynaptic density and is a novel player in neuronal plasticity and regulation of glutamatergic transmission at principal neurons. PRG1, a neuronal molecule, is highly expressed during development and regeneration processes at the postsynaptic density, modulates synaptic lysophosphatidic acid (LPA) levels and is related to epilepsy and brain injury. In the present study, we analyzed the interaction between the synaptic molecules PRG1 and LPA2R with other plasticity-related molecules the neurotrophins. The protein levels of NGF, BDNF and NT-3 were measured using ELISA in hippocampal tissue of homozygous (PRG(-/-)) and heterozygous (PRG(+/-)) PRG1 deficient mice and compared to their wild type (PRG(+/+)/WT) littermates. In the hippocampus, protein levels of NT-3 were significantly increased in PRG(-/-) mice (compared to WT-litters) while protein levels of NGF and BDNF were not affected. Since PRG1 deficiency leads to increased neuronal excitability and higher hippocampal network activity, which may well influence neurotrophin levels, we further assessed PRG1 deficient mice on an LPA2-receptor (LPA2R) deficient background, reported to normalize hippocampal over-excitability in PRG1(-/-) mice. However, on an LPA2R deficient background, protein levels of NT-3 in PRG1(-/-) mice (PRG1(-/-)/LPA2R(-/-)) were not significantly different when compared to WT animals. Since PRG1 deficient mice showed over-excitability in glutamatergic neurons. This was normalized by additional LPA2R deletion, and we conclude the increased NT3-levels were directly or indirectly attributable to increased hippocampal network activity, possibly exerting a protective effect against over-excitability.

Risperidone and haloperidol promote survival of stem cells in the rat hippocampus.

Altered neuroplasticity contributes to the pathophysiology of schizophrenia. However, the idea that antipsychotics may act, at least in part, by normalizing neurogenesis has not been consistently supported. Our study seeks to determine whether hippocampal cell proliferation is altered in adult rats pretreated with ketamine, a validated model of schizophrenia, and whether chronic administration with neuroleptic drugs (haloperidol and risperidone) affect changes of cell genesis/survival. Ketamine per se has no effect on cell proliferation. Its withdrawal, however, significantly induced cell proliferation/survival in the hippocampus. Risperidone and haloperidol supported cell genesis/survival as well. During ketamine withdrawal, however, their application did not affect cell proliferation/survival additionally. TUNEL staining indicated a cell-protective potency of both neuroleptics with respect to a ketamine-induced cell death. As RT-PCR and Western blot revealed that the treatment effects of risperidone and haloperidol seemed to be mediated through activation of VEGF and MMP2. The mRNA expression of NGF, BDNF, and NT3 was unaffected. From the respective receptors, only TrkA was enhanced when ketamine withdrawal was combined with risperidone or haloperidol. Risperidone also induced BCL-2. Ketamine withdrawal has no effect on the expression of VEGF, MMP2, or BCL-2. It activated the expression of BDNF. This effect was normalized by risperidone or haloperidol. The findings indicate a promoting effect of risperidone and haloperidol on survival of young neurons in the hippocampus by enhancing the expression of the anti-apoptotic protein BCL-2 and by activation of VEGF/MMP2, whereby an interference with ketamine and thus a priority role of the NMDA system was not evident.

Expression of mRNA of neurotrophic factors and their receptors are significantly altered after subchronic ketamine treatment.

The neurotrophic factors play an important role in the maintenance of neurone viability and neuronal communication which are considered to be altered in schizophrenia. Subchronic application of ketamine (Ket) was found to be a useful model in schizophrenia research. To further validate this model the mRNA levels of neurotrophic factors NGF, NT-3, and BDNF and their receptors TrkA, TrkB, and TrkC, respectively, were measured in different brain areas in Ket-pretreated rats subchronically dosed with the atypical antipsychotic drug risperidone (Ris). With the exception of NGF in the frontal cortex, Ket pretreatment did change NGF, NT-3, and BDNF mRNA levels in the frontal cortex, the hippocampus, the striatum, the thalamus/hypothalamus region, and in the cerebellum. These changes correspond with changes at their tyrosine kinase receptors. Ris treatment normalised altered NT-3 levels in the hippocampus and balanced BDNF levels in the same structure. It was concluded that the Ket model might reflect distinct alterations in neurotrophic factor activity as found in schizophrenic patients and, moreover, that Ris treatment rebalances disturbed neurotrophic factor activity.

Reduced number of CRF-containing neurons in the central amygdala correlated with enhanced locomotor activity following early postnatal corticosterone treatment in the Wistar rat.

In the present study, newborn rats were implanted with corticosterone (CORT) containing polymers at postnatal day 0 (releasing rate 320-80 microg CORT/kg body weight and day). Controls received a CORT-free implant. All implants were removed at postnatal day 12. At the age of 16-20 weeks, these animals were tested for emotional behavior using an elevated plus-maze and fear-sensitized acoustic startle response. On the elevated plus-maze significant differences were found between hormone treated and control animals. The CORT-group demonstrated higher numbers of entries into closed arms and all arms, and the time spent in the center of the maze was significantly enhanced. Hormone-treated and control rats showed a significant fear sensitization of the acoustic startle response. However, no significant differences were observed between the two groups. The number of CRF-immunopositive neurons in the central nucleus of the amygdala was decreased after CORT treatment, whereas the number of NPY-immunopositive neurons and total number of neurons in the amygdala did not differ significantly between both groups. In conclusion, early postnatal stress induced by CORT administration in neonatal rats led to a higher locomotor activity correlated with changes in the number of CRF containing neurons in the central nucleus of the amygdala.

Transient early postnatal corticosterone treatment of rats leads to accelerated aquisition of a spatial radial maze task and morphological changes in the septohippocampal region.

In the present study new-born rats were treated with corticosterone (CORT) between postnatal days 1 and 12. At the age of 16-20 weeks, these animals were tested for spatial learning capacity using an eight-arm radial maze. After behavioral testing, density of cholinergic fibers and sizes of the mossy fiber terminal fields in the hippocampus and number of cholinergic and GABAergic neurons in the septal area were quantified. In the radial arm maze CORT-treated animals initially showed better working memory performance than controls. However, control animals showed a significant improvement of spatial working memory in the last trials and reached similar working memory scores as compared to treated animals. At neither day of training differences in reference memory errors were found between groups. In the diagonal band of Broca, both numbers of cholinergic and GABAergic neurons were increased after corticosterone treatment. The fiber systems in hippocampus showed no significant differences between groups. In conclusion, early postnatal stress induced by CORT administration in neonatal rats results in mild, yet significant morphological and behavioral changes in later life.

Early postnatal corticosterone administration regulates neurotrophins and their receptors in septum and hippocampus of the rat.

The principal glucocorticoid in rats, corticosterone, interacts with neurons in the limbic system and leads to morphological and behavioral changes. Putative corticosterone-triggered mediators are neurotrophins. In the present study we investigated the effects of early postnatal corticosterone treatment in rats on neurotrophic factors of the nerve growth factor (NGF) family and their receptors. Newborn rats were treated with corticosterone-containing polymers until postnatal day 12. The mRNA and protein levels of the neurotrophins of the NGF family (NGF, BDNF, NT-3 and NT-4/5) and their receptors (trkA, trkB, trkC and p75) were quantified in septum and hippocampus using RT-PCR. In the septal region, we found an unchanged mRNA expression after corticosterone treatment, whereas in the hippocampus there was a general increase in mRNA. Particularly, the gene expression of NGF, NT-3, and the high affinity receptors trkA, trkB and trkC increased significantly. Quantification of the neurotrophin protein levels using an ELISA revealed significant treatment effects for NGF and NT-4/5 in the hippocampus. The present study of corticosterone treatment in young rats demonstrates interactions of steroid hormones with neurotrophic factors and their receptors in the septo-hippocampal system during the first two postnatal weeks.

Effect of early thyroxine treatment on brain-derived neurotrophic factor mRNA expression and protein amount in the rat medial septum/diagonal band of Broca.

There is evidence that morphological alterations concerning deficiency or abundance of thyroid hormones (TH) may be mediated by brain-derived neurotrophic factor (BDNF). It has been demonstrated that the mRNA-expression of BDNF is increased after TH-treatment during the first postnatal weeks. After transient treatment of newborn rats with thyroxine mRNA expression, protein concentration and number and size of BDNF-immunopositive neurons were quantified in the medial septum/vertical diagonal Band of Broca (MS/vDB). The number and size of BDNF-immunopositive neurons were estimated in young (P10) and adult (4 months). The amount of mRNA and protein are significantly increased in TH-treated rats at P10 compared to control animals. TH-treated animals showed a significant decrease of BDNF-immunopositive cell numbers in the adulthood. The results demonstrate a correlated increase of BDNF mRNA and protein in the septum at P10 which is an important stage of differentiation processes in the septohippocampal system. These results provide further evidence that BDNF is a possible candidate for the mediation the TH effects in the MS/vDB.

Anxiety-related behavior and densities of glutamate, GABAA, acetylcholine and serotonin receptors in the amygdala of seven inbred mouse strains.

The amygdala is a brain region involved in the regulation of anxiety-related behavior. The purpose of this study was to correlate anxiety-related behavior of inbred mouse strains (BA//c, BALB/cJ, C3H/HeJ, C57BL/6J, CPB-K, DBA/2J, NMRI) to receptor binding in the amygdala. Binding site densities of receptors (NMDA, AMPA, kainate, GABA(A), serotonin, muscarinergic M(1)-M(2)) were measured with quantitative receptor autoradiography using tritiated ligands. Measurements of fear-sensitized acoustic startle response (ASR; induced by footshocks), elevated plus maze (EPM) behavior and receptor binding studies showed differences between the strains except for AMPA and muscarinergic M(2) receptors. Factor analysis revealed a Startle Factor with positive loadings of the density of serotonin and kainate receptors, and the amplitudes of the baseline and fear-sensitized ASRs. A second Anxiety-related Factor only correlated with the fear-sensitized ASR and anxiety parameters on the EPM but not receptor densities. There were also two General Activity Factors defined by (negative) correlations with entries to closed arms of the EPM. Because the density of NMDA and muscarinergic M(1) receptors also correlated negatively with the two factors, these receptors had a positive effect on general activity. In contrast, correlations of GABA(A), serotonin, and kainate receptors had the opposite sign as compared to closed arm entries. It is concluded that hereditary variations in the amygdala, particularly in kainate and serotonin receptors, play a role for the baseline and fear-sensitized ASR, whereas the general activity is influenced by many neurotransmitter receptor systems.

Transient postnatal thyroxine treatment leads to variation in transmitter binding site densities in the hippocampus of rats.

Newborn rats were treated daily with thyroid hormone (TH) until postnatal day 12. In the adult animals we measured the densities of glutamatergic (N-methyl-D-aspartate (NMDA), amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA), kainic acid (KA)), cholinergic (muscarinic subtype 1 and 2 (M1, M2)), gamma-aminobutyric acid (GABA)ergic (GABA(A)) and serotonergic (5-hydroxytryptamine (5-HT(1A))) binding sites using quantitative receptor-autoradiography with tritiated ligands. In the TH-treated rats the KA binding site density was increased in the stratum oriens of cornu ammonis (CA)3, the terminal field of the infrapyramidal mossy fibers (IPMF). Densities of M2 and 5-HT(1A) were increased in the CA1 region. In contrast, binding site densities for NMDA in the entire hippocampus and for AMPA in the dentate gyrus were reduced, whereas binding site densities for M2 and GABA(A) remained unchanged. From this study we conclude that concomitant with the increase of the IPMF zone the density of the KA binding sites is specifically enhanced. In addition, we found a general shift from binding sites receiving cortical to those receiving subcortical input in the hippocampus.