Inactivation of bcl-2 results in progressive degeneration of motoneurons, sympathetic and sensory neurons during early postnatal development.

Bcl-2 is a major regulator of programmed cell death, a critical process in shaping the developing nervous system. To assess whether Bcl-2 is involved in regulating neuronal survival and in mediating the neuroprotective action of neurotrophic factors, we generated Bcl-2-deficient mice. At birth, the number of facial motoneurons, sensory, and sympathetic neurons was not significantly changed, and axotomy-induced degeneration of facial motoneurons could still be prevented by brain-derived neurotrophic factor (BDNF) or ciliary neurotrophic factor (CNTF). Interestingly, substantial degeneration of motoneurons, sensory, and sympathetic neurons occurred after the physiological cell death period. Accordingly, Bcl-2 is not a permissive factor for the action of neurotrophic factors, and although it does not influence prenatal neuronal survival, it is crucial for the maintenance of specific populations of neurons during the early postnatal period.

Retarded growth and deficits in the enteric and parasympathetic nervous system in mice lacking GFR alpha2, a functional neurturin receptor.

Glial cell line-derived neurotrophic factor (GDNF) and a related protein, neurturin (NTN), require a GPI-linked coreceptor, either GFR alpha1 or GFR alpha2, for signaling via the transmembrane Ret tyrosine kinase. We show that mice lacking functional GFR alpha2 coreceptor (Gfra2-/-) are viable and fertile but have dry eyes and grow poorly after weaning, presumably due to malnutrition. While the sympathetic innervation appeared normal, the parasympathetic cholinergic innervation was almost absent in the lacrimal and salivary glands and severely reduced in the small bowel. Neurite outgrowth and trophic effects of NTN at low concentrations were lacking in Gfra2-/- trigeminal neurons in vitro, whereas responses to GDNF were similar between the genotypes. Thus, GFR alpha2 is a physiological NTN receptor, essential for the development of specific postganglionic parasympathetic neurons.

Specific subtypes of cutaneous mechanoreceptors require neurotrophin-3 following peripheral target innervation.

Neurotrophin-3 (NT-3) is required for the development of most sensory neurons of the dorsal root ganglia. Using electrophysiological techniques in mice with null mutations of the NT-3 gene, we show that two functionally specific subsets of cutaneous afferents differentially require this factor: D-hair receptors and slowly adapting mechanoreceptors; other cutaneous receptors were unaffected. Merkel cells, which are the end organs of slowly adapting mechanoreceptors, are virtually absent in 14-day-old homozygous mutants and are severely reduced in adult NT-3 heterozygous animals. This loss of Merkel cells, together with their innervation, happens in the first postnatal weeks of life, in contrast to muscle spindles and afferents, which are never formed in the absence of NT-3. Thus, NT-3 is essential for the maintenance of specific cutaneous afferents known to subserve fine tactile discrimination in humans.

GFRalpha 1 is required for development of distinct subpopulations of motoneuron.

Glial cell-line derived neurotrophic factor (GDNF) and its relative neurturin (NTN) are potent trophic factors for motoneurons. They exert their biological effects by activating the RET tyrosine kinase in the presence of a glycosyl-phosphatidylinositol-linked co-receptor, either GFRalpha1 or GFRalpha2. By whole-mount in situ hybridization on embryonic mouse spinal cord, we demonstrate that whereas Ret is expressed by nearly all motoneurons, Gfra1 and Gfra2 exhibit complex and distinct patterns of expression. Most motoneurons purified from Gfra1 null mutant mice had lost their responsiveness to both GDNF and NTN. However, a minority of them ( approximately 25%) retained their ability to respond to both factors, perhaps because they express GFRalpha2. Surprisingly, Gfra2(-/-) motoneurons showed normal survival responses to both GDNF and NTN. Thus, GFRalpha1, but not GFRalpha2, is absolutely required for the survival response of a majority of motoneurons to both GDNF and NTN. In accordance with the phenotype of the mutant motoneurons observed in culture we found the loss of distinct groups of motoneurons, identified by several markers, in the Gfra1(-/-) spinal cords but no gross defects in the Gfra2(-/-) mutant. During their natural programmed cell death period, motoneurons in the Gfra1(-/-) mutant mice undertook increased apoptosis. Taken together these findings support the existence of subpopulations of motoneuron with different trophic requirements, some of them being dependent on the GDNF family.

Long-term use of benzodiazepines: Definitions, prevalence and usage patterns - a systematic review of register-based studies.

BACKGROUND: Numerous treatment guidelines recommend that long-term use of benzodiazepines (BZD) should be avoided primarily due to development of tolerance and a risk for BZD dependence. Despite this, long-term BZD use remains a controversial subject in clinical patient care with "for and against" debates. However, there is no explicit understanding of what is meant by long-term BZD use in real world. The aim of this study was to assess different definitions, usage patterns, prevalence and other characteristics of long-term BZD use based on published register-based studies. Synthesis of these characteristics is essential to derive a meaningful definition of long-term BZD. METHODS: Systematic review of register-based studies on long-term BZD use published in 1994-2014. RESULTS: Fourty-one studies met our predetermined inclusion criteria. The length of BZD use defined as "long-term" varied in these studies ranging from one month to several years. The most common definition was six months or longer during a year. The prevalence of long-term BZD use in the general population was estimated to be about 3%. The relative proportion of long-term BZD users (all definitions) in adult BZD users ranged from 6% to 76% (mean 24%; 95% CL 13-36%). The estimates were higher in studies only on the elderly (47%; 95% CL 31-64%). Long-term use involved typically steady treatment with low BZD doses. However, in elderly patients long-term BZD use and exceeding recommended doses was relatively common. Several characteristics associated with long-term use were found. CONCLUSIONS: Long-term BZD use is common and a clinical reality. Uniform definitions for "long-term", which is in line with population-based evidence, is needed to have more comparable results between studies. Our systematic review suggests that duration of BZD treatment over six months, the most common definition for long-term BZD use in the included studies. As also recommended previously, it is a useful starting point for further analyses on disadvantages but also potential advantages associated with long-term BZD use.

The histaminergic system in the guinea pig central nervous system: an immunocytochemical mapping study using an antiserum against histamine.

Using an antiserum against conjugated histamine we mapped the histaminergic somata and their fiber projection areas in carbodiimide-fixed guinea pig central nervous system. The neurons were large and they were found exclusively in the posterior hypothalamus, as in the rat, but in the guinea pig they were more numerous and distributed more widely in thin layer around the posterior mammillary nucleus, scattered between and within the medial mammillary nuclei, and in a dense cell cluster emerging from the caudal magnocellular nucleus and extending to the medial preoptic area. The density of histamine-immunopositive fibers was very high in the olfactory tubercle, diagonal band of Broca, nucleus accumbens, medial and cortical amygdaloid nuclei, periventricular and lateral basal hypothalamus, paraventricular thalamus, and in a region from the medial central gray to the locus coeruleus and the parabrachial nucleus. Dense fiber networks were found in the piriform and entorhinal cortex, septum, dentate gyrus, and subiculum, in most parts of amygdala, and in many areas of the hypothalamus, thalamus, substantia nigra, raphe nuclei, inferior olivary, solitary tract and medial vestibular nuclei, and neurohypophysis. Medium fiber density was observed in the internal layers of the olfactory bulb, anterior olfactory nuclei, neocortex, zone CA1 of hippocampus, and many midbrain and hindbrain regions. Low density was present in the outer layers of the olfactory bulb, other parts of hippocampus, the globus pallidus, most of the caudatus-putamen, the cerebellar cortex, and the dorsal horn of the spinal cord. The retina and most of the myelinated white matter had single or no histaminergic fibers. It may be concluded from the results that most fibers seem to follow a ventromedial route to the forebrain, reaching the amygdala ventral to the medial forebrain bundle, the hippocampus via subiculum, and the hindbrain structures via the medial central gray. As compared to the rat, the fiber projections in the guinea pig brain were denser, particularly in the hippocampus, thalamus, pons-medulla, and neurohypophysis. The fiber densities in various regions of the guinea pig brain are compared to histamine receptor densities and the possible functions of histamine are discussed.

Comparative neuroanatomy of the histaminergic system in the brain of the frog Xenopus laevis.

The distribution of the histaminergic neuronal system in the brain of the clawed frog Xenopus laevis was mapped with an antiserum against carbodiimide-fixed histamine and compared to that in mammals. The histamine-immunoreactive cell bodies were located in a small area of the posterolateral hypothalamus, close to the dorsal infundibular nucleus, which contains catecholaminergic and serotonergic neurons. This area may be homologous to the tuberomammillary nucleus in mammals. A thick process extended from each cell between the ependymal cell layer and terminated in the ventricle lumen. The number of histaminergic cell bodies in adult Xenopus brain was relatively low, as compared with the mammalian brain. Preliminary analysis of adjacent sections stained with antisera against GABA or serotonin indicated that the histamine cells were not immunoreactive for these. The pathways and distribution of histaminergic fibers in Xenopus brain showed many similarities to mammals. The densest fiber networks were present in the medial basal forebrain, particularly in the medial amygdala and septum. A distinct cluster of fibers was concentrated around the cell bodies of nucleus accumbens. In most pallial areas, the density was moderate to low. In the primordial piriform cortex and the striatum, very few fibers were seen. In diencephalon, highest fiber densities were found in the anterior and ventral thalamus and posterior and lateral hypothalamus. In hindbrain, the density was highest in the medullary central gray, as in some mammals. The results suggest that the general pattern of the histaminergic system in vertebrate brain is conserved from amphibians to mammals.

Neurotransmitters in the nervous system of Macoma balthica (Bivalvia).

The distribution of histamine-, octopamine-, gamma-aminobutyric acid- (GABA) and taurine-like immunoreactivity in the bivalve mollusc Macoma balthica was studied immunocytochemically with antisera produced in rabbits. Histamine levels in the ganglia and whole animals were also measured by high-performance liquid chromatography using a postcolumn derivatization method. Immunoreactivity for these substances, except for taurine, is found in the central nervous system of this species. The most extensive neuronal system is revealed with the antiserum against histamine. All the main ganglia contain histamine-immunoreactive cell bodies, and a dense network of nerve fibers is seen in the ganglia and nerve roots. Histamine-immunoreactive nerve fibers project to the mantle edge, lips and oesophagus. The basal part of the inhalant siphon is rich in histamine-immunoreactive fibers. Unlike histamine, octopamine- and GABA-like immunoreactivities are restricted to the central nervous system. Taurine-like immunoreactivity is not found in the nervous system of this species. In the nervous system, histamine-immunoreactive cell bodies and fibers are more numerous than those that are octopamine- and GABA-immunoreactive. The distribution of these substances in the ganglia is different. GABA-immunoreactive cells are typically smaller than most of the histamine- and octapamine-immunoreactive cells. Most GABA- and octopamine-immunoreactive cells and fibers are located in the pedal ganglion. Histamine is distributed more evenly in the ganglia and nerve roots. The biochemical measurements of histamine correlate well with the immunohistochemical findings and confirm the predominant location of the amine in the nervous tissue. These results suggest that histamine is more widespread than some other putative transmitters, and support the concept that histamine may have an important role in many physiological processes in molluscs.

Histaminergic system in the tree shrew brain.

This study mapped the histamine-immunoreactive neuronal system in the brain of the tree shrew (Tupaia belangeri) and compared its structure with that of the rat and guinea pig. The histamine-containing cell bodies lay in the posterior ventral hypothalamus in the tuberomammillary complex, as in the rodents. The morphology of this complex resembled that of the rat. The histaminergic axons projected to nearly all parts of the brain. The main ascending bundle ran ventromedially: the densest innervation was found in the ventral hypothalamus, preoptic area, septum, medial part of nucleus accumbens, and bed nucleus of the stria terminalis. High fiber densities were present in the amygdaloid nuclei and claustrum. Another pathway ran dorsomedially along the periventricular hypothalamus and sent fibers to all parts of the diencephalon. Part of these fibers followed the central gray to the midbrain and spread laterally below the inferior colliculus. Another descending pathway ran through the interfascicular and medial raphe nuclei to meet the pontine central gray. The densest fiber networks were seen in the dorsal tegmental and parabrachial nuclei, and around the locus coeruleus. Also the substantia nigra, interpeduncular and mesencephalic reticular nuclei, colliculi, and vestibular and raphe nuclei received a dense histaminergic innervation. The organization of the fibers in the tree shrew brain resembled more that in the guinea pig than that in the rat. As compared with the guinea pig, more fibers were present, particularly in the globus pallidus, central thalamus, and deep cerebellar nuclei. No fibers were seen in the outer layer of the piriform cortex. In Tupaia, a laminar organization of the fibers was evident in the hippocampus, in contrast to the rodents. Also, a dense periventricular fiber plexus was prominent.

Multiple neurotransmitters in the tuberomammillary nucleus: comparison of rat, mouse, and guinea pig.

Tuberomammillary neurons in the posterior hypothalamus are the sole source of neuronal histamine in adult mammalian brain. In the rat, these cells are reported to contain immunoreactivity for gamma-aminobutyric acid (GABA) and several neuropeptides. We compared the presence of these substances in the tuberomammillary cells of the rat, mouse, and guinea pig. In all three species, all histamine-immunoreactive neuronal cell bodies were positive for GABA. This suggests that GABAergic transmission may be important in tuberomammillary function. No cell bodies immunoreactive for thyrotropin releasing hormone (TRH) were found in the guinea pig or mouse tuberomammillary area. In contrast, about 14% of the histamine-immunoreactive tuberomammillary cells in the rat were TRH-positive. These cells were small or medium-sized and were located only in the medial part of the tuberomammillary complex. An antibody against porcine galanin stained about 45% of the tuberomammillary cell bodies in the rat and about 28% in the mouse, but none in the guinea pig. A large proportion of the cells in the rat and mouse, but none in the guinea pig, were positive for met-enkephalin-arg-phe. In contrast, all histamine-containing tuberomammillary cells in the guinea pig, but none in the rat or mouse, were immunoreactive for met-enkephalin. This may indicate a different expression of proenkephalin-derived peptides in the tuberomammillary neurons in these species. Some substance P-immunoreactive cell bodies were located in the tuberomammillary area in all three species. However, only 3% of the histamine-immunoreactive cell bodies in the rat and mouse but none in the guinea pig were substance P-positive. The neurochemical properties of the tuberomammillary nucleus that exhibited species commonality deserve to be studied neurochemically and electrophysiologically in order to determine the functional relevance of coexisting transmitters in this nucleus.

Most classes of dorsal root ganglion neurons are severely depleted but not absent in mice lacking neurotrophin-3.

During development, many neurons in the dorsal root ganglia require neurotrophin-3 for survival. However, it is not known precisely which subpopulations of sensory neurons, other than the proprioceptive afferents, are neurotrophin-3 dependent in vivo. In this study, using a battery of neurochemical markers that label different subpopulations of dorsal root ganglion neurons, we found a widespread, about 60-65% loss of cells in most subpopulations in neurotrophin-3 deficient mice. Intermediate losses were found in the heterozygous mutant mice consistent with a gene dosage effect. In agreement with this, the cell size distribution between the homozygous mutant and wild type mice was virtually identical. The loss of small neurons containing calcitonin gene-related peptide, substance P and thiamine monophosphatase activity suggests that many unmyelinated primary afferents are also lost in the mutant animals. The fact that many different sensory neuron subpopulations are lost to the same extent in neurotrophin-3 deficient mice is consistent with the proposed early role of neurotrophin-3 during neurogenesis. Interestingly, calretinin immunoreactive neurons, which contribute a minor subpopulation, were not affected suggesting that neurotrophin-3 independent regulation of neurogenesis occurs in addition to prominent neurotrophin-3 dependent mechanisms.

Histamine-immunoreactive nerve fibers in the rat brain.

A new immunohistochemical method that utilizes carbodiimide as a tissue fixative was applied to study the distribution of histamine-immunoreactive neuronal fibers and terminals in the rat brain. Immunoreactive fibers were observed in almost all major regions of the brain. They were most numerous in the different hypothalamic nuclei. Dense networks of immunoreactive fibers were also seen in the medial septum, nucleus of the diagonal band and ventral tegmental area. A moderate density of fibers was seen throughout the cerebral cortex, in some parts of the olfactory bulb and tubercle, bed nucleus of the stria terminalis, amygdala, basal parts of the hippocampus, inferior and superior colliculi, substantia nigra, lateral and medial parabrachial nucleus, and the nucleus of the solitary tract. Few histamine-immunoreactive fibers were seen in most parts of the caudate putamen, most thalamic nuclei, most pontine and ventral medullary nuclei. Histamine-immunoreactive neuronal cell bodies were found exclusively in the tuberomammillary nucleus, in agreement with previous reports. The results provide evidence for a widespread distribution of histamine-containing nerve fibers and terminals in the rat brain. Although immunohistochemical localization of histamine does not give direct evidence of a functional role of histamine in any brain area, this distribution suggests involvement in functions of the limbic system including the septal nuclei, hypothalamus and amygdala. The relatively dense histamine-immunoreactive fiber networks in the colliculi and dorsal cochlear nucleus indicate that this amine may play a role in visual functions and hearing. The paucity of immunoreactive fibers in the pontine and medullary areas suggests that the caudal projections originating from the tuberomammillary complex are minor ones compared to the major rostral projections. Several fiber projections originating from the tuberomammillary complex could be deduced from serial frontal, sagittal and horizontal sections. They contained fibers that crossed the midline at several levels of the brain. The results provide information on the target areas of the histaminergic neurons and form a basis for the examination of cellular contracts between the histaminergic neurons and other cells.

A histamine-containing neuronal system in human brain.

A well-organized network of varicose fibers was revealed throughout the frontal and temporal cortex of adult humans with specific antisera against histamine. The densest network of fibers was seen in lamina I, where varicose fibers were seen to run in parallel to the overlying pia mater. Electron microscopic immunohistochemistry revealed histamine-immunostaining in granules in a small number of nerve fibers and varicosities. Hypothalamic samples obtained from autopsy brains of adult humans revealed numerous histamine-immunoreactive nerve cell bodies in the posterior basal hypothalamus in and around the tuberomammillary nucleus. The results suggest that a histaminergic neuronal system reminiscent of that described in rodents is present in human brain.

Stroke induces widespread changes of gene expression for glial cell line-derived neurotrophic factor family receptors in the adult rat brain.

Gene expression for glial cell line-derived neurotrophic factor (GDNF) family ligands and receptors was analyzed with in situ hybridization after two focal ischemic insults of different severities. Focal ischemia was induced in rats by either 30 min or 2 h of middle cerebral artery occlusion (MCAO), causing damage to the striatum only, or involving also the parietal cortex, respectively. We found modest, transient elevation of GDNF mRNA in the dentate granule cell layer. In addition, the number of GDNF mRNA-expressing cells increased in the cortex and striatum after 2 h or 30 min of MCAO, respectively. No changes of neurturin or persephin mRNA expression were detected. Both c-Ret and GFRalpha1 mRNA levels were markedly increased in the ipsilateral cortex outside the ischemic lesion at 6-24 h after the 2-h insult, whereas GFRalpha2 expression was decreased in cortical areas both within and outside the lesion. Similar increases of c-Ret and GFRalpha1 mRNA levels were detected in the striatum, and to a lesser extent, in the cortex following 30 min of MCAO. The 2-h insult also gave rise to transient increases of c-Ret and GFRalpha1 mRNA in hippocampal subregions. Thirty minutes and 2 h of MCAO lead to elevated c-Ret, and GFRalpha1 or GFRalpha2 mRNA expression, respectively, in the ipsilateral ventroposterolateral thalamic nucleus. Both insults induced increased levels of GFRalpha1 mRNA in the subventricular zone of the lateral ventricle. Our data indicate major changes of GDNF family signaling in the forebrain, regulated mainly through altered receptor levels, in the post-ischemic phase. These changes could enhance neuroprotective and neuroregenerative responses both to endogenous and exogenous GDNF ligands.

Histamine neurons in human hypothalamus: anatomy in normal and Alzheimer diseased brains.

The anatomy of histamine-immunoreactive cell bodies in normal adult human brain was examined in detail. In addition, the distribution of these cells in three cases of Alzheimer's disease was compared to the distribution of neurofibrillary tangles. Histamine-immunoreactive cell bodies were confined to the tuberal and posterior hypothalamus, forming the tuberomammillary nuclear complex. Most of the about 64,000 histamine neurons were large and multipolar. They comprised four distinct parts: (i) a major ventral part corresponding to the classical tuberomammillary nucleus, (ii) a medial part including the supramammillary nucleus and part of the posterior hypothalamic area, (iii) a caudal paramammillary part, and (iv) a minor lateral part. The parts showed some similarity with the subgroups in rat. In human, as compared to rat, the histamine neurons occupy a larger proportion of the hypothalamus. Numerous neurofibrillary tangles were found in the Alzheimer hypothalami, concentrated in the tuberomammillary area. Most of them were of globular type and extracellular, and only a minority were histamine immunoreactive. They may represent remnants of degenerated tuberomammillary neurons.

Calbindin-D28k fails to protect hippocampal neurons against ischemia in spite of its cytoplasmic calcium buffering properties: evidence from calbindin-D28k knockout mice.

Cytoplasmic calcium-binding proteins are thought to shield neurons against damage induced by excessive Ca2+ elevations. Yet, in theory, a mobile cellular Ca2+ buffer could just as well promote neuronal injury by facilitating the rapid dispersion of Ca2+ throughout the cytoplasm. In sharp contrast to controls, in mice lacking the gene for calbindin-D28k, synaptic responses of hippocampal CA1 pyramidal neurons which are normally extremely vulnerable to ischemia, recovered significantly faster and more completely after a transient oxygen-glucose deprivation in vitro, and sustained less cellular damage following a 12 min carotid artery occlusion in vivo. Other cellular and synaptic properties such as the altered adaptation of action potential firing, and altered paired-pulse and frequency potentiation at affected synapses in calbindin-D28k-deficient mice were consistent with a missing intraneuronal Ca2+ buffer. Our findings provide direct experimental evidence against a neuroprotective role for calbindin-D28k.

Carbodiimide as a tissue fixative in histamine immunohistochemistry and its application in developmental neurobiology.

The object of this study was to develop an immunohistochemical method that could be used to study neuronal histamine, especially in nerve fibers and terminals where most previous methods have not been applicable. Three new antisera were produced in rabbits against conjugated histamine, and the fixative used in conjugation, 1-ethyl-3(3-diamethylaminopropyl)-carbodiimide (EDCDI), was used in tissue fixation and compared to paraformaldehyde. Specificity of the antisera was established with dot-blot tests on nitrocellulose, with blocking controls and affinity-purified antibodies. EDCDI appeared to be superior to paraformaldehyde as a fixative, and histamine-immunoreactive nerve cells were visualized in developing rat brain during late fetal development from embryonal day 12. By the second postnatal week, the distribution of histamine-immunoreactive neurons in rat brain had reached the adult pattern and immunoreactive nerve fibers were seen in many areas. Posterior hypothalamic neurons from newborn rat in vitro showed strong immunoreactivity for histamine and developed long varicose fibers, which covered the culture dish by the end of the fourth week in vitro. Fixation with EDCDI also allowed detection of histamine in gastric enterochromaffin-like cells and mast cells in rat. The results suggest that the histamine-containing neuron system in rat brain develops during the late fetal and early postnatal periods, and that immunoreactive neurons develop long fibers both in vivo and in vitro.

Histamine-immunoreactive nerve fibers in the mammalian spinal cord.

New sensitive antisera against histamine were used to study the distribution of histamine-immunoreactive nerve fibers in the spinal cord of several mammalian species. Tissues were fixed with carbodiimide by transcardiac perfusion or immersion. A few immunoreactive nerve fibers were found in the cervical spinal cord of the rat in the superficial laminae of the dorsal horn, around the central canal and scattered in the anterior horn. The density of immunoreactive fibers in the cervical spinal cord of the guinea pig and tree shrew was higher, but still low. The densest networks of histamine-immunoreactive fibers were seen in the cervical spinal cord of the pig. The laminar distribution of histamine-immunoreactive fibers was similar in all species. Histamine-immunoreactive fibers were densest in lamina X, followed by laminae I-II. Scattered fibers were also seen in the white matter in the lateral and posterior funiculus in the pig. In the rat and the guinea pig, no histamine-immunoreactive cell bodies were seen in the spinal sensory ganglia. The results suggest that the histamine-immunoreactive nerve fibers in the spinal cord may originate from the brain, probably from the posterior hypothalamus, and the fiber projection is more extensive in higher mammalian species. The role of histamine in the spinal cord is not known, but it may be involved in, e.g., pain sensation.

LRRTM1-deficient mice show a rare phenotype of avoiding small enclosures--a tentative mouse model for claustrophobia-like behaviour.

The LRRTM family proteins have been shown to act as synaptogenic cell adhesion molecules via interaction with presynaptic neurexins and are associated with neuropsychiatric disorders. LRRTM1-knockout mice have subtle morphological deficits in excitatory hippocampal synapses and were suggested to have impaired cognitive function. Here we report that LRRTM1-knockout mice exhibit an extraordinary phenotype of avoiding small enclosures. In the light-dark box, the knockout mice escape to dark through a standard opening as quickly as wild-type littermates but avoid escaping through a small doorway. While all wild-type mice spontaneously enter a small tube, most knockout mice do not. This apparent aversion to enter narrow space may explain other abnormalities such as increased time in open arms in the elevated plus maze and less visits through a tunnel in the IntelliCage. Moreover, LRRTM1-knockout mice show increased social interaction, reduced nest building and MK801-induced locomotion, and slower swim speed but normal water maze learning. Since LRRTM1 is predominantly expressed in thalamus, hippocampus and limbic cortex, specific synaptic defects in those areas presumably cause these behavioural abnormalities.

Determinants of mental health stigma among pharmacy students in Australia, Belgium, Estonia, Finland, India and Latvia.

BACKGROUND: Healthcare professionals commonly exhibit negative attitudes toward people with mental disorders. Few international studies have sought to investigate the determinants of stigma. OBJECTIVE: To conduct an international comparison of pharmacy students' stigma towards people with schizophrenia, and to determine whether stigma is consistently associated with stereotypical attributes of people with schizophrenia. METHOD: Students (n = 649) at eight universities in Australia, Belgium, India, Finland, Estonia and Latvia completed a seven-item Social Distance Scale (SDS) and six items related to stereotypical attributes of people with schizophrenia. RESULTS: Mean SDS scores were 19.65 (+/- 3.97) in Australia, 19.61 (+/- 2.92) in Belgium, 18.75 (+/- 3.57) in India, 18.05 (+/- 3.12) in Finland, and 20.90 (+/- 4.04) in Estonia and Latvia. Unpredictability was most strongly associated with having a high social distance in Australia (beta = -1.285), the perception that people will never recover in India (beta = - 0.881), dangerousness in Finland (beta = -1.473) and the perception of being difficult to talk to in Estonia and Latvia (beta = -2.076). Unpredictability was associated with lower social distance in Belgium (beta = 0.839). CONCLUSION: The extent to which students held stigmatizing attitudes was similar in each country, however, the determinants of stigma were different. Pharmacy education may need to be tailored to address the determinants of stigma in each country.