Brain-derived neurotrophic factor (BDNF) plays important roles in the development, maintenance, and plasticity of the mammalian forebrain. These functions include regulation of neuronal maturation and survival, axonal and dendritic arborization, synaptic efficacy, and modulation of complex behaviors including depression and spatial learning. Although analysis of mutant mice has helped establish essential developmental functions for BDNF, its requirement in the adult is less well documented. We have studied late-onset forebrain-specific BDNF knockout (CaMK-BDNF(KO)) mice, in which BDNF is lost primarily from the cortex and hippocampus in early adulthood, well after BDNF expression has begun in these structures. We found that although CaMK-BDNF(KO) mice grew at a normal rate and can survive more than a year, they had smaller brains than wild-type siblings. The CaMK-BDNF(KO) mice had generally normal behavior in tests for ataxia and anxiety, but displayed reduced spatial learning ability in the Morris water task and increased depression in the Porsolt swim test. These behavioral deficits were very similar to those we previously described in an early-onset forebrain-specific BDNF knockout. To identify an anatomical correlate of the abnormal behavior, we quantified dendritic spines in cortical neurons. The spine density of CaMK-BDNF(KO) mice was normal at P35, but by P84, there was a 30% reduction in spine density. The strong similarities we find between early- and late-onset BDNF knockouts suggest that BDNF signaling is required continuously in the CNS for the maintenance of some forebrain circuitry also affected by developmental BDNF depletion.

Brain-derived neurotrophic factor (BDNF), a neurotrophin, enhances the survival and differentiation of several classes of neurons in vitro. To determine its essential functions, we have mutated the BDNF gene. Most homozygote mutants die within 2 days after birth, but a fraction live for 2-4 weeks. These develop symptoms of nervous system dysfunction, including ataxia. The BDNF mutant homozygotes have substantially reduced numbers of cranial and spinal sensory neurons. Although their central nervous systems show no gross structural abnormalities, expression of neuropeptide Y and calcium-binding proteins is altered in many neurons, suggesting they do not function normally. In contrast with mice lacking the BDNF receptor TrkB, motor neurons appear normal in the BDNF mutant.

We evaluated the impact of early embryonic deletion of huntingtin (htt) from pyramidal neurons on cortical development, cortical neuron survival and motor behavior, using a cre-loxP strategy to inactivate the mouse htt gene (Hdh) in emx1-expressing cell lineages. Western blot confirmed substantial htt reduction in cerebral cortex of these Emx-httKO mice, with residual cortical htt in all likelihood restricted to cortical interneurons of the subpallial lineage and/or vascular endothelial cells. Despite the loss of htt early in development, cortical lamination was normal, as revealed by layer-specific markers. Cortical volume and neuron abundance were, however, significantly less than normal, and cortical neurons showed reduced brain-derived neurotrophic factor (BDNF) expression and reduced activation of BDNF signaling pathways. Nonetheless, cortical volume and neuron abundance did not show progressive age-related decline in Emx-httKO mice out to 24months. Although striatal neurochemistry was normal, reductions in striatal volume and neuron abundance were seen in Emx-httKO mice, which were again not progressive. Weight maintenance was normal in Emx-httKO mice, but a slight rotarod deficit and persistent hyperactivity were observed throughout the lifespan. Our results show that embryonic deletion of htt from developing pallium does not substantially alter migration of cortical neurons to their correct laminar destinations, but does yield reduced cortical and striatal size and neuron numbers. The Emx-httKO mice were persistently hyperactive, possibly due to defects in corticostriatal development. Importantly, deletion of htt from cortical pyramidal neurons did not yield age-related progressive cortical or striatal pathology.

To obtain insights into the expression of neurotrophin-3 (NT-3) in the mouse, we have utilized mice in which the Escherichia coli lacZ gene is integrated into the neurotrophin-3 locus (NT-3(lacZneo), Fariñas et al. [1994] Nature 369:658-661). In this mouse strain, beta-galactosidase production is under control of the NT-3 promoter in its normal chromosomal environment, and histochemical measurement of beta-galactosidase provides a simple, sensitive method to determine which cells express NT-3. Our data correlate well with previous in situ mRNA and immunocytochemical studies reporting the localization of NT-3. For example, in adult NT-3(lacZneo)/+ mice, beta-galactosidase is expressed in high amounts in limbic areas of the cortex (cingulate, retrosplenial, piriform, and entorhinal), in the visual cortex, in the hippocampal formation (dentate granule cells, CA2 cells, fasciola cinereum, induseum griseum, tenia tecta, presubiculum, and parasubiculum), and in the septum (septohippocampal nucleus and lateral dorsal septum). In contrast with other studies, our results suggest more extensive expression of NT-3 in adult and aged mouse brains with cortical expression apparent at 4.5 months. To further define the cell populations expressing NT-3 in adult mice, we have combined immunocytochemistry with histochemical staining and found that beta-galactosidase is coexpressed with a neuronal marker (NeuN) and with parvalbumin and neuropeptides, markers for GABAergic interneurons. Our studies of embryonic beta-galactosidase expression in NT-3(lacZneo)/+ mice suggest sites of NT-3 expression not previously described, including embryonic piriform cortical cells and dentate granule cell precursors.