Gamma‐protocadherins are enriched and transported in specialized vesicles associated with the secretory pathway in neurons

Gamma protocadherins (Pcdh‐γs) resemble classical cadherins and have the potential to engage in cell–cell interactions with homophilic properties. Emerging evidence suggests non‐conventional roles for some protocadherins in neural development. We sought to determine whether Pcdh‐γ trafficking in neurons is consistent with an intracellular role for these molecules. Here we show that, in contrast to the largely surface localization of classical cadherins, endogenous Pcdh‐γs are primarily intracellular in rat neurons in vivo and are equally distributed within organelles of subsynaptic dendritic and axonal compartments. A strikingly higher proportion of Pcdh‐γ‐containing organelles in synaptic compartments was observed at postnatal day 16. To determine the origin of Pcdh‐γ‐trafficking organelles, we isolated organelles with Pcdh‐γ antibody‐coupled magnetic beads from brain organelle suspensions. Vesicles with high levels of COPII and endoplasmic reticulum–Golgi intermediate compartment (ERGIC) components were isolated with the Pcdh‐γ antibody but not with the classical cadherin antibody. In cultured hippocampal neurons, Pcdh‐γ immunolabeling partially overlapped with calnexin‐ and COPII‐positive puncta in dendrites. Mobile Pcdh‐γ‐GFP profiles dynamically codistributed with a DsRed construct coupled to ER retention signals by live imaging. Pcdh‐γ expression correlated with accumulations of tubulovesicular and ER‐like organelles in dendrites. Our results are consistent with the possibility that Pcdh‐γs could have a unique function within the secretory pathway in addition to their documented surface roles.     

Association of white matter lesions and brain atrophy with the development of dementia in a community: the Hisayama Study

Aim

To investigate the association of white matter lesions volume (WMLV) levels with dementia risk and the association between dementia risk and the combined measures of WMLV and either total brain atrophy or dementia-related gray matter atrophy in a general older population.

Methods

One thousand one hundred fifty-eight Japanese dementia-free community-residents aged ≥65 years who underwent brain magnetic resonance imaging were followed for 5.0 years. WMLV were segmented using the Lesion Segmentation Toolbox. Total brain volume (TBV) and regional gray matter volume were estimated by voxel-based morphometry. The WMLV-to-intracranial brain volume ratio (WMLV/ICV) was calculated, and its association with dementia risk was estimated using Cox proportional hazard models. Total brain atrophy, defined as the TBV-to-ICV ratio (TBV/ICV), and dementia-related regional brain atrophy defined based on our previous report were calculated. The association between dementia risk and the combined measures of WMLV/ICV and either total brain atrophy or the number of atrophied regions was also tested.

Results

During the follow-up, 113 participants developed dementia. The risks of dementia increased significantly with higher WMLV/ICV levels. In addition, dementia risk increased additively both in participants with higher WMLV/ICV levels and lower TBV/ICV levels and in those with higher WMLV/ICV levels and a higher number of dementia-related brain regional atrophy.

Conclusion

The risk of dementia increased significantly with higher WMLV/ICV levels. An additive increment in dementia risk was observed with higher WMLV/ICV levels and lower TBV/ICV levels or a higher number of dementia-related brain regional atrophy, suggesting the importance of prevention or control of cardiovascular risk factors.     

Distribution of external cuneate nucleus afferents to the cerebellum: II. Topographical distribution and zonal pattern--an experimental study with radioactive tracers in the cat

Small injections of tritiated leucine and the autoradiographic method were used to demonstrate efferents from restricted portions of the external cuneate nucleus (NCE) to the cerebellum. Sites of injection were analyzed by reference to the distribution of primary muscle afferents in NCE. On transverse sections, the silver deposits form longitudinal bands that, in certain regions, are packed together and label the entire surface of the granular layer; in other parts, they are separated by empty longitudinal bands. The longitudinal deposits are not continuous in the rostrocaudal direction. On the basis of the distribution of the longitudinal bands, 14 zones have been described for lobules II-VI, and 6 zones were recognized in lobules I, VIII, and the paramedian lobule. Afferents from NCE are distributed topographically. Regions of the nucleus receiving axial and neck muscles project mainly to vermal regions of lobules I-III, and to parts of lobules VIII and IX. Regions receiving afferents from forelimb muscles send their fibers preferentially to the vermian region of lobule V, to paravermian regions of lobules IV-VI, to parts of lobules VIII and IX, and to the paramedian lobule. These distributions in several respects are in agreement with the somatotopical maps of the cerebellum. However, other features support a "mosaic" arrangement: efferents from a region of NCE are distributed over several distinct sites of the cortex and efferents from different parts of the nucleus also converge to neighboring cortical regions.     

Calcium-binding protein (calbindin-D28k) and parvalbumin immunocytochemistry: localization in the rat hippocampus with specific reference to the selective vulnerability of hippocampal neurons to seizure activity

Two neuronal calcium-binding proteins, calbindin-D28k (CaBP) and parvalbumin (PV), were localized in the normal rat hippocampus by using immunocytochemical methods to determine 1) their location and 2) whether a correlation exists between the presence of these two calcium-binding proteins and the selective vulnerability of different hippocampal neuronal populations to experimental seizure activity. CaBP-like immunoreactivity (CaBP-LI) is present in all dentate granule cells and some, but not all, CA1 and CA2 pyramidal cells. Some CA1 pyramidal cells lack CaBP-LI, and those that do are lightly stained compared to the dentate granule cells. CA3 pyramidal cells appear to contain neither CaBP- nor PV-LI, and no granule or pyramidal cells exhibit PV-LI. CaBP-LI is present in distinct populations of dentate and hippocampal interneurons but absent from others. In area dentata, CaBP-LI is present in a small number of interneurons of the molecular and granule cell layers and in a small population of presumed basket cells in or below the granule cell layer. Conversely, more presumed dentate basket cells exhibit PV-LI than CaBP-LI. In the hilus of area dentata, few cells are CaBP- or PV-immunoreactive. The hilar somatostatin/neuropeptide Y (NPY)-immunoreactive cells and hilar mossy cells, two distinct and large populations, lack CaBP- and PV-LI. In the CA3 region, CaBP-LI is present in a relatively small number of interneurons in each stratum. PV-immunoreactive interneurons in area CA3 are more numerous. In area CA1, CaBP-LI is present in many interneurons in strata radiatum and lacunosum-moleculare. Some, but relatively fewer, CaBP-positive interneurons are present in strata pyramidale and oriens. Conversely, PV-immunoreactive interneurons are numerous in strata pyramidale and oriens but rare in strata radiatum and lacunosum-moleculare. Staining with the particulate chromagen benzidine hydrochloride revealed a previously undescribed dense band of CaBP-LI in the inner dentate molecular layer, a lamina enriched with kainate-displaceable glutamate-binding sites and innervated by the apparently excitatory ipsilateral associational/commissural (IAC) pathway that originates in the CaBP-negative hilar mossy cells. Bilateral electrical stimulation of the perforant path was performed in order to destroy the hilar mossy cells and to determine if this band of CaBP-LI is normally present within the mossy cell terminals. Perforant path stimulation that destroyed hilar mossy cells throughout the dorsal portions of both hippocampi did not abolish the dense CaBP-like immunoreactivity in the inner molecular layer.     

Temporal and spatial distribution of Fos protein in the lumbar spinal dorsal horn neurons in the rat with chronic constriction injury to the sciatic nerve

The temporal and spatial expression pattern of Fos protein in spinal dorsal horn neurons was examined by immunohistochemistry in rats with chronic constriction injury (CCI) to the sciatic nerve. In normal animals, a few Fos-immunoreactive (-IR) neurons were detected in the dorsal horn of the lumbar spinal cord. Following induction of CCI, a very large number of Fos-IR neurons appeared in the spinal dorsal horn, but a significant number of Fos-IR neurons were also observed in the contralateral dorsal horn where primary afferents of the injured sciatic nerve rarely project. Sham-operated animals also had a significant number of Fos-IR neurons in the dorsal horn bilaterally. The number of Fos-IR neurons reached its maximal level 1 day following placement of the ligatures (PO 1d). The ratio of the number of Fos-IR neurons in the ipsilateral dorsal horn to the contralateral dorsal horn, however, had its peak level 3 days following CCI (3.1-fold increase compared to the contralateral dorsal horn). The number of Fos-IR neurons in the dorsal horn gradually decreased, but increased again around PO 15d. On PO 30d, the number of Fos-IR neurons decreased and became comparable to that in normal animals. The present results indicate that the induction of Fos-IR neurons in the dorsal horn caused by CCI is biphasic and reaches its maximal level on PO 3d, near the time of hyperalgesia onset.