N-cadherin localization in taste buds of mouse circumvallate papillae.

Taste buds, the receptor organs for taste, contain 50-100 taste bud cells. Although these cells undergo continuous turnover, the structural and functional integrity of taste buds is maintained. The molecular mechanisms by which synaptic connectivity between taste buds and afferent fibers is formed and maintained remain ambiguous. In the present study, we examined the localization of N-cadherin in the taste buds of the mouse circumvallate papillae because N-cadherin, one of the classical cadherins, is important for the formation and maintenance of synapses. At the light microscopic level, N-cadherin was predominantly detected in type II cells and nerve fibers in the connective tissues in and around the vallate papillae. At the ultrastructural level, N-cadherin immunoreactivity appears along the cell membrane and in the intracellular vesicles of type II cells. N-cadherin immunoreactivity also is evident in the membranes of afferent terminals at the contact sites to N-cadherin-positive type II cells. At channel type synapses between type II cells and nerve fibers, N-cadherin is present surrounding, but not within, the presumed neurotransmitter release zone, identified by large mitochondria apposed to the taste cells. The present results suggest that N-cadherin is important for the formation or maintenance of type II cell afferent synapses in taste buds.

ß-N-methylamino-L-alanine (BMAA) suppresses cell cycle progression of non-neuronal cells.

ß-N-methylamino-L-alanine (BMAA), a natural non-proteinaceous amino acid, is a neurotoxin produced by a wide range of cyanobacteria living in various environments. BMAA is a candidate environmental risk factor for neurodegenerative diseases such as amyotrophic lateral sclerosis and Parkinson-dementia complex. Although BMAA is known to exhibit weak neuronal excitotoxicity via glutamate receptors, the underlying mechanism of toxicity has yet to be fully elucidated. To examine the glutamate receptor-independent toxicity of BMAA, we investigated the effects of BMAA in non-neuronal cell lines. BMAA potently suppressed the cell cycle progression of NIH3T3 cells at the G1/S checkpoint without inducing plasma membrane damage, apoptosis, or overproduction of reactive oxygen species, which were previously reported for neurons and neuroblastoma cells treated with BMAA. We found no evidence that activation of glutamate receptors was involved in the suppression of the G1/S transition by BMAA. Our results indicate that BMAA affects cellular functions, such as the division of non-neuronal cells, through glutamate receptor-independent mechanisms.

Thrombospondin Type-1 Repeat Domain-Containing Proteins Are Strongly Expressed in the Head Region of Hydra.

The head region of Hydra, the hypostome, is a key body part for developmental control and the nervous system. We herein examined genes specifically expressed in the head region of Hydra oligactis using suppression subtractive hybridization (SSH) cloning. A total of 1414 subtracted clones were sequenced and found to be derived from at least 540 different genes by BLASTN analyses. Approximately 25% of the subtracted clones had sequences encoding thrombospondin type-1 repeat (TSR) domains, and were derived from 17 genes. We identified 11 TSR domain-containing genes among the top 36 genes that were the most frequently detected in our SSH library. Whole-mount in situ hybridization analyses confirmed that at least 13 out of 17 TSR domain-containing genes were expressed in the hypostome of Hydra oligactis. The prominent expression of TSR domain-containing genes suggests that these genes play significant roles in the hypostome of Hydra oligactis.

The nerve ring in cnidarians: its presence and structure in hydrozoan medusae.

In our previous studies of the Hydra nerve ring, we proposed the following hypothesis: "The nerve ring in the hypostome of Hydra is a central nervous system (CNS)-like neuronal structure." Related to this hypothesis, we have started to survey the nerve ring immunocytochemically using antibodies against neuropeptides throughout the whole phylum of cnidarians. In the present study, we describe nerve rings in hydrozoan medusae. We examined the medusae of five hydrozoan species belonging to three orders: Eirene sp. (order Leptomedusae), Craspedacusta sowerbyi (order Limnomedusae), Sarsia tubulosa, Turritopsis nutricula, and Cladonema radiatum (order Anthomedusae). We observed a well-developed nerve ring in all species. The nerve ring runs circumferentially around the margin of the bell. In all cases, the nerve ring was visualized by plural antibodies, suggesting that it contains different neural subpopulations. In C. radiatum, antibodies against four different neuropeptides labeled the nerve ring. We established clear (without undesirable cross-reactions) double-staining procedures with two rabbit primary antibodies. Using the double-staining method, three neural subsets visualized by three antibodies revealed completely separate neural populations. The results show that the nerve ring is a common feature in hydrozoan medusae and has a complex heterogeneous structure composed of different neural subsets.

Thiamine deficiency induces massive cell death in the olfactory bulbs of mice.

Thiamine (vitamin B1) deficiency (TD) leads to focal brain necrosis in particular brain regions in humans and in experimental animal models. The precise mechanism of the selective topographic vulnerability triggered by TD still remains unclear. We examined the distribution pattern of cell death in the brains of mice in an experimental model of TD using anti-single-strand DNA immunohistochemistry and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling methods. We found that interneurons in the olfactory bulb were sensitive to TD. The morphologic aspects of cell death in the olfactory bulb resembled those of cell death in thalamic neurons, which have previously been examined in detail. Furthermore, cell death in the olfactory bulb was partly relieved by the administration of an N-methyl-d-aspartate receptor antagonist, as was the case in thalamic lesions by TD. The superficial part of the olfactory granule cell layer seemed to be the most sensitive to TD, suggesting that differences in the afferents between superficial and deep granule cells may influence the sensitivity of these cells to TD. Our results indicate that the olfactory bulb should be considered as one of the vulnerable regions to TD.

Immunohistochemical evidence for the existence of novel mammalian neuropeptides related to the Hydra GLW-amide neuropeptide family.

The GLW-amide family is a neuropeptide family found in cnidarian species and is characterized by the C-terminal amino acid sequence -Gly-Leu-Trp-NH(2). To detect mammalian peptides structurally related to the GLW-amide family, we examined rat brain by immunohistochemistry with an anti-GLW-amide antibody. GLW-amide-like immunoreactivity (GLW-amide-LI) was observed in thin varicose fibers in some regions of the brain. Most neurons showing GLW-amide-LI were observed in the laterodorsal tegmental nucleus, pedunculopontine tegmental nucleus, and trigeminal/spinal ganglia. These results strongly suggest that the rat nervous system contains as yet unidentified GLW-amide-like peptides, and that GLW-amide-LI in the brain is a good marker for ascending projections from mesopontine cholinergic neurons.

Exposure to hexanal odor induces extraordinary Fos expression in the medial preoptic area and amygdala of Fyn tyrosine kinase-deficient mice.

Our previous study revealed that the odor of hexanal, which is derived from the wood chips used as bedding, influenced mouse maternal behavior and induced the neonatal death of Fyn-deficient (fyn(-/-)) pups born of fyn(-/-) parents. To clarify what regions of the brain are involved in this effect, we examined which brain regions of fyn(-/-) and Fyn-heterozygous (fyn(+/-)) females were activated by hexanal odor using Fos immunohistochemistry. Hexanal induced Fos immunoreactivity in the primary olfactory cortex and in the medial and central amygdala of both fyn(+/-) and fyn(-/-) mice. In fyn(-/-) females, hexanal also induced the activation of the medial preoptic area and the basolateral and posteriomedial cortical amygdala, which are known to be involved in the control of maternal and emotional behavior.

Exposure to hexanal odor influences maternal behavior and induces neonatal death in Fyn tyrosine kinase-deficient mice.

Fyn-deficient pups born of Fyn-deficient parents die because they fail to suckle within 1-2 days after birth. Here we demonstrate that the neonatal death phenotype was influenced by the genetic background and an environmental odor. The odor of hexanal (C6-aldehyde) partially impaired mouse maternal behavior and induced the neonatal death of Fyn-deficient pups born of Fyn-deficient parents. This death phenotype was first observed in the breeding environment using autoclaved chips of Douglas fir. An analysis of the volatile chemicals in the autoclaved chips revealed an approximately 10-fold greater amount of hexanal than in non-autoclaved chips. Hexanal influenced the length of time virgin female mice engage in the maternal crouching behavior. In addition, Fyn-deficient females exhibited defects in the maternal behavior of nest building and pup retrieval, regardless of exposure to hexanal. These observations provide new insights into the regulation of maternal behavior by environmental and genetic factors.