Biomedical subjects

T Terashima

Publications and source records attributed to T Terashima.

At least 55 records · Page 3Linked to original sources

PubMed1999

Cigarette smoking causes sequestration of polymorphonuclear leukocytes released from the bone marrow in lung microvessels.

Studies from our laboratory have shown that chronic cigarette smoke exposure causes a neutrophilia associated with a shortening of the mean transit time of polymorphonuclear leukocytes (PMN) though the postmitotic pool of the marrow. The present study was designed to test the hypothesis that PMN newly released from bone marrow by smoke exposure preferentially sequestered in pulmonary microvessels. The thymidine analogue 5'-bromo-2'-deoxyuridine (BrdU) was used to label dividing PMN in the marrow of rabbits; their appearance in the circulation was measured using immunocytochemistry, and their sequestration in lung tissue was determined using standard morphometric techniques. Animals exposed to 11 d of cigarette smoke (n = 6) compared with sham-exposed control animals (n = 4) showed no increase in circulating PMN counts but showed an increase in both the percentage of band cells (smoking, 9.8 +/- 1.1% versus control, 5.5 +/- 0.9%; P < 0.05) and BrdU-labeled PMN (PMNBrdU) in the circulation (smoking, 10.8 +/- 0.6% versus control, 7.5 +/- 0.3%; P < 0.05). There were more PMN sequestered in the lungs of smoke-exposed animals (51.7 +/- 3.4 x 10(7)/ml tissue) than in those of control animals (25.1 +/- 1.8 x 10(7)/ ml tissue) (P < 0.05) and a higher percentage of these cells were PMNBrdU (smoking, 16.9 +/- 2. 3% versus control, 9.6 +/- 0.4%; P < 0.05). The percentage of PMNBrdU in the gravity-independent regions (11.7 +/- 1.9%) of the lung was higher than gravity-dependent regions (7.8 +/- 1.8%) in the smoke-exposure group (P < 0.05). Transmission electron microscopy showed pulmonary capillary endothelial damage with adherent PMN in the smoke-exposure group. We conclude that younger PMN released from the bone marrow by cigarette smoking preferentially sequestered in pulmonary microvessels and speculate that these PMN may contribute to the alveolar wall damage associated with smoke-induced lung emphysema.

Animals↗

PubMed1999

Time-dependent changes of amino acids in the serum, liver, brain and urine of rats administered with theanine.

Time-dependent changes of theanine (gamma-glutamylethylamide) and other amino acids in various tissues of rats were investigated during the 24 hrs after theanine administration. When theanine (4 g/kg of body weight) was intragastrically administered to rats, the concentrations of theanine in the serum, liver and brain were significantly increased 1 hr after its administration, and thereafter gradually decreased, but reached the maximum level in the brain after 5 hrs. Theanine in these tissues had completely disappeared 24 hrs after its administration. In contrast, the administration of theanine resulted in the concentrations of theanine, urea, ethylamine and glutamic acid in the urine being significantly enhanced. These results suggest that theanine might be degraded via glutamic acid.

Amino Acids↗

PubMed1999

[Cytoarchitectonic abnormality in the facial nucleus of the reeler mouse].

Reeler, an autosomal recessive mutant mouse, is characterized by ataxic gait and tremor. In this mutant, the cerebral and cerebellar cortices and hippocampus are cytoarchitectually disorganized: neuronal components are ectopically located in these laminated structures. Since reelin, the gene responsible for the reeler mutation, was discovered by D'Arcangelo et al. (Nature 374: 719-723, 1995), remarkable progress has occurred in this field. The reelin gene encodes an extracellular protein, Reelin, that is crucial for neuronal migration. During embryogenesis, reelin is expressed in the Cajal-Retzius cells in the cerebral cortex and in the outer granule cells in the cerebellar cortex. Although non-laminated structures such as facial nucleus, inferior olivary complex, and dorsal cochlear nucleus are also cytoarchitectually deranged in this mutant, only a few studies have been done to clarify the detailed abnormalities in these non-laminated structures. In this review, we focused on the cytoarchitectonic abnormality in the facial nucleus of the reeler mouse. The branchiomotor neurons in the facial nucleus are generated from the ventricular zone of the floor of the fourth ventricle, migrate ventrolaterally, and finally settle near the ventral surface of the hindbrain. Time schedules for the generation, axon formation and migration of facial motoneurons are similar both in the normal and reeler mice, but the reeler phenotype becomes identifiable at the end of neuronal migration. Although the reason why the facial nucleus is cytoarchitectually abnormal in the reeler mouse is still unknown, the long migration of the facial motoneurons seems to be susceptible to the absence of Reelin in the reeler mouse. In spite of the cytoarchitectual abnormality, retrograde horseradish peroxidase (HRP) study confirmed that the musculotopic arrangements within the facial nucleus of the reeler mouse are still preserved, suggesting that neuronal migration and target recognition are regulated independently. More recently, other reeler-like mutants have been reported. Among them, yotari and scrambler mice arise from mutations in mdab1, a mouse gene related to Drosophila gene disabled (dab). More than 10 years ago, an autosomal recessive rat mutant, shaking rat Kawasaki (SRK), was described that exhibits a phenotype identical to reeler, but the gene responsible for this rat mutation remains unknown. Interestingly, the facial nucleus is cytoarchitectually more deranged in yotari and SRK than their reeler counterpart. Although the reason why yotari exhibits a phenotype identical to reeler in the laminated structures but not in non-laminated structures such as the facial nucleus has remained obscure, mDab1 and Reelin proteins may function as signaling molecules in a different way between laminated and non-laminated structures. Phenotypes resembling that of reeler are seen with mutations in mdab1, cdk5 and p35. Cdk5 and p35 are respectively the catalytic and regulatory subunits of a serine/threonine kinase, that could potentially operate in a common signalling pathway with mDab and Reelin. These plausible partners for Reelin and mDab1 should help us to understand how the activities of these proteins coordinate neuronal migration and rearrangement.

Animals↗

PubMed1999

Distribution and intracellular localization of a mouse homologue of Ca2+/calmodulin-dependent protein kinase Ibeta2 in the nervous system.

Ca2+/calmodulin-dependent protein kinases (CaMKs) are believed to play important roles in the development and function of the nervous system. We report here the identification and expression of mouse CaMKIbeta (mCaMKIbeta), in particular mCaMKIbeta2, an isoform of mCaMKIbeta. During embryogenesis, the mCaMKIbeta2 gene is expressed mainly in the nervous system, including brain, spinal cord, trigeminal ganglion, and retina. Within the CNS, the expression of mCaMKIbeta2 is detected in the mantle zone, but not in the ventricular zone, suggesting its possible involvement in the differentiation of neurons. In the adult brain, mCaMKIbeta2 transcripts are detected at high levels in the anterior olfactory nuclei, piriform cortex, septal nuclei, bed nuclei of the stria terminalis, hippocampal pyramidal cells, dentate granule cells, amygdala, hypothalamic nuclei, parabrachial nucleus, and nucleus of the solitary tract. The distinct gene expression pattern suggests that mCaMKIbeta2 may also be involved in different mature neuronal functions from other CaMKs. In addition, mCaMKI/beta2 proteins are localized to the cytoplasm and nuclei, but not to nucleoli, suggesting that mCaMKIbeta2 proteins might be involved in the cytoplasmic and nuclear signal transduction of the nervous system.

Amino Acid Sequence↗

PubMed1998

Glucocorticoid-induced granulocytosis: contribution of marrow release and demargination of intravascular granulocytes.

BACKGROUND: Glucocorticoid-induced granulocytosis has been attributed to enhanced release of polymorphonuclear leukocytes (PMNs) from bone marrow, delayed apoptosis, and reduced egress of PMNs into tissues. This study was designed to determine the relative contributions of PMNs released from the bone marrow and those entering the circulation from the marginated pool to the granulocytosis produced by a single dose of dexamethasone (2.0 mg/kg) in rabbits. METHODS AND RESULTS: PMN transit through the mitotic and postmitotic pools of the bone marrow and rate of release of PMNs into the circulation were measured by use of the thymidine analogue 5'-bromo-2'-deoxyuridine (BrdU) to pulse-label PMNs in the bone marrow. The shift of PMNs from the marginated to the circulating pool was measured with BrdU-labeled PMNs transferred from donor rabbits to recipients before dexamethasone was delivered. The data show that dexamethasone increased bone marrow release of PMNs and shortened their transit time through the postmitotic pool (P<0.001) but not the mitotic pool of the bone marrow (P>0.05). Dexamethasone slowed the clearance of BrdU-labeled PMNs from the circulation (P<0.05) and lengthened their disappearance (half-life) from the circulation compared with control (half-life, 4.95 versus 9. 45 hours). At 6 hours after dexamethasone, bone marrow release contributed approximately 10%, mobilization from the marginated pool approximately 61%, and a lengthened half-life in the circulation approximately 29% to the glucocorticoid-induced granulocytosis. CONCLUSIONS: We conclude that a single dose of dexamethasone causes a granulocytosis primarily by a shift of PMNs from the marginated to the circulating pool, with a minor contribution from marrow release.

Animals↗

PubMed1998

Ambiguus nucleus neurons innervating the abdominal esophagus are malpositioned in the reeler mouse.

To examine whether the migration of ambiguus nucleus (NA) neurons is affected in the reeler mouse, recombinant replication-deficient adenoviral vector carrying E. coli-galactosidase gene (lacZ) was injected into the abdominal esophagus of the reeler mouse and normal control at two months of age prior to 5 days of sacrifice of the animals. In the normal control, lacZ-positive neurons were found in the compact formation of the NA, whereas, in the reeler, they were scattered from the base of the fourth ventricle to the ventro-lateral margin of the medulla. The present study confirmed that NA neurons are malpositioned in the reeler mouse, suggesting that the migration of NA neurons is guided by the reelin-related protein (Reelin).

Abdomen↗

PubMed1998

Release of polymorphonuclear leukocytes from the bone marrow by interleukin-8.

Several studies have shown that interleukin-8 (IL-8) causes a rapid granulocytosis with the release of polymorphonuclear leukocytes (PMN) from the bone marrow (BM) partially responsible for the granulocytosis. This study was designed to quantitate the release of PMN from the BM by IL-8 and measure the transit time of PMN through the marrow after IL-8 administration. The thymidine analogue, 5'-bromo-2'-deoxyuridine (BrdU), was used to label dividing PMN in the marrow and follow their release into the circulation after intravenous IL-8. This allowed us to calculate the transit time of PMN through the mitotic and postmitotic pools of BM. BrdU was infused intravenously into rabbits 24 hours before IL-8 (2.5 microg/kg). IL-8 caused a rapid, transient granulocytopenia (5.9 +/- 0.4 at baseline v 0.2 +/- 0.06 x 10/9L at 5 minutes, P < .05) followed by granulocytosis (8.4 +/- 0.1 at 30 minutes, P < .05) associated with an increased number (0.3 +/- 0.1 at baseline v 1.2 +/- 0.6 x 10(9)/L at 30 minutes, P < .05) and percentage of band cells (P < .05), as well as a rapid increase in the number of BrdU-labeled PMN (PMNBrdU) in the circulation (0.09 +/- 0.05 at baseline to 1.5 +/- 0.6 x 10(9)/L at 60 minutes, P < .05). The transit time of PMN through both the mitotic and postmitotic pools of BM was not affected by IL-8. To determine the marrow compartment from which the PMN were mobilized by IL-8, we quantitated PMN movement from the hematopoietic and sinusoidal compartments into the circulation. The fraction of PMNBrdU in both compartments was higher than in the circulating blood (P < .05) and the fraction and number of PMNBrdU in the sinusoids decreased with IL-8 treatment (P < .05). We conclude that the pool of PMN residing in the BM venous sinusoids are rapidly released into the circulation after administration of IL-8.

Animals↗

PubMed1998

Regional differences between the immunohistochemical distribution of Ca2+/calmodulin-dependent protein kinase II alpha and beta isoforms in the brainstem of the rat.

The distribution of Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) alpha and beta isoforms in the brainstem of adult rats was investigated using an immunohistochemical method with two monoclonal antibodies which specifically recognize the alpha and beta isoform, respectively. We found that these isoforms were differentially expressed by neurons in the substantia nigra, red nucleus, dorsal cochlear nucleus, pontine nuclei and inferior olivary nucleus. Neurons in the inferior olivary nucleus express the alpha isoform, but not the beta isoform. In contrast, neurons in the substantia nigra, red nucleus and pontine nuclei were immunostained with the beta antibody, but not with the alpha antibody. In the dorsal cochlear nucleus, neurons in layers I and II were alpha-immunopositive, whereas neurons in layers III and IV were beta-immunopositive. Therefore, the distribution of the CaM kinase II alpha-immunopositive neurons is completely different from that of CaM kinase II beta-immunopositive neurons. Next we examined the possible coexistence of CaM kinase II alpha isoform and glutamate or that of CaM kinase II beta isoform and glutamic acid decarboxylase (GAD) in the single neuron by double immunofluorescence labelling using a pair of anti-alpha and anti-glutamate antibodies, or a pair of anti-beta and anti-GAD antibodies. The results indicated that neurons expressing anti-alpha immunoreactivity were also immunopositive against anti-glutamate antibody, and neurons expressing beta isoform were also immunopositive against anti-GAD antibody, suggesting that alpha-immunopositive neurons are classified as excitatory-type neurons, and on the contrary, beta-immunopositive neurons are classified as inhibitory-type neurons. In conclusion, the present study confirmed that alpha- and beta-isoforms of CaM kinase II are differentially expressed in the nuclei of the brainstem and have different roles.

Animals↗

PubMed1998

Attenuation of focal cerebral infarct in mice lacking NMDA receptor subunit NR2C.

Neuronal death following cerebral vascular occlusion may be caused in part by the action of glutamate acting through the NMDA receptor. Here we demonstrate that gene disruption of the NR2C subunit of the NMDA receptor attenuates focal cerebral ischemic injury after permanent MCA occlusion, and that a low level of NR2C is expressed and active in the cerebral cortex. NR2C-deficient mice do not show impairment of motor coordination or motor learning. Therefore the development of drugs selectively inhibiting NR2C may prove beneficial in the treatment of stroke and traumatic brain injuries.

Animals↗

PubMed1998

Effect of theanine, r-glutamylethylamide, on brain monoamines and striatal dopamine release in conscious rats.

Theanine, r-glutamylethylamide, is one of the major components of amino acids in Japanese green tea. Effect of theanine on brain amino acids and monoamines, and the striatal release of dopamine (DA) was investigated. Determination of amino acids in the brain after the intragastric administration of theanine showed that theanine was incorporated into brain through blood-brain barrier via leucine-preferring transport system. The concentrations of norepinephrine, 3,4-dihydroxyphenylacetic acid (DOPAC) and 5-hydroxyindole acetic acid (5HIAA) in the brain regions were unaffected by the theanine administration except in striatum. Theanine administration caused significant increases in serotonin and/or DA concentrations in the brain, especially in striatum, hypothalamus and hippocampus. Direct administration of theanine into brain striatum by microinjection caused a significant increase of DA release in a dose-dependent manner. Microdialysis of brain with calcium-free Ringer buffer attenuated the theanine-induced DA release. Pretreatment with the Ringer buffer containing an antagonist of non-NMDA (N-methyl-D-aspartate) glutamate receptor, MK-801, for 1 hr did not change the significant increase of DA release induced by theanine. However, in the case of pretreatment with AP-5, (+/-)-2-amino-5-phosphonopentanoic acid; antagonist of NMDA glutamate receptor, the theanine-induced DA release from striatum was significantly inhibited. These results suggest that theanine might affect the metabolism and/or the release of some neurotransmitters in the brain, such as DA.

2-Amino-5-phosphonovalerate↗

PubMed1998

Consistent injury in the striatum of C57BL/6 mice after transient bilateral common carotid artery occlusion.

OBJECTIVE: The recent availability of transgenic mice enables us to study the functional role of single gene products in cerebral ischemia. To establish an experimental murine model of transient forebrain ischemia, this study examined the temporal profile of ischemic neuronal damage in the striatum after bilateral common carotid artery occlusion. METHODS: C57BL/6 mice, which are frequently used for genetic manipulations, were subjected to 15-minute bilateral common carotid artery occlusion. Ischemic injury was examined (4, 8, 24, 48, and 96 h after reperfusion) by Nissl staining, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick-end-labeling, and nuclear staining with Hoechst 33258 dye. RESULTS: Regional cerebral blood flow was decreased to 11 +/- 6% of control values during the ischemic insult. Striatal injury was observed in 95% of animals examined after 15-minute bilateral common carotid artery occlusion. The number of small and medium-size neurons in the striatum was significantly (P < 0.05) decreased 8 hours after reperfusion and continued to decrease until 96 hours, whereas the number of large neurons remained constant. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick-end-labeling-positive cells appeared in the dorsomedial region of the striatum 48 hours after the ischemic insult and throughout the striatum 96 hours after the ischemic insult. Brain sections stained with Hoechst 33258 dye also demonstrated apoptotic nuclei 96 hours after the ischemic insult. CONCLUSION: Striatal injury after transient forebrain ischemia is reproducible in C57BL/6 mice and is a good model to study the molecular mechanisms of ischemic injury, including delayed neuronal death, using transgenic mice.

Animals↗

PubMed1998

Demonstration of putative Ca-binding domains in dentin matrix of rat incisors after daily injections of 1-hydroxyethylidene-1,1-bisphosphonate (HEBP).

In order to clarify the initial process of dentin mineralization, the inhibitory effect of 1-hydroxyethylidene-1,1-bisphosphonate (HEBP) on dentin mineralization was investigated. Rats (100 g) were subcutaneously injected with HEBP (8 mg P/kg) for 7 or 14 d, and the incisors were processed for Ca histochemistry and/or electron microscopy. HEBP-treated incisors demonstrated ladder-like alternate rows of mineralized and non-mineralized dentin at the apical end. GBHA revealed moderate Ca reactions in the non-mineralized circumpulpal dentin matrix where electron microscopy revealed rich distribution of fine mesh-like electron-dense material. Non-mineralized mantle dentin matrix was negative for Ca but contained numerous matrix vesicles (MVs) filled with crystalline and/or amorphous mineral deposits. Mineralization of circumpulpal dentin occurred independently of MV-rich mantle dentin layer in affected specimens. Our data provide histochemical evidence of possible Ca-binding property of the circumpulpal dentin matrix and its absence in the mantle dentin where MV-mediated mineralization occurs. In the mantle dentin, HEBP does not interfere with crystal growth in MVs but inhibits its outgrowth after membrane rupture. It is proposed that circumpulpal dentin matrix has a potential to mineralize independently of MV-mediated mineralization of mantle dentin, although MVs determine the initial site and timing of dentin mineralization.

Aminophenols↗

PubMed1997

Corticospinal tract neurons are radially malpositioned in the sensory-motor cortex of the Shaking rat Kawasaki.

Shaking rat Kawasaki (SRK) is an autosomal recessive mutant rat that exhibits tremor, dystonia, and ataxia and that is characterized by abnormal lamination of the cerebral and cerebellar cortices and the hippocampus. To examine whether or not layer V neurons in the mutant neocortex are malpositioned in accordance with the aberrant laminar cytoarchitecture, horseradish peroxidase (HRP) was injected into the lumbar spinal cord of SRK mutant and normal control rats to label cortical pyramids projecting through the corticospinal tract (CST). HRP-labeled CST neurons of both normal and SRK rats were found mainly in the hindlimb area of the sensory-motor cortex, indicating a normal tangential distribution of labeled CST neurons in the SRK mutant. In the radial axis, however, labeled CST neurons were spread throughout all layers of the mutant cortex, whereas those in normal rats were restricted to layer V. In the mutant, most labeled CST neurons located in the inner third of the cortex had a typical pyramidal form with an upright apical dendrite, but many of those located near the pial surface displayed abnormal shapes and could be subdivided into inverted pyramidal, horizontal, and bipolar neurons on the basis of their dendritic morphology. The abnormal distribution pattern of labeled CST neurons in the mutant was quantified using a standardized measure of their depth distribution, where 0% = the level of the white matter and 100% = the pial surface. The mean value for the SRK cortex of 47.0% was significantly greater than the figure of 40.5% for normal rats (P < 0.01, Student's t test), indicating a spread of CST neurons toward the pial surface in SRK, but even more striking was the size of the standard deviation: 30.4 in SRK compared with 7.1 in controls. The distribution pattern of CST neurons of the SRK rat was also statistically identical with that of the reeler mouse, which is a well-known mutant that also exhibits an abnormal lamination pattern in the cerebral cortex. These results indicate that neuronal components of the neocortex of the SRK mutant are intermingled along the radial axis, but not in the tangential axis, and provide further evidence for a strong similarity between this spontaneous rat mutation and the reeler malformation.

Animals↗

PubMed1997

Nuclear accumulation of HMG2 protein is mediated by basic regions interspaced with a long DNA-binding sequence, and retention within the nucleus requires the acidic carboxyl terminus.

High mobility group 2 (HMG2) protein is ubiquitously distributed in the nucleus of higher eukaryotic cells. Accumulation of an HMG2-beta-galactosidase fusion protein expressed in COS-7 cells suggested active transport of HMG2 protein into the nucleus after translation in the cytoplasm. Deletion analysis of the HMG2 sequence in the HMG2-beta-galactosidase fusion protein indicated that basic regions interspaced with the long DNA-binding sequence in HMG2, called the HMG1/2 box, are necessary for the nuclear accumulation of HMG2. The close configuration of basic regions at both ends of the DNA-binding sequence in the tertiary structure may function as the nuclear localization signal. This novel nuclear localization signal structure is different from typical ones such as the single or bipartite basic cluster in many nuclear proteins. A portion of the HMG2 molecule remained in the cytoplasm after translation. Interspecies heterokaryon assay demonstrated that the acidic carboxyl terminus of HMG2 was necessary for retention of the protein in the nucleus.

Animals↗

PubMed1997

High Critical-Current Density in the Heavily Pb-Doped Bi2Sr2CaCu2O8+delta Superconductor: Generation of Efficient Pinning Centers

Critical-current density (Jc) is a parameter of primary importance for potential applications of high-temperature copper oxide superconductors. It is limited principally by the breakdown of zero-resistive current due to thermally activated flux flow at high temperatures and high magnetic fields. One promising method to overcome this limitation is to introduce efficient pinning centers into crystals that can suppress the flux flow. A marked increase in Jc was observed in Bi2Sr2CaCu2O8+delta (Bi-2212) single crystals doped with a large amount of Pb. By electron microscopy, characteristic microstructures were revealed that probably underlie the observed enhancement in Jc: thin (10 to 50 nanometers), platelike domains having a modulation-free structure appeared with spacings of 50 to 100 nanometers along the b axis.

Journal Article↗

PubMed1997

Expression of reelin, the gene responsible for the reeler mutation, in embryonic development and adulthood in the mouse.

reelin has recently been isolated as a candidate gene, the mutation of which gives rise to the reeler phenotype in mice. In this study, we analyzed the expression of reelin during embryonic development in the mouse and in adult mouse tissues, by in situ hybridization. reelin transcripts were present on embryonic day (E) 8.5 in the somite, foregut, yolk sac, and unclosed neural plate. reelin was expressed in the brain, spinal cord, liver, and kidney throughout embryonic development, and transiently in many developing organs such as the optic cup, blood vessels, precartilage, stomach, pituitary, vibrissae, tooth germ, and in cells along growing nerve fibers. These observations indicate a role for reelin in development of organs in addition to that in neuronal migration. Furthermore, we demonstrated the existence of reelin mRNA and its cellular distribution in the adult brain, spinal cord, liver, kidney, testis, and ovary, suggesting additional roles for reelin in stabilizing the cyto-architecture and in remolding in adult organs. However, we detected no obvious phenotype of the reelin-expressing organs except for the brain in the reeler mouse, indicating the functional redundancy of this gene during the development of these organs.

Animals↗

PubMed1997

Successful treatment of an esophageal hemangioma by endoscopic injection sclerotherapy: report of a case.

We report herein the case of a 46-year-old man in whom an esophageal hemangioma was successfully treated by endoscopic injection sclerotherapy (EIS). The patient was admitted to hospital after a routine upper gastrointestinal series demonstrated a filling defect in the esophagus. Endoscopy, computed tomography (CT) images, and magnetic resonance imaging (MRI) subsequently revealed a hemangioma in the middle esophagus. EIS was performed using 99% ethanol and 1% polydocanol, and 4 weeks later, the tumor had almost disappeared. This patient has since been followed up carefully with periodic endoscopy and barium studies and additional sclerotherapy will be performed if necessary.

Esophageal Neoplasms↗

PubMed1997

Retrograde and anterograde labeling of cerebellar afferent projection by the injection of recombinant adenoviral vectors into the mouse cerebellar cortex.

Adenoviral vectors have recently been recognized as highly efficient systems for gene delivery into various tissues. We show that a reporter gene introduced into nerve terminals via an adenovirus can be used to label cell bodies retrogradely and then label the axons and nerve terminals of the infected neurons anterogradely in vivo. We injected a replication-defective recombinant adenovirus carrying the E. coli beta-galactosidase gene (lacZ) into the cerebellar cortex of the adult mouse. The first evidence of retrograde labeling was obtained at 2 days after the infection when neurons in the pontine nuclei and the reticulotegmental nucleus of the pons weakly expressed beta-galactosidase, and at 3 days post-infection when neurons in all precerebellar nuclei, known to project to the cerebellar cortex, were strongly stained with X-gal in a Golgi-like manner. Anterograde transport of lacZ gene products was recognized at 3 days post-infection; beta-galactosidase-positive axons arose from somata or dendrites of retrogradely labeled neurons, passed through the middle or inferior cerebellar peduncles, and entered the cerebellum. Anterogradely labeled mossy terminals were recognized on the injection side at 8 days post-infection, and on the contralateral side at 14 days post-infection. Beta-galactosidase expression persisted for up to two months, with a decrease in the total number of labeled cells over time. We could not find any signs of anterograde or retrograde transsynaptic labeling in the nuclei synaptically linked to the cerebellar cortex at any time point after injection up to 58 days post-infection.

Adenoviridae↗