共查询到20条相似文献,搜索用时 15 毫秒
1.
Takahashi E Miyamoto N Kajiwara N Furuya K Yanai-Taniguchi K Sugiyama F Yagami K 《Brain research. Brain research protocols》2000,5(2):159-166
To define a gene expression mechanism, it is often advantageous to use a reporter gene and transgenic mouse. The lacZ reporter gene is particularly useful for studies of the cis-regulatory element for tissue-specific expression in transgenic mice because of the ease of the enzyme assay and visualization on sections. In this report, we describe our method for examining the cis-regulatory element in transgenic mice, including choice of the lacZ gene, generation of transgenic mice, and analysis of beta-galactosidase activity. 相似文献
2.
Cloning of a new gap junction gene (Cx36) highly expressed in mammalian brain neurons 总被引:16,自引:0,他引:16
Daniele Filippo Condorelli Rosalba Parenti Francesca Spinella Angela Trovato Salinaro Natale Belluardo Venera Cardile & Federico Cicirata 《The European journal of neuroscience》1998,10(3):1202-1208
3.
Couégnas A Schweitzer A Andrieux A Ghandour MS Boehm N 《Journal of neuroscience research》2007,85(7):1515-1527
Stable tubulin-only polypeptide (STOP) proteins are microtubule-associated proteins responsible for microtubule stabilization in neurons. STOP null mice show apparently normal cerebral anatomy but display synaptic defects associated with neuroleptic-sensitive behavioral disorders. STOP null mice have therefore been proposed as an animal model for the study of schizophrenia. In the present study, the expression pattern of STOP gene in developing and adult brain has been examined by using lacZ gene inserted in the STOP locus, as a reporter gene. beta-Galactosidase (beta-gal) immunostaining was confined to neuronal cells and projections. Strong labeling was observed in the whole olfactory system, cortical layer VII, hippocampus, hypothalamus, cerebellum, habenula, fasciculus retroflexus, and interpeduncular nucleus in adults. Additionally, ventral thalamic nucleus, clusters of positive cells in striatum, and Cajal-Retzius cells of cortical layer I were labeled in young mice. The strong expression of STOP lacZ reporter gene observed in brain is confined to areas that may be involved in the schizophrenia-related symptoms observed in STOP-deficient mice. 相似文献
4.
Eiberger J Kibschull M Strenzke N Schober A Büssow H Wessig C Djahed S Reucher H Koch DA Lautermann J Moser T Winterhager E Willecke K 《Glia》2006,53(6):601-611
Using newly generated transgenic mice in which the coding region of the connexin29 (Cx29) gene was replaced by the lacZ reporter gene, we confirmed previous immunochemical results that Cx29 is expressed in Schwann cells, oligodendrocytes and Bergmann glia cells. In addition, we detected lacZ/Cx29 in Schwann cells of the sciatic nerve and in particular of the spiral ganglion in the inner ear, as well as at low abundance in the stria vascularis. Furthermore, we found lacZ/Cx29 expression in nonmyelinating Schwann cells of the adrenal gland, in chondrocytes of intervertebral discs and the epiphysis of developing bones. Electron microscopic analyses of myelin sheaths in the central and peripheral nervous system of Cx29-deficient mice detected no abnormalities. The nerve conduction in the sciatic nerve of adult Cx29-deficient mice and the auditory brain stem response as well as visually evoked potentials in 4- to 10-week-old Cx29-deficient mice were not different from wild-type littermate controls. Thus, in contrast to connexin32 and connexin47, which are also expressed in myelinating cells, Cx29 does not contribute to the function of myelin in adult mice. 相似文献
5.
Fournier B Rovira C Mailly P Fuhrman Y Mariani J 《The Journal of comparative neurology》2002,449(1):65-75
In immature rats, Purkinje cells receive synapses from multiple climbing fibers. During development, this multi-innervation regresses and only one climbing fiber innervates each Purkinje cell in the adult. The multi-innervation of immature rats is maintained in the adult if the precursors of the cerebellar granule cells are destroyed by early postnatal X-irradiation. The present study was undertaken to determine the origin of climbing fibers projecting to lobule VI-VII of the cerebellum in X-irradiated granuloprival rats. Olivary neurons were labelled by retrograde transport of wheat germ agglutinin conjugated to horseradish peroxidase, which was injected by iontophoresis in the right vermis of lobule VI-VII. Three-dimensional reconstructions of the inferior olive were made for granuloprival and control rats. No significant variation in the shape and dimension of the olive was observed between the two groups. Labeled cells were found in the middle part of the median accessory olive (MAO). In control rats, stained cells were found only in the contralateral MAO, whereas in the granuloprival rats they were located in both the contralateral and the ipsilateral MAO. Homologous zones were marked in control and granuloprival rats in the middle part of MAO. In granuloprival rats, there was a symmetry in the distribution of the stained cells in the ipsi- and contralateral MAO along the three axes. Therefore, polyinnervation involves homologous regions of both inferior olivary nuclei. 相似文献
6.
A qualitative and quantitative light microscopic study of the inferior olivary complex of normal, reeler, and weaver mutant mice 总被引:3,自引:0,他引:3
In the normal mouse (+/+; +/rl) cerebellar Purkinje cells (PCs) are aligned in a monolayer and provide the main targets for incoming olivocerebellar climbing fibers (CF). In the neurological mutants, homozygous reeler (rl/rl), homozygous weaver (wv/wv) and heterozygous weaver (wv/+), cerebellar abnormalities exist in which many PCs are either missing or displaced. Therefore, it is of interest of determine if the inferior olivary complex (IO) in these mutants is also abnormal. This report concerns results obtained from a light microscopic study of the inferior olivary complex. Counts of IO cells revealed apparent differences in the IO in homozygous reeler when compared to normal littermates. Whereas in the normal mouse there are approximately 37,000 IO cells and clearly defined olivary subdivisions, the IO of the homozygous reeler has a 22.6% reduction in IO cells (mean = 28,770) and indistinct borders between the major olivary subdivisions. With regard to the heterozygous and homozygous weaver, surprisingly the IO morphology and cell numbers are similar to that of the wildtype mouse even though the animals have only 86% (wv/+, mean = 158,155) and 72% (wv/wv, mean = 131,882), respectively, of the normal numbers of PCs (+/+, mean = 183,857). Purkinje cell counts revealed that the midline vermal region is the most affected area in the cerebellum in wv/+ and wv/wv whereas counts in the lateral hemisphere are near normal. The PC/IO ratio in the homozygous weaver is approximately 3:1 as compared to 5:1 in the wildtype mouse. Recent electrophysiological findings in wv/wv indicate that PCs are multiply innervated by CFs. Since a transient phase of multiple innervation is normal in the immature rat, the situation in the adult homozygous weaver may represent a retention of this immature state. A factor which may play a role in this is the loss of parallel fiber (PF)-PC synapses resulting from massive postnatal granule cell death. An hypothesis suggesting an intrinsic PC time-dependent mutant gene effect is presented to account for the differences in the loss of Purkinje cells between wv/wv and wv/+ and between different regions of the cerebellum. 相似文献
7.
The prion protein is central to the pathogenesis of prion diseases, although its exact function remains unclear. Although transgenic mice have been widely utilised in prion research, their PrP expression patterns have not been characterised in detail. We have studied the developmental temporal and spatial expression of a 214-bp mini human PrP promoter in transgenic mice. Transgene expression is first detected at embryonic day 12.5, a day earlier than previously reported for endogenous mouse gene by in situ hybridization. The general expression pattern closely mirrors that of the endogenous mouse PrP gene, such that this small and clearly defined transgene cassette can replace the need to use large cosmid based vectors for transgenetic modeling of human and animal prion disease. 相似文献
8.
The organization of the olivocerebellar projection in the homozygous reeler mouse (rl/rl) was studied with the use of microinjections of 3H-leucine in different regions of the inferior olivary complex (IO) or horseradish peroxidase conjugated with wheat germ agglutinin (WGA-HRP) into medial, intermediate, or lateral regions of the reeler cerebellum. The purpose of this investigation was to determine the pattern of termination of olivocerebellar climbing fibers (CFs) in the cerebellum via an anterograde tracing technique, and to determine the topographic organization of the olivocerebellar projection via both anterograde and retrograde methods. The inferior olive injections were made via the ventral (i.e., retropharygeal) approach to the IO to minimize diffusion into other brainstem precerebellar nuclei and thus to ensure accurate well-restricted, injection sites. Labeled CF terminals were seen in both the superficial Purkinje cell (PC) layer (normally positioned PCs) and around PCs in the granular layer and central masses (ectopic PCs). The pattern of labeling is suggestive of orthogonal organization, in that vertical columns of cells are labeled. This is especially apparent in the medial PC group, where at least three bands are identified. Within an orthogonal band, CF terminals are seen around both superficial and deep Purkinje cells. Our data indicate that olivocerebellar topography is generally similar in reeler and normal mice despite severe abnormalities in target cell position in the reeler. The medial cerebellar region receives input from the caudal two-fifths of the medial accessory olive (MAO). The intermediate PC cluster receives input from more rostral portions of all three olivary divisions (MAO, principal olive [PO] and dorsal accessory olive [DAO] ), while rostral portions of MAO and PO project to the lateral cerebellum. These results indicate that the zonal organization of the olivocerebellar projection in the adult reeler exhibits a pattern generally similar to that seen in normal mice. This suggests that an afferent system can develop a normal organization despite having ectopic targets. 相似文献
9.
10.
Climbing fiber projections from the inferior olive to the uvula of the cerebellum were studied in the cat by using retrograde axonal transport of horseradish peroxidase. Following large and small injections into various parts of the uvula, the distribution of labeled cells in the inferior olive was investigated. The findings indicate six longitudinal zones extending throughout the dorsal and ventral uvula: the caudal part of the nucleus beta projects to a most medially located zone (caudal beta zone) with a width of about 0.4 mm; the rostral part of the nucleus beta projects to a zone located at about 0.6 mm from the midline (rostral beta zone); the caudal part of the medial accessory olive (MAO) projects to a zone (caudal MAO zone) located lateral to the rostral beta zone; the dorsomedial cell column projects to a zone (dorsomedial cell column zone) located in the intermediate part of the uvula at about 1.2 mm from the lateral edge of the uvula; the ventral lamella of the principal olive (PO) projects to a zone (ventral lamella of PO zone) about 0.7 mm from the lateral edge of the uvula; finally, the rostral part of the MAO projects to the most lateral zone (rostral MAO zone). These conclusions are in general agreement with those of earlier studies and also provide a more detailed zonal configuration of climbing fiber projections to the uvula. 相似文献
11.
Takahashi E Miyamoto N Oki T Kajiwara N Furuya K Yanai Taniguchi K Yagami K Niidome T Tanaka I Katayama K 《Brain research》1999,850(1-2):47-54
To dissect the molecular mechanisms underlying the neuron-specific expression of the P/Q type calcium channel alpha 1A subunit gene, transgenic mice carrying a 0.5-kb, 1.5-kb, 3.0-kb or 6.3-kb 5'-upstream region of the gene fused to Escherichia coli lacZ reporter gene were produced. In transgenic mice carrying the 1.5-kb, 3.0-kb or 6.3-kb 5'-upstream region, the reporter gene was exclusively expressed in the nervous system, although those with the 0.5-kb 5'-upstream region failed to show reporter expression. Histological examinations showed that the three 5'-upstream regions induced distinct expression patterns of the reporter gene in the CNS and adrenal medulla. The 1.5-kb 5'-upstream region drove reporter gene expression in the olfactory bulb, dorsal cortex and hippocampus, while the regulatory element for the expression in the amygdaloid nucleus, septum, habenula medial nucleus, choroid plexus, substantia nigra, inferior colliculus, pontine nucleus and cerebellum was located in the 5'-upstream sequence between 1.5 kb and 6.3 kb. In the cerebellum, the expression of the reporter gene was induced by the 3.0-kb region in granule cells, whereas it was induced by the 6.3-kb region in Purkinje cells. The expression of the reporter gene in chromaffin cells in the adrenal medulla was induced only by the 6.3-kb 5'-upstream region. These results suggest that the expression of the mouse P/Q-type Ca2+ channel alpha 1A subunit gene is regulated in a complex fashion by both positive and negative cis-regulatory elements. 相似文献
12.
The purpose of these experiments was to define the topography of cuneate and spinal projections to the forelimb representation in the rostral dorsal accessory olive (rDAO). We were interested in determining whether the spinal and cuneate inputs constitute a homogeneous afferent source, and whether there is evidence that they serve different functional roles. We were also interested in determining whether the somatotopy of rDAO is the result of a point-to-point projection from its afferent sources, or whether the projection suggests a reorganization of afferents at the olive. Single unit recording was used to identify specific regions of rDAO, and the topography of inputs to the identified regions was determined by using wheat germ agglutinin-horseradish peroxidase (WGA-HRP) as a tracer. The results from retrograde tracing were confirmed by using WGA-HRP as an anterograde tracer from input sources. The cuneate and spinal neurons providing input to rDAO constitute two distinct neural populations. One consists of cells in the caudal cuneate nucleus and lamina VI of the rostral two cervical segments, the other consists of cells in the rostral cuneate nucleus. The cells in the caudal cuneate nucleus and the rostral cervical segments are large, multipolar neurons that form a single column of rDAO input cells. The column of cells projects to the contralateral rDAO in a topographic fashion with rostral regions of the column projecting to rostral rDAO, which contains cells that respond to somatosensory stimulation of the contralateral shoulder, trunk, and proximal forelimb. Caudal regions of the column project to caudal rDAO, which contains cells that respond to stimulation of the distal forelimb. Despite this topography, there is a large degree of overlap in the terminations from neighboring regions of the input column, indicating that a major reorganization occurs at the rDAO. The projection from the rostral cuneate nucleus arises from small neurons that project bilaterally to rDAO, and the input from the rostral cuneate nucleus lacks a clear topography. We propose that input from the cell column is responsible for the somatosensory sensitivity of rDAO neurons, whereas input from rostral cuneate is most likely modulatory, probably inhibitory, in nature. J. Comp. Neurol. 392:115–133, 1998. © 1998 Wiley-Liss, Inc. 相似文献
13.
14.
Olivocerebellar projections to the uvula were studied by means of retrograde axonal transport of horseradish peroxidase (HRP) in pigmented rabbits. The distribution pattern of labeled cells in the inferior olive was compared among cases following large- and microinjections of HRP into the uvula. Findings indicate topographically organized projections to longitudinally oriented zones. There are at least 6 zones in the rabbit's uvula. The caudal part of the nucleus beta projects contralaterally to a most medially located zone (caudal beta zone). The rostral part of the nucleus beta projects to a little more laterally located zone (rostral beta zone) at a distance of about 1 mm from the midline of the uvula. The caudolateral part of the MAO projects to a zone (caudolateral MAO zone) located laterally to the rostral beta zone. The dorsomedial cell column projects to a zone (dorsomedial cell column zone) located in the intermediate part of the uvula at about 2 mm from the midline. The rostrolateral part of the MAO projects to the most lateral zone (rostrolateral MAO zone) of the uvula. Finally, the ventral lamella of the PO projects to a zone (ventral lamella of PO zone) located between the rostrolateral MAO zone and the dorsomedial cell column zone. 相似文献
15.
E Wada K Wada J Boulter E Deneris S Heinemann J Patrick L W Swanson 《The Journal of comparative neurology》1989,284(2):314-335
Previous studies have revealed the existence of a gene family that encodes a group of neuronal nicotinic acetylcholine receptor (nAChR) subunits. Four members of this family have been characterized thus far; three of these subunits (alpha 2, alpha 3, and alpha 4) are structurally related to the ligand binding subunit expressed in muscle and form functional nAChRs when combined with the beta 2 gene product in Xenopus oocytes. In addition, the alpha 4 gene appears to encode two different products (alpha 4-1 and alpha 4-2) that have been proposed to arise by alternative mRNA splicing. Nine different [35S]-complementary ribonucleic acid (cRNA) probes were used in the present study to map the distribution of these nAChR subunit mRNAs throughout the central nervous system (CNS) of the rat. It was found that the beta 2 gene is expressed in most regions of the CNS, as are the alpha subunit genes as a group. However, each alpha gene is expressed in a unique, although partly overlapping, set of neuronal structures. Alpha 4 is the most widely expressed alpha gene, and the evidence suggests that mRNAs for the alpha 4-1 and alpha 4-2 products are virtually always found in the same regions, in approximately the same ratios (alpha 4-2 greater than alpha 4-1). In addition, there are several examples of cell groups that express beta 2 but none of the alpha subunit mRNAs examined here (particularly in the hypothalamus), as well as all groups that express the converse, thus suggesting that additional neuronal nAChR subunits remain to be characterized. Finally, the extensive expression of multiple alpha subunits in certain regions, particularly for alpha 3 and alpha 4 in the thalamus, suggests that there is microheterogeneity in a small population of cells or that some neurons may express more than one alpha subunit. This problem needs to be examined directly with double labeling methods but raises the possibility that some neuronal nAChRs may be composed of more than one kind of alpha subunit. The wide expression of these receptor genes suggests that nAChRs constitute major excitatory systems in the CNS. 相似文献
16.
M F Huerta T Hashikawa M J Gayoso J K Harting 《The Journal of comparative neurology》1985,241(2):180-190
Anterograde autoradiographic methods were used to determine the projection of the principal sensory trigeminal nucleus and of each of the three spinal trigeminal subnuclei to the inferior olivary complex in the cat. Our data reveal that the principal sensory trigeminal nucleus does not contribute to the trigemino-olivary pathway. Each spinal trigeminal subnucleus has a unique contribution to this pathway: pars oralis projects sparsely to the border between the dorsal accessory and principal olives (DAO-PO), pars interpolaris projects mostly to the rostral medial DAO, and pars caudalis projects mostly to the rostral medial part of the ventral leaf of PO and slightly to the caudal medial accessory olive. In the light of recent physiological and anatomical findings, our data indicate that information from each spinal trigeminal subnucleus reaches a different segment of the contralateral inferior olivary complex, which in turn distributes differentially to the cerebellar cortex. 相似文献
17.
T.M. Teune J. van der Burg C.I. de Zeeuw J. Voogd T.J.H. Ruigrok 《The Journal of comparative neurology》1998,392(2):164-178
Two different populations of projection neurons are intermingled in the cerebellar nuclei. One group consists of small, γ-aminobutyric acid-containing (GABAergic) neurons that project to the inferior olive, and the other group consists of larger, non-GABAergic neurons that provide an input to one or more, usually premotor, centers in the brainstem, such as the red nucleus, the thalamus, and the superior colliculus. All cerebellar nuclear neurons are innervated by GABAergic Purkinje cells. In this study, we investigated whether individual Purkinje cells of the C1 zone of the paramedian lobe of the rat innervate both groups of projection neurons in the anterior interposed nucleus. Two different, retrogradely transported tracers, either cholera toxin β subunit (CTb) or wheat germ agglutinin coupled to horseradish peroxidase (WGA-HRP) and a gold lectin tracer were injected into the red nucleus and the inferior olive, respectively, whereas Purkinje cell axons were anterogradely labeled with biotinylated dextran amine (BDA) injected into the paramedian lobule. Cerebellar nuclear sections studied with the light microscope demonstrated a close relation of varicosities from BDA-labeled Purkinje cell axons with both gold lectin- and CTb-labeled neurons. Branches of individual axons could be traced to both retrogradely labeled cell populations. At the ultrastructural level, synapses of labeled Purkinje cell terminals with profiles of WGA-HRP-labeled projection neurons predominated over contacts with gold lectin-containing neurons. Nine out of 367 investigated BDA-labeled terminals were observed to be presynaptic to a WGA-HRP-labeled profile as well as to a gold lectin-labeled profile. This indicates that nuclear cells that project to the inferior olive as well as those that project to premotor centers are under the influence of the same Purkinje cells. Such an arrangement would suggest an in-phase cortical modulation of the activation patterns of the inhibitory cells that project to the inferior olive and excitatory cells that project to premotor nuclei, which could explain why olivary neurons, especially those of the rostral part of the dorsal accessory olive, appear to be unresponsive to stimuli generated during active movement. J. Comp. Neurol. 392:164–178, 1998. © 1998 Wiley-Liss, Inc. 相似文献
18.
In albino rats the superior colliculus was stimulated and its evoked potentials were explored throughout the posterior vermis of the cerebellum. Climbing fiber responses were identified only in lobule VII, ipsilaterally 1.2–1.6 mm wide. In the medial accessory olive, subnucleus c, in the contralateral side both antidromically evoked potentials from lobule VII and orthodromically evoked potentials from the superior colliculus were recorded. This evidence suggests that they are tecto-olivoerebellar projections. 相似文献
19.
Dedek K Schultz K Pieper M Dirks P Maxeiner S Willecke K Weiler R Janssen-Bienhold U 《The European journal of neuroscience》2006,24(6):1675-1686
The primary rod pathway in mammals contains gap junctions between AII amacrine cells and ON cone bipolar cells which relay the rod signal into the cone pathway under scotopic conditions. Two gap junctional proteins, connexin36 (Cx36) and connexin45 (Cx45), appear to play a pivotal role in this pathway because lack of either protein leads to an impairment of visual transmission under scotopic conditions. To investigate whether these connexins form heterotypic gap junctions between ON cone bipolar and AII amacrine cells, we used newly developed Cx45 antibodies and studied the cellular and subcellular distribution of this protein in the mouse retina. Specificity of the Cx45 antibodies was determined, among others, by Western blot and immunostaining of mouse heart, where Cx45 is abundantly expressed. In mouse retina, Cx45 immunosignals were detected in both plexiform layers and the ganglion cell layer. Double staining for Cx45 and Cx36 revealed a partial overlap in the punctate patterns in the ON sublamina of the inner plexiform layer of the retina. We quantified the distributions of these two connexins in the ON sublamina, and detected 30% of the Cx45 signals to be co-localized with or in close apposition to Cx36 signals. Combining immunostaining and intracellular dye injection revealed an overlap or tight association of Cx36 and Cx45 signals on the terminals of injected AII amacrine and two types of ON cone bipolar cells. Our results provide direct evidence for heterotypic gap junctions composed of Cx36 and Cx45 between AII amacrine and certain types of ON cone bipolar cells. 相似文献
20.
The gene encoding 2′,3′-cyclic nucleotide 3′-phosphodiesterase (CNP) is one of the earliest myelin genes to be expressed in the brain. It is expressed at basal levels in some non-neural tissues but at much higher levels in the nervous system, and its relevance and mechanism are unknown. Using transgenic mice, we examined the expression pattern conferred by a 4-kilobase (-kb) 5′-flanking sequence of the mouse CNP gene coupled to the bacterial lacZ reporter gene. Here we report that this 4-kb fragment contains sufficient information to direct expression of the transgene to the tissue and/or cell type, in which CNP is normally expressed. In the central nervous system (CNS), CNP-lacZ expression was regulated in a temporal manner, consistent with endogenous CNP expression. Transgene expression was detected in embryonic brain and spinal cord in immature oligodendrocytes, and it significantly increased with age. In adult mice, β-galactosidase activity (which appeared to be oligodendrocyte specific) was found essentially in white matter areas of the CNS. Moreover, the transgene was expressed in peripheral nervous system, testis, and thymus—tissues that normally express CNP. Taken together, our results provide strong evidence that cis-acting regulatory elements, necessary to direct spatial and temporal expression of the transgene in oligodendrocytes, are located within the 4-kb 5′-flanking sequence of the mouse CNP gene. This promoter could be a valuable tool to target specific expression of other transgenes to oligodendrocytes, and may provide important new insights into myelination or dysmyelination. J. Neurosci. Res. 53:393–404, 1998. © 1998 Wiley-Liss, Inc. 相似文献