expression of cadherin-8 mrna in the developing mouse central nervous system

Expression of Cadherin-8 mRNAin the Developing MouseCentral Nervous System

KOJIRO KOREMATSU1 AND CHRISTOPH REDIES1,2*1Department of Biochemistry, Max Planck Institute of Developmental Biology,

D-72076 Tubingen, Germany2Institute of Biology III, University of Freiburg, D-79104 Freiburg, Germany

ABSTRACTThe expression of cadherin-8 was mapped by in situ hybridization in the embryonic and

postnatal mouse central nervous system (CNS). From embryonic day 18 (E18) to postnatal day6 (P6), cadherin-8 expression is restricted to a subset of developing brain nuclei and corticalareas in all major subdivisions of the CNS. The anlagen of some of the cadherin-8-positivestructures also express this molecule at earlier developmental stages (E12.5–E16). Thecadherin-8-positive neuroanatomical structures are parts of several functional systems in thebrain. In the limbic system, cadherin-8-positive regions are found in the septal region,habenular nuclei, amygdala, interpeduncular nucleus, raphe nuclei, and hippocampus.Cerebral cortex shows expression in several limbic areas at P6. In the basal ganglia andrelated nuclei, cadherin-8 is expressed by parts of the striatum, globus pallidus, substantianigra, entopeduncular nucleus, subthalamic nucleus, zona incerta, and pedunculopontinenuclei. A third group of cadherin-8-positive gray matter structures has functional connectionswith the cerebellum (superior colliculus, anterior pretectal nucleus, red nucleus, nucleus ofposterior commissure, inferior olive, pontine, pontine reticular, and vestibular nuclei). Thecerebellum itself shows parasagittal stripes of cadherin-8 expression in the Purkinje cell layer.In the hindbrain, cadherin-8 is expressed by several cranial nerve nuclei.

Results from this study show that cadherin-8 expression in the embryonic and postnatalmouse brain is restricted to specific developing gray matter structures. These data support theidea that cadherins are a family of molecules whose expression provides a molecular code forthe regionalization of the developing vertebrate brain. J. Comp. Neurol. 387:291–306, 1997.r 1997 Wiley-Liss, Inc.

Indexing terms: embryonic brain; cell adhesion molecules; cadherins; brain morphogenesis; brain

segments

Cadherins are a family of cell surface proteins thatmediate Ca21-dependent cell-cell adhesion and controlmorphogenesis in various embryonic organs including thecentral nervous system (CNS; reviewed by Takeichi, 1988,1995). In the CNS, at least 10 cadherins are expressed. Theexpression of some of these cadherins, e.g., N-, E-, R-, andB-/P-cadherin, is restricted to specific developing graymatter regions and fiber tracts (reviewed by Redies, 1995;Redies and Takeichi, 1996). Moreover, for N- and R-cadherin,it has been shown that the neuroanatomical structuresexpressing these cadherins form parts of particular neuralcircuits and functional systems (Redies et al., 1993; Arndtand Redies, 1996). In the early embryonic neuroepithelium,the expression of each cadherin is restricted to particularsegmental (‘‘neuromeric’’) subdivisions or to the boundaryregions between these subdivisions (Espeseth et al., 1995;

Ganzler and Redies, 1995; Matsunami and Takeichi, 1995;Kimura et al., 1996). Recently, the cadherin/catenin sys-tem was found to be an integral component of synapses(Beesley et al., 1995; Rose et al., 1995; Yamagata et al.,1995; Fannon and Colman, 1996; Uchida et al., 1996).

A characteristic feature of cadherins is their selectivityof binding. Cells expressing a particular type of cadherin

Grant sponsor: Max Planck Society; Grant sponsor: Land Baden-Wurttemberg Neurobiology Programme.

Dr. Korematsu’s current address is Department of Neurosurgery, Kuma-moto University Medical School, Honjo 1-1-1, Kumamoto 860, Japan.

*Correspondence to: Christoph Redies, Institute of Anatomy, UniversityHospital Essen, Hufelandstrasse 55, D-45122 Essen, Germany.E-mail: [email protected]

Received 6 March 1996; Revised 19 May 1997; Accepted 2 June 1997

THE JOURNAL OF COMPARATIVE NEUROLOGY 387:291–306 (1997)

r 1997 WILEY-LISS, INC.

preferentially bind to other cells expressing the same typeof cadherin. For example, N-cadherin-transfected cellsbind to other N-cadherin-transfected cells, but not toE-cadherin-transfected cells (Nose et al., 1988; Miyatani etal., 1989). Weak binding between cells expressing differentcadherin types has also been described (Inuzuka et al.,1991a; Matsunami et al., 1993; Nakagawa and Takeichi,1995). Based on the preferentially homotypic binding ofcadherin-expressing cells, it has been proposed that cadher-ins are involved in the sorting and aggregation of earlyneurons during CNS development and in the formation ofbrain nuclei and cortical laminae (Takeichi et al., 1990;Redies et al., 1993; Redies, 1995; Redies and Takeichi,1996). Data to support this suggestion were provided bystudies on the embryonic chicken brain. From the begin-ning of mantle zone formation, N- and R-cadherin-expressing early neurons born in the different neuromeresaccumulate in the mantle zone and sort out into differentbrain nuclei, some of which later become connected by fibertracts expressing the same type of cadherin (Ganzler andRedies, 1995).

At least three subgroups of cadherins are expressed inthe embryonic vertebrate CNS (type I cadherins, type IIcadherins, and protocadherins; Suzuki et al., 1991; Sano etal., 1993). These subgroups are distinguishable from eachother by differences in their protein sequences (Suzuki etal., 1991; Sano et al., 1993; Tanihara et al., 1994; Suzuki,

1996). Many studies have focused on the CNS expression oftype I cadherins, such as E-, R-, N-, and B-/P-cadherin(Hatta et al., 1988; Inuzuka et al., 1991b; Redies et al.,1992; Shimamura et al., 1992; Shimamura and Takeichi,1992; Redies and Takeichi, 1993; Murphy-Erdosh et al.,1994; Redies and Muller, 1994; Ganzler and Redies, 1995;Matsunami and Takeichi, 1995; Redies, 1995; Arndt andRedies, 1996). Relatively few studies (Espeseth et al.,1995; Sago et al., 1995; Kimura et al., 1996) have investi-gated the expression and role of type II (‘‘numbered’’)cadherins (cadherin-5 to cadherin-12) or of protocadherinsin the developing vertebrate CNS.

We have recently cloned a novel type II cadherin ofmouse, cadherin-8 (Korematsu and Redies, 1997). Theamino acid sequence of the mature form of this molecule is98% identical to that of human cadherin-8 (Tanihara et al.,1994). During early embryogenesis (embryonic day 9–14[E9–E14]), cadherin-8 is expressed mainly in the thymusand in the CNS. In the early embryonic brain, the expres-sion of cadherin-8 partially reflects its neuromeric organi-zation. In several areas of the developing mantle zone,distinct groups of cadherin-8-positive neurons begin toform during the neuromeric stage of brain development(Korematsu and Redies, 1997).

In the present study, cadherin-8 expression is studied indetail at relatively late stages of brain development (E18–P6) when the mantle layer has differentiated into well-

Abbreviations

3V third ventricle4V fourth ventricleI layer I of cerebral cortexII-IV layers II-IV of cerebral cortexV layer V of cerebral cortexVI layer VI of cerebral cortexVII facial nucleusXII hypoglossal nucleusAmy amygdalaap anterior pretectal nucleusAq aqueductc caudalCb cerebellumcc corpus callosumCg cingulate cortexCL centrolateral thalamic nucleusCl claustrumCNS central nervous systemcp caudoputamenCx cerebral cortexd dorsalDB nucleus of diagonal bandDG dentate gyrusDR dorsal raphe nucleusDT dorsal thalamusDTg dorsal tegmental nucleiE embryonic dayEGL external germinal layer of cerebellumEnt entorhinal cortexEP entopeduncular nucleusGP globus pallidusGr granular layer of the cerebellumHi hippocampusHT hypothalamusInt interposed cerebellar nucleusIOPr inferior olive, principal nucleusIP interpeduncular nucleusl lateralLat lateral cerebellar nucleusLD laterodorsal thalamic nucleusLDTg laterodorsal tegmental nucleusLGE lateral ganglionic eminenceLHb lateral habenular nucleus

m medialMed medial cerebellar nucleusMGE medial ganglionic eminenceMHb medial habenular nucleusMnR median raphe nucleusMoCb molecular layer of cerebellumMS medial septal nucleusOr stratum oriens of hippocampusp cerebral peduncleP postnatal dayPC paracentral thalamic nucleusPir piriform cortexPk Purkinje cell layer of cerebellumPMR paramedian raphe nucleusPn pontine nucleusPnR pontine reticular nucleusPPN pedunculopontine nucleusPrh perirhinal cortexPrH prepositus nucleus of the hypoglossal nervePV paraventricular nucleus of thalamusPy pyramidal cell layer of hippocampusr rostralRad stratum radiatum of hippocampusrtm reticular migratory stream of thalamusRtTg reticulotegmental nucleus of the ponsS subiculumSC superior colliculusSN substantia nigraSol nucleus of the solitary tractSpV spinal trigeminal nucleusSpVe spinal vestibular nucleusST striatumSTh subthalamic nucleusTe temporal cortexv ventralVL ventrolateral nucleus of thalamusVLm ventrolateral migratory stream of thalamusvm ventromedial nucleus of hypothalamusVM ventromedial nucleus of thalamusvp ventral putamenVT ventral thalamuszi zona incerta

292 K. KOREMATSU AND C. REDIES

defined neuroanatomical structures. We show that, atthese late stages of development, cadherin-8 expression isrestricted to subsets of developing brain nuclei and corticalareas throughout the CNS. For forebrain, some earlierstages (E12.5–E16) were also examined for comparison.

MATERIALS AND METHODS

Preparation of cRNA probe for in situhybridization

The cRNA probe used for in situ hybridization analysiswas generated as described (Korematsu and Redies, 1997).A plasmid, mCad8-PCR, was used for transcription of thecRNA probe. This plasmid was derived from pBluescriptSK II(1) and contains a 462-bp cDNA fragment from thethird and fourth extracellular domain of mouse cad-herin-8. Anti-sense and sense cRNA probes were tran-scribed in the presence of digoxigenin-labeled UTP (Boeh-ringer, Mannheim, Germany) after the plasmid waslinearized with appropriate restriction enzymes. Probeswere shortened to about 200 bp by alkaline hydrolysis. Bydot blot analysis, the anti-sense cRNA probe does not bindto other mouse cadherins such as N-, R-, E-, or P-cadherin(Korematsu and Redies, 1997). Anti-sense probe of mouseN-cadherin was generated in the same way, by usingplasmid bMN3sk1. This plasmid (kind gift of M. Takeichi)was derived from pBluescript vector and contains a 980-bpcDNA fragment of mouse N-cadherin (mn2 fragment;Miyatani et al., 1989).

In situ hybridization

The in situ hybridization procedure was described previ-ously (Redies et al., 1993; Redies and Takeichi, 1993).Pregnant and neonatal mice of the BALB/c strain werekilled by cervical dislocation or decapitation, in accordancewith the national and institutional guidelines on the use ofanimals in research. The brains and spinal cords of mice atembryonic day (E) 12.5, E14, E16, E18, and at postnatalday (P) 2, P4, and P6, and of adult mice were fixed inice-cold 4% (w/v) formaldehyde (FA) in phosphate bufferedsaline (PBS) for 2 hours. After immersion in a series ofsucrose solutions (12, 15, and 18% [w/v] in PBS), speci-mens were frozen in Tissue-Teq O.C.T. Compound (optimalcutting temperature 210 to 215°C; Miles, Elkhart, IN).Frozen sections of 14- to 25-µm thickness were prepared ina refrigerated microtome and mounted on glass slides.Sections adjoining those hybridized with anti-sense probewere stained with thionin for neuroanatomical orienta-tion, or hybridized with sense probe (negative control), orwith N-cadherin anti-sense probe.

After post-fixation in ice-cold 4% FA in PBS for 30minutes, sections were treated with 1 µg/ml proteinase Kfor 2 minutes at 37°C. The sections were then fixed againin 4% FA in PBS for 30 minutes. Nonspecific binding of thecRNA probe was blocked by acetylation with acetic anhy-dride (0.25% [v/v] in triethanolamine buffer). Covered bysiliconized coverslips, the sections were hybridized over-night at 50°C with sense or anti-sense cadherin-8 probe, orwith N-cadherin anti-sense probe, in a hybridization solu-tion containing 50% (v/v) formamide, 10 mM EDTA, 33standard salt citrate (SSC), 13 Denhard’s solution, 10%(w/v) dextran sulfate, and 0.5 µg/ml yeast tRNA. Afterhybridization, coverslips were removed by incubation in

53 SSC at 55°C. Subsequently, the sections were washedin 50% formamide in 23 SSC for 60 minutes at 55°C. Afterwashing in NTE buffer (10 mM Tris, 1 mM EDTA, 500 mMNaCl, pH 8.0), the sections were treated with 20 µg/mlRNAse A (Sigma, Deisenhofen, Germany) for 30 minutesat 37°C. Thereafter, the sections were washed in thefollowing solutions: NTE buffer (10 minutes at 37°C), 50%formamide/23 SSC (40 minutes at 55°C), 23 SSC (30minutes at 55°C), 0.13 SSC (30 minutes at room tempera-ture), PBS (twice for 5 minutes each at room temperature).Unspecific antibody binding sites were saturated by incu-bation with 2% (v/v) sheep serum in PBS for 20 minutes atroom temperature. The sections were incubated withsheep anti-digoxigenin Fab fragments conjugated withalkaline phosphatase (diluted 1:3,000 in 2% sheep serumin PBS; Boehringer) overnight at 8°C. After washing inPBS, bound alkaline phosphatase was visualized by incu-bation in substrate solution containing nitroblue tetra-zolium salt (0.34 mg/ml) and 5-bromo-4-chloro-3-indolylphosphate-p-toluidin salt (0.18 mg/ml) in alkaline bufferfor several hours, or overnight, if necessary. After enoughreaction product had formed, sections were dehydratedand coverslipped for observation under a light microscope(Axioplan; Zeiss, Oberkochen, Germany) or a stereomicro-scope (Stemi SV 6, Zeiss) equipped with a camera.

RESULTS

General observations

At the peri- and postnatal stages examined (E18, P2, P4,and P6), the expression of cadherin-8 is restricted toparticular developing gray matter areas throughout theCNS. A parasagittal section of an P2 brain hybridized withcadherin-8 anti-sense probe is shown in Figure 1 toillustrate this point. The intensity of the hybridizationsignal varies from region to region. In general, the cadherin-8-positive areas remain the same during the peri- andpostnatal period (E18–P6). No striking changes in thepattern of expression were observed for the different brainregions, with the exception of the cerebellum (see below).However, in most areas, signal intensity decreases asdevelopment proceeds. The in situ hybridization methodused in the present study did not detect any cadherin-8transcript in the adult brain. At earlier developmentalstages (E12.5, E14, and E16), cadherin-8 expression is alsorestricted to a subset of structures in the developingmantle zone.

Figure 2 shows adjoining horizontal sections of E18brain hybridized with anti-sense and sense probe to cad-herin-8, respectively. Several areas of dark reaction prod-uct indicating the presence of cadherin-8 transcript areseen in the section hybridized with the anti-sense probe(Fig. 2a). No specific signal can be seen in the sectionhybridized with the control (sense) probe (Fig. 2b).

The neuroanatomical structures expressing cadherin-8mRNA were identified by comparing sections hybridizedwith cadherin-8 probe and adjoining sections stained forNissl substance. For peri- and postnatal stages, 12 com-plete series of horizontal or frontal sections were analyzedin this study (E18, four series; P2, four series; P4, twoseries; and P6, two series). For earlier developmentalstages (E12.5, E14, and E16), two frontal series each wereexamined. A comprehensive list of all cadherin-8-positivestructures is given in Table 1. In the following, we willdescribe some of the cadherin-8-positive gray matter struc-

CADHERIN-8 EXPRESSION IN MOUSE CNS 293

tures in the different brain divisions in more detail andgive examples of the staining patterns observed.

Cadherin-8 expression in the peri-and postnatal forebrain

Of the cerebral cortical areas, cadherin-8 is most widelyexpressed in limbic cortical regions such as the cingulatecortex (Cg), piriform cortex (Pir), entorhinal cortex (Ent),perirhinal cortex (Prh), and hippocampus (Hi), as shown inFigures 1, 3a,c, 4c,d. In the hippocampal formation, cad-

herin-8-positive neurons are mainly found in the pyrami-dal cell layer (Py) of CA1–CA3, and in the granular celllayer of the dentate gyrus (DG), whereas few positive cellsare seen in the other layers (Figs. 1, 3b,d, 4c,d). At P6,cadherin-8 transcript has decreased to relatively low levelsin the pyramidal cell layer. At P4–6, the subiculum con-tains an area of moderate cadherin-8 expression, whereasthe pre- and parasubiculum show only low levels ofcadherin-8 transcript. The entorhinal and perirhinal corti-ces also exhibit cadherin-8 expression but only in theirupper layers (Fig. 4c,d).

Fig. 1. Parasagittal section of the postnatal day 2 mouse brain hybridized with cadherin-8 anti-senseprobe (a) and adjoining Nissl stain (b). The orientation of the sections (c, caudal; r, rostral) is shown in a.Arrowheads point at artifactual staining. For abbreviations, see list. Scale bar 5 600 µm.


Fig. 2. Horizontal sections of the embryonic day 18 mouse brainhybridized with cadherin-8 anti-sense probe (a) and sense probe (b)and adjoining Nissl stain (c). The orientation of the sections (c, caudal;

r, rostral) is shown in (b). Note that cadherin-8 expression is restrictedto particular nuclei (a) while the sense probe (b) shows unspecifichybridization signal. For abbreviations, see list. Scale bar 5 400 µm.


Apart from limbic cortical structures, cadherin-8 tran-script is also seen in some other cortical areas. Forexample, Figure 4a shows cadherin-8-positive cells in theparietal cortex at P4. Comparison with a Nissl-stainedadjoining section (Fig. 4b) reveals that signals are mostprominent in neurons of layer V. In the other cerebralcortices, such as frontal and visual cortex, cadherin-8-expressing cells are also predominantly found in thedeeper cortical layers between E18 and P4. In contrast, inthe limbic cortex, signal is more homogeneously distrib-uted. At P6, several regions of the limbic cortex expresscadherin-8 relatively strongly, whereas adjacent areasshow no or little cadherin-8 expression. For example,cadherin-8 is expressed in the upper layers of perirhinal

cortex (Prh) but not in the adjacent temporal cortex (Te;Fig. 4c,d).

The medial septal nucleus (MS), the nucleus of thediagonal band (DB), and a dorsal part of lateral septalnucleus (LS) are positive areas in the septal region (Figs.2a,c, 3a,c). The central nuclear group of the amygdala(Amy) is also stained (Fig. 2a,c). The claustrum (Cl) isvisible as a prominent cadherin-8-positive region situatedbetween the cerebral cortex and the central white matter(Figs. 1, 3a,c).

The gray matter regions constituting the basal gangliaare amongst the structures expressing cadherin-8 moststrongly. In particular, the globus pallidus (GP) shows veryhigh levels of hybridization signal (Figs. 1, 2a,c). Thestriatum (ST) displays a dorsoventral gradient of cad-herin-8 expression with higher levels of transcript seendorsally (Figs. 1, 3a,c).

The signals in diencephalic nuclei are weaker than thosein the basal ganglia. In the epithalamus, the medialhabenular nucleus (MHb) contains densely packed cad-herin-8-positive cells, whereas the lateral habenularnucleus (LHb) only contains a few scattered cells express-ing this molecule (Fig. 5a). The positive thalamic struc-tures are the anterodorsal, anteroventral, anteromedial,ventrolateral (VL; Figs. 1, 5a), ventromedial (VM; Fig.2a,c), reticular, laterodorsal (LD; Fig. 1), lateroposterior,centrolateral (CL; Fig. 5a), paracentral (PC; Fig. 5a),reuniens, and paratenial thalamic nuclei. Unlike the otherthalamic nuclei, the subthalamic nucleus (STh) showsvery high levels of cadherin-8 transcript (Figs. 1, 2a,c). Thezona incerta (zi; Fig. 2a,c), the entopeduncular nucleus(EP; Figs. 1, 2a,c), the anterior pretectal nucleus, and thenucleus of the posterior commissure also express cad-herin-8. In the hypothalamus, part of the ventromedialnucleus is cadherin-8 positive. A list of all cadherin-8-positive structures of the forebrain is given in Table 1.

The distribution of cadherin-8-positive cells in the thala-mus is different from that of N-cadherin-positive cells(Redies and Takeichi, 1993). For example, the paraventricu-lar thalamic nucleus (PV; Fig. 5) and the medial geniculatenucleus express N-cadherin but not cadherin-8. The centro-lateral (CL), paracentral (PC), and ventrolateral nuclei(VL) express cadherin-8 but not N-cadherin (Fig. 5).Similarly distinct expression patterns were found in allother divisions of the CNS, although there are also regionsthat express both molecules.

Cadherin-8 expression in the peri-and postnatal midbrain

In the midbrain, cadherin-8-positive structures (Table 1)are the dorsal tegmental nuclei (DTg; Fig. 2a,c), theinterpeduncular nucleus (IP; Fig. 6b,d), and the dorsal(DR), median (MnR), and paramedian (PMR) raphe nuclei(Fig. 6). The substantia nigra, pars reticulata (SN), con-tains dispersed cells expressing cadherin-8 (Fig. 1). Thepedunculopontine nucleus (PPN; Fig. 6a,c) contains smallclusters of positive cells. The red nucleus weakly stains forcadherin-8 and positive cells are also found in the interme-diate gray layer of the superior colliculus (SC; Fig. 6a,c).

The expression pattern of cadherin-8 in the cerebellumchanges during the perinatal period examined in thisstudy. At E18 and P2, the lateral, interposed, and medialcerebellar nuclei (Lat, Int, and Med) and narrow segmentsof the cerebellar cortex are cadherin-8-positive (Fig. 7a).The cadherin-8-positive segments in the cerebellar cortex

TABLE 1. Cadherin-8-Positive Areas in the Peri- and Postnatal (E18-P6)Mouse Central Nervous System

ForebrainCortex

Limbic cortexCingulate cortexPerirhinal cortexPiriform cortexRetrosplenial cortex

HippocampusEntorhinal cortexOther cortical areas (layer V)

Amygdala (central nuclear group)Nucleus of diagonal bandClaustrumSeptal nuclei (medial and lateral)Basal ganglia

Globus pallidusStriatum

ThalamusAnterodorsal thalamic nucleusAnteromedial thalamic nucleusAnteroventral thalamic nucleusCentrolateral thalamic nucleusLaterodorsal thalamic nucleusLateroposterior thalamic nucleusParacentral thalamic nucleusParatenial thalamic nucleusReticular thalamic nucleusReuniens thalamic nucleusVentrolateral thalamic nucleusVentromedial thalamic nucleus

Ventromedial nucleus of hypothalamusAnterior pretectal nucleusHabenular nuclei (medial and lateral)Entopeduncular nucleusSubthalamic nucleusZona incerta

MidbrainInterpeduncular nucleusLaterodorsal tegmental nucleusPedunculopontine tegmental nucleusMedian raphe nucleusDorsal raphe nucleusParamedian raphe nucleusNuclei of posterior commissureRed nucleusSubstantia nigraSuperior colliculus (intermediate gray layer)

PonsFacial nucleusPontine nucleusPontine reticular nucleusReticulotegmental nucleus of the pons

CerebellumCerebellar nuclei (lateral, interposed and medial)Granule cell layer (P6)Purkinje cell layer (P4–6)

MedullaSpinal trigeminal nucleusInferior olive (principal nucleus and cap of Kooy)Vestibular nucleusNucleus of the solitary tract (medial part)Hypoglossal nucleusPrepositus hypoglossal nucleus

Spinal cordScattered cells in gray matter


are bilaterally and symmetrically located about the mid-line. Consecutive horizontal sections reveal that the seg-ments extend parasagittally in a stripe-like fashion overseveral lobules (data not shown). At P4, cadherin-8 expres-sion can be observed in the cerebellar nuclei and inbilaterally symmetrical segment-like stripes in the Pur-kinje cell layer. Again, these stripes extend parasagittallyover several lobules (Fig. 7b,d). There were two stripes oneach side. One stripe is located as indicated by thearrowheads in Figure 7b. The other stripe is present at thelateral margin of the cerebellum and was not found at allsectioning levels. At P4, no staining is seen in the granulecell layer (Fig. 7b,d). At P6, cadherin-8 expression by cellsin the Purkinje cell layer remains restricted to parasagit-tal segments. At this stage, cells in the inner granularlayer also express cadherin-8, but these cells are homoge-neously distributed and do not form segments (Fig. 7c,e).

Cadherin-8 expression in the peri-and postnatal hindbrain and spinal cord

Nuclei expressing cadherin-8 in the hindbrain are listedin Table 1. In the pons, the pontine nucleus (Pn) and thereticulotegmental nucleus (RtTg) are strongly cadherin-8positive (Fig. 8a,b). Additionally, cadherin-8-expressing

cells are dispersed in the pontine reticular nucleus. In themedulla oblongata, small cell clusters in the spinal vestibu-lar nucleus (SpVe) and in the medial part of the nucleus ofthe solitary tract (Sol) express cadherin-8 (Fig. 8c,d).Subdivisions of the inferior olive, such as the principalnucleus (IOPr; Fig. 8c,d) and the cap of Kooy also showcadherin-8 expression.

One of the areas expressing cadherin-8 most strongly inthe hindbrain is the facial nucleus (Fig. 1; Korematsu andRedies, 1997). In the floor of the fourth ventricle, theprepositus hypoglossal nucleus (PrH) is weakly positive(Fig. 8c,d). The hypoglossal nucleus (XII) also containslabeled cells (Fig. 8e,f). In addition, cadherin-8-positivecells are observed in the motor and spinal trigeminalnucleus (SpV; Fig. 8c,d), and in the trigeminal ganglion(data not shown). In the spinal cord, cadherin-8 is ex-pressed in scattered cells of the gray matter without anyclear restriction to circumscribed areas (Fig. 9).

Cadherin-8 expression in the embryonic(E12.5–E16) forebrain

As described before (Korematsu and Redies, 1997),expression of cadherin-8 is restricted to particular develop-ing gray matter also at earlier stages of brain develop-

Fig. 3. Frontal sections through the embryonic day 18 mouse basal ganglia (a) and the postnatal day 4hippocampus (b) hybridized with cadherin-8 anti-sense probe. Adjoining sections stained for Nisslsubstance are shown in c and d. For abbreviations, see list. Scale bars 5 600 µm in a,c, 200 µm in b,d.


ment. In some brain regions, this expression relates to theneuromeric organization of the early embryonic brain. Forexample, at E12.5, cadherin-8 expression is prominent inthe medial ganglionic eminence (MGE) and in the anlageof the claustrum, whereas the lateral ganglionic eminence(LGE) shows low levels of transcript. The boundariesbetween these structures represent longitudinal bordersin the prosomeric model by Bulfone et al. (1993). At least inpart, these borders coincide with borders of cadherin-8expression (arrowheads in Fig. 10a). In the diencephalon,the borders between the pretectum and the dorsal thala-mus (DT) and between DT and the ventral thalamus (VT)also co-localize with changes in expression of cadherin-8 in

the mantle layer (arrowheads in Fig. 10b; see also Kore-matsu and Redies, 1997).

Some of the regions expressing cadherin-8 in the earlyembryonic brain can be followed to differentiate intoparticular gray matter structures that express the samemolecule at peri- and postnatal stages (Table 1). Forexample, in the telencephalon, parts of the neocortex (Cx),the claustrum (Cl), parts of the amygdala (Amy), and thenucleus of the diagonal band all derive from anlagen,which also express cadherin-8 at E12.5 (Fig. 10a,b). Theputamen (cp and vp) is also positive at E16 (Fig. 10d).Figure 10b,c,e shows that, in the diencephalon, parts of thehabenula (LHb), the anterior pretectal nucleus (ap), the

Fig. 4. Transverse sections through the postnatal day (P) 4 mouseparietal cortex (a,b) and the P6 hippocampal area (c,d) hybridizedwith cadherin-8 anti-sense probe (a,c) and stained for Nissl substance(b,d). In parietal cortex, cadherin-8 expression is most prominent inlayer V (a). At the transition from perirhinal to temporal cortex,

cadherin-8 expression falls off (arrowhead in c). The panel pairs a,band c,d represent adjoining sections. The orientation of the sectionsshown in c and d (c, caudal; l, lateral; m, medial; r, rostral) is given in c.For abbreviations, see list. Scale bars 5 50 µm in a,b, 150 µm in c,d.


ventrolateral migratory stream (VLm) of the dorsal thala-mus (DT), the reticular migratory stream (rtm) of theventral thalamus (VT; Altman and Bayer, 1978), the zonaincerta (zi), and the ventromedial nucleus of the hypothala-mus (vm) are gray matter areas that express cadherin-8 atE14 and E16. All these structures continue to express thismolecule at peri- and postnatal stages (see Table 1).

DISCUSSION

The vertebrate CNS shows a highly complex architec-ture. It consists of a number of functional units that arespecialized to process particular types of information, suchas motion, sensation, emotion, coordination, memory, etc.During development, the architecture of the CNS derivesfrom a relatively simple sheet of neuroepithelial cells. Ithas been suggested that cadherins, a family of morphoge-netic cell adhesion molecules, regulate the segregation andaggregation of early neurons and neurites during develop-ment and thereby contribute to the formation of functionalunits from a relatively homogeneous assembly of undiffer-entiated cells (reviewed by Redies, 1995; Redies andTakeichi, 1996). In the present paper, we show that, like

other cadherins, cadherin-8 is expressed by a subset ofgray matter structures in the developing mouse brain.These results suggest that cadherin-8 plays a role in theregionalization of the developing brain. At early times ofdevelopment, the cadherin-8-positive regions reflect theneuromeric organization of the embryonic mouse brain(Fig. 10; Korematsu and Redies, 1997). During laterdevelopment, cadherin-8 is expressed by a subset of differ-entiating brain nuclei, cortical layers, and their earlyembryological anlagen. Moreover, in cerebral and cerebel-lar cortex, cadherin-8 transcript is also found in specificsubdivisions of these structures.

Association of cadherin-8-positive structureswith functional systems

Previous studies have shown that the gray matter areasexpressing a particular type of cadherin can be function-ally connected to each other by fiber tracts expressing thesame cadherin (Redies et al., 1993). For example, R-cadherin is expressed by specific neural circuits that formparts of several functional systems in the embryonicchicken brain (Arndt and Redies, 1996). The existence ofcadherin-8-positive fiber tracts could not be demonstratedin the present work because the expression of this mol-ecule was studied at the mRNA level. Antibodies forimmunostaining were not available. Nevertheless, it isworthwhile to note that most of the cadherin-8-expressinggray matter structures are associated with a particular setof functional systems in the CNS. In the following section,we will discuss these associations and point out some ofthe known fiber connections between the cadherin-8-positive areas. Whether the cadherin-8-positive gray mat-ter structures are indeed connected by cadherin-8-positivefiber tracts remains to be studied in the future.

Limbic system. One of the functional systems contain-ing several cadherin-8-positive structures is the limbicsystem, which consists of the limbic cortex, hippocampus,septal area, hypothalamus, anterior part of the thalamus,amygdala, habenula, etc. These structures are closelyinterconnected to one another, as reviewed by Lopes daSilva et al. (1990), Amaral and Witter (1995), Zilles andWree (1995), and Jakab and Lerath (1995). For example,fibers from the hippocampus reach the cingulate cortex,entorhinal cortex, and the septal area. The hippocampusreceives projections from the medial habenular nucleus.The cingulate cortex, a part of the limbic cortex, receivesefferents from the anterior and midline thalamic nuclei,such as the reuniens and paracentral nuclei, and from theamygdala, septal area, claustrum, and median and dorsalraphe nuclei. Efferent fibers from the medial habenularnucleus project to the interpeduncular nucleus (Marchandet al., 1980; Shibata et al., 1986), to the medial septalnucleus, and to the median raphe nucleus. An afferentprojection to the medial habenular nucleus originates inthe nucleus of diagonal band. A projection of the dorsal andmedian raphe nuclei to the amygdala and septal nucleiwas also reported (Ma et al., 1991). All these structuresexpress cadherin-8 (Table 1). Moreover, although cad-herin-8 expression in cerebral cortex is widespread at E18to P4, highest levels of expression are consistently seen inlimbic cortex. At P6, cadherin-8 transcript is largelyrestricted to limbic cortical areas. This finding suggests apossible role for cadherin-8 in the regionalization of thecerebral cortex. The association of cadherin-8 with thelimbic system is reminiscent of the regionalized expression

Fig. 5. Adjoining frontal sections through the postnatal day 2mouse diencephalon at the level of the epithalamus hybridized withcadherin-8 anti-sense probe (Cad8) (a) and with N-cadherin anti-senseprobe (N-cad) (b). Note the differential staining of diencephalic nuclei.The arrowhead in b points at ependymal staining. For abbreviations,see list. Scale bar 5 300 µm.


of the limbic system-associated protein (LAMP), a memberof the immunoglobulin superfamily of molecules (Levitt,1984; Pimenta et al., 1995).

Basal ganglia and related nuclei. The basal gangliaand related nuclei (reviewed by Alexander and Crutcher,1990; Heimer et al., 1995) also contain a number ofcadherin-8-positive structures that are interconnected.For example, the striatum has reciprocal fiber connectionswith the substantia nigra, receives massive input from thecerebral cortex, and projects to the globus pallidus and theentopeduncular nucleus. The cadherin-8-positive cells

found in layer V of several cortical areas might possiblyparticipate in the corticostriatal projection (Akintundeand Buxton, 1992). The globus pallidus receives efferentsfrom the striatum and subthalamic nucleus, and sendsfibers to the subthalamic nucleus and substantia nigra.The subthalamic nucleus projections terminate in theentopeduncular nucleus and substantia nigra. The entope-duncular nucleus and substantia nigra send fibers to thepedunculopontine nucleus and to the thalamic nuclei, e.g.,to the ventral anterior, ventromedial, and ventrolateralnucleus. The latter nuclei, in turn, project to the cerebral

Fig. 6. Transverse sections through the embryonic day 18 mid-brain (a) and the postnatal day 2 ventral pons (b) of a mouse brainhybridized with cadherin-8 anti-sense probe.Adjoining sections stained

for Nissl substance are shown in c,d. The arrowheads in (a) point at acadherin-8-positive layer of the superior colliculus. For abbreviations,see list. Scale bars 5 300 µm.


Fig. 7. Horizontal sections of the embryonic day (E) 18 (a),postnatal day (P) 4 (b,d), and P6 (c,e) mouse cerebellum hybridizedwith cadherin-8 anti-sense probe. d,e: Nomarski optical enlargementsof the boxed areas indicated in b and c, respectively. The arrowheads in(a–c) point at segments of cadherin-8 expression in the developing

cerebellar cortex. The arrowheads in (d) and (e) point at bordersbetween groups of cadherin-8-positive and -negative Purkinje cells.The labeling of the external granular layer in (a–c) is unspecific. Seetext for a detailed description of these results. For abbreviations, seelist. Scale bars 5 500 µm in a–c, 30 µm in d,e.


cortex. In addition, a projection from the entopeduncularnucleus to the lateral habenular nucleus was reported(Herkenham and Nauta, 1977).

Cerebellum and associated nuclei. Like R-cadherinin the chicken cerebellum (Arndt and Redies, 1996), cad-herin-8 is expressed in parasagittal stripes in the E18 andP2 mouse cerebellum. At P4 and P6, these stripes arerestricted to the Purkinje cell layer. Similar stripes havebeen found by electrophysiological and histochemicalmeans and by studying the efferent and afferent connec-

tions in the vertebrate cerebellar cortex (Armstrong et al.,1974; Oscarsson, 1979; Wassef et al., 1985; Hawkes andLeclerc, 1987). Evidence for a genetic control of the devel-opment of this segmental pattern has been reported (Ober-dick et al., 1993; Millen et al., 1995). Conceivably, therestriction of cadherin-8 expression to particular parasag-ittal stripes is related to the differential expression of thismolecule in nuclei that are connected to the cerebellum, ashas been suggested for R-cadherin (Arndt and Redies,1996). For example, the afferent terminals from subgroups

Fig. 8. Transverse sections of the P2 midbrain-pontine junction(a), the E18 medulla (c) and the P2 pontine-medullary junction (e) ofa mouse brain hybridized with cadherin-8 anti-sense probe. Adjoining

sections stained for Nissl substance are shown in (b,d and f). Theorientation for all sections (d, dorsal; v, ventral) is shown in panel (c).Abbreviations are listed separately. Scale bars 5 300 mm.


of inferior olivary neurons are located in particular stripes(Armstrong et al., 1974; Apps, 1990) but their relation tothe cadherin-8-positive stripes remains to be confirmed.Interestingly, only a subset of inferior olivary neuronsexpresses cadherin-8.

As reviewed by Voogt (1995) and Ruigrok and Cella(1995), the cerebellum has a feedback system through thered nucleus and inferior olive. The pontine nucleus is alsoa source of cerebellar afferents. The cerebellar nucleiproject to the superior colliculus, to the anterior pretectalnucleus, and to the nucleus of the posterior commissure.All these structures express cadherin-8.

Hindbrain nuclei. Some of the cranial nerve nuclei ofthe brainstem, such as the facial nucleus, vestibularnucleus, spinal trigeminal nucleus, nucleus of the solitarytract, and hypoglossal nucleus also express cadherin-8, atleast in part. These nuclei have fiber connections with theother nuclei expressing cadherin-8. The facial nucleusreceives projections from the red nucleus (Travers andNorgren, 1983). The spinal trigeminal nucleus, prepositushypoglossal nucleus, and vestibular nucleus are connectedto the cerebellum and inferior olive (reviewed by Voogt,1995; Ruigrok and Cella, 1995). The hypoglossal nucleus

receives afferent fibers arising from the dorsal part of thespinal trigeminal nucleus (Borke et al., 1983).

Taken together, these results suggest the possibilitythat, like the N- and R-cadherin-expressing structures inthe embryonic chicken brain (Redies et al., 1993; Arndtand Redies, 1996), many of the cadherin-8-positive areas ofthe mouse brain are functionally connected. It should bestressed, however, that the results from the present studyare inconclusive with regard to this possibility becausecadherin-8-positive fiber tracts could not be visualized.

Comparison to cadherin-8 expressionin the early embryonic brain

In a previous study, the expression of cadherin-8 wasstudied in the mouse brain at early stages of CNS develop-ment (E9–E14) (Korematsu and Redies, 1997). Resultsshowed that, like R-, E-, and F-cadherin and cadherin-11(Espeseth et al., 1995; Ganzler and Redies, 1995; Matsu-nami and Takeichi, 1995; Kimura et al., 1996), cadherin-8expression is restricted to specific neuromeric subdivi-sions. Similar results were found in the present work (seearrowheads in Fig. 10a,b). These findings indicate thatcadherin-mediated differential adhesiveness of neural cellsplays a role in the regionalization also of the early embry-onic CNS. In the mantle zone of some of these embryonicsubdivisions, cadherin-8-expressing populations of cellswere found to accumulate and sort out from cadherin-8-negative ones. The present study shows that the cadherin-8-positive cell aggregates persist in the mantle layer atlater stages of development (E18–P6) and differentiateinto specific brain nuclei and cortical regions or partsthereof. By studying the expression of cadherin-8, severalgray matter structures can thus be followed from their(neuromeric) origins throughout most of their develop-ment. Examples are the basal ganglia, parts of the amyg-dala, the claustrum (Filimonoff, 1966), the nucleus of thediagonal band, and several diencephalic nuclei, such as theanterior pretectal nucleus, the ventrolateral and reticularnuclear complexes, the zona incerta, the habenula, and theventromedial nucleus of the hypothalamus.

Comparison with other molecules

The neuroanatomical structures expressing cadherin-8differ from those expressing N-cadherin in the embryonicmouse brain (Fig. 5) (Redies and Takeichi, 1993), althoughthere is some overlap in the expression patterns. More-over, E-cadherin and cadherin-11 also show a distinctexpression pattern in the developing mouse brain (Shi-mamura and Takeichi, 1992; Matsunami and Takeichi,1995; Kimura et al., 1996). Differential expression pat-terns were also described for N-, R-, and B-cadherin in thedeveloping chicken CNS (Inuzuka et al., 1991b; Redies etal., 1993; Murphy-Erdosh et al., 1994; Arndt and Redies,1996). These results support the idea that cadherinsconstitute a system of molecules that can be used asmarkers for functional neuroanatomical structures, suchas brain nuclei, and regions and layers of cerebral andcerebellar cortex (Redies et al., 1993). Because cadherinsare expressed during the development of these structures,they may play a direct role in regulating their formation.Other types of molecules that are expressed in a restrictedmanner by particular gray matter areas, also contribute tothese processes. These molecules include members of theimmunoglobulin superfamily of adhesion molecules (e.g.,axonin-1, BIG-1, BIG-2, and LAMP; Levitt, 1984; Zuellig

Fig. 9. Transverse section of the postnatal day 2 mouse spinal cordhybridized with cadherin-8 anti-sense probe (a) and adjoining Nisslstain (b). The orientation of the sections (d, dorsal; v, ventral) is shownin a. Scale bar 5 100 µm.


et al., 1992; Yoshihara et al., 1994; 1995; Pimenta et al.,1995; Rager et al., 1995; Redies et al., 1997), otheradhesion molecules (e.g., neuropilin; Takagi et al., 1995),and proteoglycans (e.g., Cat. 301; Hockfield et al., 1990).

ACKNOWLEDGMENTS

We thank M. Takeichi for N-cadherin cDNA, U. Schwarzfor generous support, L. Puelles for neuroanatomical sug-gestions, and V. Kastner for technical assistance.

LITERATURE CITED

Akintunde, A., and D.F. Buxton (1992) Origins and collateralization ofcorticospinal, corticopontine, corticorubral and corticostriatal tracts: Amultiple retrograde fluorescent tracing study. Brain Res. 586:208–218.

Alexander, G.E., and M.D. Crutcher (1990) Functional architecture of basalganglia circuits: Neural substrates of parallel processing. TrendsNeurosci. 13:266–271.

Altman, J., and S.A. Bayer (1978) Development of the diencephalon in therat. I. Autoradiographic study of the time of origin and settling patternsof neurons of the hypothalamus. J. Comp. Neurol. 182:945–972.

Amaral, D.G., and M.P. Witter (1995) Hippocampal formation. In G.

Fig. 10. Transverse sections through the forebrain of embryonicday (E) 12.5 (a,b), E14 (c), and E16 (d,e) embryos hybridized withcadherin-8 anti-sense probe. The orientation of all sections (d, dorsal;v, ventral) is shown in (e). The arrowheads in (a) point at borders ofcadherin-8 staining between the medial and the lateral ganglioniceminence (MGE, LGE) and between LGE and neocortex. The arrow-

heads in b point at the borders between the pretectal area and thedorsal thalamus (DT), and between DT and the ventral thalamus (VT).The arrows in b and c point at the retroflex fascicle. The stars in dindicate artifactual staining. For other abbreviations, see list. Scalebars 5 200 µm in a,b, 500 µm in c–e.


Paxinos (ed): The Rat Nervous System. San Diego: Academic Press, pp.443–493.

Apps, R. (1990) Columnar organisation of the inferior olive projection to theposterior lobe of the rat cerebellum. J. Comp. Neurol. 302:236–254.

Armstrong, D.M., R.J. Harvey, and R.F. Schild (1974) Topographicallocalization in the olivo-cerebellar projection: An electrophysiologicalstudy in the cat. J. Comp. Neurol. 154:287–302.

Arndt, K., and C. Redies (1996) Restricted expression of R-cadherin bybrain nuclei and neural circuits of the developing chicken brain. J.Comp. Neurol. 373:373–399.

Beesley, P.W., R. Mumery, J. Tibaldi, A.P. Chapman, and S.J. Smith (1995)The postsynaptic density: Putative involvement in synapse stabiliza-tion via cadherins and covalent modification by ubiquitination. Bio-chem. Soc. Trans. 23:59–64.

Borke, R.C., M.E. Nau, and R.L. Ringler (1983) Brainstem afferents ofhypoglossal neurons in the rat. Brain Res. 269:47–55.

Bulfone, A., L. Puelles, M.H. Porteus, M.A. Frohmann, G.R. Martin, andJ.L.R. Rubenstein (1993) Spatially restricted expression of Dlx-1, Dlx-2(Tes-1), Gbx-2, and Wnt-3 in the embryonic day 12.5 mouse forebraindefines potential transverse and longitudinal segmental boundaries. J.Neurosci. 13:3155–3172.

Espeseth, A., E. Johnson, and C. Kintner (1995) Xenopus F-cadherin, anovel member of the cadherin family of cell adhesion molecules, isexpressed at boundaries in the neural tube. Mol. Cell. Neurosci.6:199–211.

Fannon, A.M., and D.R. Colman (1996) A model for central synapticjunctional complex formation based on the differential adhesive speci-ficities of the cadherins. Neuron 17:423–434.

Filimonoff, I.N. (1966) The claustrum, its origin and development. J.Hirnforsch. 8:503–528.

Ganzler, S.I.I., and C. Redies (1995) R-cadherin expression during nucleusformation in chicken forebrain neuromeres. J. Neurosci. 15:4157–4172.

Hatta, K., A. Nose, A. Nagafuchi, and M. Takeichi (1988) Cloning andexpression of cDNA encoding a neural calcium-dependent cell adhesionmolecule: Its identity in the cadherin gene family. J. Cell Biol. 106:873–881.

Hawkes, R., and N. Leclerc (1987) Antigenic map of the rat cerebellarcortex: The distribution of parasagittal bands as revealed by monoclo-nal anti-Purkinje cell antibody mabQ113. J. Comp. Neurol. 256:29–41.

Heimer, L., D.S. Zahm, and G.F. Alheid (1995) Basal ganglia. In G. Paxinos(ed): The Rat Nervous System. San Diego: Academic Press, pp. 579–628.

Herkenham, M., and W.J.H. Nauta (1977) Afferent connections of thehabenular nuclei in the rat: A horseradish peroxidase study, with a noteon the fiber-of-passage problem. J. Comp. Neurol. 173:123–146.

Hockfield, S., R.G. Kalb, S. Zaremba, and H. Fryer (1990) Expression ofneural proteoglycans correlates with the acquisition of mature neuro-nal properties in the mammalian brain. Cold Spring Harbor Symp.Quant. Biol. 55:505–514.

Inuzuka, H., S. Miyatani, and M. Takeichi (1991a) R-cadherin: A novelCa21-dependent cell-cell adhesion molecule expressed in the retina.Neuron 7:69–79.

Inuzuka, H., C. Redies, and M. Takeichi (1991b) Differential expression ofR- and N-cadherin in neural and mesodermal tissues during earlychicken development. Development 113:959–967.

Jakab, R.L., and C. Leranth (1995) Septum. In G. Paxinos (ed): The RatNervous System. San Diego: Academic Press, pp. 405–442.

Kimura, Y., H. Matsunami, and M. Takeichi (1996) Expression of cad-herin-11 delineates boundaries, neuromeres, and nuclei in the develop-ing mouse brain. Dev. Dyn. 206:455–462.

Korematsu, K., and C. Redies (1997) Restricted expression of cadherin-8 insegmental and functional subdivisions of the embryonic mouse brain.Dev. Dyn. 208:178–189.

Levitt, P. (1984) A monoclonal antibody to limbic system neurons. Science223:299–301.

Lopes da Silva, F.H., M.P. Witter, P.H. Boeijinga, and A.H.M. Lohman(1990) Anatomic organization and physiology of the limbic cortex.Physiol. Rev. 70:453–511.

Ma, Q.P., G.F. Yin, M.K. Ai, and J.S. Han (1991) Serotonergic projectionsfrom the nucleus raphe dorsalis to the amygdala in the rat. Neurosci.Lett. 134:21–24.

Marchand, E.R., J.N. Riley, and R.Y. Moore (1980) Interpeduncular nucleusafferents in the rat. Brain Res. 193:339–352.

Matsunami, H., and M. Takeichi (1995) Fetal subdivisions defined by R-and E-cadherin expressions: Evidence for the role of cadherin activity inregion-specific, cell-cell adhesion. Dev. Biol. 172:466–478.

Matsunami, H., S. Miyatani, T. Inoue, N.G. Copeland, D.J. Gilbert, N.A.Jenkins, and M. Takeichi (1993) Cell binding specificity of mouseR-cadherin and chromosomal mapping of the gene. J. Cell Sci. 106:401–409.

Millen, K.J., C.C. Hui, and A.L. Joyner (1995) A role for En-2 and othermurine homologues of Drosophila segment polarity genes in regulatingpositional information in the developing cerebellum. Development121:3935–3945.

Miyatani, S., K. Shimamura, M. Hatta, A. Nagafuchi, A. Nose, M. Matsu-naga, K. Hatta, and M. Takeichi (1989) Neural cadherin: Role inselective cell-cell adhesion. Science 245:631–635.

Murphy-Erdosh, C., E.W. Napolitano, and L.F. Reichardt (1994) Theexpression of B-cadherin during embryonic chick development. Dev.Biol. 161:107–125.

Nakagawa, S., and M. Takeichi (1995) Neural crest cell-cell adhesioncontrolled by sequential and subpopulation-specific expression of novelcadherins. Development 121:1321–1332.

Nose, A., A. Nagafuchi, and M. Takeichi (1988) Expressed recombinantcadherins mediate cell sorting in model systems. Cell 54:993–1001.

Oberdick, J., K. Schilling, R.J. Smeyne, J.G. Corbin, C. Bocchiaro, and J.I.Morgan (1993) Control of segment-like patterns of gene expression inthe mouse cerebellum. Neuron 10:1007–1018.

Oscarsson, O. (1979) Functional units of the cerebellum: Sagittal zones andmicrozones. Trends Neurosci. 2:143–145.

Pimenta, A.F., V. Zhukareva, M.F. Barbe, B.S. Reinoso, C. Grimley, W.Henzel, I. Fischer, and P. Levitt (1995) The limbic system-associatedmembrane protein is an Ig superfamily member that mediates selectiveneuronal growth and axon targeting. Neuron 15:287–297.

Rager, G., P. Morino, J. Schnitzer, and P. Sonderegger (1995) Expression ofthe axonal cell adhesion molecules axonin-1 and Ng-CAM during thedevelopment of the chick retinotectal system. J. Comp. Neurol. 365:594–609.

Redies, C. (1995) Cadherin expression in the developing vertebrate brain:From neuromeres to brain nuclei and neural circuits. Exp. Cell Res.220:243–256.

Redies, C., and H.-A.J. Muller (1994) Similarities in structure and expres-sion between mouse P-cadherin, chicken B-cadherin and frog XB/U-cadherin. Cell Adhes. Commun. 2:511–520.

Redies, C., and M. Takeichi (1993) Expression of N-cadherin mRNA duringdevelopment of the mouse brain. Dev. Dyn. 197:26–39.

Redies, C., and M. Takeichi (1996) Cadherins in the developing centralnervous system: An adhesive code for segmental and functional subdivi-sions. Dev. Biol. 180:413–423.

Redies, C., H. Inuzuka, and M. Takeichi (1992) Restricted expression of N-and R-cadherin on neurites of the developing chicken CNS. J. Neurosci.12:3525–3534.

Redies, C., K. Engelhart, and M. Takeichi (1993) Differential expression ofN- and R-cadherin in functional neuronal systems and other structuresof the developing chicken brain. J. Comp. Neurol. 333:398–416.

Redies, C., K. Arndt, and M. Ast (1997) Expression of the cell adhesionmolecule axonin-1 in neuromeres of the chicken diencephalon. J. Comp.Neurol. 381:230–252.

Rose, O., C. Grund, S. Reinhardt, A. Starzinski-Powitz, and W.W. Franke(1995) Contactus adherens, a special type of plaque-bearing adheringjunction containing M-cadherin, in the granular cell layer of thecerebellar glomerulus. Proc. Natl. Acad. Sci. USA 92:6022–6026.

Ruigrok, T.J.H., and F. Cella (1995) Precerebellar nuclei and red nucleus. InG. Paxinos (ed): The Rat Nervous System. San Diego: Academic Press,pp. 277–308.

Sago, H., M. Kitagawa, S. Obata, N. Mori, S. Taketani, J.M. Rochelle, M.F.Seldin, M. Davidson, T. St. John, and S.T. Suzuki (1995) Cloning,expression, and chromosomal localization of a novel cadherin-relatedprotein, protocadherin-3. Genomics 29:631–640.

Sano, K., H. Tanihara, R.L. Heimark, S. Obata, M. Davidson, T. St. John, S.Taketani, and S. Suzuki (1993) Protocadherins: A large family ofcadherin-mediated molecules in central nervous system. EMBO J.12:2249–2256.

Shibata, H., T. Suzuki, and M. Matsushita (1986) Afferent projections to theinterpeduncular nucleus in the rat, as studied by retrograde andanterograde transport of wheat germ agglutinin conjugated to horserad-ish peroxidase. J. Comp. Neurol. 248:272–284.

Shimamura, K., and M. Takeichi (1992) Local and transient expression ofE-cadherin involved in mouse embryonic brain morphogenesis. Develop-ment 116:1011–1019.

Shimamura, K., T. Takahashi, and M. Takeichi (1992) E-cadherin expres-sion in a particular subset of sensory neurons. Dev. Biol. 152:242–254.


Suzuki, S. (1996) Structural and functional diversity of cadherin superfam-ily: Are new members of cadherin superfamily involved in signaltransduction pathway? J. Cell. Biochem. 61:531–542.

Suzuki, S., K. Sano, and H. Tanihara (1991) Diversity of the cadherinfamily: Evidence for eight new cadherins in nervous tissue. Cell Regul.2:261–270.

Takagi, S., Y. Kasuya, M. Shimizu, T. Matsuura, M. Tsuboi, A. Kawakami,and H. Fujisawa (1995) Expression of a cell adhesion molecule, neuropi-lin, in the developing chick nervous system. Dev. Biol. 170:207–222.

Takeichi, M. (1988) The cadherins: Cell-cell adhesion molecules controllinganimal morphogenesis. Development 102:639–655.

Takeichi, M. (1995) Morphogenetic roles of classic cadherins. Curr. Opin.Cell Biol. 7:619–627.

Takeichi, M., H. Inuzuka, K. Shimamura, T. Fujimori, and A. Nagafuchi(1990) Cadherin subclasses: Differential expression and their roles inneural morphogenesis. Cold Spring Harbor Symp. Quant. Biol. 55:319–325.

Tanihara, H., K. Sano, R.L. Heimark, T. St. John, and S. Suzuki (1994)Cloning of five cadherins clarifies characteristic features of cadherinextracellular domain and provides further evidence for two structurallydifferent types of cadherin. Cell Adhes. Commun. 2:15–26.

Travers, J.B., and R. Norgren (1983) Afferent projections to the oral motornuclei in the rat. J. Comp. Neurol. 220:280–298.

Uchida, N., Y. Honjo, K.R. Johnson, M.J. Wheelock, and M. Takeichi (1996)The catenin/cadherin adhesion system is localized in synaptic junctionsbordering transmitter release zones. J. Cell Biol. 135:767–779.

Voogt, J. (1995) Cerebellum. In G. Paxonis (ed): The Rat Nervous System.San Diego: Academic Press, pp. 309–350.

Wassef, M., J.P. Zanetta, A. Brehier, and C. Sotelo (1985) Transientbiochemical compartmentalization of Purkinje cells during early cerebel-lar development. Dev. Biol. 111:129–137.

Yamagata, M., J.P. Herman, and J.R. Sanes (1995) Lamina-specific expres-sion of adhesion molecules in developing chick optic tectum. J. Neuro-sci. 15:4556–4571.

Yoshihara, Y., M. Kawasaki, A. Tani, A. Tamada, S. Nagata, H. Kagami-yama, and K. Mori (1994) BIG-1: A new TAG-1/F3-related member ofthe immunoglobulin superfamily with neurite outgrowth-promotingactivity. Neuron 13:415–426.

Yoshihara, Y., M. Kawasaki, A. Tamada, S. Nagata, H. Kagamiyama, and K.Mori (1995) Overlapping and differential expression of BIG-2, BIG-1,TAG-1, and F3: Four members of an axon-associated cell adhesionmolecule subgroup of the immunoglobulin superfamily. J. Neurobiol.28:51–69.

Zilles, K., and A. Wree (1995) Cortex: Areal and laminar structure. In G.Paxinos (ed): The Rat Nervous System. San Diego: Academic Press, pp.649–685.

Zuellig, R.A., C. Rader, A. Schroeder, M.B. Kalousek, F. Von Bohlen undHalbach, T. Osterwalder, C. Inan, E.T. Stoeckli, H.-U. Affolter, A. Fritz,E. Hafen, and P. Sonderegger (1992) The axonally secreted cell adhesionmolecule, axonin-1: Primary structure, immunoglobulin-like and fibro-nectin-type-III-like domains and glycosyl-phosphatidylinositol anchor-age. Eur. J. Biochem. 204:453–463.


expression of cadherin-8 mrna in the developing mouse central nervous system

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