Rat Hair Follicle Dermal Papillae Have an Extracellular ...component cell s have an inductive effect in the genesis of hair follicles in the embryo [4]. Other evidence [5-7] has strongly

Rat Hair Follicle Dermal Papillae Have an Extracellular Matrix Containing Basement Membrane Components

J o hn R . C Oll chman, Ph.D. * l3ioscienccs D ivision, Unilcve r Resca rch, Co lworth Labo rato ry, Sharnbrook, Bcdfo rd, U . K.

Derm al papill ae are small mesenchym all y derived zones at the bases o f hai r follicles which have an important ro le in hair mo rphogenesis in the embryo and control of the hair g ro wth cycle in postn atal m ammals. The cells o f the papilla are enmeshed in a dense ex tracellular matri x which under­goes extensive changes in concert with the hair cycle. H ere it is sho wn that this matri x in anagen pelage fo llicles o f pos tnatal rats contains an abundance o f basement mem­brane components rather than derm al co mponents such as interstiti al collagens. In particular, type IV collagen, lam­inin, and basement m embrane types o f chondroitin sul fa te

The dermal papilla lies at the base o f th e mammalian hair follicle and is composed o f mesenchymal cells and ; in at least some cases, small capill aries [1,2]. A presumpti ve papill a is one o f th e ea rlies t indi cato rs o f hair fo llicle fo rmation [3] and it is pro bable th at th e

component cell s have an inductive effect in th e genes is o f hair fo llicles in the embryo [4]. Other evidence [5-7] has strongly implica ted the qerm al papill a with a role in hair gro wth and contro l o f the hair cycle. The matrix is ex tensive throu gh anagen, but by telogen is virtu all y nonexistent , the papilla being repre­sented by a small ball o f cell s at the base o f the secondary germ [8].

Littl e is known of the compositio n o f th e extrace ll lar matri x of th e dermal papilla o r the biology o f its associated cells, w hich , although presumpti ve fibroblas ts, are at leas t mo rph o log ically distin ct fro m those o f the dermis. We have previo usly shown th at the anagell dermal papill a o f rat pelage follicl es is rich ill fib ronectin (9], a widespread component o f loose connective tis­sue and many basement membranes [10-1 2]. Mo re recently in a study o f hair fo llicle mo rphogenes is [1 3], we no ted th e presence of2 basement membrane compo nents-laminin [14] and ty pe IV co ll agen [1 5]- in derm al papillae. This stud y repo rts th at the derm al papilla o f rat pela ge hair fo llicles is co n1.posed o f basement

M anuscrip t received N ovem ber 14, 1985; accepted fo r publication June II , 1986.

*Presellt address: Connect ive Tissue Labo rato ry, B. R. Boshell Dia­betes Resea rch and Ed uca tio n H ospital, Uni ve rsity o f Alabama at l3ir­millgham, Ullive rsity Station, Birmin gham, Alabam a 35294.

Reprill t requests to: Jo hn n .. C ouchm an, I'h . D . , Connecti ve Tissue Labor:tto ry, B . H. l3 0shell D iabetes r~csca rch and Educa tion Hospital, U ni ve rsity of AbbamJ at l3irmingham , Uni ve rsity Sta tion, Birmingham , Ala bam a 35294.

Abbreviatio ns: E LI SA: enzy me-linked iml11unoso rbent assay FITC: flu o rescein iso thiocyanate T RITC: tctram eth ylrhoda mine isothiocyanate

and heparan sul fate prp teogl ycans are present as m atri x components. D ermal papilla cells, when initiall y isolated fro m adult rat vibrissae and cultured in vitro , retained the potential to synthesize this spectrum of matri x co mpo­nents, but this was gradually lo~ t, to be replaced by syn­thesis of o ther components includin g type I and III co lla­gens. It seem s likely therefore that the derm al papilla cell s in vivo synthes ize a basement m embrane type of ex tracel­lul ar m atri x, although a contributio n fr o m epithelial, and in some cases ca pillary endothelial , cells cann o t be ruled o ut. J I1llJesf D en/'/a/o/ 87:762- 767, 1986

l11 embrane compo nents, with little contributio n fro m interstitial type I co ll agen, but con tainin g so me type III co ll agen. In addition, de rm al papill a ce ll s fro m rat vibri ssae synthesize these co mpo­nents in vitro durin g their first few da ys in culture, but thereafter chan ge their spectrum o f ex tracellular matri x mo lecul es to resem­ble th at o f derm al fibroblas ts, with sy nthes is o f collagen types I and III , and an apparent decrease in type IV co llagen sy nthes is.

MATERIALS AND MET HODS

Tissue Preparation and Papilla Cultures Skin fro m the mid­dorsa l regio n o f freshl y sacrifi ced newbo rn (1 - 7 days after birth) albino Wistar rats was snap-frozen in isopentane chill ed in liquid nitrogen. C ryos tat sectio ns o f 5-7 ILm thickness were cut in a pl ane compatible with achiev ing longitudinal sectio ns throug h dermal papillae. The sections were air-dried o r brieAy fi xed in 3% fres hl y hydro lyzed parafo rm aldehyde befo re appli cation of antibodies as prev io usly described \12,13].

D erm al papill ae were dissected fro m adult albino Wistar rat vibrissae as described by Jaboda and Oliver [1 6], and were cul­tured in 35-111111 plasti c Petri di shes (Co rnin g, Stone, Staffo rd­shire, U. K.) in Dulbccco's 111 0difi ed cag le 's medium (G ibco, Pais­ley , Scotl and) co ntaining 15% fetal calf serum (Flow Labo rato ries, Irvine, Scotl and). Five to 20 papillae were plated at o ne time, w ith 5-10 papillae in each dish. It was fo und th at ce ll mi g ration fro l11 isobted papillae was considerabl y enhan ced if they were mildly disrupted . C ultures w ere m aintained fo r 5 o r 20 da ys fo r iml11uno Au o rescent staining. The absen ce o f contaminatin g epi­theli al cell s fro m hair fo llicles in th e cultures was checked by stainin g with po lyclo nal antise rum to keratin. This se rul11 had the capacity to stain all hair fo lli cle epitheli al ce ll s in rat skin sectio ns.

Immunologic Reagents The 2 monoclonal antibodies rec­ognizing chondro itin 4-sulfate (9-A-2) and chondro itin 6-sulfate (3-B-3) were used at a dilutio n o f 1 : 50 o f ascites A'uid . These antibodies have been fully described [17] and have been show n to stain derm al conn ecti ve tiss ue and basement membranes , re-

0022-202X/86/S03.S0 . Copyright © 1986 by The Society fo r Inves tigative Derm ato logy, Inc.

762

VOL. 87, NO. (, DEC EMUER 19k6

spectively. Second antibodies for indirect immunofluo rescence microscopy w ere flu o rescein isothi ocya nate (FITC)-conju ga ted ra bbit antil110use IgG (fo r the 9-A-2 antibody) and FITC- or tc trarn ethylrhodamine isothiocyanate (TRITC)-conju ga ted rabbit anti mouse Ig M (for the 3-B-3 antibod y) .

R a bbit antibodies aga inst bovine fibronectin , chi cken smooth m ll scle actin , and affi nity-purifi ed antibodies agai nst bovine brain tubu lin have been described prev iously, together w ith details for irnrn Lln ofluorescent sta in ing [1 8,19]. Affinity-purifIed rabbit an­tibodies against rat type I collagen and ty pe III proco ll:!gen f201 we r e the kind g ifts of Dr. R. Timpl (Max-Planck lnstitm fUr .B ioc h emic., Martinsreid). T hey we re used at concentrHions o f 40 /kg/ m l and 100 f.L g/m l, respect ively, fo r bo th tiss ue sections and cell c ultures . T he preparation and chara cterization of an antiserum to hum an type IV co ll agen has been docu111 en ted prev io usly [1 3], and t his was used at a diluti on of 1 :30 for im/llunofl uorescence stud ies . Antiserum aga inst m urin e laminin w as a g ift from Dr. .B. L. M . H oga n and was used at a 1 : 50 dilutio n on tissue section s [21]. In additio n , murine laminin was purifi ed fro111 the Engle­breth-Ho lm-Swarm tumor by published methods 11 41 and an anti serum was rai sed in rabbits. T he antiserull1 was fo und to be lllonospecific fo r laminin by enzy m e- lin ked immunoso rbent as­say (ELISA) and iml11unoblotting techniques.

DERMAL PAP ILLA EXTHACELLULAH MATHIX 763

Rabbit antiserul11 raised aga inst heparan sul fate proteoglycan isolated from the m edium of cultured PYS-2 cells 122], a murine teratocarcinoma cell line, was also used in th is stud y. T he serum was foun d to be l11 0 nospecifi c as j udged b y E LI SA and immu­no blottin g us in g murine la l11 inin, human o r m o use type IV col­lagen , or rat fibronectin as test substrates, as well as specifi c illllllulloprecipitatio n 0[35S (sul f..1 ted)-labeled l11acI:Oll1o lecu les from PYS-2 cultures. N o reactivity w ith a species of chondroitin sul fate proteoglycan synthesized by this cell line was detectable b y ELISA, iml11unoprecip itation, or il11l11ulloblotting . Full chara cteri za tion of this antiserum w ill be repo rted elsew here (Couchman, in prep­aratIon) .

Goat antibodies to human facto r V IlI- :\ssociated protein were purcllased fro 111 Miles (S lo ugh, Buckinghamshire, U .K.) and were fo und to detect de rm al blood vessels in sectio ns of rat skin. T he serull1 was used at a 1 :30 diluti on and th e second antibody waS FIT C - conjuga ted rabbit alIti goa t IgG (Miles). Sheep ant ibodies aga inst keratin and rabbit serUlll raised agai nst vimentin were g ifts from Dr. I. Virtanen , University of H elsinki. They were used at di luti o ns of I : 5 to I : 20 and the second antibody for keratin sta ining was a FITC-co njugated rabb it 3ntishee p IgG (Miles) . A ll primary rabbit antibodies were fo l\ow'd by a FITC-conju gJted goat antirabbit IgG (Miles). Second antibod ies in all cases were

Figure L Frozen sections of newborn rat skin with h"ir fo ll icle dermal papillae stained wi th antibodies against (a) flbroncctil1 , (l» type III procol1agcn, (c) type I proco llagcn, (d) laJ11lnJl1 , (c) type IV collagcn, (I) heparan SLIlfatc proteoglycan, (~) chondrmt!l1 6-sulfatc, and (I I) chondroitin 4-sul fate. The dermal papilla is marked ill each casc (m'l'lJJIIs) and the results show an absence of type I collagen and chondroitin 4-s lll(at~, sparse type (![ collagen fibe rs and the rcmJlJ1dcr, bascmcnt membrane compon~nts, are prcsent In the papdla . L:1I11 IJ1IJ1 , type IV collagcn, and heparan sul f.1tc proteoglycan arc p articularl y heavil y stained at the boundary between epithelium and derma l papi lla matri x. a,cJ x 120; b-d X 265; g, h X 165.

764 CO UCHMAN

uscd at di luti ons of l : 30 to 1: 50 in Du lbecco's phosphate- buffered sa line (d iva len t ca ti on-free).

Specim ens were o bse rved o n a Leitz Ortho lu x II o r Nik on Optiph o t fitted with ep i-illuminat ion devices and appropriate fil­ters. Pho tographs were taken o n IIfo rd HP5 o r X Pl fi lm .

RES ULTS

InuTlunofluorescence Studies In Vivo Sccti ons of new bo rn dorsa l rat skin containing mature anagen hair fo llicles werc stained fo r a number of extracellular m atrix mol ecules, including base­ment m cmbrane co mponents. We have previo usly shown [8] that fibron ectin (Fig 1 a) is di stributed thro ugho ut the dermal papilla matri x in anagcn and pro po~ed that th e so urce of this compo ncnt was at least in part deri ved from th e co nstitu ent m esenchy mal cells [81. The papill a co ntained little type III coll agen (Fig 1 b) and even less type I coll agen (Fig 1 c), in contras t to the surro undin g dermis which was rich in both co mpo nents. H owever, the der mal papilla was positively stained by a number o f antibodies against base ment membrane components. Laminin and type IV coll agen stain the epi thelial-papilla bound ary and were distributed thro ugh th e papilla in a g ranular or punctate fashi on (Fig 1d, e, also [8]). H epai'a n sul fate pro teoglycans occur both on cel l surfa ces and in basement m embranes [23,24], and an antiserum aga inst a latter form also stained dermal papillae (Fig lj). Like lami nin , the stain­in g o f the bo undary between the papilla m atrix and hair fo llicl e epithelial cells was m ost heavily stain ed. In contras t, an intense staining of the entire dermal papill a w as generated by the 3-£-3 mOl1 oclonai antibody (Fig 19) which specifi call y binds chondro­itin 6-sulfate. This antibody has been indi cated previ ously as de­tectilig a population of chondroitin sulfate proteoglycan present in base m ent m embranes [1 7]. The findin g here that this proteo­g lycan is abundant in dermal papill ae along with laminin and type IV coll agen stron gly implies th at the extra cellul ar m atrix of the derm al papill a is of basem ent m embrane type. This was furth er confirm ed by staining with the 9-A-2 antibody which recogni zes chondroitin 4-sul fa te and wh ich, in skin , wi ll detect an abundant dermatan sulfate pro teoglyca n [17] (Fig 111). The antibod y, al­though stainin g the dermis intensely, did not sta in th e derm al papill a o f rat hair fo llicles, showin g that, in common w ith th e red uced amounts of derm al interstitial coll agens, the matri x o f derm al papillae is guite different and distin ct.

Dermal Papilla Cells in Culture The in vivo stainin g pattern s indicated that the extracellular matri x of derm al papill ae was m arkedly different from that of adja cent dermi s. AlthoLlgh so m e dermal papillae m ay contain capillary endothelial cells capable of synth es izin g basem ent membrane co nlpon ents, it seem ed unlikeiy that the matri x of the papilla as a w hole was derived from th ese cell s. Capill aries within anagen rat pelage follicl e dermal papillae are generall y extrem ely rare or absent. In order to in ves ti gate further the synthetic capabilities o f derm al papill a cells, dermal papillae from rat vib rissae were dissected and plated in culture. Cells em ergin g from the papillae were stained with antisera to fibron ectin , interstitial co ll agens, and basem ent m embrane com­ponents. This was repea ted at 5 and 20 da ys of culture. T hc results from 5-day cultures are shown in Fig 2. Alth o ugh the cell s em erg­in g from papill a explants assembled very little extracellular m a­tri x, it was clea r from intracellul ar stainin g that the cells were sy nthesizin g fibro nectin , iamin in , and type I V proco ll agcn but not type I procoll agen (not shown) , and littl e type III procoll agen (Fig 2a-d). In contrast, cell s from 20-day cultures could be sta il1 ed fo r both in terstitia l collagens and laminin, but th ere was littl e staining fo r type IV coll age!] (Fig 3a-d) altho ug h the cell s were processed in an identi ca l mann er to th ose fro m 5-day cultures. It was il o ticea ble that the apparent change in synthetic capabil ity was acco mpan ied by an altered cell m o rphology, with indica tio ns of increased spread cell area. Dermal papilla cells from 5-day cul tures were less spread (as previously reported , [1 6]) th an their

TH E JOU ltN AL OF IN VESTIGATIVE DEltMATOLOGY

Figure 2. Dcrmal papilla cells after 5 days in culture, fi xed, ex tracted, and staincd w ith antibodics to (a) fibro ncc tin , (b) iaminin, (r) type IV collagen, and (d) type III procollagcn. A poor extracellular m atri x is present consisting of a few fibroncctin containin g fibr il s (arrolll in a) , but the other co mponcnts can be sta incd intracel lubrl y, indi ca tin g synthesis. (I-C x 265, d x 165.

co unterparts fro m 20-day cultures, thou g h all cells were spread to so m e dcgree.

T o gain furth er information rega rdin g the dermal papill a cells, their cytoskeleton was displayed with antibodies to actin , tubulin , and vimentin (Fig 4a-c). In all aspects this was characteri sti c of fibrobla sts, both at 5 and 20 days . Prominent microfilam ent bun­dles terminatin g at fo ca l adhesions were visible bo th b y indirect immunofluorescence (Fig 4a) and by interferencc reflection mi­crosco p y 11 9,25] (not shown) . In addition, there were well-o r­ga nized arrays of micro tubules (Fi g 4b) and vimentin containin g 10-nIll filaments (Fig 4c). Morpho logica ll y th ese cell s did not rese mble capillary endotheli al cells, and were ncgative for a cell s·urface populatio n of factor VIII-associated protein (Fig 4d).

D ISC USSION

Fro m th e tim e o f embryon ic develo pment at whi ch a clusterin g of mesenchymal cells den otes the site of fo llicle m o rphogenesis, a dermal papilla is always associated w ith the base ofthe hair. Its stru cture changes in concert with the hair cycle, m os t noticea bl y in term s of the ex tent of extracellular m atri x, w hi ch is at its greates t in anagen and least at telogen. T he work presented here

V L. X7. NO. (, D EC EMIJ Ell 19H(,

Figure 3. Derl1l ~ 1 p~pill~ ce ll s after 20 cb ys in cu lture. fI xed. ex tracted. and stained wi th antibodies to (a) type I proco llagen, (IJ) typc III procol­lagcn, (c) "'min in , and ({~ type IV co ll agell. T he cell s have an altered profi lc of extracd lular matrix pro tein synthesis (sec text) w hich now includes type I procollagen and mu ch type III proco llagell but little or no typc IV procoll agclI. All X 265.

shows th at this extra cellular matri x is in man y W3YS unique and qu i t e di stin ct fro m that of the dermis as a w ho le. The paucity of inte r stitial co ll agens and possession o f basem ent m embrane co m­ponents arc the ke y differen ces . The use o f mo noclo nal antibodies to detect di stin ct po pulations of chondro itin sul fate pro teoglycans also clearl y shows the differences between papillae and the dermis . The fo rm er co nta in a base ment membrane type of pro teoglyca n and la ck the der lll ata n sulfate proteog lyca n so widespread in man y regions o f the dermis 11 7]. Previous electron mi croscopy studies [26] have demo nstrated stru ctures with th e m o rpho logic appea r­an ce of base ment membranes to be present w ithin the bod y of de rm al papi llae and not just restri cted to the bo undaries between papi llae and epithelial cell s o f the hair fo lli cles. Alth o ugh the tiss ue sectio ns were from new bo rn skin , essentiall y identica l res ults have been o btaincd w ith anagen fo llicles fro m th e do rsal skin of adult rats (not show n).

However, altho ugh so me basemcnt membrane co m"ponents are p r esent in derm al papillae, it appea rs no t to be the case that these structu res have an ex tra cellular matri x w hi ch in every wa y re­se n, bles the base ment membrane o f th e dermal-epiderm al junc­tio n . Stainin g for type IV coll agen is relativel y w ea k in the papi lla

DERMAL PAPILLA EXTHACELLULA R MATRIX 765

Figure 4. Cytoskeleton o f derm al papilla cells. Antibodies to (a ) actin . (Ii) tubulin . and (c) vimelltill were used . Largc numbers o f micro fil amcnt bUlldles terminat ing 3t foca l adhesions ~rc present (arrOl /ls in a). and a well -o rgani zed arra y of microtubulcs (b) alld vimentin (but no t desmin or keratill - llo t shown) cOlltainin g 10- 11111 filaments (r) arc present. (,~ T his shows the ce ll s to be lI eg~t i vc w hen s t ~i n L"d with ~l1tibodies ag~inst b cto r V III -~sso c i ~ led prote in. {I X 670; b- d X 265.

w hercas the 3-£-3 antibod y recogni zing cho nd roi tin-6-sul f.1 tc pro tcog lyca n sta ins intenscly. T hcse pattcrns arc distinct fro m interfo lli cular and fo lli cubr base ment membrane w here th e COI1-ve rse is th e case [1 3,171. It appcars, thcrcfore, that the Ill atri x of rat anagen dcrma l papillae is very hi g hl y specialized.

Our previo us d3ta also h3ve shown th:lt staining of the dcrmal papi ll a w ith antibodies aga inst fibrone ctin . laminin , type IV co l­b gen [8], and hcpa ran sul fate proteoglyca n (unpublished rcsults) ca n be so mew hat m o re concc ntrated at the very e3 r1 y stages of anagen. T here are clear indi cations that th e am o unt of basem ent membrane macromolecu les can co rrelatc with the stage of the hair g rowth cy cle 18]. T hc papillae also share som e C0 l11111 o n components with the surroundin g de rmis with which they are continu ous. Fibronectin is a very widespread cxtraccllu br matrix glycoprotein 110-12] and is found in bo th locati ons. This is C0111 -

patible w ith other wcll-docum cnted evidence th at it is present in both the dermal mat ri x and basemcnt membranes of the skin including the derm al-epidermal junctio n [9. 121. Ind eed substantial amo un ts of fibro nectin ca n be loca tcd in the dermal-epidermal juncti on of embryoni c skin as well as at the ea rl y stages of anagen 18. 121. In a simibr fas hio n the derm al papilla co ntains a few fibri ls

766 CO U CHMAN

of type III coll agen. 111 addition, it is possible that maskin g of epitopes prevents more wides pread immunologic identification of interstitial collagens in dermal papillae. Previous work has shown that some elas ti c fibrils also extend into human dermal papillae, and a clustering of elastic fibers appears to demarcate the boundary between papillae and derm.al matri ces [27]. E lastic fib ers are an important dermal constituent, but they too can be­come closely associated with the basement membranes of the dermal-epiderm al junction [28].

M any, but not all, derm al papill ae contain fine ca pill aries but it is unlikely that their endothelial cells are totally responsible for the elaboration of the basement membrane type of matrix. This is particu larly the case in rat pelage follicles where ca pillaries within the dermal papillae are rare. It is also unlikely that th e follicl e epithelial cells synthesize the entire papilla matrix, parti c­ul arly during early anagen stages . Where capillaries are present, such as in vibrissa papillae, their endothelial cells occupy a pro­portion of the anagen papill a [2], and may synthesize basement membrane components. However, these studies show that papill a cells in culture initially synthesize base ment membrane compo­nents and are negative for factor VIII-associated protein. More­over, these cells have a morphology, cy toskeleton, and mi grato ry behavior resembling that of fibroblasts [1 8,29].

Althou gh the constituent mesenchymal cells of dermal papiIJae are referred to as fibroblasts, their precise nature is uncertain. Their well-developed microfi lament bundles (Fig 4a) and syn­thesis of basement membrane components may perhaps liken them to smooth muscle cells, of wh ich some from major blood vessels lack desl!lin and are vimentin-positive [30], simi lar to fibroblasts and derm.al papilla cell s. However, prev ious data have shown the basement membrane glycoprotein entac tin to be de­monstrable in derm al connective tiss ue of newborn rats and was presumably in fibrobl as t o rigin [31]. With improvement in cul ture techniques, it will be interesting to compare th e actin isoty pes of papilla cells with fibroblasts and smooth muscle cell s [32] to assess their affinities with either cel l type.

The fun ctions of such an extracellular matrix are not clear, but there is now direct evidence of an important role for the dermal papilla and its cell s in the control of hair growth [4-7]. Indeed the cyclical chan ges in amount of matri x co rrel ate with the growth status of th e hair. The growth rate of papilla cel ls in vivo is probably very low [33,34], and we have noted before the com­parative lack of available receptors for epidermal growth factor on these cells compared with derma l fibroblas ts 135]. It is possible that the extracellular matrix of the papi ll ae influences their growth and other properties. C han ges in papilla polysaccharides with respect to hair growth have been known for many years [36], but their composition has not been elucidated. It seems likely th at the chondroitin sulfate proteoglycan of basement membrane type may be a major contributor to these previously documented phenom­ena . It is parti cularl y noteworthy that dermal papilla ceIJs in cul­ture change their phenotype with time, producing matrix com­ponents similar to dermal fibrobla sts. Ja hoda et al [37] ha ve noted that the property of cultured papi lla cells to induce hair growth is gradua ll y lost. The retention of a phenotype which can induce epithelia l proliferation may therefore be acco mpanied by the elab­oration of a unique ex tracellular matri x.

Cralejit! Ihallks are due to Dr. R . Ti/llp l (Max-Pia lick IIIS/i/111 fiir Bioche/llie, Mar/illSreid), Dr. B. L. M . l-logml (Natiollal Inslilllle Jor Medica l Research, Mill Hill , LOlldoll ), alld Dr. I. Virlallell (Ulliversily oj He/sillki) Jor killd giJ;s oj all/ibodies. The skilled assistmlCe oj Mrs. A. S loll es ill will/ring der/llal papilla cells is gralejilily ackllowledged, as we/I as jimher assistallceji-olll Ms . L. Lewis.

REFEREN CES

1. M ontagna W, Van Scott E: The anatomy of the hair fo llicle, The Biology of Hair Growth. Ed ited by W Montagna, RA Ellis. New York , Academic Press, 1958, pp 39-64

2. Cohen]: Dermis, epidermis and dermal papillae interactin g, Ad-

THE JOURNAL OF INVESTIGATIVE I)EI(MATOLOGY

vanccs in Biology o f Skin , vol IX, Hair Growth. Edited by W Montagna, Il.L Dobson. Oxford, Pergamon Press, 1967, pp 1-1 8

3. Pinkus H: Embryology of hair, The Biology of Hair Growth . Editea by W Montagna, RA E ll is. New York, Academic Press, 1958, pp 1-32

4. Kollar EJ : The induction of hair fo llicl es by em bryonic derm al pa­pillae. J Inves t Dermatol 55:374-378, 1970

5. Crounse HG, StengleJM: Influence of the dermal papi lla on survival of human scalp roots ill" an hetero logous host. J Invest Dermatol 32:477-479, 1959

6. O li ver RF: The experimental induction of w hisker growth in the hooded rat by implantation of derma l papil lae . J Embryol Exp MorphoI1 8:43- 51,1 967

7. O liver RF: The vibrissa derm al papi lla and its influence on epidermal ti ssues. Br J Dermatol 81 (suppl 3) :55-64, 1969

8. Co uchm an JR, Gibson WT: Exprcss ion of bascment membrane components through morphological changes in thc hair g rowth cycle. Dev Bioi 108:290-299, 1985

9. CouchmanJR, Gibson WT, Thom D, Weaver AC, Recs DA, Parish WE: Fibroncctin distribution in epithelial and associated tissues of the rat. Arch Dermatol Res 266:295-310, 1979

10. Yam ada KM: Cell sur f.1ce interactions w ith cxt race llular materials. Annu Rev Biochem 52:76·1-799, 1983

11. Stenman S, Vaheri A: Distribution of a major connective protein, fibronectin, in normal human tissues . J Exp Med 147:1054-1064, 1978

12. Gibson WT, Couchman In., Weaver AC: Fibronectin distribution during the development of fetal rat- sk in. J Inves t Dermatol 81 :480-485, 1983

13. Westgate GE, Shaw DA, Harrap GJ, Couchman JR: Immunohis­tochcmical loca liza tion of bascment membranc components dur­ing hair follicle morphogenesis. J Inves t Dermatol 82:259-264, 1984

14. Timpl Il., Rohde H , Robey PG, Rennard SI, Foidart JM , M artin GIl. : Laminin-a glycoprotein from basement membranes. J BioI C hem 254:9933-9937, 1979

15. Kefa lides NA, Alper R, C lark CC: Biochemistry and metabolism of basement membranes. Int Hev Cytol 61:167-228, 1979

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