STUDIES ON AMEBOID MOTION AND SECRETION OFMOTOR END-PLATES

IV. ANATomC EFFEcS oF PouoxyEuTLm ON TS NEuRoxuscuTAMEcHISM mIN TIM MoNKEY*

E J. CARE, MD.(From the Dceptmen of Axatomy, Maquette Uxbrsty School of k, ME Lkre,

Wis.)

A elimiyreport has been made1 of the observations on theearly morphologic disp of motor end-plates and early ap-pearance of projiasses of g aiing substance inthesd muscle fiber with, or folowing shortly after, the onset ofparalysis in acute poiomelt in the y. In previous sftudies byLandsteiner and Levaditi,2 Fliner and LeWiS,3 Tayr,4 Penfeld,5Fairbrotr and Hurst,' Hurst,7 Covfl,8 Toomey and Takacs,' Saliand Olitsky,1 Goodpasture and Howe and B 2 o e holgyof poliomyelitis in the monkey, att T was concentrated on thechanges in the central nervous system and on parts of the pnervous System. There have been no studies made of the alteationsin the neuromuslar mechaisms. In Hurst's ex t study on thehistology of rimental itis m them ey, his observationson muse are Imited to one statent as folows: "Muscles.everalcases of were found at aupy." On the other hand,IHass1 detectd calai nsin epimic plioyelitis(Heine-Medin's disase) in human musle that he considers moreimportant in exaining the symptms than the alterations m the cen-tral nervousThe ni of action of the virus of po -my i i producing

thed ance of the motor end-plates isunw It is aumedthat there is an abnormal e ive discharge of the secretin of hypo-1emmal axonic substance. This gold-stainig subtance of the axosof the end-plaUes is assumed to be projected into the myolasm of thBemuscle fiber. For some unko reason this sulstance projected fromthe axom does not nomaly disprse but agglutinates into masses withthe pathologic anges pduced by the virus of polio-myelitils.These masses disaear withIM the first week after theira an and are not found m all musces.The purpose of ts paper is to present the complete photomicro-

*These were carried out with the aid of grants for reserch to theDeatment of Atm of the Maqute Unity School of Me by the Cmm-tee on Sefi esearch of theA Md al Association, and the Nation Foun-tion for I Paras, iic.R led for pubiction, lDber 21, 1943.

96I

graphic evidence, revealed by the gold method, of the alterations inthe motor end-plates during the early stages of experimental polo-myelitis in the monkey. Fortunately, the photogrphs are clear enoughwithout a long, detailed description of the pathologic changes. Tneexact and dear presentation of these easily verifiable facts necessitatesthe large number of photographs.

MATERIALS AND METODS

The' present histopathologic study dealswith the results obtainedby the intrcerebral ilation of a d dose of o.5 cc. of a 5per cent virulent cordelsinito 6m eys (Macacus rhes). TheArmstrong-Lansing strain of pdiomyelitis virus was used. Addstudies were made on the c in the motor e ates of m esfrom 8 monkeys obtained from S. D. mer, Mcigan Departmentof Health, Lansig; of musdes from 5 monkys obtained from J. F.Kessel and F. J. Moore, University of Southern California, Los An-geles; of muscles from 2 monkeys o i from P. F. Clark and A. F.Rasm , Uersit ofW snin,M and of musdes from 2monkceys obained from H. E. Pearson, Scool of Public Health, Uni-versity of Michigan, Ann Arbor. We are deeply indebted to these menfor excsing the muscles and placing them imdiately into bottles sur-rounded by aacked ice in a thermos jug for shipment by air mail orexpress to Milwaukee.

It was found by eprint that little or no alteration occurred inthe motor end-plates when the muscles were cooled to a temperatureof 5 to 80 C. as soon as excised and prior to the onset of rigor mortis.Parts of each mnscle were either frozen and sectioned, or fixed andstained by various histologic tehnics. Changes in fat were studied byosmic acid, Sudan m, and scharlach R, and the neurofibrils by theBielschowsky method counterstained with iron hematoxylin and vanGieson and with toluidin blue. The best method for the demonstrationof the pathologicc M the anatmic continuity of the epilemmalaxon, hypolemmal axons that constitute the motor end-plates, and theorganization of a striped muscle fiber was a modified gold method fol-lowed by careful teasi of sa pieces (2 by 5 mm.) of the impreg-nated muscle. This report.will be limited to the findings obtaied bythe gold method. This Ranvier gold chloride method was modified byWikinson 5 and by me " and will not be descnrbed in detail in thispaper.The following brief protocols are typical:

Monkey 53 (Marquette Series)April I9, '943. Inoculated intracerebrally with o.- cc. of virulent cord emion

of the Armstrog-Lsing strain.

962 CARE9Y

EFFECTS OF POLIOMYELITIS ON NEuROMUSCULAR MECHANSMx 963

Apri 23, X943.Apr1l 26, I943.

ApcflApilApril

'9, '943.25, I943.27, 1943.

April 28, 1943.

Feb.Feb.Feb.

' 7, '943-24, I943-27, I943.3, I943.

Mar. 4, I943.

Feb.Feb.Feb.Feb.

6, i943.I8, I943.'9, 1943-20, 1943.

Feb. 21, I943.

Rise of tureuand nmti,itue ioq ; e tZ ; muscles immediately exciedaDd I tehbdmic.

onkey 54 (Maquee Seris)Inoculated intracerebrally with the Armstrong.an g straiRise of temperatre.Fime geral tremor of weakened musdes of lower extremities andparys of those of the upper extrmities.Com te palysis of arms, legs and tunk. Diffiulty in respira-tion. KilI with eal M es i a excied andrun through gold techic.-

MoxkeY 490 (Krmes Seies)Inoculated m aly with the Armsrong-Lansing strain.Rise of temperaturWeakness of right flimb.Copet paralysis of ars and pa l paralysis of legs. Exsan-

tion chloroform for cardiac e Musces x-csed mediately and pl in bottles within iced tmosjandshippedby airexpresMusdes received and run through gold technic.

Monkey C-89 (COrk's Series)Inoculated intracermbrafly with the Kessel's McK. strain.Ataxia and tremorMed l paresu

Complete fiaccid p of as, legs and tnmk. Et iedand uses i dtely exdsed and placed inbMstts icdbyergc

Muscles d by exrp uss ad run t1 II d lord tehic-

Monkey K-334 (Pensox's Series)Dec. I8, 1942. Illated intraerIt 2aly and itasaily with the Armstng-

Ia straLDec. 21, 1942. R ld ie ebrally nd mtaslly with a stool sus-

Dec. 30, 1942. Complede laccidp s of bothlsDec. 31, 1942. Ethei Mu of ght bw leg excised and immediately put

in botles w iti ced thermosju . D_ed by air express.Jan. 2, I943. Musdes eived anr techic

Monke K-38 (Pewsoues Seies)ApR I, 1943. Iiocuated with 2 cc. of a 20 per cent

of the Amstrong-Lang strain.April 2, I943. Rise oft t to 102.20F.Api i8, I943. Temperture, I06° F.; tremors, ataxia, ruiffled fur, paralysis of

the right foelg, weakne of the bahc and both hind legs.Apri 19, 1943. TempeatuM 9740 F. F.CetApril 20, 1943. Tempatue, 90o F.April 21, 1943. Ethenzed Musc IFs t excised aid put in bottes wiin

iced t11rmos jug; shippd to by exprss.April 22, 1943. Muscles cve and atn through the gold chloril

tecni.

At the a psies, the usual finding of intense congestion of themeningeal vessels and of the gray matter, especially of the anteriorhorns of the spinal cord, was noted; on sectio the nervous tissue wasmore moist than rmal and the cut surface g ing. The affectedmuscles were intensely hyperemic and more fragile than the normalones.

Thirty selected muscles from both fore and hind limbs in 2 normkeyS wre prepared by thee gold mthod a ied to the para-

lyzed muscles and- used as controls. More than 3000 teased prepara-tions of the innervation of dfferent mus were used this study.Tle auripous masses of material projeed from the degenerating

axon were found in rtain musces of 8 of the 23 monkeys studied.These dislacd axonic masses were found in greatest abundance i thefollowing musces: biceps brachii, intercostals, quadriceps extensorfemoris, biceps femoris, gastrcnemius, tibialis anterior, sternodeido-mastoid, trapezius, pe als major and deltoid.

ERIMENTAL REsuLTSHistopatkdogy of Motor EAd-Plates

The motor nerve plates in the relatively mal muscle, quadicepsfemoris, in the monkey, varied from the rated state with widefronds;5 to2o,in d (Fig. I) totheea state30to40is in diaer. The raed motor end-plates were usually rlated tocoarse, widely spaced cross striations found in the narrow muscle fibers.ITere were variations, however, in relationship. The ndedplates were uually related to fine, closely spaced striations found inthe wide muscle fibers. There was a losely graded series of transi-tioal stag in the itenal structure of the muscle fibers in relationto motor end-plates of various sizes. Tlhe acted motor end-platehad hypolemmal axon teminals sounded by an accumulation ofKiihne's granules. This mantle of Kie's granules was either di-minid in size or completely depleted in tbe e ed nerve plates.

Wiith progressive increase of ameboid e nsion of the motor nerveplat there was a dimiution of Kiibne's granules and a correspondingincrease i nmber and fineness of closely spaced cross striations. Thestrngly cotracted muscle fibers had large motor nerve plates withtwo or more miliform projections separated by wide spaces. Withprogressive increase nm size of the motor nerve plate and icrease offineness of the dark and light transverse striations, the contractedmuscle fiber took a differential stain. This indicated either a physicalor c ical change in the contracted muscle fiber. There were many

964 CAM

EFFECTS OF POIOMYELIS ON NEUROMUSCULAR aANIx 965

transitional stagsa and gradations between the arsely and finely stri-ated muscle fiber. There was a substntial a id o of theprocesses of the nerve plate in various dections n the muse sub-stance which eased the surface area of the nerve pate and vari-abiblty of the musce striae. This was evidence tat during life themuscle stdrae were not constant and rigid menmranes.

In some muscle fibers there was a periodic alternation of fine, closelyspaced striations and coarse, widely spaced ones. In other places themuscle substance was atd into m s in which no striationswere visibe with the highest powers of the miroscope. This compactmuscle subsitance resembled Zenk7rs waxy Frm e-

perimental evidence, Wells I conduded that these opaque hyalinemasses repressent a local a of lactic acd. By apling lacticadd to striated muscle ix itro and ix vivo he sed strucural changesresembling erwaxy d r . Te intnalstrcure of themuscle Ofber, therfore, appeared to be itimately rated to the vari-able biohemical piti The variation in the nrmal sructureof the living muscle fiber appeared to be an ssion of the changinchem reactions. There may be a clo associat between histologic structure and chemical c ti than is noW reized.The diffetial fiber tpes were present in the normal muse (Fig.

I) but they were absent in the paralyzed musce (Fig. 2). Many Ofthe end-plate on the first day of pawalyse retracted into smalball-ikem (Flg. 2) with rtened s cylders and itenseaffinity for gohl Likewise, on the first day of paralysis, there were afewnl g ar end-plate that had awk affinity for gold.Between 20 and 25 per cent of the motor end-plates we absent in thep lyzed muse onthefist day (Fig. iI). Within 2 to 4 days afterthe onset of pralsis about So per cent of the end-plates had disp-peared (Figs. 6, 12 to I6, and 27 to 33). During the first 24 hours theparalyzed musce had defliite passive hyperemia of the musular cap-illaries and veins. The muscular fibers were prominenty identified bythe presence of congested tramuular vessels (Figs. 3 and 4, 7 toIO, and 17). During the first 48 hours the parazed musce had treesof end-plates with variable degrees of retraction and granlar frag-mentation (Figs. 3 to 5).Many retracted tes of axons were completely depleted of end-

plates by the third or fourth day of paralysis (Fig. 6) and had sharpor bud-like epilemmal teminals in the triceps muscle. The rates ofnerve degeneration were unequal in different muscles of the same ani-mal and in different fibers of the same muscle. The wekened biceps

brachii musce had an innervating tree of retracted ball-like motor end-plates (Fig. Io) wherea the paralyzed tricep muscle had, on the firstday of paralysis, an inervating tree with prctically complete absenceof all of the motor end-plates (Fig. iI).

Observations were made of an unusual histologic condition charac-terized by masse of gold-staining material dosely assoatedwith thedegenerating tree of inervation. These masses were located both ex-ternally and internally to the muscle fibers (Figs. 12 to i6). Theyvaried in length from 5 to I05 p. They were found in 8 of the 23monkeys studied from I to 4 days after the onset of paralysis. Theywere not found after the seventh day of paralysis. The gold-stainingmasses were found in the following muscls: biceps brachii, intercos-tals, quadriceps etensor femoris, biceps femoris, st s, tbialisanterior, sternocleidomastoid and trapezius. These gold-staining m-dusion masses were composed of granules O.I to 2 , ad globod bodies2 to 4.5sp indiameter (Figs. 18 tO 26). hof these bodies was fre-quently surrounded by a clear halo-like space (Fig. 26). In some placesthese masses were uniformly and densely stained. In others they had adear-cut arrangement into cross striations, whereas in still others theperiphery of the auripus masses was indented in a festoon-like man-ner by the normal cross striations of the musce fiber. In many placesthe granules of the gold-staining mes were aligned in a manner cor-resoding to that of the normal cross striations of the muscle fiber.In other locations the cross striations of the inclIon mlssesad aperiodicity of their own which did notc to that of the imme-diately lated cs striations in the mu fiber.

it is assumed that these gold-staining inclusion mses were pro-jected from the terminaton of the d ating axon into the musclesubstance. The assumed reti ip of these masses to the axonicsubstance is based upon two histologic facts; namely, (I) the similarityof the staining acity of the inclun masses to that of the axon,and (2) the apance of the inclusionm s out in the muscle sub-stance time to the attenuation of the axc terminalsand diminution of their content of gold-staining substance. In otherwords, these inclusion masses were assumed to represent an abnormalsecretion of the substance of the motor end-pate and a failure of nor-mal dispersal of this Product of axonic secretion throughout the musclefiber.These inclusion masses had a more tense staining capacity for gold

than that of the normal cross striations in the musle fiber. SinceHurst 13 claimed that he found sporidiosis in some of the muscle rs

966 CAREY

EFFECTS OF POLIOMYELIrIS ON NEURtOMUSCULAR MCANISX 967

of the monkey, care was taken to differentiate histologically these gold-staining inclusion masses from sporidia. In I8,246 muscle fibers of theleftq extensor femoris muscle (monkey 490, Kamer's series)these inclusion masses were found in 14,64 fibes in close a iwith the zone of rapidly disintegrating innervation. Tlese inclusionmasses had not been found ino muscles of 2 normal control monkeys,nor m those eained more than i week after the onset of paralysis.These ephmeral inclusion masses at the degenerating myoneural jumc-tion appeared to be characteristic, as far as this limited evidence goes,of crtain unk chemical reactions that occur at certain times dulr-ing the early stages of e al piomyelis withim and near thedegmmt innervation of muscles in so monkeys. There may bean affiniy of the virus of poliomyelitis or its products for acetyllineor some related substances delivered by the end-plates into the muscefiber.The degenerative process resulting in either the dimnution or com-

plete ion of gold-staining subtance from within the terminalaxons of motor nerves appared to begin at the end-plates in the musclefiber a then progressed in a Centripetal dieCtion (Figs. 27 to 33).The was first a dissolution of the hypolemmal axons and a relatedvariable quantity of the granules of Kiihne that constitute the end-plates. At the beginning the epilemmal axons were engorged, in manyla, with gold-staining sulbtan (Figs. 6 and iI) but denuded of

end-plates. Within 48 to 96 hours some of the intramuscular nervetrees showed their epilemmal axons with decreased amoimts or com-pletely depleted of axonic gold-staining substance. The ghost-likeoutline of some of the axons had either scattered granules of the frag-mented axons or complete abce of granules peripherad, whereasmore Centrad (Figs. 27 to 33) the same epilemmal axons had a tongaffnity for gold because of the persistence of the axonic subtane.

ITere is no adquate cemical anation as yet for either themorphogic dissolution of the end-plates or the centripetal directionof deplion of the peripheral terminals of the epiemmal axons of theirgold-staining substance produced by poliomyelitis. Manye tswere suggted by these findings and more facts must be establisdbefore an adequate theory may be advanced to explain these observedfacts of the patholo of the motor end-plates.

Passive hyperemia of the capilaries was prominent in some musclesduring the pre-paralytic and early paralytic stages of muscle weakness.Each muscle fiber was clearly delimited because of the greatly con-gested cpillaries, the content of which had abnormally increased af-

finity for gold (Figs. 3, 4, 7, 8, 9, and 17). In some places there wasa beginning proliferatio of the pervascular reticulo-endotheial mesen-chymal ceis m the weakened m c E h vascular sprouts ofthe b ng of vasculgenesis were observed in some places (Fig. 9).

S^rI. The early histoolg chang that ccurred in the neuromuscularmcan of tal m monys were studied by

the gold inm arson wih the muscles from 2 normal controlanils. A quanti anasis of the anatomic changes was aided byboth lo-wer adhig-oer

2. The earliest ptologica was h emia and beginningiL ar infiltation of ertam ares of the intu r blood

vessels, wich had moe granules with strong affinity for gold in thelumen th did the noaL This yperem was detected in weakenedbut paralyzed m in certan placs pnor to alteratinsi thestructure of the motor endplates.

3. On the first day of paalysis sone of the end-plates were re-tracted in baille mases and others werehy t and granular.AboUt 20 per cent of the mtor plates we aent the paralyzedmuscle on the first day, and within 2 to 4 days about so per cent of theend-plates iap The rates of denervation and deenationwere not onlyu l i difet E of thes anial but indifferent groups of musle fibers iin the se musle.

4. PrOjected masses of gd-sting anc subtance were foundin cloe relationship to the d tree of innetvation from i to4 days after the onset of pa s. They wre absent after 7 daYs ofparalysis. They were found in certai muscs of 8 of the 23 monkeysstudied. 'Ilhese o mass were sed of granules andspheroid bodies that varied from o.I to 4. i diatr and in somepaces were aranged in cross which had a more itense af-fnity for gold than the ordinary cross stons of the muscle fiber.Tiese e gold-staining were not found in 30 muscles of2 n l contrd m nr m usces after i week of paralysis.These projeed maes of axoni subnce at the degrating neuro-mus rmechaim a ared to be characteristic, as far as this limitedevidence goes, of certain unkwn cemical reactions that obcur atcertain times during the early stages of experintal polio elitiswithi and near the g- mnervation of some muscles in somemonTeys.

5. Thle degeneratio appeared to begin in the motor end-lates and

968 cl

EFFECTS OF POLIOMYELITI ON NEUROMUSCULAR ECHANS 969

then to extend in a centripetal diection though the epilemmal axonsin many of the nerve trees observed.

6. Ietntative hypothesis is advanced that the his h cges observed at the nerus lar mechaism during the early

stages of pomyelitis are the result of an abrmal excitation of thesecretory mechanism of the motor end-plates which results in the pro-gressive ustion of the gold-sta g axonic s n leading todenervation at the m ju i. s early material e tonof the teminals of motor nerves in skeletal musle by the virus ofpolomyelitis will be studied in relation to the ical secretion ofacetylchone. This l study will be extended to include acorrelation of the change i the neuromuscular. i with thosein the spinal cord and m ll oblongata i the se animaL

I wish to express gratitude to Mr. ILo Maseust, DItor of the Departmentof Art and Photography, for aid with the pnicoapbs; to Dr. G. KastenTafmadme, Assistant Professor of Anny, for ding the manucipt; and toMesss Eugene Haushalter, John Sdkmitz, James Keyes, Joseph Hamel and RobertJeub for technial aid in the tg of musde and nerve plates.

ADDENDUX

Subsequent to the acceptance of this paper for publication evidencehas been obtained which supports the satement that metabolic sub-stances such as latic and other acds may ac ulate abnormally dur-ing anoa locally produced in the muce and hereFby destroy themotor end-lates. The abnormal a of epem axonicsubn extrinsic to the end-plates undergoing liquefaction are ob-served in some of the muscle fibers. This is due either to an abnormalpermeability of the end-plates and secretion of axonic material or adelay in its dissolution after this material is secreted from nerve tomuscle."

It has been established that abnormal am of lactic acid appearin the blood in hemorrhagic, traumatic and "gravity" shock.The pathology of the end-plates leadmng to their destruction and theevanescent appearance of axic nervous material in the muscle durgthe incipient stages of hr shock is quite c le tothat of the early effect of lactic acid and poliomyelitis. Gesell and hiscolleagues found that lactic acid caused a delay in the destruction ofphysiologically deposited acetylcholine at the synapses.2 5 Whetheror not the abnormal amount of nervous material histologically ob--served in muscle im poliomyelitis, in the lactic acid series of experi-ments, and in experimental shock are the morphological expression of

970 CARE

the temporary a an of, and the delay in, the ssolution ofcetyicholine i musce annot be answered conclusively at the present

time. CES

i. Carey, E. J. Mo ic effects of polonyelitis virus upon motor end platesin the monkey. Proc. Soc. Etxer. Biol. & Med., I943, 53. 3-5.

2. Landst, K, and Levaditi, C. La tra de la parlysie finantie auxsnges. Compt. rexd. Soc. de bid., i9o9, 67, S9294.

3. Flemnr, S, and Lewis, P. A. erimental epidemic pdiomyelitis in m es.J. Epe. Med., x9IO, 12, 227-255.

4. Taylor, ]E D. Blood counts in ental poliomyelitis in the monkey.J. Esper. Med., 1919, 29 97-i4.

S. Penfield, W. A method of staining and microglia (cambinedmethod). Am. . Path., 1928, 4, 1S3-I57.

6. Fairrother, R W., and Hurst, E. W. 'Te p of, and propagationof the v in, e rental poJomyelitis. I Path. & Bact., 1930, 33,'7-45.

7. Hurst, E. W. A further to the sis of experm talpoiomyeitis: in tion into the sciatic nerve. J. Patk. & Bact., I930,33, 1133-1143.

8. Covell, W. P. Nudear dcges of nerve acl in acute polomyelitis. Proc. Soc.Exper. Bid. & Med., 1929-30, 27, 927-929.

9. Toomey, J. A., and Takacs, W. S. Furthe e rents with poli ev in cotton rats. Proc. Soc. Ezper. Bid. & Med., 194I, 46, 319-321.

I0. A AB., and Ofitsky, P. K. The olfactory bulbs in Elxpeta polio-mYc 1iis. A. M. A., 1937, I08, 21-24.

II. G e, E. W. 'Tbe at and Pathogenesis of Poliomyeitis. In:Infanie Paralysis. A Sym n l red at Vanderbilt University, April,1941. National Foudation for nfantile Paralysis, New York, I9I,Pp. 85-125.

12. Howe, H AL, and Bodian, D. Neual Mechanisms in Poliomyelitis. The Com-mo elth Fund, London, 1942, PP. 4 and 78.

13. Hurst, E. W. 'The istlo f ental po yeitis. . Path. & Ba.,1929, 32, 457-477.

14. Han G. B. Landry's Paralysis: its dinical and p o c features. J. Newo-p. & Ezper. Nerdl., 1943, 2, 293-300.

I5. Wilkinson, H. J. The i min of siated muse. M. J. Austraia, 1929, 2,76-793. Expenmental studies on the in vation of striated musce. J.Comp. Newel., 1930, 51, 129-I5I.

I6. Carey, E. J. Studies on a motion of motor vpa. II. Pathoogiceffects of C)2 and el on thve a d moti in motornerve platin t l muse. Am. J. Path., 1942, 3S, 237-289.

17. Zen, F. A. Ueber dieVg derwkdiicn Museln im Typhusabdoxninalis. A. E. JuDg, Edang, 863; F. C. W. Vogel, Liig, 1864.

i8. Wells, H. G. e p of waxy degenertion of striated musces(Zenker's d t ). . Ezper. Med., I9o9, II, I-9.

REFERENCES FOR ADDENDUMI9. -Carey, E. J., and , L. n d uction of motor end plates by

lactic aid Proc. Soc. Ezper. Bid. & Med., 1944, 55, 194-197.

EFFECTS OF POLOMYELrIS ON NEUROMUSCULAR MECHANISM 971

20. Govier, W. M., and Greer, C. M. The effect of certain anestetics on blooket acid levels. J. Pharncd. & Esper. Therap., I941, 73, 441-445.

21. Gutmann, H., Olson, W. ]L, Kroal, ]EL H, Levinson, S. O., and Nechele, HLChemical studies in tamatic shock. Am. J. Physio., i94i, 133, 308-309.

22. Cole, W. H., Allison, J. B., TLeatln, J. H, Nastuk, W. L., and Anderson, J.A. Addois n the diabit duing "grvt shock." Ant. Rec., 1942, 84,48.

23. Carey, E. J., Mss, L. C., Zeit, W., Hauhalter, E., and Sdchitz, J. Acuteanatomic breakdown of motor end plates i hm hagic shock. Proc. Soc.EzPe,. Bid. & Med., I944, 56, II5-II8.

24. Gesel, R., Braseld, C. R, and Hansen, E. T. Possbe r6les of cH in neuro-phsiology. Proc. Fed. Am. Socs. Ezper. BioM., 1942, I, 29.

25. Hansen E. T., Worziak, J. J-, and GeselL R- Physiolcal effects of artifi-ay ministered acetylchoine and eserne. Proc. Fed. Am. Socs. Exper.Bio., 1942, I, 36.

[lustr.tions f]olow]

DES(31P'flON OF P1TIS

nta 172

Ixd 2. 16 teadil nmml t the ift_SMdrkqm faunas(F I) in the -u i thst in the let pm s fam is

MP (F 2) Omthe tddof IaLy Iathemm (Fr. I) the isaw,tim te of the intw ajt and tieuepw t

hdthat awe 4 d l s ed. Oatheitdiy ofi(2) sum Of SC ni h stDa dq lyw gu,awe am-foinuly ad Ste i a haintekfstKyfiibes-Tre u _m hy of the of the efibs (Fi 2).X ISO.

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Carey Effects of Poliomyeitis on Neuromuscular Mechanism

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PIATI 172

PLATE 173

FIGs. 3 and 4. Ihe teased innervation of the nonparalyzed but weakened gas-trocnemius musde from a monkey in which the upper extremities were para-lyzed for i day. Tnere is definite enlargeent and hyperemia of theintramuscular blood vessels. The motor end-plates are in variable degrees ofretraction, and some are in a state of granular fragmentation. The epilemmalaxons are beaded and in some places fragmented. X iSo.

974

AMucAN JoutwAL or PArHOLoGY. VOL. XX PLATE 17,

t~~~~~~~~~~~~~~~~~~tf

&7

3

it IL- ;

L..4 '-Cae

t /

. L t v%II;.

Effects of Poliomyelitis on Neuromuscular Mechanism

97S

PLATE 174

FIGs. 5 and 6. The teased innervation of the nonparalyzed but greatly weakenedtriceps musde (Fig. 5) in contrast to the paralyzed biceps brachii muscle(Fig. 6) from the same monkey (no. 490, Kramer's series). The hypolemmalaxons of the motor end-plates are fragmented into granules which have astippled pattern and take a deep stain with gold (Fig. 5). The epilenmmalaxons are irregularly beaded. The motor end-plates have completely disap-peared in the biceps brachii muscle (Fig. 6) except in a few locations wherea diffuse granulation is found. The epilemmal axons are definitely enlarged,shortened and beaded, and terminate in rounded- or ovoid bulbs which areenlarged and are stained deeply with gold. In many places the cross striationsof the musde fiber have disappeared and a diffuse granulation is present.X I50.

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s,-VA

Carey Effects of Polomyelitis on Neuromuscular Mechanismn

977

PLAEIM 174

PLATE 175

Fics. 7, 8 and 9. The teased motor innervation from the nonparalyzed but weak-ened bicePs femoris muscle (Figs. 7, 8 and 9) i day after the onset of theparalysis of both upper ext ities. Ihere was muscle weakne assoated witha striking passive hyperemia and larent of the capillare and the intra-muscular vessels, with begning perivascular infiltration of leukocytes and afew lymphocytes. There is increased vlsibility in the nuclei of some musclefibers (Fig. 8) arranged in longitudinal rows. A majority of the motor end-plates show retraction and increased staiing capacty. There is beading ofthe epilemal axons. Ihe content of the sted blood vessels has increased

inin aii for gold. X 80.

978

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FIGs. io and iI. Teased innervation of the nonparalyzed but weakened bicepsbrachii musde (Fig. io) of the monkey. There is definite ular dgea-tion of some muscle fibers and passive hyperemia of the intramuscular capil-laries. The mal axons are rtened and beaded, and the hypoenmalaxon of the end-plate is retracted into rn, d in terminals. Themotor innervation of the parabzed triceps musce (Fig. iI) is from the oppo-site limb to that lustrated above (Fig. io). lTere is a koss of the majority ofthe motor end-plates and the faint outine of those present is occupied bygranules d y arrang. The e mal axons are retracted, beaded, en-larged, and have intense affinity for gold. X 8o.

980

AMUKAN JouRNAL OF PATHOLOGY. VOL. XXC

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PLATE 177

FIwS. 12 and I3. Ihe motor innervation of the paralyzed right biceps brachii muscle(Fig. 12) of monkey 490, Kramer's series. There is a loss of motor end-platesbut there are present multiple masses, both inside and outside of the musclefiber, that have the same inten affinity for gold as that of the normal axon.It is assed that these masses were projected from the termination of theaxon into the musde substance. TIhe relationship of these masses to the axonicsubstance is based upon two factors; namely, (i) similarit of staining capacityand (2) the attenuat of the teminals of the amn smultaneo with thepresence of the gold-staining masses in close relationship to the motor innerva-tion. The motor innervation of the left paralyzed biceps brachii muscle (Fig.13) in the same monkey likewsie has these gold-staining inclusion masses indose relatiohip to the attenuated terminals of the axons. There are endo-thelial sprouts of new blood vessels in a state of passive hyperemia (Fig. I3).X 80.

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FIGs. 14 and IS. The teased motor innervation of the paralyzed right gastrocnemiusmusde (Fig. 14) of monkey 338, Pearson's series. There is a dose relationshipof the projected gold-staining masses to the attenuated terminals of the axons.These indusion masses vary in size from rounded, globoid bodies, I to 2 IL indiameter, to masses of these granules that vary from 5 to IO5 E in length. Theteased motor innervation of the paralyzed left tibialis anterior muscle (Fig.15) in the same animal as above (Fig. 14) not only possesses retracted deep-staining motor end-plates but dosely related gold-staining iclusion masseswhich appear simultaneously with the attenuation and, in some instances, withcomplete depletion of the gold-stainig substance of the axons. X 8o.

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FIG. i6. The teased motor innervation of the right sternocleidonastoid muscle ofmonkey 53, Marquette series, had projected gd-stainin masses which variedfrom 5 to Io5 i& in length and which were related to the degenerating innerva-tion. The absence of the motor end-pates concomitant with the appearanceof the gold-staining projection masses in the musde fiber is dearly evident.X 8o.

FIG. 17. The teased fibers of the weakened right trapezmsmusce of monkey53 had enlaged and congested intamuscular blood vessels. Tlere is evidenceof a beginning perivascular infiltration of polymorphoudear leukocytes andsome lymphocytes. Passive hypeamna with enl agement of the blood vesselsis one of the earliest sgns of patholc chan in the involved muscle. X I50.

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FIG. I8, ig and 20. Elongted projection masses of axonic substce in the para-lyzed stromis muscle (Figs. I6, I 7 and i8). IJhse projectio masseshave a greater affiity for gold than the dark cross striations of the musdefiber. These masses are c d of rouded bodies I to 4 p in diameter, andin some places a a d in crss striatio. In other places the peripheryhas a festooned border, the convexties of which c d with the ordinarydark cross striations of the musde fiber. (Fig. I8). X 750.

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FIGS. 21 to 26. Elongated projection masses of axonic substance in the paralyzedgastrocnemius musde 2 days after the onset of paralysis. Some of these pro-jection masses have a deep uniform stain, the border of which is festooned(Fig. 21). In others there is a definite arrangement of the globoid graulesinto cross striations. Some of these globoid granules arranged in cross stria-tions are surrounded by a light circular border (Fig. 26). In some places thesegranules are aligned to correspond with the normal cross striations of themuscle. In other locations they have a periodicity of their own which doesnot correspond to the immedtely related cross striations in the muscle fiber(Fig. 26). lTese axonic projection masses have a more intense affinity forgold than that of the cross striations of the muscle. In some places (Fig. 25)there is a diffuse aranent of the projected axonic granules in relationshipto others which are dnitely cross striated. X 750.

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FIGs. 27 to 3I. Elongated projection masses of axonic substance in the paralyzedbiceps brachii muscle. It may be noted that correpoding with the presenceof the axonic masses projected into the substance of the muscle fiber there is adecrease of gold-staining material in the axons, 4 days after the onset ofparalysis (Figs 27 and 28). There is an absnce of gold-staining axonic massesin the paralyzed triceps muscle (Figs. 29, 30 and 31), 4 days after the onsetof paralysis, taken from the same limb (Figs. 27 and 28). There is a diminu-tion or complete depletion of the substance in the axonic terminals. llTispathologic chang appears to progress in a centripetal direction from theoriginal location of the motor end-plates. Ihe terminal epilemmal axons insome places (Figs. 29, 30 and 31) have merely a faint ghost-ke outline withno affinity for gold, or a greatly decreased staining capacity for gold. X 8o.

992

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FIGS. 32 and 33. here is a decrease, or a complete exhaustion, of the gold-stainingsubstance in the terminal axons of the motor innervation of the paralyzedtriceps musde, 4 days after the onset of paralysis. This pathologic changeappears to progress m a centripetal direction from the original location of themotor end-plates. The terminal epilemmal axons in some places (Figs. 32 and33) have merely a faint ghost-like outline with no affinity for gold or a greatlydecreased staining capacity for gold. Corresponding to the complete depletionof the substance in the terminal axons there is an eventual disappearance ofthe axonic inclusion masses that were originally projected into the substanceof the muscle fiber. In some places, the cross striations of the musde fibersare replaced by a disorganizd mass of granules and in others by a more hyalineappearance comparable to Zenker's waxy degeneration. X 250.

994

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