RATE OF SEBTJM EXCRETION FROM THE GLANDS TO THE SKIN SIJRFACE*

KIMIO 1KM, M.D.

Although many reports which suggested theexistence of the secretory innervation of thesebaceous glands have been ignored by manyof those concerned with the concept of inter-dependence of lipids and sweat on the skinsurface, the last doubt has not been eliminatedowing to the difficulty in observing the excretionfrom individual sebaceous and sweat glands(1) simultaneously before emulsification. Theauthor has attempted to investigate the presenceof secretory innervation of the sebaceous glandswhich have been a subject of this controversy.

It has been postulated by Rothman, that todetermine the amount of sebum excretion, it isnecessary to check the amount of accompanyingsweat or to atropinize the skin area in order toexclude accompanying perspiration (1). For thepurpose of this investigation, however, it wasdetermined not to first atropinize the skin area.The author has developed a method to absorbboth sweat and sebum at the same time butseparately onto the starch paper upon theirdelivery to the skin surface, allowing them noopportunity of emulsification. Using this method,observations were made on the rate of sebum ex-cretion which is not influenced by perspiration,when intracutaneous administration of the auto-nomic drugs (Part I) or emotional stimulation(Part II) were applied.

METHOD5 AND RE5ULT5

Part I. Intracutaneous Administration of theAutonomie Drugs

The upper back was chosen as test skin areawhere considerable amount of sebum excretionand no "emotional" perspiration could be ex-pected. The rectangle skin area (5 cm x 7.5 cm)centering along the vertebral line (shown infigure 1) was first cleansed by swabbing withacetone. The upper half of the rectangle was

* From the Department of Physiology (Pro-fessor Hatsuo Nitta, Director), Nagoya CityUniversity Medical School, Nagoya, Japan.

Presented at the Thirty-fifth Annual Meetingof the Physiological Society of Japan, Kanazawa,Japan, May 4, 1958.

Received for publication May 9, 1958.

painted with iodine tincture (U.S.P.), and thelower half was merely cleansed with 70% alcohol.

0.05 cc of each of two different autonomic drugs(a and b) were then administered intracutane-ously at the left and right parts of the upper andlower division of the rectangle (figure 1: a anda', b and b'). 0.05 cc of physiological saline wereintracutaneously injected in the median line ofthe upper and lower division of the rectangle(figure 1: s and s').

Before, immediately alter, 20 minutes after,40 minutes after and 1 hour after the adminis-tration of the drugs, the rectangular starch paperwas placed on the test skin area, being presseddown evenly with the use of foam-rubber forexactly 3 minutes. Both sweat and sebum wererespectively absorbed into the starch paperwithout having any opportunity of emulsification.

Sweat droplets penetrated the upper half ofthe starch paper were indicated as dark purplespots of iodostarch. Sebum absorbed into thelower half of the paper was then exposed to osmicacid vapor by placing the paper over the mouthof the osmic acid bottle (1 cm in diameter) fora period of 1 minute, and was indicated as darkbrownish dots in circlets.

Just before each application of the starchpapers, wipings were taken for the amount ofsebum which was replaced during each test. Alsoin order to eliminate the influence of stimulationby acetone, it was necessary to wait about fifteenminutes after the first swabbing by acetone untilthe blind application was recorded before injec-tion as control.

The autonomic drugs which were used for thisexperiment and their contents in solution wereas follows: epirenamin chloride (epinephrine)1:1000, acetylcholine chloride 1:200, pilocarpinehydrochloride 1:1000, physostigmine 1:2000,carbaehol 1:1000, mecholyl chloride 1:1000,scopolamine hydrobromide 1:100, pituitan(pituitrin) 10 units/cc, ergotamine tartrate1:100, chlorpromazine 1:200, etc. The experi-ment was undertaken during the period of June1956 to May 1957, (room temperature of 20°to 25°C) on 27 healthy subjects aged 20 to 43and including 1 female subject.

27

28 THE JOURNAL OF INVESTIGATIVE DERMATOLOGY

Fia. 5 FIG. 6

A part of the results obtained is shown inFigures 3 to 8. Survey of the results showed:

1. There was no significant change in theamounts of sebum excretion before and afterthe administration of the autonomic drugs,and between the two antagonistic drugs.

2. Dark brownish dots of sebum which wereindicated on the lower half of the starch paperswere found not to be parallel with dark purplespots of sweat on the upper half of the papers.

3. On the lower half of the starch papers, sweatdroplets were also stained by osmic acid vapor

with light greyish or dark pinkish tint. Thesespots of sweat were easily differentiated from thedark brownish dots of sebum in the same circlets.

Part II. Response to "Emotional" Stimulation

The forehead was chosen as the test skin areabecause "emotional" perspiration and highamounts of sebum excretion could be expected inthis site (figure 2).

Electrodes were fastened to palms of thesubject lying in a supine posture, in order torecord the psychogalvanic reflex. When heat-pain

S.

A A

FIG. 1 Fin. 2

Fio. 3 Fia. 4

S.

2o.:

SEBUM EXCRETION FROM GLANDS TO SKIN SURFACE 29

FIG. 7 FIG. 8

was added by Hardy's pain machine (2) with theapproximate intensity of 10 dols, the out-pouringsweat and sebum were absorbed by the starchpaper for 1 or 2 minutes, with no opportunity foremulsification.

Pre-procedures were performed in tbe same wayas described in Part I. This experiment wasundertaken during the period of May 1957 toMarch 1958 on 32 male subjects aged 33 to 64,at a room temperature of 20° to 28°C.

Some of the results obtained are shown thFigures 9 to 12.

1. Generally, no perspiration stained thestarch paper as long as the subjects relaxed inroom temperatures of 20° to 25°C, nor was therea psychogalvanie reflex response. However, whenthe psychogalvanic reflex appeared in responseto the heat stimulation, the amount of per-spiration indicated on the starch papers oftenincreased (not always—figure 12), and was re-garded as mental perspiration. On the other hand,there was no significant difference between theamount of sebum excretion before and afteremotional stimulation regardless of the presenceof mental perspiration. This relationship was inagreement with that as observed in Part I con-cerning the administration of the autonomicdrugs.

2. Amount of sudoriferous spots and sebaceousdots indicated on the starch paper were alsofound not to be parallel in this experiment.

3. On very rare occasions, when no responsewas observed in psyehogalvanie reflex and thepaper was stained with perspiration, it was re-garded as perpetual perspiration in the roomtemperature below 28°C and as thermal perspi-ration in the room temperature over 28°C.

DISCUSSION

Since Miesher and Schoenberg's experimenta-tions, it has been considered necessary to dif-ferentiate casual level (saturation level), produc-tion capacity (excretion rate) and retained level(residual level). The term production capacityis also known as the replacement sum or theexcretion rate. This seems to be dependent uponboth 1) the rate of sebaceous flow from the glandsto the skin surface and 2) 11w rate of sebaceous spreadover the skin.

The amounts of sebum excretion which havebeen postulated to be parallel with the amountsof accompanying perspiration (3, 4) could beeither saturation level (total level) or "productioncapacity (excretion rate) in the general use ofthe term" which include both the rate ofsebaeeous spread over the skin and the rate ofsebaceous flow from the glands to the skin sur-face. On the other hand, the amounts of sebumexcretion which were dealt with in this experi-ment were not influenced by perspiration sinceboth sweat and sebum were adsorbed onto thesame starch paper at the time of their deliveryto the skin surface, with no opportunity foremulsification. Therefore these amounts shouldbe "production capacity (excretion rate) in thespecific use of the term," that is, the rate ofsebaceous flow from the glands to the skin surfaceonly. It would seem useful, therefore, to divideand clarify the term "excretion rate" as follows:

excretion excretion in the specifl°rate from or rate in use of the term

excrctioo the glands the ductrate spreading excretion. . . .. in the general

rate over or rate on use of the termthe skin the skin

C 3,Thysoitrte

30 THE JOURNAL OF INVESTIGATIVE DERMATOLOGY

FIG. 10

I -- .I —- - *41 .t9

-

/ , -—---I,

____ -- -

J_' - ii -

FIG. 9

'___.____._____.—_ .___,, ----—--...__—, J - '_IlIS'J •'%.##

. r'r°")

SEBTJM EXCRETION FRO1'I GLANDS TO SKIN SURFACE 31

FIG. 12

%UZ 4tn.Jatn

Ada tn-Ut#I •-..

:2k.

..!• • -

- --. ..—'., . ,- •.

F'.-.

I—.

(1w pwp.-rat'on)

c--•-•

-F

S(,,.a..tJ .b*ip;r.t.on)

Fio. 11

• --:4ffT.-.-

r-.

£

——4.

satntI'

' ••'-' ! 'It

'I

. —-—/r

• .4:.

(no pas,;,at..s)

32 THE JOURNAL OF INVESTIGATIVE DERMATOLOGY

This amount of sebum excretion (excretion ratefrom the glands) was observed not to parallel theamount of available perspiration. Also in con-trast to the amount of perspiration, the amountof sebum was not influenced by the intracutane-ous administration of the autonomic drugs norby emotional stimuli. These results present evi-dence of an absence of autonomic innervation ofthe secretory activity of the sebaceous glands.

Lipids in sweat which were indicated in thelower division of the starch paper by beingstained with light greyish or dark pinkish tintshould not be overlooked. Herrmann, Prose andSulzberger (3, 4) found that small but neverthe-less significant amounts of ether soluble materialson the palms where there arc no sebaccous glands.After a period of latency during which the freshlyoutpouring sweat droplets remained unstainedwith osmic acid vapors, a black dot usually ap-peared in the center of their base, followed bymore or less complete darkening of each of theindividual droplets. This observation lead themto the conclusion that lipids were attracted bythe sweat from underlying stratum corncum andwere supplied by the keratinizing epidermal cells.However, the amount increased in ether solublematerials which paralleled that of perspirationhas been explained with the interdependence ofsweat and lipids, not with the amount of lipidsin sweat. Takagi and Nitta (5) reported that thelittle amount of the small fat particles from thecccrinc glands histologically stained by osmicacid, resulted in a light greyish tint, not in adark brown or a black tint as were the cases ofneutral fat.

As previously mentioned, the staining resultsof the lipid indicated on the starch papers byosmic acid vapor, resembled that of the smallfat particles found by Takagi and Nitta. How-ever, whether fats are preformed components ofsweat, or merely are found to be in sweat beingderived from other sources, the question remainsunanswered in this paper.

In conclusion, the results obtained here havenever disagreed with the concept of Herrmannat al concerning parallelism in the amount ofsebum and sweat, but merely were observed ina different period of excretion, before the oppor-tunity for emulsification. Bc that as it may, inorder to investigate the existence of the secretoryinnervation of the scbaccous glands, it has beenregarded necessary to consider the excretion rate

from the glands, i.e., the excretion rate in thespecific use of the term.

SUMMARY

1. A method was developed to investigate theexistence of secretory innervation of the scbacc-ous glands, by absorbing sweat and scbumseparately and simultaneously onto starch paperupon their delivery to the skin surface, allowingthem no opportunity for emulsification.

2. With this method, the rate of scbaccous flowfrom the glands to the skin surface was observed,and was differentiated from the rate of sebaccousspread over the skin.

3. There were no significant changes in theamounts of scbum excretion which were not in-fluenced by perspiration before and after theintracutaneous administration of the autonomicdrugs, as compared with the amounts of perspira-tion on the back.

4. The same relationship was found on theforehead when the mental perspiration due toheat-pain was indicated on the starch paper.

5. The differentiation of types of perspirationon the forehead (mental, perpetual, and thermal)were tried by the reference to psychogalvanicreflex.

6. No parallelism was found to exist betweenthe amount of sebum excretion (the rate ofscbaccous flow from the glands to the skin sur-face) and that of perspiration.

7. The lipids in sweat were indicated on thestarch paper by osmic acid vapor.

AcKNowLEDGMENT

The author is indebted to Prof. Hatsuo Nittafor his constant interest and encouragement.

ADDENDUM

After submitting this manuscript for publica-tion, it was suggested to the author by Dr.Stephen Rothman, that the scbum which pene-trated the lower half of the starch paper couldhave been gravimctrically measured. In thelatest issue of the J. Invest. Dermat., 30: 99,1958, which the author read also after submittingthis manuscript for publication, Doctors Kligmanand Shelley reported that the absorption ofscbum by papers are sensitive enough to detectthe tiniest traces of lipoid and small differencesamong individuals, although 15% of lipoid re-mained on the skin. Yet it is obvious that gravi-

SEBUM EXCRETION FROM GLANDS TO SKIN SURFACE 33

metrical measurement could have made theresult more accurate, and the author expresseshis gratitude for Dr. Rothman's advice.

Drs. Kligman and Shelley also pointed out thatthe amount of sehum which reaches the skin sur-face in a given period of time is not necessarilythe amount produced during that time. Part ofit is pre-formed sebum pooled in the sebaceousfollicles. This old but important concept whichthe author had not considered shook the basisof this investigation. However they also haveobserved and reported that emotion had noeffect on lipoid excretion with a cup methodapplied on the forehead of two subjects. Inci-dentally they submitted the term "lipoid de-livery" for "production capacity", and theauthor preferred the term "excretion rate" to"production capacity".

REFERENCES1. ROTHMAN, S.: The problem of innervation, p.

303—305. Physiology and Biochemistry of theSkin. Chicago, University of Chicago Press,1953.

2. HARDy, J. D., WOLFF, H. G. AND GOODELL, H.:Pain Sensations and Reactions. Baltimore,Williams & Wilkins, 1952.

3. HEREMANN, F. AND PE05E, P. H.: Studies onthe ether soluble substances on the humanskin. I. Quantity and "replacement sum".J. Invest. Dermat., 16: 217, 1951.

4. HERRMANN, F., PRosE, P. H. AND SULZBERGER,M. B.: Studies on sweating. \T. Studies ofquantity and distribution of thermogenicsweat delivery to the skin. J. Invest. Der-mat., 18: 71, 1952.

5. TAKAeI, S. AND NITTA, H.: Kansen no soshiki-seirigaku teki kenkyu. (5) Jintai-kansen noshibo-bunpitsu. (Histological studies onsweat glands. (8) Fat secretion in humansweat gland. J. Physiological Society ofJapan. 9: 504, 1944 (in Japanese). Cited inKuno's Human Perspiration, p. 240—41.Springfield, Ill., Charles C Thomas.

94 THE JOURNAL OF INVESTIGATIVE DERMATOLOGY

24. Wynn, C. H. and Iqbal, M.: Isolation of ratskin lysosomes and a comparison with liverand spleen lysosomes. Biochem. J., 98: lOP,1966.

25. Olson, R. L. and Nordquist, R. E.: Ultramicro-scopic localization of acid phosphatase inhuman epidermis. J. Invest. Derm., 46: 431,1966.

26. Rowden, C.: Ultrastructural studies of kera-tinized epithelia of the mouse. I. Combinedelectron microscope and cytochemical studyof lysosomes in mouse epidermis and eso-phageal epithelium. J. Invest. Derm., 49: 181,1967.

27. Prose, P. H., Sedlis, E. and Bigelow, M.: Thedemonstration of lysosomes in the diseasedskin of infants with infantile eczema. J. In-vest. Derm., 45: 448, 1965.

28. Hall, J. H., Smith, J. G., Jr. and Burnett, S.C.: The lysosome in contact dermatitis: Ahistochemical study. J. Invest. Derm., 49:590, 1967.

29. Pearse, A. C. E.: p. 882, Histochemistry Theo-retical and Applied, 2nd ed., Churchill, Lon-don, 1960.

30. Pearse, A. C. E.: p. 910, Histacheini.stry Thea-retscal and Applied, 2nd ed., Churchill, Lon-don, 1960.

31. Daniels, F., Jr., Brophy, D. and Lobitz, W. C.:Histochemical responses of human skin fol-lowing ultraviolet irradiation. J. Invest.Derm.,37: 351, 1961.

32. Bitensky, L.: The demonstration of lysosomesby the controlled temperature freezing sec-tion method. Quart. J. Micr. Sci., 103: 205,1952.

33. Diengdoh, J. V.: The demonstration of lyso-somes in mouse skin. Quart. J. Micr. Sci.,105: 73, 1964.

34. Jarret, A., Spearman, R. I. C. and Hardy, J. A.:Histochemistry of keratinization. Brit. J.Derm., 71: 277, 1959.

35. De Duve, C. and Wattiaux, R.: Functions oflysosomes. Ann. Rev. Physiol., 28: 435, 1966.

36. Waravdekar, V. S., Saclaw, L. D., Jones, W. A.and Kuhns, J. C.: Skin changes induced by

UV irradiated linolenic acid extract. Arch.Path., 80: 91, 1965.

37. Nicolaides, N.: Lipids, membranes, and thehuman epidermis, p. 511, The EpidermisEds., Montagna, W. and Lobitz, W. C. Aca-demic Press, New York.

38. Wills, E. D. and Wilkinson, A. E.: Release ofenzymes from lysosomes by irradiation andthe relation of lipid peroxide formation toenzyme release. Biochem. J., 99: 657, 1966.

39. Lane, N. I. and Novikoff, A. B.: Effects ofarginine deprivation, ultraviolet radiationand X-radiation on cultured KB cells. J.Cell Biol., 27: 603, 1965.

40. Fukuyama, K., Epstein, W. L. and Epstein,J. H.: Effect of ultraviolet light on RNAand protein synthesis in differentiated epi-dermal cells. Nature, 216: 1031, 1967.

41. Daniels, F., Jr. and Johnson, B. E.: In prepa-ration.

42. Ito, M.: Histochemical investigations of Unna'soxygen and reduction areas by means ofultraviolet irradiation, Studies on Melanin,Tohoku, J. Exp. Med., 65: SupplementV, 10, 1957.

43. Bitcnsky, L.: Lysosomes in normal and patho-logical cells, pp. 362—375, Lysasames Eds.,de Reuck, A. V. S. and Cameron, M. Church-ill, London, 1953.

44. Janoff, A. and Zweifach, B. W.: Production ofinflammatory changes in the microcircula-tion by cationic proteins extracted from lyso-somes. J. Exp. Med., 120: 747, 1964.

45. Herion, J. C., Spitznagel, J. K., Walker, R. I.and Zeya, H. I.: Pyrogenicity of granulo-cyte lysosomes. Amer. J. Physiol., 211: 693,1966.

46. Baden, H. P. and Pearlman, C.: The effect ofultraviolet light on protein and nucleic acidsynthesis in the epidermis. J. Invest. Derm.,43: 71, 1964.

47. Bullough, W. S. and Laurence, E. B.: Mitoticcontrol by internal secretion: the role ofthe chalone-adrenalin complex. Exp. Cell.Res., 33: 176, 1964.