Quantitative Histochemistry of Human Skin

Abstract

Ite('eived for publication Jttne 1, 1959 StU(ly of the enzymes of skin an(l l)articuiarlY of el)i(lerlflis has i)eeli llan(licappe(l 1)y its histological complexity. The micro methods devised by I)r. 0. H. Lowry have pernlitte(l accurate Samfling of various (('11 ty)es in brain, an(l other structures. In this a jer, (lata are l)resentecl testing :tIuI vali(lating nlinor nlodiflcations of these lilethO(15 that are useful for the studs' of various ('elI types in litlIllali skin. Study of the enzynles of skin :tiicl especially of ('j)i(lerIllis has 1)een Ilalldicapped also by iifficulty in se)arating and honiogenizing elji(lerl1lis, an(I at't'urate sanl)ling of the at'tive layers an(l structures. At'curate sanl)hng is necessary 1)ecaUSe the sul)jacent (lernus is relatively inactive. Soine of tllese difficulties have l)een circulllveflte(l so that sn all 5itIlll)les of particular tell tVl)es (all 1)e analvze(l. 'lilt' methods are sufficiently l)reeiSe and setisitive to l)('rlllit conll)arison of nlicrogr:ull 5aI1 l)les. 'i'his)a)er rel)orts nleasurenlent of five emizvines fromii different 11letal)Olic l)atll\\'aYs in 5-20 'y sahlil)les (wet weight) of honlogellates of hunlan ('l)i(lerlllis, or 0.5 to 5 y sanll)les fronl freeze (lried sections. ihe activity of four of these enzvnu's (It) not a i ear to llave 1)eel1 reported in ('l)i(lerluis I)efOi(. I)ata slll)wing the distril)UtiOn of these enzvnles in el)i(lernlis 1(11(1 (lernlis of the sole of the foot and skill of the groin and mdivi(lual samples of hair follicles, sel)aceous aIl(l sweat glands are presented. These may serve as baseline observations for studies for carcinogenesis, healing wounds and other physiological or)atllOl0gical l)ro('esses. Ml:TIlol)s To Ol)taih! tile ol)timt!m cOn(lit ions for the metho(1s human epidermal homogenates were used. The epidermis was separated by our modifi (at it)!! (2) of t lie ''stretch met hod'' of \an Scott From the I)epart

Fine Structure and Enzyme Histochemistry of Stratified Squamous Epithelium in the Human Uterine Cervix: With Reference to the Epidermis in the Female Thigh

“…The presence of isocitric acid dehydrogenase in the human epidermis has been described quantitatively (Cruickshank et al, 1958). By the same method, the quantity of lactic acid (LDH), malic acid (MDH) and g:ucose-6-phosphate dehydrogenase (G6PDH) in human skin has also been demonstrated (Hershey et al, 1960).…”

Metabolic Processes in Skin.

“…The activities of the dehydrogenases required for the initial steps have heen measured hy Hershey and Mendle (1954) and Hershey et al (1959) and further evidence for the existence of this pathway was obtained hy measurement of the differing rat^es and which ^^C^ and ^^G^ lahelled glucose were converted into "CO2 (Freinkel, 1960). Finally, Yardley and Godfrey (1963) demonstrated the presence of the intermediates sedoheptulose-7-phosphate, sedoheptulose diphosphate and rihose-5-phosphate in skin incuhated in ^^P-orthophosphate and/or glucose-U-^*0.…”

“…Isocitric dehy^irogenase has since been shown to he present in epidermis (Cruickshank et al, 1958) and several intermediates of the cycle have been isolated (Pqmerantz-and Ashornsen, 1960). In addition the activities of most of the enzymes involved have heen measured hy fluorometric technique (Hershey et al, 1959).…”

“…Finally, Yardley and Godfrey (1963) demonstrated the presence of the intermediates sedoheptulose-7-phosphate, sedoheptulose diphosphate and rihose-5-phosphate in skin incuhated in ^^P-orthophosphate and/or glucose-U-^*0. Hershey et al (1959) comment that enzymes of this cycle are considerahly more active than those of the tricarhoxylic cycle. Halprin and Chow (1961) present evidence that there is a relative hlock of the tricarhoxylic cycle at the isocitric dehydrogenase stage and it may he that " direct " oxidation of glucose is the major oxidative pathway.…”

Subcellular distribution and partial characterization of gingival mitochondrial and soluble malate dehydrogenases

“…though malate dehydrogenase is ktiown to Introduction 1^^ associated with oxidative phosphorylation Malate dehydrogenase (E.C.I.1.1.37) has (Fine et al 1976) in isolated rat oral mucobeen previously studied within the various sal mitochondria, recent evidetice also sugcell layers in skin (Hershey et al 1960, gests that M-MDH, and S-MDH isoenzymes Adachi et al 1969) and within keratinized act as important carriers of reducing equivoral mucosa (Gerson et al 1966(Gerson et al , 1970, alents from the mitochondria to the cytosol Bergquist 1972). However, this data was via the "malate shuttle" (Krebs et al 1967, confined to total enzymatic activities of the Rognstad & Katy 1973, Wheatley 1974.…”