Summary
Hepatocytes from 12-day-old rats, pre- and post-natally exposed to alcohol, together with those from pair-fed controls, were isolated and subfractionated in six cell subpopulations on Percoll density gradients. These cells were characterized using a combination of biochemical and stereological methods. The low density cells (F2) mainly showed biochemical and stereological features of perivenous hepatocytes, whereas the heavier cells (F6) were primarily periportal hepatocytes. The alcohol-metabolizing enzymes, alcohol dehydrogenase and aldehyde dehydrogenase (high and low Km) showed more activity in the F2 fraction. Alcohol-altered mitochondria and Golgi apparatus occurred mainly in F2 cells, whereas the endoplasmic reticulum and lysosomes appeared to be more altered in the F6 hepatocytes. Alcohol also induced the appearance of some small hepatocytes, with a well-developed rough endoplasmic reticulum and an increased number of mitochondria. Biochemical data indicated that glutamate dehydrogenase and alanine aminotransferase were more affected in F2 cells from alcohol-treated rats, and that the activity of the ethanol-metabolizing enzymes was also reduced in these hepatocytes. Our results indicate that alcohol exposure during zonal development in the liver could have a selective effect on specific cell components depending on the acinar zone, and that the perivenous hepatocytes appear to be more damaged under these conditions.
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Bengtsson BG, Kiessling K-H, Smith-Kielland A, Morland J (1981) Partial separation and biochemical characteristics of periportal and perivenous hepatocytes from rat liver. Eur J Biochem 118:591–597
Bernaert D, Wanson J-C, Mosselmans R, De Paermentier F, Drochmans P (1979) Separation of adult rat hepatocytes into distinct subpopulations by centrifugal elutriation. Morphological, morphometrical and biochemical characterization of cell fractions. Biol Cell 34:159–174
Cruz-Orive LM, Weibel ER (1981) Sampling designs for stereology. J Microscopy 122:235–257
Drochmans P, Wanson J-C, Mosselmans R (1975) Isolation and subfractionation on Ficoll gradients of adult rat hepatocytes. Size, morphology and biochemical characteristics of cell fractions. J Cell Biol 66:1–22
Guerri C, Grisolía S (1982) Effects of prenatal and postnatal exposure of rats to alcohol: Changes in (Na+,-K+)ATPase. Pharmacol Biochem Behav 17:927–932
Guerri C, Sanchis R (1986) Alcohol and acetaldehyde in rat’s milk following ethanol administration. Life Sci 38:1543–1556
Guerri C, Wallace R, Grisolia S (1978) The influence of prolonged ethanol intake on the levels and turnover of alcohol and aldehyde dehydrogenases and of brain (Na + K)ATPase of rats. Eur J Biochem 86:581–587
Gumucio JJ, Miller DL (1981) Functional implications of liver cell heterogeneity. Gastroenterology 80:393–403
Gumucio JJ, Miller DL (1982) Liver cell heterogeneity. In: Arias I, Popper H, Schachter D, Shafritz DA (eds) The liver: biology and pathobiology. Raven Press, New York, p647
Idéo G, DeFrancis R, Ninno ED, Dioguardi N (1972) Decrease of rat liver glutamate dehydrogenase after chronic administration of ethanol. Enzymology 43:245–251
Ji S, Lemasters JJ, Christenson V, Thurman RG (1983) Selective increase in pericentral oxygen gradient in perfused rat liver following ethanol treatment. Pharmacol Biochem Behav 18:439–442
Jungermann K, Sasse D (1978) Heterogeneity of liver parenchymal cells. TIBS Sept 198–202
Kanamura S, Kanai K, Oka M, Shugyo Y, Watanabe J (1985) Quantitative analysis of development of mitochondrial ultrastructure in differentiating mouse hepatocytes during postnatal period. J Ultrastruct Res 93:195–204
Katz N, Teutsch H, Jungermann K, Sasse D (1976) Perinatal development of the metabolic zonation of hamster liver parenchyma. FEBS Lett 69:23–28
Koivula T, Koivusalo M (1975) Different forms of rat liver aldehyde dehydrogenase and their subcellular distribution. Biochim Biophys Acta 397:9–23
Koivula T, Lindros KO (1975) Effects of long-term ethanol treatment on aldehyde and alcohol dehydrogenase activities in rat liver. Biochem Pharmacol 24:1937–1942
Lemasters JJ, Ji S, Stemkowski CJ, Thurman RG (1983) Hypoxic hepatocellular injury. Pharmacol Biochem Behav 18:455–459
Lieber CS (1983) Alcohol and the liver. In: Arias IM, Frenkel M, Wilson JHP (eds) The liver annual. Elsevier, Amsterdam, New York, p 106
Lieber CS, DeCarli LM (1976) Animal models of ethanol dependence and liver injury in rats and baboons. Fed Proc 35:1232–1236
Lindahl R, Clark R, Evces S (1983) Histochemical localization of aldehyde dehydrogenase during rat hepatocarcinogenesis. Cancer Res 43:5972–5977
Lindros KO, Bengtsson G, bc]Salaspuro M, Väänänen H (1986) Separation of functionally different liver cell types. In: Thurman RG, Kauffman FC, Jungermann K (eds) Regulation of hepatic metabolism. Plenum Publ Corp, New York, London, p 137
Loud AV (1968) A quantitative stereological description of the ultrastructure of normal rat liver parenchymal cells. J Cell Biol 37:27–46
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275
Ma MH, Biempica L (1971) The normal human liver cell. Cytochemical and ultrastructural studies. Am J Pathol 62:353–372
Meihuizen SP, Blansjaar N (1980) Stereological analysis of liver parenchymal cells from young and old rats. Mech Ageing Dev 13:111–118
Morrison GR, Brock FE (1967) Quantitative measurement of alcohol dehydrogenase in the lobule of normal livers. J Lab Clin Invest 70:116–120
Morrison GR, Brock FE, Karl IE, Shank RE (1965) Quantitative analysis of regenerating and degenerating areas within the lobule of carbon tetrachloride-injured liver. Arch Biochem Biophys 111:448–460
Popper H, Lieber CS (1980) Histogenesis of alcoholic fibrosis and cirrhosis in the baboon. Am J Pathol 98:695–716
Radford J, Bhathal PS (1985) Purification of fetal rat hepatocytes. Cell Biol Int Rep 9:677–688
Rappaport AM, Borowy ZJ, Lougheed WM, Lotto WN (1954) Subdivision of hexagonal liver lobules into a structural and functional unit. Anat Rec 119:11–34
Renau-Piqueras J, Gómez-Perretta C, Guerri C, Sanchis R (1985 a) Qualitative and quantitative ultrastructural alterations in hepatocytes of rats prenatally exposed to ethanol with special reference to mitochondria, Golgi apparatus and peroxisomes. Virchows Arch [Pathol Anat] 405:237–251
Renau-Piqueras J, Guerri C, Miragall F, Gómez-Perretta C, Báguena-Cervellera R (1985 b) Alterations in the cytochemical activity of several phosphatases in hepatocytes from rats exposed prenatally to ethanol. Virchows Arch [Cell Pathol] 49:249–259
Renau-Piqueras J, Miragall F, Guerri C, Báguena-Cervellera R (1987) Prenatal exposure to alcohol alters the Golgi Aparatus of newborn rat hepatocytes. A cytochemical study. J Histochem Cytochem 35:221–228
Renau-Piqueras J, O’Connor JE, Báguena-Cervellera R, Grisolia S (1986) Ammonium chloride-induced alterations in growth kinetics and ultrastructure of murine neuroblastoma cells. A flow cytometric and stereologic study. Virchows Arch [Cell Pathol] 50:271–283
Rubin E, Lieber CS (1967) Early fine structural changes in the human liver induced by alcohol. Gastroenterology 52:1–13
Salinas M, Wallace R, Grisolía S (1974) Comparative studies in vivo and in vitro of rat-liver enzymes. Eur J Biochem 44:375–381
Sanchis R, Guerri C (1986) Chronic ethanol intake in lactating rats: milk analysis. Comp Biochem Physiol 85:107–110
Schmucker DL, Mooney JS, Jones AL (1977) Age-related changes in the hepatic endoplasmic reticulum: A quantitative analysis. Science 197:1005–1008
Schmucker DL, Mooney JS, Jones AL (1978) Stereological analysis of hepatic fine structure in the Fischer 344 rat. J Cell Biol 78:319–337
Segal HL, Matsuzawa T (1970) L-Alanine aminotransferase (Rat liver). In: Tabor H, Tabor CW (eds) Methods in Enzymology, vol XVII A. Academic Press, Inc, New York pl53
Seglen PO (1976) Preparation of isolated rat liver cells. Meth Cell Biol 13:29–83
Shank RE, Morrison G, Cheng CH, Karl I, Schwartz R (1959) Cell heterogeneity within the hepatic lobule (quantitative histochemistry). J Histochem Cytochem 7:237–239
Sturgess JM, de la Iglesia FA (1972) Morphometry of the Golgi apparatus in developing liver. J Cell Biol 55:524–530
Thurman RG, Ji S, Lemasters J (1986) Lobular oxygen gradients: Possible role in alcohol-induced hepatotoxicity. In: Thurman RG, Kauffman FC, Jungermann K (eds) Regulation of hepatic metabolism. Plenum Publ Corp, New York, London p 293
Tonda K, Hasegawa T, Hirata M (1983) Effects of phenobarbital and 3-methylcholanthrene pretreatments on monooxygenase activities and proportions of isolated rat hepatocyte subpopulations. Mol Pharmacol 23:235–243
Väänänen H, Salaspuro M, Lindros K (1984) The effect of chronic ethanol ingestion on ethanol metabolizing enzymes in isolated periportal and perivenous rat hepatocytes. Hepatology 4:862–866
Wanson JC, Drochmans P, May C, Penasse W, Popowski A (1974) Isolation of centrolobular and perilobular hepatocytes after phenobarbital treatment. J Cell Biol 66:23–41
Weibel ER (1979) Stereological methods, vol 1. In: Weibel ER (ed). Academic Press, New York
Welsh FA (1972) Changes in distribution of enzymes within the liver lobule during adaptative increases. J Histochem Cytochem 20:107–111
Williams M (1977) Stereological techniques. In: Glauert AM (ed) Practical methods in electron microscopy, vol 6, part 2, chap 2. North Holland/American Elsevier, Amsterdam, p 1
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Sancho-Tello, M., Renau-Piqueras, J., Báguena-Cervellera, R. et al. A biochemical and stereological study of neonatal rat hepatocyte subpopulations. Virchows Archiv B Cell Pathol 54, 170–181 (1987). https://doi.org/10.1007/BF02899209
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DOI: https://doi.org/10.1007/BF02899209