TANAKA, SEISUKE

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TitleElectron Histochemical Demonstration of Acid PhosphataseActivity in the Motoneurons of the Cervical Spinal Cord of Ratduring Axonal ReactionAuthor(s)TANAKA, SEISUKECitation日本外科宝函 (1965), 34(4): 829-832Issue Date1965-07-01URL http://hdl.handle.net/2433/206516RightType Departmental Bulletin PaperTextversionpublisherKyoto University原著Electron Histochemical Demonstration of Acid Phosphatase Activity in the Motoneurons of the Cervical Spinal Cord of Rat during Axonal Reaction by SE!SUKE TANAKA From the Dep"rtment of OrthopwJ1c沿urgery・K、＜＞l<>Univ町、1¥'Schcxil of Mediにltlt'(Directけr；ドnd, Dr. T1;Tsuo ho) and the Dep:irtment of Anat川町， IふけtoUniversity School of Medicme (Director : Prof. Dr. M1rn10 OK，、MOTO)Received for Publication Apr. 10, 1965 829 Acid phosphatase activity in terms of lysosomes in the central nervous system has been investigated at the level of light microscopic histochemistry by several authors (BARRON，’611) , BECKER & BARRON，’614) ; BARRON & SKLAR，’6l 2l; BARRON & TUNCBAY, ’623) ; 0GA w A etal.，’6Q9> ; 0GA WA et al.，’6110)). NOVIKOFF （’59) 7> found enlargement of lysosomes in the cells of the proximal tubu-les in experimental renal hydronephrosis, and for such enlarged lysosomes he designated the term "cytolysome”. Barron attempted to study cytochemically response of lysosomes to injury of nervous tissue and reported that enlarged droplets rich in acid phosphatase activity are demonstrated in the necrotizing neurons in the brain injured by diphtheria toxin (BARRON，’611>). The similar findings in the neurons have also been observed during the early phase of both postmortem autolysis and necrobiosis in the anoxic and anoxic-ischemic encephalopathy (BECKER & BARRON，’61 4l) and also during axonal reaction (BARRON & SKLAR，’612>). In the present investigation attempts were made to demonstrate electron histochemi-cally response of acid phosphatase activity in the motoneurons of the cervical spinal cord to axonal reaction. The cervical spinal cords of Wistar rats, weighing 120 to 200g, were used. Sだctionof the brachial nerve was performed through posterior paraspinal approach. Animals were sacrificed seven and fourteen days after operation. Go MORI’s lead acetate method for acid phosphatase (GOMORI，’5261) was essentially used in cold formoトcalciumfixed frozen sections. Substrate-free incubating media were taken for controls. After incubation specimens were postfixed in Palade’s buffered osmium tetroxide and processed according to the routine procedure for the observ-ation by the ele-ctron microscope (0GA w A etal.，’6211)). In the present study deposits of electron opaque lead phosphate indicating acid phos-phatase activity were positive in dense bodies (lysosomes) in the perikaryon (Fig. 1). In some dense bodies precipitates of lead phosphate were found in their limiting membrane and also scatteredly in the bodies. The smooth membranes, presumably the membrane 830 日本外科宝函第34巻第4号components compnsmg the Golgi apparatus wer己 alsopositive for enzymatic act1v1ty. However, no mitochondria showed enzymatic activity. NOVIKOFF & EssNER （’62) S) described that lysosomes are produced by dilatation and separation of the Golgi cisternae. OsrNCHAK (964) 12> also reported that acid phosphatase activity is localized to certain Golgi cisternae, surrounded by a single membrane. Though size of enzymatically active dense bodies in the motoneurons did not show any appreciable change during axonal reaction, number of dense bodies seemed to be in-creased markedly (Figs. 2 & 3) . The electron density of dense bodies itself was also much higher in axonal reaction than in normal controls. This finding is due to abundance of lead phopshate precipitates indicating the increased enzymatic activity within dense bo-dies. Occasionally vacuolated dense bodies were also observed. Enlarged lysosomes (cyto・lysome) observed under the light microscope by BARRON & SKLAR （’61) 2> were not seen in the present investigation carried at the level of electron microscope. In the present investigation it was observed that dense bodies markedly increased in number in cervical motoneurons during axonal reaction. This finding may indicate the increased acid phosphatase activity, which is in agreement with the result of biochemical study by BoorAN & MELLORS （’45) 5>. The increased enzymatic activity may be indicative of increased hydrolytic activity as suggested by NOVIKOFF （’59) 7>, however, direct involve-ment of the enzyme with cellular metabolic activity in the motoneurons during the process of axonal reaction回 nnot be excluded. This study "'"' cJrried out under the helpful 川 gc,l1<n by Dr. KAzuo OcAwA, Proftで判》rof Anatomy, Kansai Mt'dical Sch川 I,Mリriguchi，。日ka,Japan REFERENCES I J Barron, K. D. Histochemical 叶udyof neuronal necro'i' produced by intracerebral injection of diphtheria toxin. ¥i’ぞurology.1 : 714, 1961. 2) Barron, K. D. and Sklar, S. : R白ponseof lysosom引 ofbulb＜》spinalmotoneurons to axon section. Neuro-logy, 11 : 866, 1961. 3) R<rron, K. D. and Tuncbay, T. 0. : Hist<)c:1emistry of acid phosphatase and thiamine pyrophcsphatase during axon r白ction.Arn. J. Path., 40 : 637, 1962. 4) Becker, I¥. H. and B1rron, K. D. ・ The cytochemistry of anoxic and anoxic-ischemic encephalopathy m rats. Am. J. Path .38 : 161, 1961. 5) Bodian, D. and ¥1elors, R. C. : The regenerative cycle of motoneurons, with 'P"-'C、1al町長renceto phosphatase activity. J. Exp. Med., 81 . 469, 1945. 6) Gomnri, G. : Microscopic hi>t•x:hemistry. Principles and practi町 Chicago,University of Chicago Pr出 1195'.2 7) :¥'ovikof, A. B. : The proximal tubule cel in experimental hydronephrosis. J. Biophys. Biochern. Cytol., 6 : 136, 1959. 8) Novikoff, A. B. and E州ner,E. : Pathological changes in cytoplasmic organelles. Federation Pr田町dmgs,21 : 1131, 1962. 9) Ogawa, K.. Mizun<入 0!.,Hashimoto, K.. Fuji, S. and Okamoto, K. : Cytochem1'try of cultured neural ti地 U白 IL Acld1tional staining techniques with special reference to ly臼泊ornes.Proc. Dept. An川 Kyoto Univ. Sch. Med., 4: I, 1960. 10) Ogawa, K. Millmo, N. and Okamoto M. ・ Iλ＇o".imes in cultured cels. J. Histochem. Cytochem., 8 202, 1961. 11) 0日awa.K., Shinonaga, Y. and Suzuki, T. : Electron histochemical demonstration ,f acid phosphatase activity in the rat cerebellum. J. Electronmicr., 11 : 111, 1962. 12) （九ll・hak,J. : El町tronmicroscopic localizallon of acid phosphatase anオthiaminepyrophosphatase activity in hypothalamic neurosecretory el Is of the rat. ]. （ぞI.Biol. 21: 35, 1964. ELECTRO:¥ HIST()CHEMICAL DEMONSTRATIO:¥ OF ACID PHOSPHATASE ACTIVITY 831 Fig. Fig. 2 Fig.Ji 3 832 日本外科宝函第34巻第4号Fig. 1 Dei~＞sib of electron opaque lead phosphate showing the acid phosphatase 出：tivity are seen in dense 民di白 inperil泊ryonof cervical motoneurons in the normal adult rat. 26000 x Figs. 2 & 3 Cervical motoneurons. Fourteen d3ys after the 同Ctli>nof brachia! nerve. Note an increase m number of dense bodies. The electron density of』di白 isalso highly increased. Fig. 2, 26000× Fig. 3, 29000 x 和文抄録軸索反応、の際l乙見られるラ ット脊髄前角細胞内酸性フオスファターゼ活性の電子顕微鏡学的研究京都大学医学部整形外科学教室（指導：伊藤鉄夫教授）京都大学医学部第I解剖学教室（指導：岡本道雄教授）BJ 中 清 介ラットの上院神経を切断し， 7日及び14日後におけ ムのれが楢し同時にリ ソゾーム内酵素活性の上昇がる頚髄前角細胞を用いた。材料は冷フォルモール・カ 認められた．時々空胞をもったリソゾームの出現もみルシウムで固定し，これを凍結切片とした｛も Gomori られた．の錯酸鉛法で酸性フオスファターゼ活性を検出した． これらの事実は軸索反応の際に前角細胞内におい後固定には Paladeの緩衡4酸化オス ミウム液を使用 て，酸性フオスファーターゼ活性が上昇することを示しp 通常の電子顕微鏡標本と同じ過程を経てエポン包 している．この酵素活性の上昇は細胞内加水分解作用埋を行なった． の冗進を示すかもしれないが，一方酸性フオスファタ酸性フオてファターゼは正常脊髄前角細胞内におい ーゼが軸索反応の際に， 直接前角細胞内物質代謝に関てもリ ソゾー ム 「ly""''me）および Gomori装置を含む 与していることを全〈否定し去るわけには行かない．滑函小胞体にも認められた．神経切断例ではリソゾー

軸索反応の際に見られるラット脊髄前角細胞内酸性フオスファターゼ活性の電子顕微鏡学的研究

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軸索反応の際に見られるラット脊髄前角細胞内酸性フオスファターゼ活性の電子顕微鏡学的研究

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