Histochemical studies of several oxidative enzymes of the muscle spindles in mouse limbs were carried out. Observations elucidated the existence of the three types of intrafusal fibers which can be distinguished from each other by the difference in the activity of oxidative enzymes as in the muscle fibers, i. e., the first one is the &#34;red intrafusal fibers&#34; and has the most intense activity of oxidative enzyme, the second one the &#34;white intrafusal fibers&#34; with a minimal activity, and the third, &#34;medium intrafusal fibers&#34; being distinguished by their oderate activity of oxidative enzymes from the others.</p

Mori, Masahiko

Ogata, Takuro

Okayama University Scientific Achievement Repository

Acta Medica OkayamaVolume 16, Issue 6 1962 Article 4DECEMBER 1962Histochemical Demonstration of the ThreeTypes of Intrafusal Fibers of Muscle Spindles,a Study on Oxidative EnzymesTakuro Ogata∗ Masahiko Mori†∗Okayama University,†Osaka University,Copyright c©1999 OKAYAMA UNIVERSITY MEDICAL SCHOOL. All rights reserved.Histochemical Demonstration of the ThreeTypes of Intrafusal Fibers of Muscle Spindles,a Study on Oxidative Enzymes∗Takuro Ogata and Masahiko MoriAbstractHistochemical studies of several oxidative enzymes of the muscle spindles in mouse limbswere carried out. Observations elucidated the existence of the three types of intrafusal fiberswhich can be distinguished from each other by the difference in the activity of oxidative enzymesas in the muscle fibers, i. e., the first one is the “red intrafusal fibers” and has the most intenseactivity of oxidative enzyme, the second one the ”white intrafusal fibers” with a minimal activity,and the third, ”medium intrafusal fibers” being distinguished by their oderate activity of oxidativeenzymes from the others.∗Copyright c©OKAYAMA UNIVERSITY MEDICAL SCHOOLActa Med. Okayama 16, 347-350 (1962)HISTOCHEMICAL DEMONSTRATION OF THE THREETYPES OF INTRAFUSAL FIBERS OF MUSCLESPINDLES, A STUDY ON OXIDATIVE ENZYMESTakuro OGATA and Masahiko MORI*Department of Surgery, Okayama University Medical SchoolOkayama, (Director: Prof. D. ]innai)*Department of Oral Surgery, Osaka University, OsakaDirector: Prof. K. KawakatsuReceived for publication, Di'Cember 1, 1962In the previous papersH it has been reported that the striated muscle fibersof mammals can be divided into three types from their activities of the his-tochemically demonstrable oxidative enzymes, i. e. the first one having thehighest activity is the smallest in diameter, which corresponds to the red musclefibers, the second one is the lowest in activity, larger in size, and the thirdhaving the moderate enzyme activity and being called "medium fibers" by theauthors, medium in size as compared with the other two. Correlated with thesethe authors have studied the oxidative enzyme activity of intrafusal fibers of themuscle spindle and it has been demonstrated that the intrafusal fibers can alsobe classified into three types from their activities of the oxidative enzymes as inthe striated muscle fibers of mice.MATERIALS AND METHODSThe striated muscles of limbs in healthy adult mice served as materials.The muscles were removed after sacrificying the animals under ether anesthesia,and cut into sections 20-30l-l thick in a cryostat at -16°C and then mountedon a slide glass. For the histochemical demonstration of succinoxidase, lacticdehydrogenase, malic dehydrogenase, a-glycerophosphate dehyrogenase, andDPNH-diaphorase and TPNH-diaphorase were employed, as in the methoddescribed by PEARSEll• After the incubation at 37°C for 30 minutes, the slideswere mounted by glycerin without dehydration.RESULTSThe intrafusal fibers of mouse muscle spindles were divided into threetypes according to the activities of oxidative enzyme reaction as the case withstriated muscle fibers (Fig. 1). The smaller intrafusal fibers showed a higheractivity of succinoxidase and contained a greater amount of formazan, while3471Ogata and Mori: Histochemical Demonstration of the Three Types of IntrafusalProduced by The Berkeley Electronic Press, 1962348 T. OGATA and M. MORIFig. 1 Fig. 2Fig. 1. The Muscle Spindle of M. Gastrocnemius in Mouse ()( 400).Showing succinoxidase reaction. Note the muscle spindles composed of the red intra-fusal fibers (intense activity, RF) and the medium intrafusal fibers (intermediate acttivity,MF) and the left side muscle spindle is composed of the white intrafusal fibers (minimalactivity, WF).Fig. 2. The Muscle Spindle of M. Gastrocnemius in Mouse ()(400)Showing succinoxidase reaction. Note the muscle spindle composed of the white intra-fusal fibers only.the larger intrafusal fibers a lower actIvIty. And the medium sized intrafusalfibers showed an intermediate activity between the smaller intrafusal fibers andthe larger fibers. The activity of succinoxidase of the smaller fibers corres·ponded to that of the red muscle fibers, and the medium and larger intrafusalfibers showed the same degree of activity as the medium and white musclefibers.Thus the smaller intrafusal fibers with a marked activity were designatedas "red intrafusal fibers", the larger ones as "white intrafusal fibers" and themedium ones as "medium intrafusal fibers" in the present paper.Most intrafusal fibers had shown such a relationship as mentioned abovebetween the size and succinoxidase reaction, but some intrafusal fibers revealedno such a relationship, e. g. some white intrafusal fibers were small in size.Some of the muscle spindles were composed of only one type of the intra·fusal fibers, while the others were of different types of the intrafusal fibers.Similar findings were observed in the histochemical reactions of lactic de-hydrogenase, malic dehydrogenase, a-glycerophosphate dehydrogenase, DPNH·diaphorase and TPNH·diaphorase. i. e., the red intrafusal fibers showing ahigher activity of these enzymes, the medium intrafusal fibers a moderate ac-tivity and the white intrafusal fibers the lowest.2Acta Medica Okayama, Vol. 16 [1962], Iss. 6, Art. 4http://escholarship.lib.okayama-u.ac.jp/amo/vol16/iss6/4Intrafusal Fibers of Muscle Spindles 349DISCUSSIONMuscle spindles were known as the sensory receptors in the striated muscle(CooPER)8. Recently, KONOO (1957)7 reported that two types of intrafusal fiberswere distinguished in cat muscle spindles by Sudan black B staining. In ourprevious papers (OGATAl-'l, 1958; OGATA and MORI\ 1961, and TSUKAMoToand MORI8, 1962) it was reported that three types of muscle fibers, namely,the red, white and medium muscle fibers were distinguished in mammalianmuscle fibers. In the present study, the three types of intrafusal fibers aredifferentiated in the mouse muscle spindles by oxidative enzyme reactions. Andthe three types of intrafusal fibers have been designated as "red intrafusal"medium intrafusal" and "white intrafusal fibers", respectively.The metabolism of intrafusal fibers has been left almost unclarified. KARLand BRZENSKI (1961)11 stated that the spindle space contained a high amount ofhyaluronic acids and it showed no lymphatic system on either PAS or Hale-PASstam. But the present study revealed that the three types of intrafusal fibersshowed the same oxidative enzymatic activity as that of the three types of themuscle fibers already described. From the present results, it may reasonablybe assumed that the metabolic activity in intrafusal fibers is similar to that ofmuscle fibers. Since mitochondria contain a larger amount of oxidative andreducing enzymes, it may be safe to assume that the red intrafusal fibers con-tain a considerable number of mitochondria, while the white intrafusal fiberscontain a little or none of them. As the other vertebrates possess three types ofmuscle fibers, it is likely that they may have three types of the intrafusal fiberas well. This problem, however, needs further investigation.SUMMARYHistochemical studies of several oxidative enzymes of the muscle spindlesin mouse limbs were carried out. Observations elucidated the existence of thethree types of intrafusal fibers which can be distinguished from each other bythe difference in the activity of oxidative enzymes as in the muscle fibers, i. e.,the first one is the "red intrafusal fibers" and has the most intense activity ofoxidative enzyme, the second one the "white intrafusal fibers" with a minimalactivity, and the third, "medium intrafusal fibers" being distinguished by theirmoderate activity of oxidative enzymes from the others.REFERENCES1. OGATA, T.: A histochemical study of the red and white muscle fibers. Part 1. Activity ofthe succinoxidase system in muscle fibers. Acta Medicinae Okayama 12, 216, 19582. OGATA, T.: A histochemical study of the red and white muscle fibers. Part n. Activity3Ogata and Mori: Histochemical Demonstration of the Three Types of IntrafusalProduced by The Berkeley Electronic Press, 1962350 T. OGATA and M. MORIof the cytochrome oxidase in muscle fibers. ibid. 12, 228, 19583. OGATA, T.: A histochemical study of the red and white muscle fibers. Part III. Activityof the diphosphopyridine nucleotide diaphorase and triphosphopyrindine nucleotide diaphorasein muscle fibers. ibid. 12, 233, 19584. OGATA, T., and MORI, M.: A histochemical study on the muscles of various animals. Ac-tivities ot oxidative enzymes and phosphorylase in muscle fibers. Proc. fap. Histochem.Assoc. 2nd Ann. Meeting, 19615. PEARSE, A. G. E.: Histochemistry. Theoretical and Applied. I and A. Churchill, London(&cond Edition), 19606. COOPER, S.: Muscle spindles and other muscle receptors. The structure and function ofmuscle. Vol. 2, edited by G. H., Bourne, Academic Press, N. Y., London, 19607. KONDO, M.: Functional differences of skeletal muscle (in Japanese), 10th Meeting Jap. Elec-tromyogram Soc.8. TsuKAMoTo, S., and MORI, M.: Spatial distribution of skeletal muscle fibers based onsuccinic dehydrogenase acttivity. (in prepar)9. KARL, D., und VON BRZEZINSKI, v. D. K.: Untersuchungen zur Histochemie der Mus-kelspindeln zur Topochemie und Function des Spindleraumes und der Spindelkapsel. ActaHistochem. 12, 277, 19614Acta Medica Okayama, Vol. 16 [1962], Iss. 6, Art. 4http://escholarship.lib.okayama-u.ac.jp/amo/vol16/iss6/4

Histochemical Demonstration of the Three Types of Intrafusal Fibers of Muscle Spindles, a Study on Oxidative Enzymes

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