Intersegmental reflex actions from a joint sensory organ (CB) to a muscle receptor (MCO) in decapod crustacean limbs

18  Download (0)

Full text

(1)

With 10 figures Printed in Great Britain

INTERSEGMENTAL REFLEX ACTIONS FROM A JOINT

SENSORY ORGAN (CB) TO A MUSCLE RECEPTOR (MCO)

IN DECAPOD CRUSTACEAN LIMBS

BY B. M. H. BUSH,* J. P. VEDEL AND F. CLARAC

Ddpartement de Neurobiologie comparde, C.N.R.S. - INP. 10,

31 chetnin Joseph-Aiguier, \iz~j\ Marseille cedex 2

(Received 21 June 1977)

SUMMARY

In the walking legs of decapod crustaceans, intersegmental reflex actions originate from various joint proprioceptors. The activity of the ' accessory flexor' (AF) muscle, which with the myochordotonal organ (MCO) con-stitutes a muscle proprioceptor for the mero-carpopodite (M-C) joint, is modulated by the sensory discharge of a joint receptor (CB chordotonal organ) for the more proximal, coxo-basal (C-B) joint. Selective mechanical stimulation of the CB organ also reflexly modifies the motor activities of the main M-C flexor and extensor muscles (recorded as EMGs).

1. Dynamic CB stretch (as would occur during a dorso-ventral C-B movement - i.e. 'depression' of the limb) stimulates motor discharge to the M-C extensor muscle, while dynamic release of CB (as during a ventro-dorsal C-B movement - or leg ' elevation') excites the accessory flexor as well as the main flexor muscle.

2. Successive M-C muscle responses to repetitive sinusoidal changes of CB length differ quantitatively according to the direction (stretch or release) of the first CB movement, in some cases increasing but more commonly 'adapting' with repetition.

3. Reflex discharge frequencies of the extensor, flexor and accessory flexor motoneurones increase with velocity of CB movement.

4. Eye illumination, and spontaneous or other sources of increased central excitability, generally increase the CB reflex drive to the flexor and accessory flexor muscles and, in parallel, decrease the reflex action on the extensor

muscle.

The results are discussed in terms of the role of proprioceptive reflexes in intersegmental co-ordination of the leg joints. In particular the significance of the reflex regulation of the myochordotonal receptors, and thereby the gain of the M-C resistance reflexes, is considered in the light of the observed 'co-activation' of main flexor and receptor muscle motoneurones.

INTRODUCTION

In the walking legs of decapod crustaceans, the 'myochordotonal organ' (MCO) (Barth, 1934) constitutes with the 'accessory flexor' (AF) muscle a neuromuscular-sensory system whose functions can be compared to those of the thoracico-coxal

(2)

48 B. M. H. BUSH, J. P. VEDEL AND F. CLARAC

organ (Alexandrowicz & Whitear, 1957; Bush, 1976), the muscle receptor organ of the crustacean abdomen (Alexandrowicz, 1951, 1967; Fields, 1976), and also the mam-malian muscle spindle (Matthews, 1972). In all these systems, specialized muscle fibres transmit to the associated sensory cells the mechanical strains consequent upon joint movements, but can also act directly on the sensory discharge by their own contraction. The crustacean myochordotonal organ has been extensively studied, both anatomically (Clarac & Masson, 1969; Alexandrowicz, 1972) and physiologically (Cohen, 1963; Evoy & Cohen, 1971; Clarac & Vedel, 1971, 1975). Its role in mediating 'resistance reflexes', analogous functionally to vertebrate stretch reflexes, has also been well established (Bush, 19656; Cohen, 1965; Evoy & Cohen, 1969; Vedel, Angaut-Petit & Clarac, 1975 a; see also Evoy, 1976; Clarac, 1977).

The existence of a single, discrete muscular structure, exclusively involved in the control of the activity of a sensory organ (MCO), allows a study of the reflex organiza-tion and physiological significance of its motor command. The accessory flexor muscle is supplied by only one tonic excitatory motoneurone, whose discharge is modulated by spontaneous or induced central influences (Vedel et al. 1975 a). It is also subject to resistance-reflex-like control, by MCO itself and by the chordotonal organs (MC 1 and MC 2) of its own mero-carpopodite (M-C) joint (Bush, 19656; Evoy & Cohen, 1969, 1971; Vedel et al. 1975 a).

This paper considers the 'intersegmental reflex' actions exerted on the AF muscle by a joint proprioceptor organ located in a more proximal segment of the same leg, namely the coxo-basal chordotonal organ (CB), which monitors movement of the coxo-basipodite (C-B) joint (Bush, 1965 a). These reflex effects are compared with those exerted by the same CB organ on the extensor and flexor muscles producing M-C joint movements.

Thus the present study characterizes the reflex modulation, by one particular joint receptor, of the activity of the single excitatory motoneurone innervating a specific neuromuscular sensory organ. It also aims to extend our present understanding of the overall role of one proprioceptor in intersegmental motor co-ordination within a walking leg. This work represents part of a general study of the CB organ influences on all the joint musculature in the same limb (see Clarac, Vedel & Bush, 1978). Preliminary accounts of some of these results have appeared previously (Bush & Clarac, 1975; Vedel, Clarac & Bush, 19756).

MATERIAL AND METHODS

Four decapod crustacean species were used in this study: the crayfish, Astacus

leptodactylus (Astacura); the rock lobsters, Palinurus vulgaris and Jasus lalandii

(Palinura); and the shore crab, Carcinns mamas (Brachyura). Work on the three macrurans was carried out in Marseilles, that on the brachyuran in Bristol. The intact animal was strapped dorsal side up in a Perspex dish filled with cooled sea water maintained at about 10 °C throughout the experiment.

(3)

49

Fig. i. Diagram of the organization of the joints of a rock lobster walking leg, with the muscles and joint receptor organs of the coxopodite and meropodite segments also shown. In the proximo-distal direction the joints are, successively: thoracico-coxal (T-C), coxo-basal (C-B), ischio-meropodite (I-M), mero-carpopodite (M-C), carpo-propodite (C-P), and propo-dactylopodite (P-D).

The muscles are the levators (lev.) and depressors (dep.) of the C-B joint, and the extensor

(ext.), flexor (flex.) and accessory flexor (ace. flex.) of the M-C joint. The chordotonal organs

(CB and MC) of the C-B and M-C joints, and the myochordotonal organ (MCO), are represen-ted by stippled areas. The intersegmental reflex influences of CB upon the extensor (e), flexor (/), and accessory flexor (af) motoneurones, demonstrated in this paper, are also indicated schematically (these are not intended to imply direct neural connexions).

extracellular EMG activity of the limb muscles in the meropodite, were as described in the preceding paper (Clarac et al. 1978).

Intracellular recordings from single muscle fibres of the distal head of the accessory flexor muscle, exposed in situ in the posterior leg of Carcinus, were made using 3 M - K C I rilled glass micropipettes. In these experiments, ramp-function CB length changes of 1-1-2 mm (12-16% of its mid-resting length) were applied by means of a servo-controlled puller unit (Bush, Godden & Macdonald, 1975).

Functional morphology of the pereiopod

Four muscles produce the movements of the C-B joint which levate and depress the distal part of the leg, namely the anterior and posterior levator and depressor muscles (Fig. 1). Between the two levator muscles lies the coxo-basal chordotonal organ (CB), whose sensory cells respond to levation and depression movements of the C-B joint (Bush, 1965 a).

Two muscles are involved in producing the M-C joint movements, the extensor muscle inserting dorsally and the main flexor muscle lying ventrally (Fig. 1). In addi-tion, the accessory flexor (AF) muscle, whose tendon is attached in parallel to that of the main flexor, is characterized by its small size and the fact that it is composed of two separate parts linked by a very thin rod-like tendon. The proximal part is spindle-shaped and runs longitudinally within the meropodite, while the distal part is flattened and obliquely orientated. This is not a true 'effector' muscle, since it does not contribute any significant flexor power in the limb. Instead it is essentially a receptor

(4)

50 B. M. H. BUSH, J. P. VEDEL AND F. CLARAC

ext ' ' l l l l i m i l l l l l I " M l l l l l l l I I ' 'I1 1 1!1 1

ace. flex I I I I | | || | || | I l l l l H I ' H I H ||| || | I l l l l l l l l l 1 1 1 I'I I | I | I l l l l l l l l l l l ] 11 III 1 I l l ' l l l l l l l I I II I I I I I

CB

i i i l i l l l l l l l l l l M i l 1 "

I il i n n I I n i 11 i I i i 11 11 nl l i n II11 I i 11 i i n 11 i i ii I ii i I ii i 111 hi i l i n

I I I I | | | I I 1 1 1 ! ' M M 1 * I I I I I I I I M i l l

11 1 1 i 1 1 1 ii 1 1 n 1 1 1 I I I I i 1 1 11 11 i i i i i i i i l l i l i i 11 n i n i n i 1 1 1 111 i i i 11 I I I I

3s

Fig. 2. Electromyograms (EMGs) recorded from the extensor and accessory flexor muscle of the M-C joint in Palinurus vulgaris, showing reflex effects of stretch and release of the CB chordotonal organ. CB dynamic stretch excites the active extensor motor unit, and releasing CB increases the accessory flexor motor discharge (A-C). The extensor unit is generally more sensitive to the resting CB length than the accessory flexor activity (B). A, Four successive sinusoidal CB length changes, starting and ending in a released position. B, One CB stretch, and C, one release.

In this - and subsequent EMG figures - the imposed CB length changes (about 2 mm in

Palinurus) are monitored in the lower traces (or upper traces in Fig. 7); CB stretching upwards

and releasing (shortening) downwards.

proximal end. The whole complex is therefore aptly termed a 'myochordotonal' organ, the only one of its kind in the leg (Cohen, 1963; Clarac, 1968). The myo-chordotonal receptor organ (MCO) is associated functionally in series with the proximal part of the AF muscle. Distally in the meropodite near the distal head of the AF muscle are two ordinary 'chordotonal organs' (joint receptors), MC 1 and MC 2. MC 1, MC 2 and MCO all respond to passive flexion and extension of the M-C joint (Wiersma, 1959; Clarac & Vedel, 1971, 1975).

The M-C extensor and flexor muscles are respectively innervated by two and four excitatory motoneurones, the AF muscle by only one (Wiersma & Ripley, 1952). Each of these muscles, including the accessory flexor, also receives a peripheral inhibitory innervation. The extracellular EMG electrodes used in this study provided a reliable monitor of any activity occurring in the excitatory motoneurones supplying the muscles under observation. For technical reasons considered in the previous paper, however, discharges of the inhibitory motoneurones could not be monitored in this way, so that no conclusions can be drawn about their activity in the reflexes studied here (see Clarac et al. 1978).

RESULTS

(5)

I l||l

\t i l l ' I •• I

nrr flex \ Hill III I i n i n i i l I H I n i i| I l||l nlilll II . i | | i | i » | i M A

Q 111 1 111 1 111 i n n m i K H H I l 1 1 1 i n 1 n u n 1 m 1 n 1 1 1 • 11 1 1 1 1 1 1 1 1 1 1 1 n m i n i m i H I M I M I I I I I 1 1 1 1 1 1

I. Ill HI!

r

i n i m i l i i i i i i i i I I ii i i i ii i i m i n i l I I H I I I H I M I I I I I I I I I I I I 11 i l l i i n i i i i n i I i 11 I I i l I i i l l

D

nlilll II 1 1 n i l 1 11 1111 1 • 11 11 i i i i 1 i n 1 it il

• M l , { | M II 1 i l l i l l I I I l l l l l l d I II ,1 ,1 I

3s

Fig. 3. Reflex responses of the accessory flexor and main flexor musoles of the M-C joint to sinusoidal changes of CB length, starting from either stret;hed or relaxed lengths (Palinurus). Two different flexor units are activated by dynamic release of CB (A-C), but show opposite responses to static CB length (B-D). Accessory flexor unit frequency is also increased by releasing CB, but does not stop completely during CB stretching, nor is it significantly affected by resting CB length.

species, except where otherwise indicated. Likewise, the results were identical for the different thoracic limbs, but all results illustrated here were obtained from the posterior two walking legs or, in Astacus kptodactylus, from the cheliped.

Reflexes evoked by CB in M-C muscles

Extensor muscle. In all reptantian Decapoda this muscle is innervated by two

excitatory motoneurones (Wiersma & Ripley, 1952). Stretching CB in the stationary limb excites a single extensor motor unit, and releasing CB inhibits it (Fig. 2). This unit often discharges tonically, at a frequency which depends partly upon the degree of flexion of the M-C joint. For any given M-C position, this tonic discharge is generally somewhat greater when CB is held stretched than when it is at a shorter length.

Flexor muscle. This muscle is exceptional among the leg muscles in receiving four

(6)

B. M. H. BUSH, J. P. VEDEL AND F. CLARAC

A B

ace. flex.

CB

L ^ ^ - > .

D

ll

I s

Fig. 4. Reflex effects of CB length changes on intracellular activity of the accessory flexor muscle in Carcinus maenas. Records A, B, C and D are from the same muscle fibre at different times. In each sequence, trace 1 is a control recording of the tonic activity in the muscle when CB is held released (horizontal CB trace), and recordings 2, 3 and 4 show the responses at 10 s intervals of the same fibre when CB is stretched at constant velocity, held, and then released again at the same velocity.

Accessory flexor muscle

This receptor muscle is innervated by a single excitatory motoneurone, together with a peripheral inhibitory motoneurone (Wiersma & Ripley, 1952; Angaut-Pettt, Clarac & Vedel, 1974). Like the two tonic units innervating the main flexor muscle, the single excitatory accessory flexor unit (AF) is reflexly excited by CB shortening (Figs. 2-5; see also Bush & Clarac, 1975). Its generally pronounced tonic discharge, however, commonly is little if at all influenced by the resting length of CB. Nor does this tonic discharge vary consistently with the resting position of the M-C joint. Rather it appears to depend upon unknown factors which affect the degree of central excitability. Stretching CB sometimes inhibits this tonic discharge, though often only slightly and sometimes not at all. Thus the most consistent influence on the accessory flexor muscle seen in the present experiments is that of CB shortening in exciting it.

Intracellular recording from accessory flexor muscle fibres

(7)

53

ace. flex. Ml ' M l flex. Mil Illl

CB

«

U

II Illl II

• • • ' ' '

I.I.I H I M inn 11

1 U I I I I I I 111111111111 1 M I I 1

I I I II11HI l l 444- M i l l MM I Ifl'l'llll"" II M" I

i i i 11 i 11 I I I I 11 i i n i l illllllllllllllli in i n i m i in m i n i l i m i 11 i n i n i l i

, I 1 1 1 11 i n ! 11 I I I I I I *I|I||IIIHH 11 mi 11111 I m i 11111 i i i i i i i M

II II ' I " ' " D

I I I I I I I I I I I 11 I I I I M l I I I I l l l I I I I I I I I I I I II I

3s

Fig. 5. Effects of different starting directions of CB length changes on the flexor and accessory flexor motor activity (Palinurus). When the CB movement begins with a release (A), the first response of both muscles is stronger than those to the subsequent releases. If a stretch precedes the first release, the succeeding responses are quite similar. C and D show close temporal correlation between the occurrence of spikes in one flexor unit, and those in the accessory flexor unit (recorded in this preparation directly from the motor axon).

glass micropipettes. Some of these preparations showed only weak or inconsistent activity, probably due to damage during the dissection, but in the more healthy, reflexly active ones the response to CB length changes was clearcut.

Stretching CB generally inhibited any ongoing spontaneous excitatory junctional potential (e.j.p.) activity, while releasing CB caused a somewhat variable but definite increase in e.j.p. frequency (Fig. 4). The resulting facilitation and summation of e.j.p.s. led to a small plateau of depolarization, with peak e.j.p. amplitudes of 10 mV or more at the relatively low frequencies obtained. The fact that they were always depolarizing e.j.p.s, with no indication of a reversal potential near resting potential (which was usually around - 5 0 to - 6 0 mV in the more responsive muscle fibres), indicates that they were indeed the intracellular, postsynaptic responses to impulses in an excitatory motoneurone. No evidence was in fact obtained in these experiments of any activation of the peripheral inhibitory motoneurone. Thus it can safely be concluded, on the basis both of these intracellular recordings and of the foregoing extracellular EMGs, that CB shortening reflexly activates the single excitatory motoneurone of the accessory flexor muscle.

Coupling between accessory flexor and main flexor impulses

(8)

54 B. M . H . BUSH, J. P. VEDEL AND F . CLARAC

and not precise, and in many records appears to be largely absent. Nevertheless almost synchronous firing of these units occurs sufficiently often to suggest a definite func-tional, central coupling between these two (or three) motoneurones, albeit a rather loose one. Such coupling has been noted previously between the accessory flexor unit and a tonic flexor unit in Astacus leptodactylus (Petit et al. 1974; Angaut-Petit & Clarac, 1976). In the present study it has been seen in Palinurus as well as

Astacus. No comparable degree of coupling is ever seen between the accessory flexor

and extensor excitor units (see Fig. 2).

In preparations showing clear coupling between these two tonic units, the degree of coupling does not appear to be significantly modified by the resting length, or change of length, of CB. During a 23 s period with CB held stretched in the same preparation as that represented in Fig. 5, for example, 90 flexor and 128 accessory flexor impulses occurred, and 73 (i.e. 8 1 % of the former) were closely coupled-i.e. to within ca. 10% of the flexor inter-spike interval. The corresponding activity for a similar 24 s period with CB relaxed, to a length 3 mm shorter than before, were 115, n o , 102 and 89%, respectively. Nor do the 'spontaneous' bursts of activity occasionally encountered in these preparations (see below) disrupt this coupling, when present. In some such cases, in fact, the amount of coupling may even increase, due in part to a rise in the discharge frequency of the tonic flexor unit to near that of the accessory flexor unit (Fig. 9D). Even close pairs of impulses in this condition may be fairly well coupled between the two units, though they are rarely absolutely synchronous.

When a second flexor unit was tonically active, this too showed some tendency to fire roughly simultaneously with the accessory flexor unit (Fig. 3). In this case the coupling was more in evidence when CB was held stretched than when it was at a short resting length (Fig. 3D). Here too, however, this difference may have been more apparent than real, owing to the very low discharge frequency when CB was relaxed; there were insufficient occurrences of these flexor motor impulses overall to allow quantitative comparison, as was possible for the smaller flexor unit.

Occasionally a further, much looser form of 'coupling' was observed in some of these experiments. A tendency towards gentle rhythmic discharge can sometimes be seen in the accessory flexor motor unit (Fig. 3 C, D; see also Fig. 6 C, D in the previous paper, Clarac et al. 1978). In such cases the tonic flexor unit may fire single (or groups of) impulses roughly in phase with the AF 'bursts'. When active, the same can be said of the second, larger, tonically discharging flexor unit (Fig. 3 D). Again, however, too few recordings are available to permit any quantitative analysis of this phenomenon.

Responses to repeated sinusoidal CB movements

(9)

55

ace. flex.

^ 2 0

-3 10

Q.

B

i l

flex.

D

Fig. 6. Histograms of the accessory flexor (A, B) and flexor (C, D) muscle responses to four successive sinusoidal CB length changes (in Palinurus), showing their dependence upon the direction of the first C6 movement (release in A and C, stretch in B and D). The tonic activities (mean muscle potential frequencies), with CB at a fixed length, are represented by stippled areas, activity during CB movement by hatched areas. CB sinusoidal movement (period, 4 s) is indicated by the triangular waveform beneath each histogram (r, release;

s, stretch).

first stretch in each case having started in the mid-position rather than in the fully released position; cf. Fig. 2 A.

With a long series of sinusoidal CB length changes, the reflex discharges of all responding units, but particularly the accessory flexor unit, commonly continue to adapt over several cycles (Fig. 8 A). The curves in Fig. 8 A represent the mean number of impulses per excitatory half-cycle, in each of ten successive cycles, averaged over five (or nine) velocities of CB movement (cf. Fig. 8 B). A small ' reverse adaptation', i.e. a progressive increase in successive responses, is sometimes seen in the units responding to the second movement direction of a series, particularly the extensor unit. However, this can probably be accounted for partly by the different starting position of CB, relative to its normal length range in situ.

Relation between CB velocity and reflex discharge frequency

(10)

CB

ace. flex, flex. exl.

B. M. H. BUSH, J. P. VEDEL AND F. CLARAC

• I l l

_

111 1 1II 1

1 1 1 1 1

III III

1 1 1

- . — _ - - —

' ' illll 1'

^ ^ - ^

i t i l l

•IMII

i M i l ' i II ' i l l 1 1 1 1

-^_ ..— ^ -^-^

i i i i

i ' 1 1 1 1 1

M l 1

1 ' '

i i I N I 1 i i i

B

D

i i iii i i

3s

Fig. 7. Effects of sinusoidal CB movement at four different rates on the accessory flexor, flexor and extensor muscles of the M-C joint in the cheliped of Astacus leptodactylus (CB movement amplitude approx. 1 •$ mm).

frequencies vary directly with stretch or release velocity. This has been described previously for the classical intra-segmental resistance reflexes (see Bush, 1962, 19656), but it also clearly applies to the intersegmental reflexes studied here, particularly those of the three M-C joint muscles.

If the sinusoidal stretch is changed smoothly during one cycle from a cycle period of ca. 12 s to one of ca. 3-5 s, the amplitude of the instantaneous frequency (i.e. reciprocal of inter-impulse interval) envelope, and the peak values of the AF motor frequency, increase markedly at the higher CB velocity. Furthermore, they adapt with cycle repetition at the higher velocity to a mean peak level still well above the adapted level at the lower velocity. Comparison of the responses of all three muscles in the meropodite, at four different velocities of sinusoidal CB length change (Fig. 7), reveals a clearcut positive relationship between their reflex discharge frequencies and CB velocity.

(11)

57

I4

12

8. 8

.§ 6

io

I i i

AF2

0 1 - 2 3 4 5 6 7

Cycle number

8 9 10 0-5 1 I S

Average CB velocity (mm/s)

Fig. 8. Variation of accessory flexor ( A F i , AF 2), flexor (F) and extensor (E) muscle responses to ten successive sinusoidal CB length changes (A), and (B) with velocity of sinusoidal CB movement (Astacus). Only the 'excitatory' half-cycle is considered in each case, i.e. CB release for AF and F and CB stretch for E.

In A, the ordinate represents the mean number of impulses for each of the ten successive cycles, averaged over the five (AF 2, F, E) or nine (AF 1) velocities used (indicated in B). In B, the mean frequency values for each velocity were obtained by dividing the total number of impulses during the appropriate half-cycle by the half-cycle duration, and averaging this value over ten successive cycles. Curves AF 2, E and F were all from the same set of record-ings; AF 1 was from the same preparation 3 h earlier.

In any event, it may be concluded that in the passive, quiescent animal, the reflex response frequencies of these three tonic motor units vary directly and fairly systema-tically with velocity of CB length change, in addition to showing a rather less readily denned or consistent 'adaptation' to successive sinusoidal length changes. It should be noted that although increased velocity of CB movement does result in an increase in the mean frequency of the AF (and F and E) motor impulses, the total number of impulses per half-cycle remains roughly constant, or may even decrease slightly with the decrease in cycle duration. However, it seems highly probable that the observed differences in AF as well as F and E mean frequencies will, as in other crustacean neuro-muscular systems, result in corresponding differences in AF (and F and E) tension development, and hence also in the afferent feedback from MCO (see Dis-cussion).

Effect of light falling on the eyes

(12)

B. M. H. BUSH, J. P. VEDEL AND F. CLARAC

ace. flex.. flex..

ext. n

niiimiilliilllllll i linimlimmili I II il n ilnlmlii'"'I > I " i nmlmln I u

Light

M H I ! HfH—I H-H 1

Y-Light

ace. flex, flex.

3s

Fig. 9. Effect of eye illumination (A-C), or 'spontaneous' central activation (D), on the accessory flexor, flexor and extensor muscle responses to CB movements (Astacus). The accessory flexor and flexor discharges increase during light (A, C) or central activation (D), while extensor activity decreases (A) or stops completely (C). (B continues on from A, D is from a different preparation.)

commonly i-2—1*8 s, indicating a polysynaptic pathway. The intensity of the effect varies directly with the change in intensity of the light falling on the eyes. Responses to sinusoidal changes in CB length summate with the light response so as to modulate the high frequency discharge (Fig. 9 A-C). This enhanced discharge of the accessory and main flexor units following illumination declines progressively over several CB cycles, often lasting for 10 s or more before the discharge frequencies return more or less to their 'normal' pre-illumination levels.

(13)

ace. flex, in H | 1 i 11 n n i | | j | i i i [ i | | HH|||||||[||||||||| I I'll I'HUnit |' | ' I | | | " I 11 n II i i i i JLJI n . i I i i "

11 1111 j 1 11 M I 11 mill 11111 nun 1 nn 1111 1 11 1 111 11

flex. •

CB

11 1 11 1 1 IWIIIIIIIIIlllUllllllllllMlllllllllHlilllKlllllllhllllll III 111 III I It III II II I; II

1 1 1 1111 nun iiiiMiininmniiiniiiiiiiiiiiiniiiii 1111 in 1 1111111 111 1 111

3s

Fig. 10. An example of 'spontaneous' central modulation (B) of the accessory flexor, extensor (downward spikes on the upper traces), and flexor responses to CB length changes, compared with the 'normal' reflexes (A) (Palimtrus). The central drive increases accessory flexor and flexor activity, and strongly depresses the extensor reflex response.

Central influences on reflex responsiveness

As in most experiments on 'lively', viable crustacean preparations with intact CNS, sporadic, apparently spontaneous, bursts of motoneurone activity occasionally occurred in the present study (Fig. 9D). Such centrally originating activity often tends to override, and sometimes completely block, the normal reflex responses to proprioceptive stimuli. An example of this is illustrated in Fig. ioB, where both the flexor and the accessory flexor units were evidently centrally activated while the extensor unit was inhibited (cf. Fig. 10A). This might have represented part of a general ' arousal' reaction, similar to that referred to above in response to exterocep-tive stimuli.

In contrast to the other two muscles, the accessory flexor unit still appeared to be modulated by CB length changes in a more or less normal manner. This again is reminiscent of the situation during a strong light reaction (e.g. Fig. 9 C), when the accessory flexor unit was more clearly modulated than the main flexor units or the totally inhibited extensor unit. Slight modulation of the main flexor unit by CB was nevertheless still discernible, although owing to its relatively low discharge frequency, it was less obvious here than in the accessory flexor. It would seem, then, that the reflex action of CB upon the accessory flexor motoneurone in particular, does con-stitute a fairly persistent influence upon this receptor muscle, an observation which must have important functional significance.

DISCUSSION

(14)

60 B. M. H. BUSH, J. P. VEDEL AND F. CLARAC

movement also reflexly influences the more tonically active excitatory motoneurones of the main flexor and extensor muscles of the same, M-C joint (see Clarac et al. 1978). Evidence is also presented that, as would be expected, all these reflexes - in-cluding those of the AF motoneurone - can be overridden or substantially modified by 'central drive' and other, 'extrinsic' factors, including illumination of the eyes and general tactile stimulation.

Reflex and central 'co-activation' of AF and F

Of particular interest in the present results is the broadly parallel nature of the dynamic reflex actions of CB length changes upon the AF and tonic main flexor (F) motoneurones. Thus both AF and one or two of the four excitatory motoneurones of the flexor muscle are reflexly excited by CB shortening and inhibited by stretching CB, while the tonic extensor unit (E) is excited during stretching. Moreover, both AF and F units respond at frequencies which increase together with velocity of CB shortening; they both also adapt to repetitive sinusoidal movements with a similar ' time-course' (Fig. 8, cf. AF 2 and F). Further, central drive and other extrinsic factors also tend to affect the AF and F neurones similarly, and the extensor unit in opposite sense.

The 'static' reflex actions on these motor units of different resting CB lengths, in contrast to the dynamic reflexes referred to above, are less predictable. Firstly, the AF motoneurone's tonic background frequency is not consistently related to CB length, being greater at times with CB held stretched and at others with CB relaxed, even over quite short recording periods. Possibly this reflects the variety of influences evidently impinging upon the AF motoneurone (cf. Evoy & Cohen, 1971; Vedel et al. 1975 a). More surprising is the observation that in most cases where two main flexor (F) units are tonically active, the resting length of CB evidently affects them in opposite ways (Fig. 3C, D). The smaller, most tonic flexor unit is more active when CB is relatively short than when it is held stretched, as would be expected by analogy with the intrasegmental reflexes, whereas the reverse applies to the larger of these two units. The functional significance of this reciprocal influence of CB length upon two units of the same muscle is as yet obscure.

Coupling between AF and F impulses. Apart from the general parallel activation of

(15)

smaller unit. Since CB length appeared to have a partially antagonistic effect on this unit (see above), additional interneurones may be involved here.

Motor effects on MCO afferents and reflexes

Since the accessory flexor is demonstrably a ' receptor muscle' rather than a power muscle, any variation in AF motor discharge frequency, whether central or peripheral in origin, is likely to influence the sensory signal emanating from the associated myochordotonal receptors (Clarac & Vedel, 1971; Cohen, 1965; Evoy & Cohen, 1971). The strength of any such efferent influence on the afferent signal will presumably depend upon the magnitude of the frequency changes. Thus the increase in AF frequency with velocity of CB shortening, or on eye illumination, for example, or its progressive adaptation with repetitive CB movements, will probably result in corresponding changes in feedback from the MCO.

Like the two M-C joint chordotonal organs, MC 1 and MC 2, the myochordotonal (MCO) receptors contribute to the intrasegmental resistance (or 'myotatic') reflexes of the main flexor and extensor muscles resulting from passive M-C joint movement, albeit in a more labile manner (Bush, 19656; Evoy & Cohen, 1969; Vedel et al. 1975 a). Any variation in AF motor activity might therefore be expected to affect the 'gain' of the intrasegmental MCO reflex loop. Under appropriate experimental con-ditions, isometric contraction of the accessory flexor muscle tends to cause reflex contraction in the main flexor muscle (Clarac & Vedel, 1975), and accordingly to enhance the reflex resistance to any imposed M-C extension - i.e. to increase the gain of this myotatic reflex. Thus the intersegmental reflex excitation of AF by CB short-ening will tend to reinforce any concurrent intrasegmental flexor resistance reflex, in addition to the direct, intersegmental reflex excitation of the main flexor muscle caused by CB shortening.

In vivo, when variations in discharge rate of the flexor and extensor motoneurones

result in overt movements at the M-C joint, co-activation of the AF and F unit would tend to adjust the AF length to that of the flexor muscle. That is, such AF activity could serve essentially to maintain the gain of the MCO receptors, and thereby of the whole MCO -> M-C servo loop, similar to the role postulated for the motor control of the thoracico-coxal muscle receptor (Bush, 1977). Whether this servo loop gain was actually increased or decreased, rather than simply maintained at a more or less constant level, would depend quantitatively upon the relative degrees of excitation (or inhibition) of the AF and F motoneurones. Certainly where the variation in flexor and extensor motor activity results, not in overt M-C movement but simply in tonus changes, any variation in AF motor discharge might indeed be expected to modulate the loop gain accordingly. From the present experiments it is clear that, although the CB reflex drive to AF often does appear to exceed the flexor drive, particularly in

Astacus (Figs. 7, 8), this can vary, even in the same preparation (Fig. 5, cf. A, B, C).

Different stimulating factors, moreover, can affect AF and F to different extents (Fig. 9, cf. A, D).

Chordotonal organs and intersegmental co-ordination

The results presented in this and the preceding paper strongly suggest that the CB organ, and very probably each chordotonal organ in the leg, contributes to the regula-tion and co-ordinaregula-tion of posture and movement, not only of its own joint but of the

(16)

62 B. M . H . BUSH, J. P. VEDEL AND F . CLARAC

entire leg. An important element in this regulatory control is the specific reflex modulation by CB of the motor output to, and hence probably of the proprioceptive re-afference from, the myochordotonal muscle receptor organ in the meropodite. The apparently greater 'flexibility' of this reflex, compared to the accompanying reflexes from CB to the main M-C power muscles, probably reflects its susceptibility to other interacting or potentially overriding influences, both peripheral and central.

In these terms, a plausible general hypothesis is that the MCO might be critically involved in determining the optimal overall load distribution, and hence posture, of the leg for a particular stance (cf. Evoy, 1976). The 'pure' chordotonal organs like CB (i.e. those without efferent control) might then provide, through the various intra-and intersegmental reflexes described, a 'fine adjustment' of the position intra-and move-ment of each joint in the leg. Such an hypothesis would be consistent both with previous results from behavioural studies, e.g. those involving receptor ablation or limb immobilization (Evoy & Cohen, 1971; Clarac, 1977), and with the present evidence of highly specific (yet possibly widespread) proprioceptive regulation of AF motor activity. Further experiments are planned to test this hypothesis, and to seek answers to the other questions raised by this investigation.

REFERENCES

ALEXANDROWICZ, J. S. (1951). Muscle receptor organs in the abdomen of Homarus vulgaris and Palinurus

vulgaris. Q. Jl microsc. Sci. 92, 163-199.

ALEXANDROWICZ, J. S. (1967). Receptor organs in thoracic and abdominal muscles of Crustacea. Biol.

Rev. 42, 288-301.

ALEXANDROWICZ, J. A. (1972). The comparative anatomy of leg proprioceptors in some decapod Crustacea. J. mar. biol. Ass. U.K. 52, 605-634.

ALEXANDROWICZ, J. S. & WHITEAR, M. (1957). Receptor elements in the coxal region of Decapoda Crustacea. J. mar. biol. Ass. U.K. 36, 603-628.

ANGAUT-PETIT, D. & CLARAC, F. (1976). A study of a temporal relationship between two excitatory motor discharges in the crayfish. Brain Res. 104, 166-170.

ANGAUT-PETIT, D., CLARAC, F. & VEDEL, J. P. (1974). Excitatory and inhibitory innervation of a crustacean muscle associated with a sensory organ. Brain Res. 70, 148-152.

BARTH, G. (1934). Untersuchungen uber Myochordotonalorgane bei dekapoden Crustaceen. Z. viiss.

Zool. 145, 576-624.

BUSH, B. M. H. (1962). Proprioceptive reflexes in the legs of Carcinus maenas (L.). J- exp. Biol. 39, 89-105.

BUSH, B. M. H. (1965 a). Proprioception by the coxo-basal chordotonal organ, CB, in legs of the crab,

Carcinus maenas. J. exp. Biol. 42, 285-297.

BUSH, B. M. H. (19656). Leg reflexes from chordotonal organs in the crab, Carcinus maenas. Comp.

biochem. Physiol. 15, 567-587.

BUSH, B. M. H. (1976). Non-impulsive thoracic-coxal receptors in crustaceans. In Structure and

Function of Proprioceptors in the Invertebrates (ed. P. J. Mill), pp. 115-152. London: Chapman and

Hall.

BUSH, B. M. H. (1977). Non-impulsive afferent coding and stretch reflexes in crabs. In Identified

Neurons and Behavior of Arthropods (ed. G. Hoyle), pp. 439-460. New York: Plenum Press.

BUSH, B. M. H. & CLARAC, F. (1975). Intersegmental reflex excitation of leg muscles and myochordo-tonal efferents in decapod Crustacea. J. Physiol., Lond. 246, 58-60P.

BUSH, B. M. H., GODDEN, D. H. & MACDONALD, G. A. (1975). A simple and inexpensive servosystem for the control of length or tension of small muscles or stretch receptors. J. Physiol. Lond. 245, 1— 3P. CLARAC, F. (1968). Proprioception by the ischio-meropodite region in legs of the crab Carcinus

mediterraneans C. Z. vergl. Physiol. 61, 224—245.

CLARAC, F. (1977). Motor coordination in crustacean limbs. In Identified Neurons and Behavior of

Arthropods (ed. G. Hoyle), pp. 167-186. New York: Plenum Press.

(17)

CLARAC, F. & VEDEL, J. P. (1971). Etude des relations fontionnelles entre le muscle ftechisseur accessoire et les organes sensoriels chordotonaux et myochordotonaux des appendices locomoteurs de la Langouste Palinurus vulgaris. Z. vergl. Physiol. 72, 386-410.

CLARAC, F. & VEDEL, J. P. (1975). Proprioception by chordotonal and myochordotonal organs in the walking legs of the rock lobster Palinurus vulgaris. Mar. Behav. Physiol. 3, 157-165.

CLARAC, F., VEDEL, J. P. & BUSH, B. M. H. (1978). Intersegmental reflex co-ordination by a single joint receptor organ (CB) in rock lobster walking legs. J. exp. Biol. 73, 29-46.

COHEN, M. J. (1963). The crustacean myochordotonal organ as a proprioceptive system. Comp. Biochem.

Physiol. 8, 223-243.

COHEN, M. J. (1965). The dual role of sensory systems: detection and setting central excitability. Cold

Springs Harb. Symp. quant. Biol. 30, 587-599.

Evoy, W. H. (1976). Modulation of proprioceptive information in Crustacea. In Neural Control of

Locomotion (ed. R. M. Herman, S. Grillner, P. S. G. Stein and D. G. Stuart), pp. 617-645. New

York: Plenum Press.

Evoy, W. H. & COHEN, M. J. (1969). Sensory and motor interaction in the locomotor reflexes of crabs.

J. exp. Biol. 51, 151-169.

Evoy, W. H. & COHEN, M. J. (1971). Central and peripheral control of arthropod movements. Adv.

comp. Physiol. Biochem. 4, 225-266.

FIELDS, H. L. (1976). Crustacean abdominal and thoracic muscle receptor organs. In Structure and

Function of Proprioceptors in the Invertebrates (ed. P. J. Mill), pp. 65-114. London: Chapman and

Hall.

MATTHEWS, P. B. C. (1972). Mammalian Muscle Receptors and their Central Actions. London: Edward Arnold.

VEDEL, J. P., ANGAUT-PETIT, D. & CLARAC, F. (1975a). Reflex modulation of motoneurone activity in the leg of the crayfish Astacus leptodactylus. J. exp. Biol. 63, 551-567.

VEDEL, J. P., CLARAC, F. & BUSH, B. M. H. (19756). Coordination motrice proximodistale au niveau des appendices locomoteurs de la Langouste. C.r. hebd. Seanc. Acad. Sci., Paris, D 281, 723-726. WIERSMA, C. A. G. (1959). Movement receptors in decapod Crustacea. J. mar. biol. Ass. U.K. 38,

I43-I52-WIERSMA, C. A. G. & RIPLEY, S. H. ^952). Innervation patterns of crustacean limbs. Physiologia comp.

Oecol. 2, 391-405.

(18)

Figure

Fig. i. Diagram of the organization of the joints of aand joint receptor organs of the coxopodite and meropodite segments also shown
Fig. i. Diagram of the organization of the joints of aand joint receptor organs of the coxopodite and meropodite segments also shown p.3
Fig. 5. Effects of different starting directions of CBcorrelation between the occurrenceresponse of both muscles is stronger than those to the subsequent releases
Fig. 5. Effects of different starting directions of CBcorrelation between the occurrenceresponse of both muscles is stronger than those to the subsequent releases p.7
Fig. 8. Variation of accessory flexor (AFi, AF 2),to ten successive sinusoidal CB length changes (A), and (B) with velocity of sinusoidal CBmovementfor AF and F and CB stretch for E
Fig. 8. Variation of accessory flexor (AFi, AF 2),to ten successive sinusoidal CB length changes (A), and (B) with velocity of sinusoidal CBmovementfor AF and F and CB stretch for E p.11
Fig. 10. An example of 'spontaneous' central modulation (B) of the accessory flexor, extensor(downward spikes on the upper traces), and flexor responses to CB length changes, comparedwith the 'normal' reflexes (A) (Palimtrus)
Fig. 10. An example of 'spontaneous' central modulation (B) of the accessory flexor, extensor(downward spikes on the upper traces), and flexor responses to CB length changes, comparedwith the 'normal' reflexes (A) (Palimtrus) p.13

References

Related subjects : Chordotonal organ