Experimental Brain Research 1, 338--358 (1966)

Convergence of Excitatory and Inhibitory Action

on Interneuroncs in the Lumbosacral Cord*
T. HONGO, E. JANKOWSKA** and A. LUNDBEtgG
Department of Physiology, University of G6teborg, Sweden

Received December 27, 1965

Summary. Intracellular recording has been made in spinal cats from more
t h a n 100 interneurones in the dorsal horn and intermediary region of the lum-
bosacrM spinal cord. The majority of interneurones receive not only EPSPs but
also I P S P s from primary afferents. The I P S P s are evoked from three different
systems, group I muscle afferents (probably Ib), low threshold cutaneous affcrents
and the FI{A. The shortest central latency of the I P S P s indicates a disynaptic
linkage from primary afferents. Interneurones with monosynaptic EPSPs from
group I muscle affercnts m a y receive I P S P s from all the above mentioned afferent
systems. Interneurones with monosynaptic E P S P s from cutaneous afferents
receive their inhibition from the two latter afferent systems. Convergence of
EPSPs and I P S P s from the FI~A m a y occur on the same interneurone. The results
are discussed mainly with respect to inhibitory interaction between spinal reflex
pathways.

Key Words: EPSPs - - I P S P s - - Interneurones - - Spinal cord

Introduction
There are m a n y studies on the effect from primary afferents on interneurones
in the spinal cord. Several investigators have aimed at finding cells t h a t m a y be
part of a given neuronal pathway such as the recurrent inhibitory p a t h w a y to
motoneurones (EocLxs, FATT and KOK]~TSU, 1954a) and the reciprocal I a inhibi-
tory pathway to motoneurones (EccL~s et al., 1956). Other investigators have
studied the eleetrophysiological properties of interneurones (F~A~K and FIzO~T~S,
1956; HAA~A~X~ et al., 1958; KOLMODIN and SKOGLUXD, 1958; HIZ~T and K u ~ o ,
1959a, b; WALL, 1959; KOSTYUK, 1959; EccL~s et at., 1960) or more directly
aimed at classifying different types of interneurones. For example KOT,MODIN
(1957) made a systematic study of adequate activation from muscle receptors of
interneurones located in different parts of the gray matter, KOL~ODIN and SKOG-
LUND (1960) analysed interneurones activated b y exteroeeptive stimulation and
EocLxs et al. (1960) classified interneurones on the basis of the monosynaptic
EPSPs they receive from primary afferents.

* This work was supported by the Swedish Medical Research Council (Project No 14X--
94--02A).
** IBl%O-Unesco fellow. Present address: Department of Neurophysiology, Nencki Insti-
tute of Experimental Biology, Warsaw, Poland.
 Postsynaptic Inhibition in Interneurones 339

These classifications were based largely on extracellular recordings b u t with

i m p r o v e d t e c h n i q u e (ef. below) it is now possible to record i n t r a c e l l u l a r l y fl'om a
larger n u m b e r of i n t e r n e u r o n e s . This was required for the present s t u d y because
the p r i m a r y aim was to investigate i n h i b i t o r y processes i n interneurones. Our
i n t e r e s t i n i n t e r n e u r o n a l i n h i b i t i o n arose from recent findings suggesting t h a t
there is a n i n h i b i t o r y i n t e r a c t i o n b e t w e e n spinal reflex p a t h w a y s (L~J~I) et al.,
1965; AND]~N et al., 1964; JANKOWSKA et al., 1965). Several a u t h o r s have described
t h a t volleys i n p r i m a r y afferents (or receptor s t i m u l a t i o n ) m a y i n h i b i t the dischar-
ges i n i n t e r n e u r o n e s ; special a t t e n t i o n to these i n h i b i t o r y effects has been given
b y KOLMODIN (1957) a n d KOLMODIN a n d SKOGLU~D (1960). I t will be shown i n
this paper t h a t a great m a j o r i t y of i n t e r n e u r o n e s i n the dorsal h o r n a n d inter-
m e d i a r y region receive b o t h e x c i t a t o r y a n d i n h i b i t o r y p o s t s y n a p t i c potentials
from p r i m a r y afferents. A p r e l i m i n a r y report has been given (Ho~Go et al., 1965a).

Methods
Most of the experiments were performed on unanaesthctized spinal cats, operated under
ether and subsequently decerebrated or anaemically decorticated as described by VOORHOEVE
(I960). Some experiments were made on spinal cats anaesthetized with ehloralose (60 mg/kg)
and no difference was noticed in the two kinds of preparations. A few findings on eats under
chloralose that were not made spinal will also be reported. The cats were paralyzed with
Flaxedil and artificially respirated. The blood pressure was measured throughout all the
experiments and prevented from falling below about 90 mm tIg. The spinal cord was transected
in the lower thoracic region, the dorsal column removed for a length of two segments and both
spinal halves dissected and mounted for stimulation in the descending direction. The lower
lumbar cord was also exposed and the S1, L7 and sometimes L6 ventral roots sectioned. The
microelectrode was inserted from the cord dorsum. Cells were classified as interneurones when
they could not be antidromically activated from either the dissected spinal halves or from the
ventral roots. In most of the experiments the microelectrodes were filled with a solution of
2 M potassium citrate but electrodes filled with 3 iV[ potassium chloride were sometimes
employed to demonstrate the reversal of IPSPs. It proved favourable not to use microelcc-
trodes with too fine tips; generally they were pulled to have a long narrow shaft with a very
fine tip, which, however, was broken away to obtain a tip diameter of about 1/~. Intraeellular
recording was facilitated by the employment of a new motor-driven manipulator. The micro-
electrode was usually forwarded in steps of 10/l and this movement took 50 msec. It is our
definite impression that the yield of cells was much higher with this manipulator than with
the hand.driven manipulator previously used. Nevertheless it is much more difficult to obtain
good intracellular records from interneurones than from motoneurones and for unknown
reasons very few cells were recorded from in some cats. The synaptic potentials were recorded
with an AC amplifier with a time constant of 0.8 sec and the membrane potential with a paral-
lel DC channel connected to a separate oscilloscope. Initially after penetration spike discharges
were often evoked and under these conditions it was difficult to analyse the synaptic poten-
tials. Sometimes the discharge was stopped with a hyperpolarizing current through the record-
ing electrode but it often proved more favourable to wait until the membrane potential
became lower, between 20~40 mV, and the spike mechanism of the cell had deteriorated. It
is realized that through this procedure a somewhat distorted picture of the synaptic poten-
tials was obtained in that at the low membrane potential the EPSP would be smaller and the
IPSP larger than at the normal membrane potential. In some neurones extraeellular records
of spike discharges were taken before penetration or else the discharges were taken soon after
impalement and those records could be compared with a later record disclosing the synaptic
potential. After withdrawal from the cell extracellular records were always taken so that the
intraeellular potential could be compared with the extraeellular field potential. The angle of
the manipulator and the depth from the cord dorsum were measured for the units recorded
from, and after tracking in one region of the cord the microelectrode was left and its position
identified in later histological controls, whereupon the location of the interneurones found in
340 T. Ho~oo, E. JANKOWSK• and A. LUNDBERG:

this region could be determined. For this purpose the cord was fixed in 10 ~ formalin, embed-
ded in paraffin and the electrode tracks identified in serially cut unstained or stained sections.
Due precaution was taken to the shrinkage which was about 15 ~
The following hindlimb nerves were dissected and mounted for stimulation, the abbre-
viations employed in the paper are given in parenthesis: quadrieeps (Q), posterior biceps-
semitendinosus (PBSt), anterior biceps-semimembranosus (ABSm), gastrocnemius-soleus
(G--S), plantaris (P1), flexor digitorum and hallueis longus (FDL), deep peroneal (DP), sural
(Sur), superficial peroneal (SP), the posterior joint nerve (J), eontrMateral hamstring (co H),
contralateral sural (co Sur). Only those interneurones in which all the nerves or most of them
were tested are included in the material presented. Other abbreviations: excitatory post-
synaptic potential, EPSP; inhibitory postsynaptie potential, IPSP; postsynaptie potential,
PSP; primary afferent depolarization, PAD; flexor reflex afferents, Ft~A.

Results
1. Interneurones influenced/tom group I muscle a~erents
I n t e r n e u r o n e s with m o n o s y n a p t i e E P S P s from group I muscle afferents m a y
receive e x c i t a t o r y a c t i o n also from o t h e r afferent s y s t e m s a n d in t h e m a j o r i t y of
t h e m I P S P s are evoked. The convergence p a t t e r n onto these i n t e r n e u r o n e s is
i l l u s t r a t e d in Fig. 1 - - 6 a n d s u m m a r i z e d a t t h e e n d of this section in Tables 1 - - 3 .

2rnV 2msec
E Tibl,7 F 1.~ G SP2,3 H ]

b-mY ~

I l? 1.3 J 3.3 K 13.3 L FDL3.3

20 msec ~ a ~ , , * ,.,,:~
:Fig. 1. Interneurone receiving a monosynal~tic E P S P /tom I a a~erents and an I P S P Jrom the E R A . Intraeellular
recording (upper traces) with a citrate electrode from a cell in L7 located 2.6 m m from the cord dorsum. The
lower traces in D, G, It, K and L were recorded after withdrawal of the micro-electrode to a j u s t extracellular
position. The middle traces in the latter records and the lower traces in A - - F , I and J are from the L7 dorsal root
entry zone. Stimulus strengths in this and all other figures are indicated in multiples of the threshold for each
nerve. There is separation in I a and I b volleys from D P and a nearly m a x i m a l E P S P evoked in C a t a s t r e n g t h
t h a t is m a x i m a l for the I a b u t subthreshold for the I b volley. I n I there is no effect by a m a x i m a l group I volley
from Q b u t an I P S P is evoked when the stimulus s t r e n g t h is increased in J a n d :K to activate high threshold
muscle afferents. The intracellular traces in A - - D were taken at higher gain (upper calibration) and in I - - L a t
the lower gain b u t all the extraeellular records in D, G, tI, K and L were taken at the higher gain. The upper
time calibration refers to A - - D , the lower to E L. All records in this and the following figures i f n o t otherwise
stated consist of superimposed traced. Unanaesthetized spinal eat

W e h a v e confirmed t h a t i n t e r n e u r o n e s with a m o n o s y n a p t i c E P S P from group I

muscle afferents m a y receive a p o l y s y n a p t i e E P S P from t h e F I ~ A (EccLws et al.,
1956, 1960, for i n t r a c e l l u l a r records ef. Fig. 7 LTJNDBERG, NORRS]~LL a n d VOOR-
~oEv~, 1962). The previous r e p o r t t h a t t h i s m a y occur in i n t e r n e u r o n e s mono-
 Postsynaptie Inhibition in Interneurones 341

synaptically activated from group I a affcrents (EccLES et al., 1956, 1960) has been
confirmed and is supplemented b y our finding t h a t also interneurones with a
monosynaptie E P S P exclusively from Ib afferents m a y receive an E P S P from
the FRA.
Many of the interneurones with group I E P S P s receive I P S P s from the F R A
and this has been found with interneurones in which the E P S P is evoked from I a
afferents as well as in those in which E P S P is evoked exclusively from Ib afferents.

A L21,65 ~ ~ ~-~D G-S1.55

E G-S15.5 F PBStIB,Z .G sur20.7 2msec

Fig. 2. Interneurone receiving a monosynalgtic E P S P #ore I b agerents and an I P S P ]rom the F R A . Intracellular
recording (upper traces) w i t h a citrate electrode from a cell located 2.5 m m from the cord dorsum. The lower
traces in C - - G were recorded after withdrawal o f the microelectrode to a j u s t extracellular position. The lower
traces in A and B a n d the middle traces in C - - G are from the L7 dorsal root entry zone. There is graded stimu-
lation of the Q nerve in A - - C , a m a x i m a l I a volley has no effect in A b u t a monosynaptie E P S P is evoked in B
and C when the s t i m u l u s s t r e n g t h is increased to evoke a I b volley. The I P S P s in E a n d F are evoked from high
threshold muscle afferents. The upper time calibration refers to A - - D , the lower to E - - G . Voltage calibration
refers to the microelectrode recording. Unanaesthetized spinal cat

The former type is illustrated in Fig. 1. The monosynaptie E P S P in this inter-

neurone is evoked exclusively from the DP nerve and clearly from I a afferents. A
particularly large I P S P is evoked from the mixed tibial nerve (E, F) but stimu-
lation of the cutaneous SP nerve (G), high threshold muscle ( J - - L ) and joint (H)
afferents also gives IPSPs. I n the interneurone of Fig. 2 the monosynaptic E P S P
is evoked from Ib afferents in the quadriceps nerve (A--C), the only other group I
E P S P is from the G - - S nerve in D. High threshold muscle afferents give an I P S P
as is particularly well shown in F, and there is also a large I P S P evoked from the
sural nerve in G.
Fig. 3 illustrates a reversal of the I P S P evoked from the F R A with increased
intracellular chloride. A monosynaptic group I E P S P from F D L is shown in A - - C
and at slower speed in D with some later synaptic actions, which, as those evoked
from high threshold joint afferents in E, very well m a y be mixed I P S P s and EPSPs.
A large monosynaptic E P S P from the mixed tibial nerve is followed b y an I P S P
and an I P S P is also evoked from the SP nerve (G, H). The lower row shows
the expected reversal of the I P S P from the tibial nerve (J) and from the SP nerve
(K, L). A depolarization is evoked from the joint nerve, and since the time
course in E and I are different it is possible to conclude t h a t the response consists
of mixed I P S P s and EPSPs.
I t is of interest t h a t m a n y interneurones receive a convergence of monosynaptie
EPSPs from group I muscle afferents and from cutaneous afferents. Such a con-
vergence has been observed in cells with polysynaptic excitation from high thres-
342 T. HONGO, E. JAXKOWSI(A and A. LUNDBERG:

hold muscle afferents and is found in many cells that receive IPSPs from the FleA.
This is illustrated in Fig. 4 for a cell with a group I E P S P from P1 (A--E), G--S
( F - - J ) and F D L (K) and a monosynaptic E P S P from the cutaneous SP (L--N).

FIlL
A 1,5

5 rnsec 20 msec
E~j F~ fib 6 SP H
i
I
11~ ~ # ~ f o r A.B.C

hyperpolt~~, d
for ZT.E.H
2.10_e,- ~ ~ _ ,~,_...,._.._ lg L
20 msed 10msec 20msec
Fig. 3. Reversal o / I P S P s in an interneurone. Intracellular recording (upper traces) w i t h a KC1 electrode from a
cell located a t a depth of 2.05 m m from the cord dorsum. The lower traces in I - - L are obtained after withdrawal
of the microelectrode to a j u s t extracenular position. Lower traces in A - - H and middle traces in I - - L are from
the L7 dorsal root entry zone. There is a monosynaptic E P S P from the F D L nerve ( A - - D ) and from the Tib
nerve (F). The following pairs of records were taken simultaneously a t different speeds: C and D, G and H, K and
L. Records E ~ were taken before and the corresponding lower records I - - L during passage of a hyperpolarizing
current (2 9 10 -9 A) t h r o u g h the recording microelectrode. ICeversal of the I P S P is particularly clear w i t h the
effect from SP, compare records G and K or the slower records H and L. l~esting membrane potential 45 nlV.
Unanaesthetized spinal cat

6-S
...... ~ /~~i"~
~ZI
2 msec
SP LTP
0 9.1

2mYI ItL_I =

2 msec 4 msec 20 msec

Fig. 4. Intcrneurone recewing monosynaptic E P S P s ]rom group I muscle agerents and cutaneous agerents and I P S P s
]rom the F R A . Intracellular recording (upper traces) w i t h a citrate electrode from a cell at a d e p t h of 2.55 m m
from the cord dorsum. Upper records are intracenular, lower records in N and O are extraee]lular obtained after
withdrawal of the electrode to a j u s t extraccllular position. Lower records in A - - ~ [ and middle records in N and 0
are from the L7 dorsal root entry zone. There is graded s t i m u l a t i o n of the Pl nerve i n A--JE and of the G - - S nerve
in F--ft. The records in lV[ and N were t a k e n simultaneously at different sweep speed. Observe the I P S P in N and
the smaller I P S P in O evoked from high threshold afferents of the D P nerve. Voltage calibration in ff refers to
A I K , ill iV[ tO L and M and ill O to Iq and O. Time calibration below M refers to L and 5~, below O to N and O.
Spinal cat under ehloralose
 Postsynaptic Inhibition in Int~erneurones 343

At the stronger stimulation in M and N the latter is followed b y an I P S P which in

all likelihood is part of a FleA response. I n interneurones with a group I E P S P
from a nerve displaying separation in I a and -[b volleys it was found t h a t a mono-
synaptie E P S P m a y be evoked from cutaneous afferents in interneurones t h a t
receive a monosynaptic I a excitation. This was not found in any of the inter-
neurones t h a t when tested with the same criteria received their excitation exclu-
sively from I b afferents. However, a monosynaptic E P S P from cutaneous afferents

PBSt
A 1.22 B 134 C 142
Fig. 5. Interueurone receiving monosynaptic E P S P s
and disynaptic I P S P s ]tom group I muscle afferents
and in addition E P S P from the ipsilateral and contra-
lateral F B A . Intracellular recording (upper traces)
~p 2m'~c
w i t h a citrate electrode f r o m a neurone located at a ,ct&t.U~
d e p t h of 2.4 m m f r o m the cord dorsum. Extracellular
traces obtained a f t e r withdrawal of the microelee-
trode to a j u s t extracellular position arc s h o w n in ~-S
D, H, L, P a n d T. Middle traces in the latter records / 1,1~ J 1.22 K 1,43 Z 2,37
a n d lower traces in all other records are f r o m the . & ~
L7 dorsal root e n t r y zone. I n A P is shown the
effect o f graded s t i m u l a t i o n of the P B S t , D P , G - - S ~ "
a n d P1 nerves as indicated. There is a m o n o s y n a p t i c ]91
E P S P f r o m :P:BSt (A D), D P ( E - - H ) , A:BSm a n d
F D L a n d a small effect f r o m G - - S a n d plantaris, M 1,40 N 1.51 0 1.,;I P, 3.45
which are best seen in records L a n d I~ in com-
parison w i t h the extracellular traces. D i s y n a p t i c ~ ,,, ~-~ -3,
I P S P s are e v o k e d f r o m D P ( F - - H ) , G - - S (,I--K),
PI ( N - - P ) a n d Q (T). The later E P S P in t t and R ABSm ~F~~ O
is e v o k e d b y group I I muscle afferents a n d the 02.6 ' T 1.3
E P S P f r o m c o i l is also evoked f r o m h i g h threshold
muscle afferents as illustratedin U - - Y . The s t i m u l u s ~.., -r
s t r e n g t h in U is m a x i m a l for group I b u t the in- coH.
4 msec co.sur
c o m i n g volley is recorded a t lower amplification
t h a n t h a t used in u a n d X . T i m e calibration for
A - - D is shown below C a n d time calibration for
U
~..~x~
1.~ V 4,5X~
E - - Y below S. U - - Y were t a k e n a t the slower
speed indicated below. U n a n a e s t h e t i z e d spinal eat

2Omsec

was found in several interneurones with monosynaptie excitation from high thres-
hold group I afferents of nerves t h a t did not display separation in Ia and Ib
volleys (cf. Fig. 4, A - - E and F - - J ) . I t is therefore likely t h a t also interneurones
with a I b E P S P can be monosynaptieally excited from cutaneous afferents.
An additional feature in Fig. 4 is the unit contributions to the E P S P in A - - C
and F - - J . I f each unitary potential is evoked from a single afferent there are at
least 8 group I afferents from G - - S converging on this cell to give the E P S P in J.
These unit contributions are considerably larger than the average E P S P t h a t is
evoked in motonenrones from a single I a afferent and t h a t represents the release
of only one quantum (Kv~o, 1964).
I n some interneurones an I P S P is evoked from group I muscle afferents. The
interneurone in Fig. 5 receives monosynaptic EPSPs from PBSt (A D), DP
( E ~ H ) , ABSm (Q), F D L (g) as well as small effects from G - - S and P1 (cf. intra-
and extracellular traces in L and P). The effect of graded stimulation in A - - H
suggests t h a t the E P S P is evoked both from I a and Ib afferents of the PBSt nerve.
There is a disynaptic group I I P S P evoked from G - - S (I--L), P1 (M--P) and Q
(T), and there is probably also a disynaptie I P S P superimposed on the mono-
344 T. HONGO,E. JANKOWSKAand A. LUNDBERG:

synaptic E P S P evoked from DP in F--I-I. I n addition there is an E P S P evoked

from high threshold muscle affcrents in H and R and from the SP nerve in S. Also
volleys in the contralateral F R A give an E P S P (V--Y). The group I I P S P ( J - - L
and N - - P ) is probably evoked from Ib affcrents but there is no separation in Ia
and Ib volleys and hence some uncertainty remains.
I n Fig. 6 is shown that the same interneuronc m a y receive IPSPs both from
group I muscle afferents and from the FRA. There is a monosynaptic E P S P from
plantaris (A--D) and from the F D L (E). The disynaptic group I I P S P in G is

2mVl i-" ....... -"'--'-"-

0
F 1.3 6 2.0 ~ ,~1 d 6.2

4mVl 4 msec
Sill
20 msec
6-S N

"- XE
I(

,4-
PBSt
18.7 Z
2-0-,7 0 ]

". 7~;.• ......

Fig. 6. Interneuror~e receivi~g a monosynaptie E P S P from group 1 muscle a~erents and I P S P s / t o m group I muscle
a~erents and 1tom the F R A . Intracellular recording (upper traces) w i t h a citrate electrode from an interneurone
located at a d e p t h of 3.0 m m from the cord dorsum. The lower traces in D, E, J, L, N and 0 are microelectrode
recordings obtained after withdrawal to a just extracellular position. The middle traces in those records a n d the
lower traces in the other records are from the L7 dorsal root entry zone. Nonosynaptie E P S P s are evoked b y
volleys in group I afferents from P1 and F D L . There is graded s t i m u l a t i o n of the P1 nerve in A - - D . The quadriceps
nerve is stim~dated in ~ ' - - I , the s u b m a x i m a l I a volley in F is ineffective b u t when the stimulus strength is raised
to give a I b volley there is a disynaptic I P S P in G. Low threshold group I I afferents are s t i m u l a t e d in I-I a n d there
is a later I P S P which is seen better in record I taken simultaneously at lower speed. The I P S P s in K - - N were
evoked when the s t i m u l u s s t r e n g t h was raised above t h a t needed to evoke m a x i m a l group I volleys. The initial
effect in L shown at faster speed in K is probably a disynaptie group I I I P S P . The high amplification was employed
in A - - D , all the other records were taken at the lower amplification. I m m e d i a t e l y after i m p a l e m e n t of the cell the
group I E P S P from P1 was as large as t h a t from F D L b u t the series i n A - - D were taken later when the membrane
potential was lower. Unanaesthetized spinal cat. Time calibration for A - - H is shown below H and for the other
records below J

probably evoked from Ib afferents and with stronger stimulation a later I P S P is

evoked, which is shown at slower speed in I and J. Large IPSPs evoked from high
threshold affercnts of other muscles are shown in L and M, the effect from cuta-
neous afferents in N and from high threshold joint affercnts in O.
On recording from interneurones supplied with excitation from nerves which
displayed separation in Ia and Ib volleys we have found that a group I I P S P m a y
be evoked in interneurones receiving an E P S P exclusively from I a afferents and
probably also in those with excitation from Ib afferents because the E P S P was
evoked from high threshold group I muscle afferents (A D, Fig. 6). The con-
vergence of group I excitation and inhibition from different nerves was investi-
gated in 18 interneurones. I n 8 of them the E P S P was evoked from one muscle or
from synergie muscles and the I P S P from an antagonist. The convergence from
 Postsynaptic Inhibition in Interneurones 345

group I afferents in the remaining 10 neurones is shown in Table 1. In two of

these cells (2 and 9) there is excitation from a flexor and inhibition from other
extensors than the antagonist, 6 cells receive both excitation and inhibition from
extensor muscles (1, 3, 6, 7, 8, 10) and the remaining cells are more complex since
they receive convergence of the same modality of synaptie action from both
flexors and extensors (4 and 5).
The excitatory and inhibitory convergence from different afferent systems on
interneurones influenced from group I muscle afferents is summarized in Table 2
and 3. Table 2 shows all the interneurones in which volleys in group I muscle

Table l
Convergence of monosynaptie group I E P S P s and disynaptie group I I P S P s on 10 interneurones.
Only interneurones with more complex convergence pattern are included in the table (see text).
Cells 1 4 from experiments on spinal cats are represented in Table 3. Cells 7--10 are from
experiments on cats in which the ipsilateral spinal half was intact to permit investigation of
e~ects from the rubrospinal tract

Nerves giving gr I EPSP ( + ) and IPSP (--)

Q PBSt ABSm G-S PI I FDL DP
i
1 0 0 0 0
2 o +0 +0 0
3 +0 0 0 + + 0
4
5 0 r +0
....
-
§
+ i
+
+
+--
+
6
+ ] + 0
7 0 0 + , + 0
8 0 0 - 0 I 0 0
9
10
+o
0 +0
0
0
- +0 [ 0 0
0
- --1 +
Table 2
Convergence of E P S P s (Ira) and I P S P s ( D ) on interneurones receiwng monosynaptic E P S P s
but no 1 P S P s from group I muscle a]erents. Disynaptic E P S P s from cutaneous a~erents are
indicated only if the volley in high threshold muscle afferents did not evoke E P S P s
A B C D E F G

Cut. monos.
GrI f ~ ; ; ~ i~m I
Cu~. dis.
FRA ] []
Nr of cells 2 8 4 1 1 3 7

afferents evoked monosynaptie EPSPs but not disynaptic IPSPs. Table 3 summa-
rizes the convergence on the interneurones with disynaptie IPSPs from group I
afferents. All the interneurones in Table 2 and 3 receive synaptic effects also from
other afferents than group I muscle afferents. Of 40 interneurones with mono-
synaptie EPSPs from group I muscle afferents 20 receive monosynaptie EPSPs
also from eutaneous afferents and 7 interneurones an EPSP from the FRA. The
inhibitory convergence is even more impressive; in all but 2 of these 40 inter-
neurones IPSPs were evoked from some source, in no less than 32 from the FRA
346 T. Ho~Go, E. JANKOWSKAand A. LV~DBERG:

an d in 14 f r o m group I muscle afferents. I n Table 3 t h e r e are 7 i n t e r n e u r o n e s w i t h

d i s y n a p t i c g ro u p I I P S P s t h a t do n o t receive a group I E P S P . D i s y n a p t i c group I
I P S P s are, however, r e l a t i v e l y rare in i n t e r n e n r o n e s t h a t lack m o n o s y n a p t i c
group I E P S P s , t h e 7 cells in F - - J Table 3 c o n s t i t u t e 11% of t h e cells in our
m a t e r i a l w i t h o u t m o n o s y n a p t i c g r o u p I E P S P s (eft Table 4 an d 5). B y comparison
35 % of t h e i n t e r n e u r o n e s with g r o u p I E P S P s received a d i s y n a p t i c group I I P S P .

Table 3
Convergence of EPSPs ( I ) and I P S P s ([]) on interneurones receiving disynaptic group I
I P S P s from group I muscle a]erents. All group I EPSPs in A - - E are monosynaptic
A B C D E F G H I J
Gr.I [] []

lo
Cut. monos.
Cut. dis.
FRA .
Nr of cells 2 3 5 1 3 1 3 1 1 1

I t should be n o t ic e d t h a t t h e r e m a y be c o n v er g en ce of I P S P s from different

afferent sources. A c o m m o n convergence p a t t e r n is t h a t of B an d C, Table 3.
These i n t e r n e u r o n e s with m o n o s y n a p t i e E P S P s from g r o u p I muscle afferents an d
from cutaneous afferents receive I P S P s from g r o u p I muscle afferents an d f r o m t h e
I~RA. The cells in C in a d d i t i o n h a v e a d i s y n a p t i c I P S P from cu t an eo u s afferents
that has been listed as a separate effect. Cutaneous afferents are part of the FRA
but the disynaptic IPSP from low threshold cutaneous afferents may not be part
of the FleA effect since it can be found in cells in which EPSPs are evoked from
high threshold muscle afferents (D, Table 3, cf. also next section). A comparison
of Table 2 and 3 reveals that disynaptic cutaneous IPSPs are more common in
interneurones with group I IPSPs (Table 3) than in the interneurones in Table 2,
which receive EPSPs but not IPSPs from group I muscle afferents.
ECCLES et al. (1960) classified interneurones as receiving Ia or Ib EPSPs. It has later
been reported (EccLEs, 1964a) that convergence from Ia and Ib afferents may occur. Apart
from findings as those in Fig. 5 suggesting convergence from Ia and Ib afferents we have
confirmed it in recordings from interneurones in which the group I EPSP was evoked from a
nerve with perfect separation in Ia and Ib volleys. ECCL~,s et al. (1960) found that some inter-
neurones receive a disynaptic group I EPSP. This has been confirmed in two interneurones.
One of them is represented in A, Table 3. This interneurone received monosynaptic group I
EPSPs from P1 and FDL and also a disynaptic EPSP from these nerves which is not indicated
in the table. The other interneurone, which is not included in any table received no group I
monosynaptic EPSP but a disynaptic group I EPSP from G--S, P1 and FDL; there was also
a late IPSP from the FleA. ECCLESet al. (1960) correlated these polysynaptic group I effects
on interneurones with a polysynaptie EPSP that sometimes can be evoked in motoneurones
from very low threshold afferents in muscle nerves. We do not now believe that this polysyn-
aptie effect in motoneurones is evoked from Ia afferents. The reason is that it can only be
evoked from the FDL nerve, and when the interosseous nerve (of. HUNT and MeI~TYR]~, 1960)
was separated from the muscle nerves proper a polysynaptic EPSP was evoked in motoneu-
rones from the former but not from the latter nerve (unpublished observations).
The monosynaptic groupI EPSP in interneurones has almost invariably a shorter time course
than the Ia EPSP in motoneurones and this is probably due mainly to the depolarized state of the
interneurones. The longlasting EPSP in Fig. 7 is exceptional, and is probably recorded from an
interneurone and not from a motoneuronal dendrite. Recording was made in the ventral horn
 P o s t s y n a p t i c I n h i b i t i o n in I n t e r n e u r o n e s 347

w h i c h also c o n t a i n s i n t e r n e u r o n e s a c t i v a t e d f r o m p r i m a r y afferents (WILHS a n d WILLTS, 1966).

T h e m o n o s y n a p t i c E P S P is e v o k e d f r o m I a afferents o f t h e Q a n d s a r t o n i u s (Sart) n e r v e s .
I n a d d i t i o n a late E P S P w a s e v o k e d f r o m h i g h t h r e s h o l d m u s c l e afferents as is i l l u s t r a t e d in
L a n d P for t h e effect f r o m Q. T h e I a E P S P h a s a t o t a l d u r a t i o n o f m o r e t h a n 50 msec. A pro-
longed t r a n s m i t t e r a c t i o n h a s b e e n p o s t u l a t e d to a c c o u n t for p r o l o n g e d E P S P s in m a n y t y p e s

Fig. 7. Interneurone receiving longlasting

monosynaptie EPSPs #ore fa agerents. sart. 3.90
Intracellular recording (upper traces) with
a citrate electrode from an interneurone in
L6 located in the ventral horn at a depth
of 4.1 mm from the cord dorsum. The lower
traces are from the L6 dorsal root entry
zone. Corresponding upper and lower re-
cords in A--H and I--P were obtained
simultaneously at different sweep speeds.
There is graded stimulation of the sar- ~ ~ ~~.
torious nerve (8art) in A--H and of the
Q nerve in I--P. A maximal monosynaptie /Omsec
EPSP from Q is evoked in J at a strength
that is subthreshold for Ib but gives a a l, la A 90z
maximal Ia volley. A later EPSP in L and
P is evoked from high threshold muscle
afferents. A H were taken immediately
after impalement and I--P later. This
difference probably explains the smaller
afterhyperpolarization that follows the
spike potential in the latter records. No
spontaneous spikes were observed in the
period when I--P were recorded but ob-
serve that after the late spikes in 0 the
potential returns to the same level as after 20 msec
the spikes evoked on the rising phase of
the EPSP. Hence there is no evidence for survival of transmitter and rebuilding of the EPSP after the early spikes.
The spike potential was 65 mV but the resting membrane potential was not measured. The ipsilateral spinal half
(except dorsal coluum) was in intact connection with higher centres. Choralose anaesthesia

o f cells (cf. ECCLES, 1964a). H o w e v e r , w h e n a spike is e v o k e d on t h e s u m m i t of t h e E P S P in

t h e i n t e r n e u r o n e of Fig. 7 t h e l a t e r p h a s e of t h e E P S P is e n t i r e l y r e m o v e d a n d n o t r e b u i l t
a f t e r w a r d s . T h i s i n d i c a t e s ~hat t h e r e is no s u r v i v a l of t h e t r a n s m i t t e r b e y o n d t h e rising p h a s e
of t h e E P S P b u t t h a t t h e ] o n g l a s t i n g E P S P p r o b a b l y is c a u s e d b y a long t i m e c o n s t a n t o f t h e
m e m b r a n e . T h e late d i s c h a r g e s in M a n d 0 are p r e s u m a b l y c a u s e d b y a local r e s p o n s e b e i n g
a d d e d to t h e p e r s i s t a n t d e p o l a r i z a t i o n of t h e E P S P . T h e slow r a t e o f rise ( I - - L ) a n d t h e slow
d e c a y o f t h e E P S P m a y indicate either t h a t a d e n d r i t e w a s i m p a l e d or if t h e s o m a w a s i m p a l e d
t h a t t h e s y n a p t i c i m p i n g e m e n t w a s on dendrites, b u t w h a t e v e r t h e m e c h a n i s m t h e r e is t h e
i m p l i c a t i o n t h a t a d i s c h a r g e is p r o d u c e d a f t e r a long l a t e n c y b y a m o n o s y n a p t i c E P S P e v o k e d
f r o m I a afferents.

2. Interneurones receiving monosynaptic E P S P s ]rom cutaneous agerents

In many of the interneurones described in the last section monosynaptic
EPSPs were evoked from cutaneous afferents in addition to the effect from group I
muscle afferents. The present group of interneurones resemble those but were
neither excited nor inhibited from group I muscle afferents. IPSPs from the FRA
are common as has already been illustrated in preliminary publications (Ho~Go
et al., 1 9 6 5 a ; L U N D B E R G , 1 9 6 6 ) a n d i s f u r t h e r i l l u s t r a t e d in the interneuronc of
F i g . 8. T h i s cell r e c e i v e s a m o n o s y n a p t i c EPSP f r o m S P ( B , C) a n d T i b ( D ) . A p a r -
t i c u l a r f e a t u r e i s t h e l a r g e I P S P e v o k e d f r o m S P i n A b y a s t i m u l u s t h a t is s u b l i -
minal for the EPSP. At higher stimulus strength a later IPSP appears in B and
i n c r e a s e s i n C. T h e s e t w o w a v e s a r e a l s o e v o k e d f r o m t h e m i x e d T i b i n D w h e r e a s
stimulation of the sural nerve and muscle nerves gives only the later IPSP (E--H
at slower speed). The central latency for the IPSP evoked f r o m S P i s 1.3 m s e c
Exp. Brain Res. Vol. I 23
348 T. HOl~GO, E. JANKOWSI~Aand A. LUNDBERG:

indicating a disynaptic linkage. Disynaptic IPSPs from cutaneous afferents were

found in the majority of the cells monosynaptically activated from cutaneous
afferents (Table 4) but in the large majority of the cells the monosynaptie EPSP
is evoked at slightly lower or the same strength of stimulation as the IPSP. The

~_ ~ C 2.5

E F 010.7 0 PBSt 54 H P~91

"--- lOmV
20msec
Fig. 8. Interneurone receiving a monosynaptic E P S P ]rom cutaneous agerents, a disynaptic I P S P #ore cutaneous
agerents and an I P S P #ore the IeRA. Intrace]lular recording (upper traces) with a citrate electrode from an inter-
neurone located at a depth of 1.9 toni from the cord dorsum. The lower traces in C H were obtained after with-
drawal of the mieroelectrode to a just extracellular position. The middle traces in these records and the lower
traces in A and B are from the L7 dorsal root entry zone. Observe t h a t with graded s t i m u l a t i o n of the St' nerve
an I P S P is evoked in A and an E P S P only when the stimulus strength is raised to 1.5 times threshold in B. I n B
there also appears a later I P S P , which increases with stronger s t i m u l a t i o n in C. The I P S P from the muscle nerves
in F - - H are evoked from high threshold afferents. Voltage calibration in D is v a l i d for A - - D and the calibration
in H for E - - t I . A - - D are taken at the faster speed, E - - t t at the slower, l{esting membrane potential 35 inV.
Unanaesthetized spinal eat

d i s y n a p t i c I P S P from c u t a n e o u s afferents is f o u n d in m a n y of t h e interneurones

in which I P S P s are also e v o k e d from high t h r e s h o l d muscle afferents, b u t is n o t
necessarily a p a r t of the F R A response. This is i n d i c a t e d b y t h e records in Fig. 9
i l l u s t r a t i n g t h a t a large d i s y n a p t i c I P S P m a y be e v o k e d from cutaneous afferents

Table 4
Convergence of EPSPs ( I ) and 1PSPs ( 0 ) on interneurones receiving monosynaptic EPNPs
from cutaneous agerents but no P S P from group I muscle a]erents
A B C D E F

.monos
Cut. dis.
r r
FRA [] Ol []
Nr of cells 5 2 9 6 2 4

(D, E) in an i n t e r n e u r o n e t h a t receives a n E P S P from high t h r e s h o l d muscle a n d

joint afferents (B, C). A similar convergence p a t t e r n has also been o b s e r v e d in
neurones of t h e spinocervical t r a c t (unpublished). Table 4 s u m m a r i z e s t h e con-
vergence p a t t e r n onto neurones of this section. Of 28 i n t e r n e u r o n e s 17 receive
I P S P s from t h e F R A , 16 d i s y n a p t i c I P S P s from c u t a n e o u s afferents a n d t h e r e is
convergence of b o t h effects in 11 cells (B, C). Six cells receive a n E P S P from
t h e F R A b u t no I P S P from a n y source, a n d t h e 5 cells in A are of t h e t y p e illu-
s t r a t e d in Fig. 9 with E P S P s from t h e F R A a n d a d i s y n a p t i c I P S P from cutaneous
 Postsynaptic Inhibition in lnterneurones 349

afferents. I n 4 interneurones (B, E) both E P S P s and I P S P s are evoked from

the FRA. One cell of the latter group is illustrated in Fig. 10. The upper row
shows extracellular records t a k e n before impalement. A train of spikes is evoked
from SP, Q and F D L hut there is no discharge evoked from high threshold affe-

A 0].5 B ]5,3 ~ ~ ~ Lc stir]9

$-10 ITISeC5mY ~ ~ ~"~i " ' ' ' ' ~ , - ~

Fig. 9. Interneurone receiving a monosynaptic E P S P #ore cutaneous aUerents, a polysynaptic E P S P from the F R A
and a disynaptie I P S P from cutaneous agere.nts. Intracellular recording (upper traces) with a citrate electrode from
an interneurone a t a d e p t h of 1.4 m m from the cord dorsum. The lower traces in B - - E are extracellular obtained
after withdrawal o f the microelectrode to a j u s t extracellular position. The middle traces in those records and the
lower trace in A are from the L7 dorsal root entry zone. There is a monosynaptic E P S P in n from SP followed by
a disynaptic I P S P and a later wave of E P S P . I n A a group I volley from Q has no effect b u t an E P S P is evoked
from high threshold muscle afferents of this nerve in B. Spinal cat anaesthetized with chloralose

rents of PBSt (E) and only one spike from Sur (D). The corresponding lower
records show the expected E P S P s from SP, Q and F D L but also a large I P S P
from PBSt (J) and Sur (I). Moreover the E P S P s from Q (G) and F D L (H) are cut
short b y an I P S P . I t is noteworthy t h a t the I P S P dominates in I despite the fact

SP
A 14.9, 0 FDL sur PBSt
B 12,8 C 26,7 0 49.2 E
- -
29.9-
12mV

20msec
Fig. 10. Comparison oJ extra- and intracellular records Jrom an interneurone, l%cords A - - E are extraeellular ob-
tained before i m p a l e m e n t of an interneurone located at a d e p t h of 2.0 m m from the cord dorsum. The lower traces
in F - - K are from the L7 dorsal root e n t r y zone. The records in ]0" J are intracellular and evoked b y the same
volleys t h a t are indicated in the corresponding upper records. I n K there is a j u s t suprathreshold s t i m u l a t i o n o f the
SP nerve showing a n E P S P t h a t does not evoke discharge. Single traces in A - - F . Unanaesthetized spinal cat

t h a t there is an extracellular discharge in D and t h a t there is a hyperpolarization

following the spike in F, whereas the same volley in A gives a long train of impulses.
A probable explanation is t h a t in the depolarized state after impalement the
EPSPs are decreased and the I P S P s increased. Hence in I an E P S P m a y be con-
cealed b y the IPSP, the same m a y hold true in F, although there is also the after-
hyperpolarization to consider. Generally there has been a very good agreement
between extra- and intracellular records in t h a t EPSPs found after impalement
with regard to size and time course could explain the extraeellular discharge. I f it
is assumed t h a t the cell bodies are impaled it follows t h a t there is no need to
postulate a localized synaptie impingement with impulse initiation in remote
dendritic regions.
23*
350 T. Ho~Go, E. JANKOWSKAand A. LUNDBERG:

3. Other types o/interneurone8

I n t h e last g r o u p of cells no m o n o s y n a p t i c a c t i o n was f o u n d from a n y nerve
tested. Seven of t h e 30 neurones in T a b l e 5 receive an E P S P from t h e F l e A a n d
two cells b o t h an E P S P a n d a n I P S P from t h e F R A (Fig. 14). H o w e v e r , t h e
largest group is F with 13 cells t h a t
A SPlg,8 B surlT,3 C TiblS.5 receive a n I P S P from t h e F R A
b u t no E P S P from a n y source as
i l l u s t r a t e d in Fig. l l. There are
large I P S P s from cutaneous ner-
ves in A a n d B a n d t h e effect
from high t h r e s h o l d muscle affe-
D DP2.17 E 5.45 F 1g.9 r e n t s is exemplified in D - - G
w i t h s t i m u l a t i o n of D P a n d Q as
i n d i c a t e d . I n this e x p e r i m e n t t h e
t i b i a l nerve was n o t cut in order
G LI 12.4 H coil 20.7 / co.sur11.6 to p e r m i t a d e q u a t e s t i m u l a t i o n of
t h e foot. W i t h a n y k i n d of mani-
p u l a t i o n of t h e foot (pressure,
pinching a n d b e n d i n g of toes) no
- 20 msec 5 mV u n i t a r y E P S P s were o b s e r v e d b u t
d resting K pressure L o n l y I P S P s as shown for pressure
~ ,,..v-v-vtv',~
of t h e toes in K a n d L. H e n c e i t
was n o t possible to o b t a i n evi-
I0 msec dence t h a t a n E P S P was concea-
5 mV
led b y t h e large I P S P e v o k e d
Fig. 11. lnterneurone receiving an I P S P ]rom the t~RA. Intra-
cellular recording (upper traces) with a citrate electrode from a from Tib in C. H o w e v e r , i t is ob-
ceil at a depth of 2.3 m m from the cord dorsum. Lower traces
in A - - C , F and G are obtained after withdrawal of the micro-
vious t h a t c a u t i o n m u s t be exer-
electrode to a just extracellular position. The middle traces in cised in judging these cells because
those records and the lower traces in H - - L are from L7 dorsal
root entry zone. The I P S P s from D P and Q are evoked from
t h e resting p o t e n t i a l in m o s t cells
high threshold afferents, observe t h a t the m a x i m a l group I volley was b e t w e e n 2 0 - - 4 0 m V a n d t h e
from D P has no effect (D). The tibial nerve was dissected, b u t
in intact connection with the foot ; K and L show the I P S P s evo- resulting increase of t h e I P S P
ked adequately on pressure of the toes and should be compared could v e r y well obscure an E P S P
with the resting record in G. The resting membrane potential
was 45 mV. Single traces in J - - L . Unanaesthetized spinal cat i f m i x e d actions were e v o k e d
from one nerve (cf. last section).
There is also t h e p o s s i b i l i t y t h a t some of these i n t e r n e u r o n e s received e x c i t a t i o n
from a hip nerve t h a t was n o t dissected for s t i m u l a t i o n . H o w e v e r , t h e r e m a y well
be i n t e r n e u r o n e s t h a t receive o n l y I P S P s from p r i m a r y afferents a n d are supp-

Table 5
Convergence of E P S P s ( l l ) and I P S P s (rl ) on interneurones that neither received monosy-
naptic E P S P s from cutaneous a#erents nor P S P s from group I muscle a#erents. Disynaptic
cutaneous EPSPs are indicated only for interneurones that did not receive E P S P s front high
threshold muscle a#erents
A B C D E F
Cut. dis.
12111 []
Nr of cells 1 5 3 7 1 13
 Postsynaptic Inhibition in Interneurones 351

lied with excitation from descending pathways. I n some of the interneurones of

column F, Table 5 a monosynaptic E P S P was evoked by a descending volley in
the ipsilateral spinal half.

4. E~ect ]rom contralateral nerves

The effect of volleys in the contralateral hamstring and sural nerves was tried
in many of the interneurones. I n no case was there any effect from contralateral
group I muscle afferents but volleys in the contralateral F R A evoked I P S P s and
more rarely EPSPs in some of the interneu-
roues. I t is of particular interest that inter-
neurones that receive an E P S P from the
ipsilateral F R A m a y receive an I P S P from
the contralateral FleA. This is illustrated in
Fig. 12. The EPSPs evoked from ipsilateral
cutaneous afferents and high threshold
muscle afferents are shown in A D and
E - - F illustrate the I P S P s from the contra- B , ~ " " sur 17.4 E co. sur 11.6
lateral nerves. The effect from contralateral
nerves was tested in 15 of the interneurones
that received EPSPs from the ipsilateral
F R A and in 8 of them there was an I P S P
evoked from the contralateral FRA. I n two
of these 15 interneurones volleys in the
contralateral F l e a evoked an E P S P and in
the remaining 5 cells there was no effect
from the contralateral side. The effect of
volleys in the contralateral F R A were tested
on 21 interneurones that received an I P S P
from the ipsilateral FRA. Only in one of 20msec
these interneurones there was an E P S P /Omsec
Fig. 12. Interneurone receiving an E P S P #ore the
from the eontralateral side whereas I P S P s ipsilateral Ft~A and an I P S P #ore the contra-
were evoked in 9 of them. lateral t~RA. Intracellular recording (citrate elec-
trode) from an interneurone located a t a d e p t h of
1.6 m m from the cord dorsum. The upper traces
5. Location o/interneurones are intracellular, the lower traces are extra-
cellular obtained after withdrawal o f the micro-
The position of the interneurones re- electrode to a j u s t extracellular position. The
middle traces are from the ipsilateral L7 dorsal
corded from was determined ss described root entry zone. A n E P S P was evoked from high
threshold muscle affcrents of m a n y muscle
under methods. All interneurones in A, nerves but only the effect from D P is shown
B and C, Fig. 13, received monosynaptic (n). The I P S P from the contralateral H is evoked
from high threshold muscle afferents. Unauaes-
EPSPs from group I muscle affcrents. thetized spinal cat
There is a comparison of the location of
interneurones in A with and without group I IPSP, in B with and without a
monosynaptic E P S P from cutaneous afferents and in C of group I interneurones
in which EPSPs or I P S P s were evoked from the FRA. Except for a few group I
interneurones located in the dorsal horn all these interneurones are in the region
of the intermediate nucleus where the focal potential from group I afferents is
large (EccL]~s et al., 1954b), and there is no evidence for specific locations within
352 T. HONGO, E. JANKOWSKA and A. LUNDBERG:

this nucleus of interneurones with different convergence patterns. The interneu-

runes in D are those represented in Table 4, they all receive a monosynaptic E P S P
from cutaneous afferents but are not influenced from group I muscle afferents.
There is no evidence for specific location of those interneurones that in addition
receive polysynaptic E P S P from the F R A as compared with those with I P S P

C D

A 9 gr I EPSP without gr I IPSP

x and
B x gr Z EPSP without cut.monus.EPSP
9 ": ~nd
s x gr I EPS'P and FRA EPSP
9 IPSP
Z7x cut.monos. s and FRA IPSP
9 " EPSP
•
E x FBA EPSP
E 9 IPSP

Fig. 13. Location o/interneurones with digerent connections/rum primary a~erents. The drawings show the location
of those 75 interneurones in Table 2--5 which were located as described under Methods. All interneurones in A C
received monosynaptic E F S P s from group I muscle afferents and a comparison is made of the location of cells
with and w i t h o u t a group I IPSP(A), w i t h and w i t h o u t a monosynaptic E P S P from cutaneous afferents (B) and
of cells receiving E P S P s or I P S P s from the FRA(C). The cells in D received monosynaptic E P S P s from cutaneous
afferents (Table 4) and the comparison refers to cells with E P S P s or I P S P s from the FleA. E shows the position
of cells w i t h o u t a monosynaptic E P S P from p r i m a r y afferents (Table 5). The ce]ls in C - - E t h a t received mixed
excitatory and inhibitory effects from the F R A have been classified receiving EPSPs. All recordings were made
in acute spinaI cats and more ventral regions in the grey m a t t e r (which also contains m a n y interneurones t h a t can
be influenced from p r i m a r y afferents) were not explored in these experiments

from the FRA. There is some tendency for a double localization of the interneu-
roues in D, the majority of the cells are clearly more dorsally located than the
interneurones in A--C, but a smaller group is found more ventrally in the inter-
mediate nucleus. E shows the wide dispersion of interneurones influenced from
the FRA.
 Postsynaptic Inhibition in I n t e m e u r o n e s 353

6. T h e e ~ e c t o / N e m b u t a l
I t is surprising t h a t the large I P S P s t h a t can be evoked from primary afferents in inter-
neurones has not been observed in previous experiments with intracellular recording (cf.
ECCL~s et al., 1960). This m a y possibly be connected with the fact t h a t the animals were
anaesthetized with Nembutal. I n one interneurone we have observed t h a t the I P S P from the
F R A was considerably reduced b y a small dose of N e m b u t a l as is shown in Fig. 14. The corres-
ponding upper and lower records were t a k e n before and after the intravenous injection of
5 mg/kg of Nembutal. The comparison of record B and F in particular shows a considerable
reduction in the I P S P after N e m b u t a l and the decreased hyperpolarization in E, G and H is
presumably also caused b y a reduction in the I P S P although this is more difficult to judge

A 0 12,8 ABSm10.2
B r PBSt C~

Nembutal~
5 mg/kg
lerrl

2Omsec 5mYI
Fig. 14. The egect o] ~embutal on an interneurone receiving mixed excitatory and inhibitory e~ects ]rom the FRA.
Intracellular recording (upper traces) with a citrate electrode from an interneurone located at a depth of ].5 mm
from th~ cord dorsum. Lower records in E--~ are obtained after withdrawal of the microelectrode to a just extra-
cellular position. Corresponding upper and lower records were evoked by the same volleys before and after intra-
venous injection of 5 mg/kg Nembutal. The t)S1)s in A--C and E--G are evoked from high threshold muscle
afferents. Throughout the recording the resting membrane potential was 65 mY and the spike potential 70 mY.
Spinal unanaesthetized cat

because of the spike potentials evoked in these records. There is evidence of a decreased rest-
ing excitatory b o m b a r d m e n t after N e m b u t a l b u t there was no change of the resting potential
(60 mV) or the spike potential (75 mV). Hence it is unlikely t h a t the decreased I P S P is caused
b y a repolarization of the m e m b r a n e and more likely t h a t Nembutal has depressed trans-
mission in the inhibitory pathway. However, further experiments are required for a more firm
establishment of the hypothesis t h a t the inhibitory pathways are more sensitive to N e m b u t a l
t h a n the excitatory.

Discussion
I n t r a c e l l u l a r r e c o r d i n g h a s b e e n m a d e f r o m m o r e t h a n 100 i n t e r n e u r o n e s
located in the dorsal horn and intermediary region. IPSPs (reversed with increased
intracellu]ar chloride) are evoked by volleys in peripheral nerves in the great
m a j o r i t y o f t h e i n t e r n e u r o n e s . T h e s e I P S P s a r e e v o k e d f r o m a t l e a s t t h r e e diffe-
rent afferent systems: group I muscle afferents, low threshold cutaneous afferents
and the FRA. There has been no indication that monosynaptie IPSPs can be
evoked from primary afferents in interneurones, the shortest central latency for
t h e I P S P s i n d i c a t e s t h a t t h e m i n i m a l l i n k a g e is d i s y n a p t i c . T h i s f i n d i n g s u p p o r t s
t h e g e n e r a l i z i n g p o s t u l a t e t h a t p r i m a r y a f f e r e n t s a r e a l w a y s e x c i t a t o r y (ECcL~S,
1960, 1964a). T h e s e I P S P s m a y a c c o u n t f o r i n h i b i t i o n f r o m t h e p e r i p h e r y o f
d i s c h a r g e s i n i n t e r n e u r o n e s (KOLMODIN a n d SKOGLUND,1954, ] 9 5 8 , 1 9 6 0 ; FRARK
a n d FUO~TES, 1955; KOLMODIN, 1957 ; H V ~ T a n d K v ~ o , 1959a, b) b u t p r e s y n a p t i c
i n h i b i t i o n c a u s e d b y a P A D ( E c c L E s , 1 9 6 4 b ) m a y also c o n t r i b u t e .
354 T. HONGO,E. JANKOWSKAand A. L[rNDBERG:

The excitatory and inhibitory convergence is summarized in Tables 1--5. I n

Tables 1--3 no attempt has been made to classify effects from Ia and Ib afferents
separately because it was in many eases difficult to decide from which of these
afferents an effect was evoked. I t was, however, observed that I P S P s are evoked
from the F R A in interneurones receiving monosynaptie Ia EPSPs as well as in
those receiving monosynaptie Ib EPSPs. Likewise in both types of interneurones
disynaptic group I IPSPs were found. The latter effect is evoked mainly from
rather high threshold group I afferents suggesting a Ib origin. An inhibitory con-
vergence from group I muscle afferents and from the F R A is very common (Fig. 6,
Table 3) but the group I effect cannot be classified as part of the F R A response
because a group I I P S P has been found also in interneurones that receive exci-
tation from the F R A (Fig. 5 Table 3). Group I IPSPs are more common in inter-
neurones that receive a monosynaptic group I E P S P than in other types of inter-
neurones, the percentages being 33 and 11 respectively. Furthermore the nerves
to many hip muscles were not dissected and it cannot be excluded that interneu-
rones of the latter group did receive a monosynaptie E P S P from group I muscle
afferents. I t should also be noted that disynaptie IPSPs from low threshold
cutaneous afferents (ef. below) is found in interneurones with monosynaptic
group I EPSPs and much more often if there also is a group I IPSP.
Internenrones that are monosynaptieally activated from cutaneous afferents
form a large part of the present material as could rather be expected because of
the size of the extraeellular monosynaptic focal potential that can be evoked from
cutaneous afferents (CooM~S et al., 1956). Volleys in the F R A evoke IPSPs in
many of these interneurones. However, from cutaneous afferents there seems to
be a two-fold inhibitory effect. A disynaptic I P S P is evoked from very low thres-
hold cutaneous afferents and a later I P S P at somewhat higher strength of stimu-
lation. I t is possible that only the later wave of the I P S P is a part of the FI~A
response and that the disynaptie I P S P from the lowest threshold cutaneous affe-
rents is transmitted via a pathway that is not shared with high threshold muscle
afferents. The suggestion that the low threshold cutaneous afferents form a sepa-
rate afferent system in contributing inhibition to interneurones is based mainly
on the finding that a disynaptic I P S P can be evoked from these afferents in inter-
neurones that receive an E P S P from high threshold muscle afferents (Fig. 9).
Similar findings have been made with neurones of the spinoeervieal tract (Ho~TGo,
JA~KOWSKA and LUnDBErG, to be published).
Most of the interneurones recorded from were located as described under
methods. The great majority of the interneurones in which a monosynaptie E P S P
is evoked from group I muscle afferents are located in the intermediate nucleus in
REX~D'S (1954) layer V and VI where the focal potential evoked from group I
muscle afferents is large (EccL~s et al., 1954b). Most of the cells with a monosy-
naptic EPSP only from cutaneous afferents are located more dorsally in the region
where the extraeellular monosynaptie focal potential from these afferents is large
(CooMBs et aI., 1956), but there is a tendency for a double distribution in that some
interneurones of this type were found more ventrally in the intermediate nucleus
and the interneurones with a monosynaptic E P S P not only from cutaneous affe-
rents but also from group I muscle afferents are located within the latter region.
There was no evidence for a differential location of interneurones receiving predo-
 Postsynaptic Inhibition in Interneurones 355

minantly E P S P s or I P S P s from the F R A or for the interneurones with group I

EPSPs t h a t depend on whether they receive a group I I P S P or not (Fig. 13).
Before discussing the functional significance of the different types of interneu-
rones it must be emphasized t h a t the intracellular recording technique m a y be
selective in t h a t some types of interneurones m a y be more easy to impale than
others. There is a wide variation in size of the cell bodies of interneurones. Could
it be t h a t interneurones with an integrative function receiving extensive conver-
gence of excitatory and inhibitory convergence from primary afferents have larger
cell bodies (and therefore are easier to record from) than interneurones with a
simple input functioning mainly as relay cells ? However, in considering this pro-
blem it must be recalled t h a t interneurones of reflex pathways also receive effects
from descending pathways; for example the interneurones transmitting the reei-
procal I a inhibition receive effect from at least two descending pathways (LI~NI)-
BEI~G and VOOI~HOEVE, 1962; HONGO et al., 1965b). I t is possible t h a t a certain
degree of integration occurs in all interneuronal stations and t h a t there are no
intcrneurones with a simple relay function but nevertheless the possibility remains
t h a t the recording technique involves a selection. Even apart from these considera-
tions the excitatory and inhibitory convergence on interneurones cannot be dis-
cussed in more than general terms because of the difficulty in ascribing any inter-
neurone to a particular neuronal pathway. Several of the afferent systems are
already now known to have a multitude of connections in the spinal cord. With I a
afferents, for example, relatively few connections are known but there m a y be
others so far not discovered. Further experiments are required to find out if the
excitatory convergence from different afferents can be correlated with an occur-
rence of spatial facilitation between these different afferents in some neuronal
paths (el. I~. M. EcclmS and LU~DB~RC, 1958). Of interest in this connection is not
only the excitatory and inhibitory paths to motoneurones but also the paths to
primary afferents and ascending pathways. For example, there are pathways to
cutaneous afferents from cutaneous afferents, Ib afferents and the F R A (EccL~,S
et al., 1963b ; CARP~NTEI~et al., 1963) and to Ib afferents from Ib afferents and the
F R A (EccLES et al., 1963a). Convergence of inhibition from group I muscle affe-
rents and the F R A is found in neurones of the ventral (OsoAI~SSON, 1957; ECCLES
et al., 1961) and dorsal (Ho~Go and OKADA, unpublished) spinocerebellar tracts.
The existence of group I inhibition and of inhibition from the FleA in so m a n y
of the interneurones offers challenging possibilities for further experiments. There
is at present only a limited knowledge about the inhibitory interactions t h a t m a y
occur at an interneuronal level between neuronal pathways but our findings
suggest t h a t such interactions are very common. No inhibition from group I affe-
rents of a group I path, be it to motoneurones, to ascending pathways or primary
afferents, has so far been described t h a t cannot be explained b y interaction at a
primary afferent level. With respect to the inhibitory effect from the F R A atten-
tion should be drawn to the fact t h a t the pathway from I a to I a afferents can be
very effectively inhibited at an interneuronal level (LuND et al., 1965). The time
course of this inhibition led to the suggestion t h a t the mechanism is presynaptic,
caused b y depolarization of the terminals of interneurones, but postsynaptic
inhibition of interneurones could also contribute.
356 T. HONGO,E. JANKOWSKAand A. LUNDBERG:

The inhibitory effects from the FRA should also be considered in relation to
the possibility of inhibitory interaction between alternative reflex pathways from
the FRA. There is evidence suggesting excitatory and inhibitory reflex paths
from the F l e a to both flexor and extensor motonenrones (R. M. ECGLES and
LUND:aERG, 1959; I~OLMQVISTand LUNDB]mG, 1961; I-[OLMQVIST, 1961; WILSON
and KATO, 1965) and these alternative paths are probably controlled not only
from higher centres but also by inhibitory interactive processes. A dramatic
example of alternative reflex pathways is provided by the spinal reflexes evoked
from the FRA after the administration of DOPA (L-3,4-dihydroxyphenylalanine).
This drug depresses the short-latency reflexes normally evoked from the FRA in
the acute spinal cat and releases an entirely different reflex pattern from these
afferents (AND~x et al., 1964; JANKOWSKAet al., 1965). It is likely that the path-
way transmitting the latter response normally is closed at an interneuronal level
through inhibition from the short-latency reflex paths. Hence the release of the
new pattern after DOPA may be a consequence of a decreased inhibitory action
from the short-latency reflex path that results when transmission in this reflex
path is depressed (LUNDBERG, 1966). This interpretation presupposes the exi-
stence of interneurones receiving both inhibition and excitation from the FRA
and it is therefore of special interest that convergence of EPSPs and IPSPs from
the FRA now has been found in a number of interneurones.
Some interneurones are influenced not only from ipsilateral but also from con-
tralateral nerves. I t has been a rather common finding that interneurones acti-
vated from the ipsilateral Ft~A receive an I P S P from the eontralateral FRA.
Reciprocal effects from the two hindlimbs are known to exist in the interneurones
that transmit the late discharge from the Ft~A to motoneurones after the short-
latency FRA paths have been depressed by DOPA (JANKOWSKAet al., 1965). The
present findings suggest a similar reciprocal organization of the short-latency
pathways from the FRA, but again there is the uncertainty that it is not known
to which pathways the interneurones belong, and because of the bilateral PAD
evoked from the FRA (g. M. EccLEs et al., 1964a, b) it is extremely difficult to
investigate if there is a reciprocal relationship at an interneuronal level between
the short-latency FRA pathways from the two hindlimbs to motoneurones.
By definition a FRA pathway receives the same synaptic action from cuta-
neous afferents, high threshold muscle and joint afferents (R. M. ECGLES and
LtrND~ERG, 1959). In many interneurones an E P S P is evoked from low threshold
cutaneous afferents but not from high threshold muscle and joint afferents. Some
of these interneurones may belong to paths to primary afferents or ascending
pathways (CARPENTER et al., 1963; LUNDBERG and OSCARSSON, 1961), but it is
possible that some of them belong to pathways to motoneurones. There is increa-
sing evidence that there are special paths from cutaneous afferents to motoneu-
rones that are not part of the FRA pathways. For example, there is a pathway
from pressure receptors of the central plantar cushion to the motoneurones of toe
extensors (E~G]~E~G, 1964). Differential effects from the rubrospinal tracts on
synaptie actions evoked in motoneurones from low threshold cutaneous afferents
and from the FRA also suggest that specialized pathways may exist also from
other cutaneous areas (Ho~Go et al., 1965b). I f so the inhibitory action in these
interneurones may have a very special significance. The inhibitory effect from
 Postsynaptic Inhibition in Interneurones 357

l o w t h r e s h o l d c u t a n e o u s a f f e r e n t s m a y g i v e s p a t i a l s e l e c t i v i t y while i n h i b i t o r y
effects f r o m t h e F R A on t h e i n t e r n e u r o n e s m a y assist t h e p r i m a r y a f f e r e n t d e p o -
l a r i z a t i o n e v o k e d f r o m t h e F R A in l o w t h r e s h o l d c u t a n e o u s a f f e r e n t s ( E c c L ~ s
et al., 1963b) in s u p p r e s s i n g d e l i c a t e reflexes f r o m t h e s k i n d u r i n g t h e flexor reflex.

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Prof. ANDERS LUNDBERG

Department of Physiology
University of GSteborg
Medicinaregatan 11
G6teborg SV, Sweden

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