Exp Brain Res (1995) 104:275-286 9 Springer-Verlag 1995

D. Boi r e 9 R. Morri s 9 M. Ptito - F. Lepor e 9 D. O. Frost

Effects of neonatal splitting of the optic chiasm on the development
of feline visual callosal connections
Received: 15 April 1993 / Accepted: 13 December 1994
Ab s t r a c t Duri ng nor mal post nat al devel opment , t here is
an over pr oduct i on and subsequent part i al el i mi nat i on of
t he callosal proj ect i ons of cort i cal areas 17 and 18 in t he
cat. In t he present study, we i nvest i gat ed how neonat al
splitting of t he opt i c chi asm affect s this process. Our re-
sults i ndi cat e that neonat al splitting of t he opt i c chi asm
exaggerat es t he nor mal l y occurri ng partial el i mi nat i on of
i mmat ur e cal l osal proj ect i ons: it causes a si gni fi cant
r educt i on in t he t ot al number of neurons in the supra-
granul ar l ayers that send an axon t hr ough the corpus cal-
losum. It does not, however, cause a si gni fi cant change
in t he number of cal l osal l y proj ect i ng neurons in t he
i nfragranul ar layers. These dat a suggest that in addi t i on
to ot her fact ors pr evi ousl y descri bed, the l evel or spatial
distribution of correl at ed bi nocul ar i nput to visual corti-
cal neurons may i nf l uence t he st abi l i zat i on/ el i mi nat i on
of i mmat ur e cal l osal connect i ons.
Ke y wor ds Corpus cal l osum 9 Vision - Cor t ex 9
Pl ast i ci t y. Cat
I nt r oduct i on
Visual exper i ence affect s t he devel opment of visual cal-
losal connect i ons. Reari ng in the dark (Frost and Moy
1989), wi t h bi l at eral eyel i d suture or bi l at eral enucl e-
ation (Innocent i and Frost 1980; Innocent i et al. 1985) or
D. Boire 9 R. Morris 1 9 M. Ptito 9 E Lepore
Groupe de Recherche en Neuropsychologie Expdrimentale,
D6partement de Psychologie Universit6 de Montr6al, C.R 6128,
succursale A, Montrdal, Qu6bec, Canada H3C 3J7
D. O. Frost ( ~ )
Department of Pharmacology and Experimental Therapeutics,
University of Maryland School of Medicine,
655 West Baltimore St., Baltimore, MD 21201, USA;
Fax no: (410) 706-0032, e-mail: dfrost@umabnet.ab.umd.edu
Present address:
1 Department of Neurology and Neurosurgery,
Montr6al Neurological Institute, 3601 University, Montrdal,
Qu6bec, Canada H3A 2B4
wi t h al t ernat i ng monocul ar occl usi on (Frost et al. 1990)
all resul t in a subnormal number of cal l osal l y proj ect i ng
neurons (callosal neurons) in areas 17 and 18; enucl e-
ation (and, marginally, al t ernat i ng monocul ar occl usi on)
pr oduces an abnor mal l y wi de distribution of callosal
neurons in areas 17 and 18, whi l e dark reari ng and lid
suture pr oduce a slightly nar r ower t han normal distri-
bution. In kittens rai sed wi t h conver gent or di vergent
strabismus, monocul ar enucl eat i on or monocul ar eyel i d
suture, cal l osal neurons in areas 17 and 18 have a some-
what mor e wi despread distribution t han in nor mal ki t t ens
(Innocent i and Frost 1979; Ber man and Payne 1983), but
the rel at i ve number of cal l osal neurons is unknown. In
kittens rai sed wi t h strabismus, callosal axon t ermi nal s
are also abnor mal l y wi despread ( Lund et al. 1978). Vi-
sion appears to exert its effect s on the di st ri but i on of cal-
losal neurons by modul at i ng t he devel opment al el i mi na-
t i on of some i mmat ur e callosal axons (Innocent i 1981;
Koppel and Innocent i 1983; Innocent i et al. 1986; Ber bel
and Innocent i 1988), whi ch nor mal l y leads to a charac-
t eri st i cal l y rest ri ct ed t angent i al distribution of callosal
neurons (Innocent i et al. 1977; Innocent i and Cami ni t i
1980).
In the present experi ment , we sought to cl ari fy the role
in callosal devel opment of bi nocul ar i nput to the visual
cort ex. The reari ng condi t i ons that pr oduce an abnormal -
ly wi despread distribution of callosal neurons - monocu-
lar eyel i d suture, monocul ar enucl eat i on, strabismus
(and, marginally, al t ernat i ng monocul ar occl usi on) - all
vi rt ual l y abol i sh the normal bi nocul ar responsi veness of
cort i cal neurons in area 17 (revi ewed in Frost et al.
1990). However, t he i nt erpret at i on of t hese fi ndi ngs is
compl i cat ed by t he fact that all of t hese mani pul at i ons
di ffer wi t h respect to t hei r combi nat i on of several ot her
fact ors that mi ght also be expect ed to i nfl uence the dis-
t ri but i on of cal l osal neurons (see Di scussi on).
We suspect ed t hat neonat al splitting of t he opt i c chi-
asm at t he mi dl i ne mi ght be a usef ul mani pul at i on f or
f ur t her i nvest i gat i on of t he i nf l uence of bi nocul ar vi-
sion on cal l osal devel opment . In nor mal cats all of t he
vi sual cor t ex t hat r epr esent s t he bi nocul ar vi sual fi el d
276
receives correlated input from both eyes via the thal-
amocortical pathway; in addition, that part of the corti-
cal binocular field representation that is the target of
callosal axons can also receive correlated input from
both eyes via the corpus callosum. In cats reared with
monocular enucleation, monocular eyelid suture, stra-
bismus or alternating monocular occlusion, visual corti-
cal neurons do not receive correlated input from both
eyes, either because of the absence of effective input
from one eye (monocular enucleation or lid suture) or
because corresponding regions of the two eyes do not
receive the same pattern of visual stimulation (strabis-
mus, alternating monocular occlusion). By contrast, in
the case of optic chiasm splitting (OCS), the region of
termination of callosal axons is the only region to re-
ceive correlated input from both eyes. Information from
the nasal part of the visual field of the contralateral eye
is transmitted to the cortex via the corpus callosum and
is restricted to the callosal recipient zone; both in the
callosal recipient zone and in the remainder of the bin-
ocular field representation, the thalamocortical pathway
supplies input only from the nasal part of the visual
field of the ipsilateral eye. Thus, unlike the other ma-
nipulations previously tested, OCS maintains correlated
input from the two eyes, but reduces it in amount and
spatial extent, and alters the way in which it is relayed
to the cortex.
When the optic chiasm is split in adult cats, binocu-
larly responsive neurons in area 17 are restricted to the
region of termination of the corpus callosum, where
they constitute about one third of the visually respon-
sive population, rather than over 80% as in normal cats
(cf. Hubel and Wiesel 1962; Berlucchi and Rizzolatti
1968; Lepore and Guillemot 1982). Among the binocu-
lar neurons, the receptive fields through the two eyes
are located in heteronymous parts of the nasal visual
field and are always bordered on their medial side by
the vertical meridian (Berlucchi and Rizzolatti 1968).
Although in earlier studies the receptive field properties
of binocular neurons as tested through the two eyes in-
dividually were characterized as being identical (on the
basis of their orientation and direction tuning; Berluc-
chi and Rizzolatti 1968; Lepore and Guillemot 1982), it
has more recently been shown that the response to the
contralateral eye, which is mediated through the corpus
callosum, has lower contrast sensitivity and less spatial
and temporal resolution than the response to the ipsilat-
eral eye, mediated by the thalamocortical pathway
(Berardi et al. 1987). At the behavioral level, following
OCS at maturity, only the ability to discriminate high-
contrast, low-spatial frequency stimuli transfers from
one hemisphere to the other (Berardi et al. 1988; Cenni
et al. 1988)
The effects of neonatal OCS are somewhat different.
Following OCS at age 3 weeks (Bisti et al. 1990) or 6-8
weeks (Yinon et al. 1988), the proportion of binocularly
driven cells, though lower than normal, is higher than
following OCS at maturity. The type of information
transmitted through the corpus callosum also differs de-
pending on the time of OCS: Following OCS at age 3
weeks, unlike following OCS at maturity, the responses
to the contralateral eye of single neurons in the callosal
recipient zone are not reduced in contrast sensitivity and
spatial and temporal acuity, and the interocular transfer
of pattern discrimination can occur at high spatial fre-
quency and low contrast, not just at low spatial frequen-
cy and high contrast as in cats subjected to OCS as
adults (Bisti et al. 1990). OCS at 3 weeks of age causes
reductions in monocular and binocular acuity and also
elevates monocular and binocular thresholds for depth
perception, although the later deficit may be less severe
than for OCS performed at maturity (Timney and Lans-
down 1989). Because of the less severe behavioral defi-
cits following neonatal OCS than following OCS at ma-
turity, it has been suggested that neonatal OCS may
cause a rearrangement of callosal connections (Bisti et
al. 1990), although it is unclear how changes in the ana-
tomical arrangement of callosal connections might be re-
lated to the behavioral data.
Methods
A total of four experimental cats were used in this study. The cats
were reared normally until 12 days (OCS-2, 9 and 10) or 13 days
(OCS-3) of age, when they underwent a chiasmotomy using the
transbuccal approach described by Myers (1955). Four normal
adult cats used in previous studies (Frost and Moy 1989; Frost et
al. 1990) served as controls. All husbandry and manipulations
were carried out in accordance with the guidelines of the Canadian
Council on Animal Care and the United States Department of Ag-
riculture.
For splitting of the optic chiasm, the kittens were anesthetized
with halothane ca. 1.5% in a mixture of N20/O 2 (70:30). They
also received atropine (0.25 rag, IM), dexamethasone (2 mg/kg,
IM) and 5% dextrose in Ringer solution (60 ml/kg, sub-cutaneous-
ly administered at the end of the surgery). Surgery was performed
aseptically; EKG and temperature were monitored continuously.
At the end of the surgery, the kittens were given antibiotics (ampi-
cillin) and, when completely awake, were returned to their moth-
ers, who generally accepted them without problems. They were
then raised in the animal colony until they were 808, 710, 796 or
796 days old (OCS-2, 3, 9 and 10, respectively).
When the cats were mature, they received large, unilateral in-
jections of horseradish peroxidase (HRP) in the visual cortex so
as to retrogradely label the callosally projecting neurons in the
opposite hemisphere. Cats were initially anesthetized with ket-
amine hydrochloride (20 mg/kg, IM), then intubated and main-
tained on halothane ca. 1.5% in a mixture of 2 parts N20 and 1
part 02. They also received atropine (0.25 mg, IM), dexametha-
sone (2 mg/kg, IV) and 5% dextrose in Ringer solution (60 ml/kg,
IV, administered continuously during surgery). Surgery was per-
formed aseptically; EKG and temperature were monitored contin-
uously. A craniotomy was made to expose the lateral and postlat-
eral gyri. We made 10-12 0.5 gl injections of 40% HRP (Sigma
type VI) in H20 in these gyri using a Hamilton syringe with a 27-
G needle inserted into the cortex through small holes in the dura.
Injections were spaced ca. 1.5 mm along the anterior-posterior
axis and alternated between the medial and lateral sides of the gy-
ri. This procedure completely fills areas 17 and 18 with HRP
(Innocenti and Frost 1980). At the completion of surgery, the
craniotomy was closed with Silastic polymer and the cats re-
ceived dexamethasone (2 mg/kg, IV) and penicillin G procaine
(45 000 U/kg, IM). After reanimation, the cats were returned to
their cages.
277
Forty-eight hours postoperatively, the cats were deeply anes-
thetized with a lethal dose of pentobarbital sodium and transcardi-
ally perfused with the following sequence of solutions (all in
0.1 M PO 4 buffer, pH 7.4): (1) 0.9% NaC1 (ca 1000 ml), (2) 1%
paraformaldehyde, 2% glutaraldehyde (ca. 2000 ml), (3) 10% su-
crose (ca. 2000 ml), (4) 20% sucrose (ca. 2000 ml). Solutions 1
and 2 were at ambient temperature, 3 and 4 were at 4~ The
brains were then dissected, stored overnight in 30% sucrose in
P O 4 buffer at 4~ and sectioned while frozen in the coronal plane
at 80 ~tm. A one in two series of sections was then reacted for
HRP histochemistry using Mesul am' s (1978) tetramethyl benzi-
dine (TMB) technique and mounted on chrome-alum treated slides
(preincubation and incubation were at ambient temperature in the
dark; all other steps were at 4~ After drying for 1-2 h at ambi-
ent temperature, then overnight at 4~ the sections were dehy-
drated rapidly in graded alcohols at 4~ coverslipped with Per-
mount containing 1% BHT and stored at 4~
Al l of the sites of HRP injection were similar in size, location
and density, and resembled those obtained previously using simi-
lar techniques (Innocenti and Frost 1980; Innocenti et al. 1985;
Frost and Moy 1989; Frost et al. 1990). The HRP reaction product
fills most of the grey and white matter of the lateral and postlateral
gyri; occasionally a lighter precipitate extends into the suprasylvi-
an gyms. The completeness of the injections in areas 17 and 18
was confirmed by retrograde and anterograde transport of HRP to
all of the dorsal nucleus of the lateral geniculate body (LGd), ex-
cept, in some cases, the caudolateral extremity.
In all brains, HRP-l abel ed callosal neurons in a series of
sections spaced 320 gm apart and encompassing the caudal
13-15 mm of the cortex were charted at 250x using a computer
microscope. Additional sections between those used for the recon-
structions were inspected. Criteria for the identification of labeled
neurons were similar to those previously described (Innocenti and
Frost 1980). The significance of differences in the mean number
of callosal neurons per section in differently reared animals was
determined using a two-tailed Mann-Whitney U-test.
Using the computer, we made flattened reconstructions of the
distributions of labeled callosal neurons (Innocenti et al. 1985;
Frost and Moy 1989; Frost et al. 1990). Briefly, in each coronal
section, the labeled neurons were projected onto a contour running
400 gm below the pial surface. The contour was straightened and
divided into 100-gm segments and the number of neurons project-
ed onto each segment was indicated by vertical lines whose
lengths were proportional to the number of neurons. The rows of
lines representing the sections were aligned using the convexity of
the lateral gyms as a landmark. The cytoarchitectonic border be-
tween areas 17 and 18 was determined on selected sections count-
erstained with toluidine blue, using the criteria of Otsuka and
Hassler (1962).
In each brain, we measured the mediolateral width of the corti-
cal volume containing callosal neurons (callosal efferent zone,
CZ) at the three levels that divide the rostral-caudal extent of area
17 into four equal parts. (If one or two callosal neurons lay at a
great distance from the main body of callosal neurons, they were
not considered in calculating the width of the CZ). The mean of
these three widths was taken as an index of the width of the callos-
al zone in each animal. We then used the Mann-Whitney U-test to
determine the significance of differences in the width of the cal-
losal zone in normal cats and cats whose optic chiasma had been
split.
For each case, a second series of sections was stained with cre-
syl violet. These sections were used to confirm the completeness
of the split of the optic chiasm in two ways. First, the chiasm itself
was examined and the presence of a midline incision confirmed.
Second, the dorsal lateral geniculate nuclei were examined bilater-
ally for the transneuronal atrophy of the A-l ami nae that results
from the removal of crossed retinal input as a consequence of
the neonatal chiasm split. In all the cases reported in this study,
comparison of the A-laminae with the Al - l ami nae (which receive
uncrossed retinal input that is not affected by neonatal chiasm sec-
tion) revealed severe atrophy throughout the full extent of the A-
laminae bilaterally.
Results
Nu mb e r o f c a l l o s a l n e u r o n s i n a r e a s 17 a nd 18
o f n o r ma l cat s
The di s t r i but i ons o f l a b e l e d c a l l os a l ne ur ons i n nor ma l ,
a dul t cat s we r e as p r e v i o u s l y r e p o r t e d us i ng t he TMB and
d i a mi n o b e n z i d i n e t e c hni que s ( I nnoc e nt i 1980, 1986;
I nnoc e nt i a nd Fr o s t 1980; Se a gr a ve s a nd Ro s e n q u i s t 1982;
I nnoc e nt i et al. 1985). The y ar e s u mma r i z e d her e f or pur -
pos e s o f c o mp a r i s o n wi t h t he di s t r i but i ons i n OCS cat s.
I n n o r ma l cat s , c a l l o s a l n e u r o n s l i e i n a b a n d r u n n i n g
r o s t r o c a u d a l l y a l ong t he b o r d e r b e t we e n a r e a s 17 a nd 18
a nd e x t e n d i n g 1- 3 mm e i t he r s i de o f t he b o r d e r ( Fi gs . 1,
5). Thi s c a l l o s a l z o n e i s f l a n k e d b y u n l a b e l e d ( a c a l l o s a l )
r e g i o n s c o r r e s p o n d i n g t o mo s t o f a r e a 17 a nd t he l a t e r a l
pa r t o f a r e a 18. Thus , t he b a n d o f c a l l o s a l n e u r o n s s pa n-
ni ng t he 17/ 18 b o r d e r i s di s t i nc t f r o m t he c a l l o s a l z one s
i n a r e a 19 or i n t he s p l e n i a l s ul cus ( I n n o c e n t i 1980;
Se a g r a v e s a nd Ro s e n q u i s t 1982) . Ho we v e r , at di f f e r e nt
r o s t r o c a u d a l l e ve l s i n di f f e r e nt a n i ma l s , one t o t hr e e
b r i d g e s o f c a l l o s a l n e u r o n s s t r et ch a c r o s s t he f ul l me d i o -
l a t e r a l e x t e n t o f a r e a 18 t o j o i n t he c a l l o s a l z o n e i n a r e a
19. Fo r c o u n t i n g p u r p o s e s , t he l a t e r a l b o r d e r o f t he cal -
l os a l z o n e i n a r e a s 17 a nd 18 wa s e x t r a p o l a t e d a c r os s
t he s e b r i d g e s ( Fi g. 5).
I n a r e a s 17 a nd 18, c a l l o s a l n e u r o n s ar e d i s t r i b u t e d i n
t wo r a d i a l l y s e pa r a t e d, s u p e r p o s e d l a mi n a e i n l a y e r s I I I
a nd I V ( s u b z o n e a) a nd l a y e r VI ( s u b z o n e c) ( Fi g. 1).
Ne u r o n s i n t he t wo s u b z o n e s we r e e a s i l y d i s t i n g u i s h e d ;
t he f e w c a l l o s a l n e u r o n s i n l a y e r I I we r e a t t r i but e d t o
s u b z o n e a a nd t he f e w c a l l o s a l n e u r o n s i n l a y e r V t o t he
n e a r e s t s ubz one . Th e r e ar e ma n y mo r e c a l l o s a l n e u r o n s
i n s u b z o n e a t ha n i n s u b z o n e c ( Fi g. 4, Ta bl e 1; s e e al s o,
I n n o c e n t i 1980) .
Th e b o u n d a r y b e t we e n a r e a s 17 a nd t 8 i s n o t o r i o u s l y
di f f i c ul t t o d e t e r mi n e p r e c i s e l y i n t he cat ; t he r e f or e , s e p-
ar at e c ount s o f t he c a l l o s a l n e u r o n s i n e a c h a r e a we r e not
a t t e mpt e d. Th e n u mb e r o f l a b e l e d c a l l o s a l n e u r o n s di s -
t r i but e d a r o u n d t he a r e a 17/ 18 b o r d e r i n e a c h s e c t i on
d e c r e a s e s f r o m c a u d a l t o r os t r a l l e ve l s ( Fi gs . 2 - 3 , 5). Su-
p e r i mp o s e d on t hi s t r e nd a r e one or mo r e p e a k s t hat cor -
r e s p o n d a p p r o x i ma t e l y t o t he a r e a c e nt r a l i s r e p r e s e n t a -
t i on and, l es s r e l i a bl y, t o t he b r i d g e s c r o s s i n g a r e a 18.
Th e a v e r a g e n u mb e r o f n e u r o n s p e r s e c t i o n i n s u b z o n e s a
a nd c o f our n o r ma l cat s wa s 263. 7 a nd 39. 1, r e s p e c t i v e -
l y. As we p r e v i o u s l y o b s e r v e d ( I n n o c e n t i et al . 1985) ,
t he r e wa s s i g n i f i c a n t i n t e r a n i ma l v a r i a t i o n i n t he me a n
n u mb e r o f c a l l o s a l n e u r o n s p e r s e c t i on ( Fi g. 4, Ta bl e 1).
Nu mb e r o f c a l l o s a l n e u r o n s i n a r e a s 17 a nd 18
o f opt i c c h i a s m- s p l i t cat s
OCS i n ki t t e ns doe s not a l t e r t he r a d i a l d i s t r i b u t i o n o f
c a l l o s a l n e u r o n s i n a r e a s 17 a nd 18. I n OCS cat s as in
n o r ma l cat s , c a l l o s a l n e u r o n s i n- a r e a s 17 a nd 18 a r e di s -
t r i but e d i n t wo di s t i nc t , r a d i a l l y s e p a r a t e d , s u p e r p o s e d
l a mi n a e i n l a y e r s I I I / I V a nd l a y e r VI ( Fi g. 1).
17
N4
Fi g . 1 Computer-microscope plots of the distribution of HRP-la-
bel ed callosal neurons in coronal sections through areas 17 and 18
of one normal (N-4) and one OCS (OCS-2) cat. Ar r owheads indi-
cate the 17/18 border as determined by cytoarchitectonic criteria
after Nissl counterstaining. I ns et bet ween brai ns shows dorsal
view of the left hemisphere of N4 (caudal is up, lateral is to the
right); t he l i ne across the hemi sphere indicates the coronal level
from which these sections were taken
Th e c a l l o s a l e f f e r e nt z o n e i n a r e a s 17/ 18 o f OCS cat s
c o n t a i n s 29. 2% o f t he n o r ma l n u mb e r o f l a b e l e d c a l l o s a l
n e u r o n s i n s u b z o n e a a nd 94. 3% o f t he n o r ma l n u mb e r i n
s u b z o n e c ( Fi g. 4, Ta bl e 1). I n OCS as i n n o r ma l cat s ,
t he r e i s i n t e r a n i ma l v a r i a b i l i t y i n t he n u mb e r o f l a b e l e d
c a l l o s a l n e u r o n s p e r s e c t i on. Th e r e f o r e , we t e s t e d t he
s t a t i s t i c a l s i g n i f i c a n c e o f our r e s ul t s u s i n g t he Ma n n -
Wh i t n e y U- t e s t a p p l i e d t o t he me a n n u mb e r o f n e u r o n s
p e r s e c t i o n i n e a c h a n i ma l , wi t h n e x p r e s s i n g t he n u mb e r
o f a n i ma l s . Thi s t es t s h o we d t hat OCS c a u s e s a s i gni f i -
c a nt r e d u c t i o n i n t he me a n n u mb e r o f l a b e l e d c a l l o s a l
n e u r o n s i n s u b z o n e a ( P=0 . 0 2 0 9 ) but not i n s u b z o n e c
( P =I . 0 ) . Th e n u mb e r o f l a b e l e d c a l l o s a l n e u r o n s i n OCS
cat s wa s a l s o r e l a t i v e l y mo r e v a r i a b l e t ha n i n n o r ma l
cat s. Wh e r e a s t he r a t i o o f t he s t a n d a r d d e v i a t i o n t o t he
me a n wa s 0. 195 a nd 0. 128 i n s u b z o n e s a a nd c o f n o r ma l
cat s , r e s p e c t i v e l y , i n OCS cat s t he c o r r e s p o n d i n g r a t i os
we r e 0. 572 a n d 0. 391.
Al t h o u g h t he r e d u c t i o n s i n t he n u mb e r o f c a l l o s a l
n e u r o n s o c c u r at al l r o s t r o c a u d a l l e ve l s , we ha ve t he i m-
p r e s s i o n t hat t he r e d u c t i o n i n s u b z o n e a ma y b e s o me -
wh a t g r e a t e r i n t he c a u d a l 3 mm or s o o f a r e a s 17/ 18
( wh i c h r e p r e s e n t t he c o n t r a l a t e r a l u p p e r h e mi f i e l d ; Tu s a
et al . 1978 1979) t ha n e l s e wh e r e . Da t a on t he r o s t r o c a u -
17
9 " ; %
278
DORSAL
1 mm LATERAL
OCS 2
Ta bl e 1 Summary of data used in statistical comparisons of nor-
mal (iV) and OCS cats. From left to right, columns indicate case,
mean number of callosal neurons per section in subzone a of areas
17/18, mean number of callosal neurons per section in subzone c
of areas 17/18 and mean width of subzone a at the three rostrocau-
dal levels that divide areas 17/18 into four equal parts
Case Subzone a Subzone c CZ width
neurons/section neurons/section (ram)
N3 269.7 39.2 7.39
N4 333.8 44.4 6.98
N5 218.8 40.4 4.78
N6 232.3 32.4 4.68
Mean+SD 263.7+51.5 39.1+5.0 5.96+1.43
OCS-2 46.27 47.22 5.14
OCS-3 41.47 24.27 2.46
OCS-9 136.9 51.32 5.68
OCS- 10 83.46 24.63 4.76
Mean_+SD 77. 04_+44. 14 36. 86_+14. 43 4. 51_+1. 42
da l d i s t r i b u t i o n o f c a l l o s a l n e u r o n s ar e s u mma r i z e d i n
Fi gs . 2 a nd 3, wh e r e we ha ve c o mp a r e d t he d i s t r i b u t i o n s
i n f our pa i r s o f n o r ma l a nd OCS cat s : (a) t he n o r ma l a nd
OCS cat s ( N- 4 / OCS - 3 ) wi t h, r e s p e c t i v e l y , t he mo s t a nd
l e a s t c a l l o s a l n e u r o n s p e r s e c t i o n i n s u b z o n e a o f t he i r
gr oups ; t hi s c o mp a r i s o n p r o v i d e s a ma x i mu m e s t i ma t e o f
t he ef f ect s o f n e o n a t a l OCS; (b) t he n o r ma l a nd OCS
cat s ( N- 4 / OCS - 9 ) wh i c h i n t he i r r e s p e c t i v e g r o u p s ha d
t he mo s t c a l l o s a l n e u r o n s p e r s e c t i o n i n s u b z o n e a; (c)
t he n o r ma l a nd OCS cat s ( N- 5 / OCS - 3 ) wh i c h i n t he i r r e-
s pe c t i ve g r o u p s ha d t he l e a s t c a l l o s a l n e u r o n s p e r s e c t i o n
i n s u b z o n e a; (d) t he n o r ma l a nd OCS cat s ( N- 5 / OCS - 9 )
wh i c h we r e mo s t n e a r l y ma t c h e d i n t he n u mb e r o f cal -
Z 8(x7-
9
g -
(,.) 6o0
a' )
" " 400
Z
9
~:~ 20o
aa
Z o
A
SUBZONE A
oo o ~ 9 OCS3
o
o N 4
o
0 0 O^ 0
0
0
0 0 0 ~ O0 000
0 O0 0 0
0 0 0 0 0 0
0 000
rl O_
9 o* 9 % 0 ~
9 : i ,
2000 4000 6000 8000 10000 12000 14000 16000
LEVEL (btm)
Z 8o0
O
Q.~ 600
O 0
" ~ 4 0 0
z
0
20o
z o
B
279
o % " OCS9
oO o
o N4
0
0 o o
9 o o 0 o o o 000
o ~ 0 0 0 ~ 0o0 0 00o
" ; - ' . 0 o % 0 0 o
~ ~ o ~ o~ 9 ~ ~ o O~ ~ Oo
9 OON O o
u i u u u i u n
2000 4000 6000 8000 lfXXX) 1200( 7 14{XX.) 16(XX)
LEVEL (btm)
Z 5 0 0 "
_ o
b- 4O0-
.
m
c~ 3 0 0 -
Z 2o0-
9
a<
10o
Z 0
o
o 9 OCS3
o o ~ o N5
0 o
o
o OO o 0 0 o ~
Oo o Oo o Oo 0
0 0 0 0 0
0 0
e ~ e ~ b ~ 1 7 6 ~
. 0 , 9 " o o l b o ~ 1 7 6 1 7 6 o ~ o o O o ~ 1 7 6 % o o ~
i i i i i i i i
2(X)0 4000 6000 8000 10000 12000 14000 16f)00
C LEVEL (btm)
F i g . 2A-D Rostrocaudal distributions of labeled callosal neurons
in subzone a in two normal cats (N-4, N-5) and two OCS cats
(OCS-3, OCS-9). Instructive pairings of normal and OCS cases are
described in text. The horizontal axis indicates positions of coronal
sections in micrometers rostral to the caudal extremity of the neo-
cortex. The vertical axis indicates the number of HRP-labeled cal-
losal neurons in subzone a. Open circles represent data points from
normal cats, whereas filled circles represent data points from OCS
cats. The distributions represent: A The normal and OCS cats (N-
4/OCS-3) with the most and least callosal neurons, respectively, of
their groups. B The normal and OCS cats (N-4/OCS-9) which in
their respective groups had the most callosal neurons per section. C
The normal and OCS cats (N-5/OCS-3) which in their respective
groups had the least callosal neurons per section. D The normal
and OCS cats (N-5/OCS-9) which were the most nearly matched in
the number of callosal neurons per section in subzone a
losal neurons per section in subzone a. (N-5 and OCS-9
were also the normal and OCS cats which had, respec-
tively, the least and most callosal neurons per section of
their groups; the difference between them thus provides
a mi ni mum estimate of the effects of neonatal OCS). In
all four normal cats, the number of subzone a callosal
neurons per section rises rapidly passing rostrally from
the caudal tip of the cortex, hits a relatively high peak,
then plateaus, or declines slowly (Fig. 2); in all the OCS
cats, the number of subzone a callosal neurons per sec-
tion rises more slowly to peak at about 3 mm, then pla-
teaus, or declines slowly (Fig. 2). We have previously
observed a similar effect in cats reared with binocular
eyelid suture (Innocenti and Frost 1980), with dark rear-
ing (Frost and Moy 1989) and with alternating monocu-
Z 5 0 0 -
-
[-.. 400-
o o 3 0 0 "
Z 2 O0
O -
~oo-
z o
o
o
o 9 OCS9
o o ~ o N5
'~ o
e o o 9 o
9 0
0 0 00
0 ~0 O00 0 O0 0 000
O O 0 9 00 00
9 O 0 O 0 O 0 0 9 0 _ _ _ 0
g O 9 9 e N p O 9 w w ~ 0 ~ 0
9 ~ o 0 0 ~ o o
o
n 1 I I I t I I
2(X)O 4( X) O 6( X) O 8 O0 0 I ( X) ( X) 120(X) 1400~) 160O0
D LEVEL (pro)
lar occlusion (Frost et al. 1990). Extreme cases N-4 and
OCS-3 clearly differ most in the caudal 3 mm of the cor-
tex. The rostrocaudal distributions of callosal neurons in
subzone c of both normal and OCS cats resemble the
corresponding distributions in subzone a (Fig. 3). We
have not tested the possibility that, within either sub-
zone, specific depths or neuronal types may be preferen-
tially affected.
Tangential distribution of callosal neurons in normal
and chiasm-split cats
In normal, adult cats, both the mediolateral width of sub-
zone a and its position with respect to the crown of the
lateral and postlateral gyri show significant individual
variations (Table 1, Fig. 5). The position of subzone a
reflects that of the border between areas 17 and 18 as
determined cytoarchitectonically. Few callosal neurons
extend medially into area 17 as far as the suprasplenial
sulcus; this happens mainly rostrally.
In OCS as in normal cats, the density of callosal neu-
rons peaks near the 17/18 border and diminishes progres-
sively with increasing distance from the border (Figs. 1,
6). In both normal and OCS cats, the callosal zone is rel-
atively wide in the caudal 3 mm of the cortex and pro-
gressively narrows further rostrally. In OCS cats, the me-
diolateral width of subzone a appears to be somewhat
less than in normal animals, particularly caudally. This
280
SUBZONE C
Z 2(X)
9
r/?
~" 100"
Z
9
e-/
Z
A
0
O0
9 OCS3
o N4
o ooo
9 0 0 O 0
~ 1 4 9 o % 0 o o
~ 0 0 o O o
lb o e ~ - ~
9 "
l l l n I l U l l
0 2000 4000 6000 8000 10000 12(XX) 14000 16000 18000
LEVEr, ( gm)
Z 2(X)-
9 9 OCS9
9 ~ o N4
Oo
"~" l f f d
c ~ o c x ~
Z
O " ^ ~ ~ 1 7 6 o
o o 0 n ~ o o . o ~ 9 9 r, O O ~ . ~ -
LT-I
Z o- - oo , 9 I l I l l l l l
0 2000 4(100 6 ( X) O 8( X) O 10000 12(X)O 14000 160(X) 180fX)
B LEVEL ( gm)
Z 200-
9
O~
100"
C13
z
9
z 0-
9 OCS3
o N5
O
0 0 o
o
o o 9
O0
O O 0 ~ 9 9 o 0 o 0
9 O o O o ~ % o ~ . , _ % -
9
' I ' I ' I ' J ' I ' I ' I 0 I [
2{1OO 4(X)0 60(0) 8f X) O IC)O{X) 120(10 14000 I60(X) 180(X)
C LEVEL (btm)
t endency is mos t pr onounced in cat OCS- 3, whose cal -
l osal zone is r el at i vel y uni f or m al ong its r ost r o- caudal
extent. I n OCS cats, the mean of our i ndex of t he wi dt h
of the cal l osal zone was 75. 7% of nor mal (Tabl e 1);
however, this nar r owi ng was not st at i st i cal l y si gni fi cant
( Mann- Whi t ney U-test; P=0. 3865) .
Mor phol ogy of cal l osal neur ons
Mos t cal l osal neur ons are pyr ami dal cells, t hough s ome
are spi ny st el l at es or spi ndl e- shaped ( I nnocent i 1980).
TMB r eact i on pr oduct per mi t s t he vi sual i zat i on onl y of
neur onal s omat a and pr oxi mal dendri t es, whi l e still
per mi t i ng t he as s es s ment of neur onal si ze and br oad
mor phol ogi cal class. Usi ng the TMB t echni que, we di d
not not i ce any abnor mal i t i es in neur onal si ze or f or m
among l abel ed cal l osal neur ons in OCS cats, al t hough no
quant i t at i ve meas ur ement s wer e at t empt ed.
D i s c u s s i o n
Rel i abi l i t y of resul t s
The number and t angent i al di st ri but i on of l abel ed cal l os-
al neurons in areas 17/18 of cat s r ear ed accor di ng to the
s ame par adi gm show s ome vari abi l i t y. We have t aken
meas ur es to r educe t he cont ri but i on of t echni cal fact ors
Z 2 0 0 -
9
E-
L)
C/?
1(~)'
Z
9
Z 0
0
9 OCS9
9 9 o N5
oj
0 o 0
o
0 o 9 9 9 O~ O0 _ _ 9 9
o 9 9 ~ ' ~ ~ , ~ 8 ~ ' ~ o ~ - o
w O CO ~ 0 ~ 0 ~ ~O000D 9
I ' I ' I ' I ' I ' ] ' I 0 n I
2000 4(X)0 6(W}0 8000 100(X) 120(X) 140fX) 160(X) 180(~,)
D LEVEL (urn)
F i g . 3A-D Rostrocaudal distributions of labeled callosal neurons
in subzone c for the same cats as illustrated in Fig. 2. The same in-
structive pairing of the normal and OCS cases as described in text
and illustrated in Fig. 2 are shown. The same conventions as in
Fig. 2 have been used, except that the vertical axis represents the
number of labeled callosal neurons in subzone c
to this vari abi l i t y: (a) we st andardi zed t he i nj ect i on pro-
cedure and hi st ol ogi cal pr ocessi ng of t he t i ssue f r om one
case to the next; (b) we chart ed and count ed l abel ed neu-
rons wi t h the same opt i cs in si mi l ar l y spaced sect i ons
di st ri but ed over a compar abl e sect or of cort ex in each
case; (c) by chart i ng a f ew sect i ons several t i mes, we es-
t i mat ed our i ndi vi dual or i nt eri ndi vi dual count i ng error
to be on the order of +3%. Our cri t eri a for i dent i f yi ng
l abel ed neur ons and f or di fferent i at i ng t hem f r om ot her
l abel ed el ement s are di scussed el sewher e (Innocent i and
Fr ost 1980).
I n i ndi vi dual ani mal s, t he st andar d devi at i on of t he
mean numbe r of cal l osal neur ons per sect i on is l ar gel y
due to r ost r ocaudal var i at i ons in t he numbe r of t hese
neur ons. I mpor t ant i ndi vi dual var i at i ons in t he di st ri bu-
t i on of cal l osal neur ons have al so been r epor t ed in nor-
mal rat s ( Cus i ck and Lund 1981) and monke ys (Van
Essen et al. 1982). I n t he cat , i ndi vi dual var i at i ons in
r et i not opi c maps in vi sual areas ( Tusa et al. 1978) ma y
cor r el at e with, and pos s i bl y cause, t he var i at i ons in
numbe r and di st r i but i on of cal l osal connect i ons. Ther e
are al so i mpor t ant i ndi vi dual di f f er ences in t he t ot al
numbe r of cal l osal axons count ed by el ect r on mi cr os co-
281
500"
Z
0 400
300'
[] SUBZONE A
[] SUBZONE C
100 i
0
r ~ r.~ ra~ ~ Z
EXPERI MENTAL
9 ~ It3
z
NORMAL
when the stabilization/elimination of immature callosal
connections is complete (Innocenti and Caminiti 1980;
Berbel and Innocenti 1988). (2) Postnatally, subzone a
is the principal source of transient callosal projections
and undergoes much more severe natural reduction
postnatally than does subzone c (Innocenti and Cami-
niti 1980); therefore, subzone a might be expected to be
more affected by visual experience, an expectation
I borne out in the present data on OCS cats. We have pre-
vi ousl y discussed other possible reasons for the differ-
ential sensitivity of subzones a and c (Innocenti et al.
i 1985). Rearing kittens with OCS also marginally de-
creases the width of the callosal zone.
We have presented (Innocenti and Frost 1980; Inno-
centi et al, 1985) multiple observations that make it un-
likely that difficulties in HRP transport or visualization
would account for the effects of early binocular eyelid
suture on the number and distribution of callosal neurons
in areas 17/18; similar arguments apply to the present re-
sults in OCS cats, although a precise estimate of the
z
OCS-induced loss of callosal axons originating from ar-
] eas 17/18 awaits direct axon counts in the corpus callo-
s um.
Fi g. 4 Hi s t ogr ams showi ng t he me a n n u mb e r of HRP- l abel ed cal -
l osal neur ons per sect i on i n areas 17/18 of each cat i n t hi s study.
Nor mal and OCS cat s are gr ouped separ at el y al ong t he hor i zont al
axis. The vertical axis i ndi cat es t he numbe r of HRP- l abel ed neu-
r ons per sect i on. Empty bars r epr es ent count s f or s ubzone a; filled
bars r epr es ent count s f or s ubzone c. Vertical lines r epr es ent 1
st andar d devi at i on i n t he count s f r om i ndi vi dual cats. Horizontal
lines bet ween t he s ubzone a hi s t ogr ams i n each gr oup i ndi cat e t he
r aean numbe r of l abel ed s ubzone a cal l osal neur ons per sect i on for
t he gr oup
py in both cats (Koppel and Innocenti 1983; Berbel and
Innocenti 1988) and monkeys (Lamantia and Rakic
1990).
Effects of chiasmotomy on callosal connections
Rearing kittens under conditions of OCS reduces the
number of callosal neurons in subzone a to 29.2% of its
normal value, and is without a significant effect on the
number of callosal neurons in subzone c. As di scussed
previously for binocular eyelid suture (Innocenti and
Frost 1980; Innocenti et al. 1985), the OCS-i nduced re-
duction in the number of callosal neurons in areas
17/18 seems to be related to the natural postnatal re-
shaping of callosal connections (Innocenti and Caminiti
1980), i.e., the elimination of axons that cortical neu-
rons transiently send through the corpus callosum
(Innocenti 1981; Koppel and Innocenti 1983; Berbel
and Innocenti 1988). OCS may exaggerate this normal
elimination of callosal axons. This interpretation is sup-
ported by two other observations: (1) While the end of
the period during which OCS has its effects on callosal
connections has not been determined, OCS clearly has
a significant effect when initiated prior to the time
Effects of chiasm splitting on the organization
of the visual system
It is important to comment briefly on the effects of neo-
natal optic chiasm split on visual system organization.
As explained in the Introduction, the most significant ef-
fect of OCS on visual system organization is to reduce
the amount and spatial extent of correlated input to the
cortex from the two eyes, and to alter the way in which
that input is relayed to the cortex. The relevance of this
effect to callosal development is discussed below.
OCS has additional effects. We have not examined the
retinae in our neonatally chiasmotomized cats and know
of no other studies involving neonatal or adult chiasmo-
t omy in which this has been done. However, following
chiasmotomy in adult cats, retinal ganglion cells in the
nasal retina presumably degenerate as a consequence of
axotomy, as do all retinal ganglion cells following optic
nerve section (Stone 1965, 1978). However, since neona-
tal optic tract section causes abnormal partial stabiliza-
tion of a normally transient uncrossed retinal projection
originating in the nasal retina (Jacobs et al. 1984), in ne-
onatally OCS cats there may be a weak, abnormal retinal
input from the ipsilateral nasal retina to the LGd and vi-
sual cortex, although the projection may be too sparse to
be functionally significant. The LGd in neonatally OCS
cats shows marked transneuronal degeneration of the
laminae deprived of retinal input by OCS (see Methods),
as has been previously demonstrated following neonatal
or adult monocular enucleation (Guillery 1973); proba-
bly, as for enucleation, the rate of degeneration is faster
following neonatal manipulation. The geniculate laminae
receiving uncrossed retinal input appear normal at the
light microscopic level.
282
N 6
. . . . . . . . . i . . . . . . . . , . . . . . . . . . . . . . . .
N 3
, * * 2 r a m
. . . . 9 C
9 9
o e
" ' oo
e
M
; 'i ............ II I . . . . . . . 9 9 * l , ~ l ] ] l ~ l , I i . . . . .
. . . . . . . . . . . , i , , m . . . . 9 e
ee . . . . . . . . . . . . . . . l l l ~, t . . . . . . 9 . . . . . . . . . . . . n, . m, ~ . . . . . . . 9
~ ~ : ~,,~:iiii!i'.!!':l]!i!!]iii.!!!!?::::: . . . . . . . "
Fi g, 5 Dorsal view computer reconstructions of a part of callosal
subzone a in four normal adult cats. Flattened representations of
posflateral and lateral gyri represent hatched areas in correspond-
ing insets of dorsal view of brains (based on photographs). Dotted
lines represent, from left to right (lateral to medial), the fundi of
the lateral, postlateral and supraspenial sulci. The asterisks mark
the boundary between areas 17 (medially) and 18 (laterally). The
neurons in subzone a of each section were projected onto a line
running parallel to the pial surface and 400 gm deep; the line was
divided into bins of 100 gm and the number of neurons in each bin
was represented by a line segment whose length was proportional
to the number of neurons in the bin; the number of l abel ed neu-
rons represented by a unit of line segment height is the same in
each case. Each row of line segments represents one section.
Cross-hatching indicates regions of areas 18 and 19 that are con-
tinuous with the reconstructed parts of the callosal zone and with-
in which there is a high density of labeled callosal neurons; parts
of these regions form "bridges" (described in the text) and show a
great deal of individual variability in both normal and chiasm-split
cats. Neuronal counts used for histograms and statistics exclude
the hatched regions and correspond to the parts of subzone a re-
constructed with line segments. Solid lines caudolaterally in each
reconstruction indicate that at the levels where the lines are pres-
ent the reconstruction could not be extended further laterally, since
the cortex lateral to the lines was sectioned obliquely. (Scale lines
represent 2 mm; M medial, C caudal)
To our k n o wl e d g e , t he r e ha ve b e e n no qua nt i t a t i ve
s t udi e s o f t he h i s t o l o g y o f t he v i s u a l c o r t e x i n cat s s ub-
j e c t e d t o OCS as n e o n a t e s or a dul t s , t h o u g h t he f e a t ur e s
t hat d i s t i n g u i s h a r e a s 17 a n d 18 a r e s t i l l r e c o g n i z a b l e . I t
i s p o s s i b l e t hat t he c h a n g e s d e s c r i b e d i n t he p r e c e d i n g
p a r a g r a p h c a u s e mi n o r a l t e r a t i ons o f t he c or t i c a l r e pr e -
s e n t a t i o n o f t he vi s ual f i e l d i n n e o n a t a l l y OCS cat s . As
s u mma r i z e d i n t he I n t r o d u c t i o n , t he p r i n c i p a l f u n c t i o n a l
c o n s e q u e n c e o f OCS i n a dul t cat s i s t o r e d u c e t he c on-
t r a s t s e ns i t i vi t y, s pa t i a l r e s o l u t i o n a nd t e mp o r a l r e s o l u -
t i on o f v i s u a l i n f o r ma t i o n t r a n s mi t t e d t h r o u g h t he c o r p u s
c a l l 9 an e f f e c t t hat a p p e a r s t o be a me l i o r a t e d f ol -
l o wi n g n e o n a t a l OCS. I t i s di f f i c ul t t o r e l a t e t he s e f unc -
t i o n a l c h a n g e s t o our o b s e r v a t i o n s on c h a n g e s i n t he
n u mb e r o f c a l l o s a l ne ur ons . Al s o , s i nc e OCS wa s pe r -
f o r me d i n our s t u d y at a ge 1 2 - t 3 d a y s a n d i n t he f unc -
t i ona l s t udi e s at a ge s 3 - 8 we e k s , c a u t i o n s h o u l d be e xe r -
c i s e d i n c o mp a r i n g t he t wo set s o f r e s ul t s s i nc e t he mo r -
p h o l o g i c a l a n d f u n c t i o n a l c o n s e q u e n c e s o f OCS a r e a g e -
d e p e n d e n t .
Na t u r e o f vi s ua l c o n t r o l o f c a l l o s a l d e v e l o p me n t
The s u p e r n o r ma l r e d u c t i o n s i n t he n u mb e r o f c a l l o s a l
n e u r o n s t hat o c c u r as a c o n s e q u e n c e o f n e o n a t a l b i n o c u -
0 o
OCS 2
w
: ~ , i : . ~ , : : ! l l l i i l i l i l i ~ l J I : ! l L i l l i l I I l i ! l ~ : i ; i ; : : ~ ' " : ~
. . . . . . . . , - - ~ , o m , b , i i , H , i , . + . . . . . . . . . . . . .
OCS 9
2 mm
~ M
Fig. 6 Dorsal view computer reconstructions of a part of callosal
subzone a in four adult OCS cats. All conventions are as in
Fig. 5
lar eyelid suture, binocular enucleation or dark rearing
(Innocenti and Frost 1980; Innocenti et al. 1985; Frost
and Moy 1989) suggested to us that patterned visual ex-
perience was necessary for the selective stabilization
(Changeux and Danchin 1976; Bear et al. 1987) of the
normal complement of callosal neurons in areas 17/18
[although visual experience could have stabilizing and
destabilizing effects on immature callosal connections at
different stages of development (Innocenti et al. 1985)].
The present data on the consequences of OCS rearing
demonstrate a supernormal developmental reduction in
the number of callosal neurons, which is even greater
than that we have previously observed following rearing
with binocular eyelid suture (Innocenti et al. 1985) and
alternating monocular occlusion (Frost et al. 1990), the
deprivation paradigms that had, until the present study,
produced the most severe effects on the number of cal-
losal neurons (cf. Innocenti and Frost 1980; Frost and
Moy 1989). This result is extremely surprising, since
whereas binocular eyelid suture causes severe reductions
in the vigor and selectivity of cortical neuronal responses
to visual stimulation (reviewed in Sherman and Spear
283
9 9 . . . . . . i i ' i i i i i i i , i i
m
OCS 3
OCS 10
' 1 1 1 , 1 i 8 4 1 8 4 i i i 7 .
. . . . . . . . . , / , i : 1 2 , , , : i : ? , : 1 1 1 1 ' : '
. . . . . . . . . . q . . . . . . . . . . . . . .
1982; Fregnac and Imbert 1984), the neurophysiological
consequences of neonatal OCS are relatively benign, be-
ing mainly a severe loss of binocular responsiveness
among individual cortical neurons, which is somewhat
mitigated in the immediate vicinity of the representation
of the vertical meridian (Bisti et al. 1990).
It might be suggested that the greatly reduced abso-
lute level of visually evoked activity reaching the cortex
is the principal cause of the exaggerated developmental
loss of immature callosal neurons following neonatal
OCS. Three previous observations argue against this hy-
pothesis: (a) Neonatal binocular eyelid suture (Innocenti
et al. 1985) and neonatal alternating monocular occlu-
sion (Frost et al. 1990) appear to equally exaggerate the
loss of immature callosal neurons, although the effect of
both manipulations is less than we obtained following
neonatal chiasmotomy. The demonstration that alternat-
ing monocular occlusion and binocular eyelid suture
have approximately equal effects suggests that the great-
ly decreased absolute level of visually induced neuronal
activity in the optic nerve and lateral geniculate nucleus
of binocularly eyelid sutured cats, as compared to alter-
nating monocular occluded cats, cannot fully account for
the effects of binocular eyelid suture. (b) Bilateral neona-
tal enucleation causes a less severe reduction in the final
number of callosal neurons than does bilateral neonatal
eyelid suture (Innocenti and Frost 1980) even though vi-
284
sually evoked activity persists among thalamocortical
afferents following bilateral lid suture but not following
bilateral enucleation. Thus, the greatly reduced absolute
level of visually evoked activity reaching the visual cor-
tex in neonatally chiasmotomized cats is unlikely to fully
explain the exaggerated developmental loss of immature
callosal neurons in those animals. (c) Even following
rearing with monocular eyelid suture, monocular enucle-
ation or strabismus, paradigms that cause a widening of
the callosal zone (and thus maintain some callosai effer-
ents that would otherwise be eliminated), most of the
callosal projection present at birth is eliminated (Inno-
centi and Frost 1979).
Although the results were not quantified, we have
previously demonstrated that rearing with monocular
eyelid suture, monocular enucleation or strabismus caus-
es a clear widening of the callosal zone and, thus, the
maintenance of callosal efferents which would otherwise
be eliminated (Innocenti and Frost 1979 and unpublished
data). [Alternating monocular occlusion causes a mar-
ginal widening of the callosal zone (Frost et al. 1990).]
Since all these rearing conditions disrupt the normal bin-
ocular responsiveness of cortical neurons in area 17, we
had expected that OCS rearing, which also disrupts bin-
ocular responsiveness, would cause a similar widening of
the callosal zone. However, following neonatal OCS, the
callosal zone is marginally narrowed. This finding might
be explained in two ways:
(1) While OCS, alternating monocular occlusion,
monocular enucleation, monocular eyelid suture and
strabismus all disrupt the binocular responsiveness of
area 17 neurons, they differ with respect to their combi -
nation of several factors that might be expected to in-
fluence the distribution of caltosal neurons: the number
of eyes simultaneously stimulated, the t ypes of stimula-
tion simultaneously applied to the two eyes, and the
effects that the rearing paradigm has on the shape of
the ocular domi nance distribution of area 17 neurons.
In alternating monocul ar occlusion and monocular enu-
cleation, onl y one eye at a time is stimulated. In OCS,
monocular lid suture and strabismus, bot h eyes are
stimulated simultaneously, but since the eyelids act as
diffusers and strabismic cats are alternating ambl yopes,
the stimulation is well focused in one eye and diffuse
(for lid suture) or poorl y focused (for strabismus) in the
other. In cats subj ect ed to neonatal OCS, the acuities of
the two eyes are equal, but are about half of normal, al-
though they are about t wi ce the acuity of the t wo eyes
in cats subj ect ed to OCS as adults (Ptito et al. 1990 and
unpubl i shed data). This is significant since deprivation
by absence of stimulation and by diffuse or defocused
stimulation differ (Tieman et al. 1983; Christen and
Mower 1987, inter alia). In monocularly lid sutured
(Wiesel and Hubel 1962, 1965) and monocularly enu-
cleated cats, the ocular dominance of area 17 neurons is
sharply skewed in favor of the normally stimulated eye,
whereas in alternating monocul ar occl usi on and strabis-
mic cats, the ocul ar dominance distribution is U-shaped
(Hubel and Wiesel 1965; Tieman et al. 1983), and for
cats raised with OCS, the ocular dominance distribution
is skewed in favor of the ipsilateral eye (Bisti et al.
1990). Thus, it is possi bl e that by virtue of their unique
combinations of these factors, the five rearing para-
digms differ with respect to the amount by which they
cause the CZ to widen, a possibility that can only be
tested by comparing much larger groups of cats, all
processed by the same histological methods, than are
currently available.
(2) Perhaps the tendency toward widening of the cal-
losal zone, which might be caused by reduced binocular
responsiveness, is swamped in OCS reared cats by the
supranormal developmental reduction in the number of
callosal neurons; the exaggerated elimination of callosal
neurons would most severely affect the edge of the cal-
tosal zone in peripheral area 17, where catlosal neurons
are always less dense than further centrally (Figs. 5, 6).
[A similar interaction between two effects of abnormal
visual experience may occur in cats reared with binocu-
lar eyelid suture and alternating monocular occlusion. In
cats reared with binocular eyelid suture, despite a reduc-
tion in binocular responsiveness (reviewed in Sherman
and Spear 1982; Fregnac and Imbert 1984), there is a
narrowing of the callosai zone, perhaps owing to the su-
pranormal developmental elimination of callosal neurons
(Innocenti et al. 1985)].
The present results on the effects of neonatal optic chi-
asm split indicate that the influence on callosal develop-
merit of binocular input to the visual cortex, may be more
complex than previously appreciated. Our data suggest
that both the amount and spatial distribution of correlated
binocular input to visual cortical neurons may influence
callosal development, and that these parameters and their
effects may be graded rather than all-or-nothing. In nor-
mal cats all of the visual cortex that represents the binocu-
lar visual field receives correlated input from both eyes via
the thalamocortical pathway; in addition, that part of the
cortical binocular field representation that is the target of
callosal axons can also receive correlated input from both
eyes via the corpus callosum. In cats reared with monocu-
lar enucleation, monocular eyelid suture, strabismus or al-
ternating monocular occlusion, visual cortical neurons do
not receive correlated input from both eyes, either because
of the absence of effective input from one eye (monocular
enucleation or lid suture) or because corresponding re-
gions of the two eyes do not receive the same pattern of
visual stimulation (strabismus, alternating monocular oc-
clusion). However, in the case of OCS, the region of ter-
mination of catlosal axons is the only region to receive
correlated input from both eyes: infon~aation from the
nasal part of the visual field of the contralateral eye is
transmitted to the cortex via the corpus callosum and is re-
stricted to the callosal recipient zone; both in the callosal
recipient zone and in the remainder of the binocular field
representation, the thalamocortical pathway supplies input
only fi om the nasal part of the visual field of the ipsilater-
al eye. Thus, unlike the other manipulations previously
tested, OCS maintains correlated input from the two eyes,
but reduces it in amount and spatial extent and alters the
wa y i n wh i c h i t i s r e l a y e d t o t he cor t ex. Th e s e c ons i de r -
at i ons s ugge s t t hat t he a mo u n t or s pat i al di s t r i but i on o f
c or r e l a t e d b i n o c u l a r i nput t o vi s ual c or t i c a l ne ur ons mi g h t
i nf l ue nc e t he n u mb e r or di s t r i but i on o f ne ur ons t hat ma i n-
t ai n p e r ma n e n t c a l l os a l pr oj e c t i ons .
Th e p r e c e d i n g d i s c u s s i o n o f h o w n e o n a t a l OCS p r o -
duc e s i t s e f f e c t s on t he c o r p u s c a l l o s u m e mp h a s i z e s t he
f act t hat t he p o s t n a t a l d e v e l o p me n t o f t he c o r p u s c a l l o -
s u m i s u n d e r t he c o n t r o l o f mu l t i p l e , i nt e r a c t i ng i nf l u-
e nc e s wh i c h di f f e r i n t he ma g n i t u d e a nd q u a l i t y o f t he i r
e f f e c t s ( I n n o c e n t i a n d F r o s t 1980) . A c h a l l e n g e f or f u-
t ur e r e s e a r c h i s t o u n d e r s t a n d wh y a nd h o w t he s e i nf l u-
e nc e s ha ve t he e f f e c t s t hat t he y do.
Acknowl edgement s This research was supported by grants EY-
03465 from NIH and MH-40157 from NIMH to D. O. F and by
grants from the National Science and Engineering Research Coun-
cil of Canada and from FCAR (Qu6bec) to M. E and F.L. The
authors also wish to thank Wendy West for excellent technical
assistance and Lauren Ptito for assistance with the manuscript.
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