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The effects of differential environments on

plasticity of cortical pyramidal neurons in adult
rats

Article in Experimental Neurology · January 1979

DOI: 10.1016/0014-4886(78)90276-5 · Source: PubMed

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EXPERIMENTAL NEUROLOGY 62, 658-677 (1978)

Effects of Differential Environments on Plasticity of

Dendrites of Cortical Pyramidal Neurons
in Adult Rats

H. B. M. UYLINGS,* K. KUYPERS,* M. C. DIAMOND,-) AND

W. A. M. VELTMAN*~ l

*Netherlands Imtitute for Brain Research, P.O. Box 41850, 1009 DB Amsterdam,
The Netherlands and tDeparfme& of Physiology and Anatomy, University of
California, Berkeley, California

Received March 9.1978, revision received July 31,1978

Thickening of the frontal cortex and especially the occipital cortex was
observed in adz& rats after exposure to the “enriched” condition. An
increase in branching and in length of terminal segments was found in the
dendritic tree of pyramidal cells in layers II ad III of the visual cortex of
the adulf rat after exposure to “standard” and enriched conditions. These
exposures began at day 112 and continued 30 days. The increase observed in
the basal dendritic tree of pyramidal cells in the superficial layers was sig-
nificantly greater in the enriched conditions than in the standard condition.
It appeared, furthermore, that branching occurred predominantly on basal
terminal segments of all orders at a considerable distance from the tip. This
mode of growth is similar to that observed in the cortex of normal immature
rats. The differential conditions did not influence the bifurcation angles.
The dendritic and cortical changes and changes reported in the literature
indicate that the effects of differential experience are not limited to a short,
“critical” period.

INTRODUCTION
In studies of the ontogenetic development of dendrites it was frequently
assumed that no further outgrowth of dendrites occurs in adulthood (10).
1 We are indebted to Dr. P. D. Coleman for the use of his dendrite tracking system,
to Dr. A. Miodonski, Miss R. E. Johnson, and Mrs. B. M. Przybylski for their expert
histological assistance, to Drs. E. L. Bennett and J. G. Parnavelas for the most help-
ful comments, and to Mrs. N. L. A. de Klerk for typing this paper. H. B. M. IJ.
was supported by a fellowship of the Netherlands Organization for the Advancement
of Pure Research (Z.W.O.). Please send reprint requests to Dr. H. B. M. Uylings
in Amsterdam.

0014-4886/78/0623-0658$02.00/O
Copyright 6 ld78 by Academic Pkss, Inc.
All rights of reproduction in any form reserved.
 PLASTICITY OF CORTICAL DENDRITES IN ADULT RATS 659

Morest (IS), however, suggested the possibility of a limited degree of

neuronal growth in adulthood. A study of Smit ct al. (25), furthermore,
indicated that a potency for further outgrowth of basal dendrites of pyram-
idal cells could still be present at the terminal segments of cells in the
superficial half of the visual cortex in the young adult rabbit. They spec-
ulated that, by this potency, basal dendrites of pyramidal cells could
possibly retain an ability for plastic changes in adulthood. In 1974, Ru-
ledge et al. (24) observed a further outgrowth in apical dendrites after
daily electrical stimulation of the cortex in six adult cats. Those investi-
gators detected an increase in dendritic branching and in the number of
spines on apical dendrites of layer II and III pyramidal cells in the cortex
contralateral to the stimulated side. That increase in dendritic branching
and spines, however, was artificially induced by the long-term electrical
stimulation of implanted electrodes. It would be of interest, therefore, to
examine whether or not environmental conditions can influence the geom-
etry of nerve cell processes in adult brain. We studied rats exposed to
various environmental conditions to answer the following questions. (i)
Does dendritic outgrowth occur in pyramidal cells in adult rats (naturally,
or by induction of differential experience) ? (ii) If so, which parts of the
dendritic tree grow ? (iii) How does the dendritic outgrowth in adulthood
compare with its growth during postnatal development as described in the
literature ?

MATERIALS AND METHODS

Envirowlental Conditions. Twenty-four sets of three littermates of pig-
mented male rats of the Berkeley S1 strain, each matched for body weight,
were housed in standard laboratory cages from weaning until Day 112.
At that time, each set of three littermates was distributed so that each
member of a litter was assignedto one of three groups. The first group of
24 animals, one from each litter, was killed at the onset of the experimental
period (Day 112) to provide baseline material (i.e., B group). The second
group was divided into eight subgroups of three animals each and were
housed separately in a standard laboratory cage (32 X 20 x 28 cm) to
study the effect of age (i.e., SC group). The third group was divided into
two subgroups of 12 animals each. These two subgroups were housed sep-
arately in a large “enriched” cage (70 X 70 X 46 cm) and were provided
each day with six or more toys from a standard pool to study the effect
of a relatively enriched environment [i.e., EC group, see (23)]. The SC
and EC groups lived in their respective conditions for 30 days (from Day
112 to Day 142).
Histological Procedwes. At Berkeley, the brains of all animals were
removed after intraperitoneal injection of Nembutal. The brains of 12 sets
660 UYLINGS ET AL.

were placed into 10% formol-saline and were stained with thionin. The
brains of the remaining 12 sets were immersed in Golgi-Cox fluid prepared
as described by Van der Loos (35). In Amsterdam, when impregnation
was found to be complete, the brains were processed according to the Van
der Loos’ (35) modification of the Golgi-Cox technique and sectioned 150
p thick in the coronal plane [see Fig. 1 in (30) 1. Alternate sections were
Nissl-counterstained (34). As in previous studies [e.g., (23) ] all animals
were coded to prevent experimenter bias and the code was broken upon
completion of the measurements.
Analysis of Cortical Thickness. The coronal sections stained with thio-
nin were used by Diamond for thickness measurements of the frontal
(M), somesthetic (S) , and occipital (V) cortices (Fig. 1) . Each section
was divided into dorsal and dorsolateral strips (B and C) and lateral
strips (D and E) of the cortex. The positions of the sections can be related
to Krieg’s areas (16) as follows. The M sections represent parts of areas
10 and 8 ; the S sections encompass area 4 and areas 3, 2, and 2a. For the
V sections (just anterior to the splenium), strip B is in area 18, strip C
in areas 17 and 18a, D in areas 39 and 41, and E in area 41. For further

FIG. 1. A schematic view of the rat brain, indicating the cortical areas examined for
cortical thickness. M, S, and V represent sections taken from, respectively, the frontal,
somesthetic, and occipital cortices. Each section is divided into dorsal and dorsolateral
strips (B and C) and lateral strips (D and E) of the cortex. See (8) for specific
localization.- = positions of cortical thickness measurements, and hatched areas
are positions of dendritic measurements.
 PLASTICITY OF CORTICAL DENDRITES IN ADULT RATS 661

v terminal

‘IntermedIate

ma,” shaft
terminal
tuft

main
wth
oblique
dendrites
shaft

FIG. 2. A schematic drawing of a cortical pyramidal cell, showing the subdivision

of dendrites.

procedural details the reader is referred to (8). The cortical thickness

measurementsare obtained from eleven sets of three rats each.
Analysis of Dendrites. Six sets of brains (18 animals) stained with the
Golgi-Cox method were used for dendritic measurements. The remaining
sets were discarded for a variety of technical reasons. Seventeen to 20
pyramidal cells per animal that appeared to be completely impregnated
were selected semirandomly for analysis. These cells were in layers II
and III of the visual cortex, mainly area 17, caudal to the splenium [Fig.
1, and see Fig. 10 in (16)]. Pyramidal neurons in layer II which
resembled the modified superficial pyramids of O’Leary (19, 20) were not
analyzed in this study. A total of 333 cells was measured with the semi-
automated tracking system of Dr. P. D. Coleman [University of Rochester,
New York ; for a detailed description see (6) 1. A 100~ oil-immersion
objective with a numerical aperture of 1.3 and a long working distance of
0.26 mm in a Leitz Orthoplan microscope was used for the dendritic
measurements. After tracking and measuring all dendrites of each pyram-
idal cell, the Data General 1220 computer calculated the order of each
dendritic segment as well as its curvilinear length, the distance of its
termination (bifurcation or end point) from the cell body center, and the
bifurcation angles. These data were used to calculate the following param-
eters with the aid of an IBM 1130 computer : (i) The number of dendritic
segments per neuron for each order. (ii) The length of intermediate and
terminal segmentsfor each order (Fig. 2). (iii) The distance of the bifur-
 662 UYI,IK(~S ET AL.

cation and tenninal points to the cell body-center. (iv) The intermediate
angle, the absolute difference of pairs of side angles, and the flatness of a
bifurcation (26, 31, 33).
The number of dendritic segments is an index of the branching frequency
of dendrites. Ordering of the dendritic segments was done by denoting the
segments arising directly from the cell body as being of the first order,
and the order number was raised by one beyond each bifurcation [ (32) ;
p. 73 in (35) 1. For this parameter no correction was made for the cutting
of dendrites, which is inherent in the use of 150-pm sections. The cal-
culated percentage of cut segments per order level is about the same for the
EC, SC, and B groups. The cut segments were, therefore, included in the
counts. In addition to order, we distinguished the dendritic segments with
respect to type in intermediate and in terminal segments (Fig. 2). The
bifurcations are categorized according to the types of the daughter segments
into four groups (29). In this report we present mainly the results on
basal dendrites ; for a more detailed report on apical dendrites we refer to
(29, 30).
Statistical Methods. In testing the significance of differences for each
variable, the parametric Student’s t-test was applied. If conditions for
applying the t-test were not fulfilled, nonparanletric tests were applied
(the Kruskal-Wallis test and/or the Mann-Whitney U-test). A paired
t-test was applied in testing the cortical thickness measurements. The per-
centage variation in terminal segment length for each rat was estimated
to examine whether or not the sample size of six rats for each environ-
mental condition was sufficient. For this purpose, the percentage variation
in terminal segment length per order was calculated among rats for each
condition. The F-test was applied to demonstrate whether or not the
existence of the added variance component among rats (from different
litters) was significant. This could be applied because the terminal segment
lengths were normally distributed and the sample was a random one. The
best fitting theoretical frequency distribution was determined for each set
of observations forming a frequency distribution. The theoretical frequency
distributions applied were the normal, the log normal, and the gamma-
distribution. The normal and lognormal distributions are characterized by
the two parameters, the n1ean and the variance of n and 1~ x, respectively.
The gamma-distribution is characterized by the three parameters, 01,p, and
y. The probability density function of the gamma-distribution is

j-(x; a, p, y) = -& e-‘S-‘l’/B “;2 ?-I, x > a, /3 > 0, and y > 0.

( >

a is the origin parameter, i.e., the lowest possible value of x; p is the

scale parameter and determines the scale of both f(~) and X; and y is the
shape parameter of the distribution. For 1 < y < 2, the density function
 PLASTICITY OF CORTIC’AL 1~1’Sl~RITES lri ~U1UJ.T RATS 663

is unimodal, skewed toward higher values of X, and shows an inflection

point at the right side of the mode. For y > 2, the density function is uni-
modal, skewed toward higher values of .r and has two inflection points, one
to the right and one to the left side of the mode. For y > 8, the density
function approximates the density function of a normal distribution (IS).
In an iterative fashion, values for p and y were selected so that the goodness
of the fit was the best. The ~olmogorov-Sminiov test was applied as the
test for goodness of fit between the obserl-ed and the theoretical frequency
distributions (27).
The measuring error of the tracking system and the operator was esti-
mated by tracking the basal dendrites of a pyramidal cell repeatedly ten
times. The “standard error” in the length -of 4.5 dendrite segments- and
in the distance calculations of this repetition was determined.

RESULTS

Thichrss of Ncocortex

The thickness measurements of the frontal, somesthetic, and occipital

cortices are given in Table 1. In these cortical regions no significant
difference was observed between the 142-day-old SC group and the 112-
day-old baseline (B) group, except in the 5’ section, strip D where a
significant decrease was fom7d in the SC group relative to the B group at
P < 0.05. However, the cortex of the EC group was significantly thicker
in the frontal and the occipital regions (Table 1) compared to the B group,
with the largest increase occurring in the visual area (strip B, 7%).
Therefore, age had a small decreasing effect on cortical thickness, whereas
the EC condition resulted in a significant increase in cortical depth.

The segments counts of the basal dendrites are given in Table 2. It is

seen that the number of first-order segments is the same for all three
groups, which is equivalent to the statement that the mean number of basal
dendrites per neuron did not change under either experimental condition
(SC, EC). The segment counts of higher order, however, showed signifi-
cantly higher values for the EC and SC groups compared to the B group,
with the EC group showing a somewhat higher value for all higher orders
than the SC group. When the total number of segments per neuron was
counted irrespective of order, the branching frequency for the EC group
was found 8% higher than was the frequency for the SC group.
This overall difference is significant according to Stuclent’s t-test (P <
0.05). There seems, therefore, to be a relatively large effect of age on
 a
TABLE 1 E
Cortical Thickness (Layer I Excluded)

Cortical Bb SCb ECb Difference

area” (CT)
Mean SD Mean SD Mean SD EC vs. EC SC vs. Bd EC vs. SCe
(rm) (rm) bm) (4 (pm) (w-4

M
B 1749 50 1727 48 1803 48 3t -1
c 1807 63 1803 44 1838 64 2 0

S
B 1629 64 1598 73 1674 52 3 -2
C 1669 85 1656 57 1714 67 3 -1
D 1465 88 1492 82 1510 68 3 2

V
B 1083 25 1079 42 1159 29 7f 0 7J F
C 1114 39 1114 35 1163 37 4” 0 4”
D 1270 56 1230 37 126.5 39 0 -39 30
E 1239 60 1203 87 1239 72 0 -3 3

0 For explanation of M, S, and V, and B, C, D, and E, see Fig. 1.

6 N = 11 animals.
c [lo0 x (EC - B)/B] $&.
d Cl00 X (SC - B)/B] YO.
e Cl00 X (EC - SC)/SC] $&.
f P < 0.001 (according to paired t-test, df = 10).
9 P < 0.05.
hP < 0.01.
PLASTICITY OF CORTICAL DENDRITES IN ADULT RATS 66.5
666 UYLINGS ET AL.

Li
Ii

+- -+-- + +
 PLASTICITY OF CORTICAL DENDRITES IN ADULT RATS 667

13asalDendrites: Parameters of Rest Fit, Gamma-Distribution (u = 0)

Order H

Pa B (m) Y No. P b’ (rm) y No

Intermediate segment length

1 <O.OS 7.8 2.4 432 0.44 7.7 2.7 4.51 0.12 9.0 2.5 4i0
2 0.21 12.1 1.7 457 0.17 1.l.Y 1.4 551 0.35 1S.S 1.4 567
3 0.91 16.1 1.5 25.5 0.x3 li.3 1.4 370 0.86 19.0 1.3 405
4 0.95 13.0 1.i 83 O.Y7 17.6 1.5 121 0.81 20.3 1.2 1.~6
5 - - 15 0.Y4 24.8 1.3 26 O.YS 25.6 1.1 30
6 1 - 2 - 6

Terminal segment lengtl~

1 10 - - 8 - - -
2 O.Y4 15.8 7.2 113 &ii 22.7
-- 5.4 110 0.81 16.3 8.6 112
3 O.Y4 1Y.l 5.4 253 0.50 24.2 4.7 303 O.Y6 18.2 6.9 288
4 0.80 18.6 5.5 162 0.87 17.2 6.2 262 0.99 17.4 6.5 269
5 0.x4 21.Y 3.8 60 0.93 11.0 9.2 10.1 0.x0 16.0 6.6 112
6 - 13 O.Y8 14.4 6.5 2X - 1X
7 - 0 - - 3 - 4

neuronal branching accompanied by an additional smaller environmental

effect.

The intermediate and terminal lengths of the basal dendrites and of the
apical main shaft, terminal tuft, and oblique dendrites were measured
separately. Only the measurements of the segment lengths of basal den-
drites and of the apical main shafts are reported here. Some data on
oblique dendrites are reported in Uylings et al. (30).
Intermediate Scpcnts of Basal Dendrites. The frequency distributions
of these segments of the cells measured from the three environmental condi-
tions (B, SC, EC) were unimodal and skewed toward higher values (Fig.
3). These observed frequency distributions were fitted to a normal, a
lognormal, and a gamma-distribution function. A gamma-distribution func-
tion had the best fit except for the first-order intermediate segments of
the B and EC groups, for which a lognormal distribution function gives
a better fit. The parameters of the best fitting gamma-distribution func-
tions are given in Table 3. The P values which indicate the goodness of the
fit are also tabulated, and show that only one fit deviates significantly from
the observed frequency distributions. The mean and SE values of the inter-
mediate segment lengths for the three conditions are presented in Table 4
and Fig. 4. These values cnmpare favorably with measurements reported
by other investigators (17, 25). The one-tailed Mann-Whitney U-test
(Table 4) showed that the segment length of all orders, except order 1, did
668 UYLINGS ET AL.

not deviate significantly among the three environmental groups. The inter-
mediate segment length of the first order for the EC rats was significantly
larger than that of the SC, and the SC was in turn significantly larger than
that of the B group. According to the Kruskal-Wallis test, the inter-
mediate segment lengths of all orders for each condition could not be
treated as one population (P < 0.05). This result could be attributed to the
lower values of order 1 and 2 intermediate segments. Orders 3, 4, 5, and 6
intermediate segments, however, did not differ significantly for each
condition.
Terr&naZ Segments of Basal Dendrites. The frequency distributions of
the terminal segment lengths were unimodal and almost symmetrical (Fig.
5). These frequency distributions did not deviate significantly from a
normal distribution function, and the gamma-distribution function showed
the best fit for the distributions of the terminal segment lengths. The
parameters of the best fitting gamma-distribution function and the P values
are shown in Table 3.
The mean and the SEM values of the terminal segment lengths for the

SEGMENT LENGTH
Pm

.O B
7
175 - A A SC
. q EC

150 -

125 -

100 -

75 -

50 -

25 -

FIG. 4. Plots of the mean and SEM values versus order of lengths of intermediate
segments (open symbols) and terminal segments (solid symbols).
 TARLE 4
Length of Intermediate Segments of Basal Dendrites

Order B SC EC Mann-\Vhitney U-test P value <

Mean SEM No. Mean SEM NO. Mean SEM No. EC > B SC > B EC > SC
(ml (rm) Cm) (km) (m) bm)

1 19.1 0.6 432 20.8 0.6 451 22.7 0.7 470 0.001 0.01 0.05
2 20.0 0.8 467 19.7 0.8 551 21.1 0.8 567 NS NS NS
3 24.2 1.3 255 24.8 1.1 370 25.3 1.1 40.5 NS NS NS
4 22.6 1.9 83 26.2 1.9 122 24.9 1.8 1.56 NS NS NS
5 26.5 5.0 1.5 32.8 4.9 26 28.1 4.5 30 NS NS NS
6 - - 1 - - 2 - - 6 - - -
670 UYLINGS ET AL.

three environmental conditions are given in Table 5 and Fig. 4. As for the
lengths of terminal segments in the B rats, they compare favorably with
measurements reported by Lindsay and Scheibel (17) for superficial
pyramidal cells in the visual cortex of adult albino rats. It appears from
Table 5 that the lengths of all terminal segments of the EC rats are signifi-
cantly larger than those of the B rats, except in order 6 (Student’s t-test).
Terminal segments of all orders, and particularly of orders 1, 2, and 3,
were longer in the EC than the SC animals. The difference in length
between the EC and the SC groups was larger than that between the SC
and the B groups (Table 5). Thus, the environmental effect on termina1
segment lengths is greater in adult rats than is the age effect.

TERMINAL SEGMENT LENGTH

nr. B SC. EC.
10

FIG. 5. Frequency distribution of lengths of basal terminal segments in B, SC, and

EC rats. Class width is 1 pm.
PLASTICITY 01; CORTICAL DENDRU’ES IN ADULT RATS 671
672 UYLINGS ET AL.

According to the Kruskal-Wallis test (P < O.Ol), it is not justified to

treat the lengths of all terminal segments irrespective of order as one pop-
ulation, as has been done by other investigators [p. 84 in (5) ; (17) ; (14) 1.
Only the terminal segments of orders 4, 5, and 6 were not significantly
different for each group, except order 5 in the B group. The F-values
showed (Table 6) that the presence of the “added variance component
among” rats is significant only for order 3 of the EC group and order 4
of all three (B, SC, EC) groups. This table also shows that the percentage
variation among rats from different litters is always smaller than lo%,,
which indicates that the variance of terminal segment lengths for every
environmental condition is, for the most part, determined by the variation
of lengths within individual rats. This indicates that the number of rats
used for each environmental condition, six, was sufficient.
Intermediate Segments of Apical Main Shaft. The mean and SEM values
of these segments for the three environmental conditions are reported in
Uylings et al. (30). The range of mean values for all orders for every
condition was larger than that of the intermediate segments of the basal
dendrites (23 to 50 F) . The intermediate segment lengths of the apical
main shafts did not differ significantly between the three conditions. The
only exceptions were found in the 2nd order between the EC and the B
groups and in the 4th order between the SC and the B groups.
Terminal segments of apical main shaft. The number of these segments
was too small to test differences between conditions. However, the data sug-
gest that they are considerably longer than the intermediate segments of
the apical main shafts. The average length of a terminal segment was about
150 @n.

TABLE 6

Variation in Terminal Segment Length among Rats per Condition

Order B SC EC

F. Variation F* Variation F* Variation

among among among
animals animals animals
(%I (%I (%I

2 1.05 0.3 0.87 0.0 2.18 6.0

3 0.96 0.0 1.59 1.2 4.5.5- 7.1
4 2.39b 4.9 5.3oa 9.1 3.908 6.2
5 0.89 0.0 1.47 2.8 0.87 0.0

aP < 0.01.
bP < 0.05.
 PLASTICITY OF CORTICAL DENDRITES IN ADULT RATS 673

Standard Meantring Error

Length measurements of basal dendrites showed that nearly all inter-

mediate segments were shorter than 40 pm, and nearly all terminal seg-
ments were longer than 40 pm (Figs. 3, 5). A ten-times repetition of length
measurements of 45 basal dendritic segments showed that 7S70 of the
segments with a mean smaller than 40 pm had a standard measuring error
smaller than 2 pm, and that 78% of the segments with a mean larger than
40 f*rn showed a standard error smaller than 3 pm.

Distance fro11~ Cell Ceftter

The mean distance from the cell body center to bifurcations to order 5
of basal dendrites was significantly smaller than the distance from the body
center to the terminal tips of all orders. Thus, the bifurcation zone is
located within the ending zone. The mean distance between terminal tips
and cell body center was longer in the EC rats. Similar to the lengths of
terminal segments, the differences in mean distance of the terminal tips to
cell body center between the EC and the SC groups were larger than those
between the SC and the B groups (30).

Bifurcation Angles
The analysis of the bifurcation angles showed that they did not differ
significantly for the three environmental groups. A detailed analysis has
reported in Uylings (29).

DISCUSSION
Mode of Grozulth of Basal Defzdrites ill Adulthood
The present quantitative analysis revealed that the basal dendrites of
layers II and III pyramidal neurons of the EC and SC rats showed length-
ening in the terminal segments and an increase in branching relative to
the B animals. The lengths of the intermediate segments were approx-
imately equal in the three experimental groups. If the observed increase in
branching occurred at the IeveI of the intermediate segments, one would
expect a shortening in the length of these segtnents in the EC and the SC
groups. This, however, was not observed in the basal dendrites (Fig. 3,
Table 4). (Herewith, it is assumed that lengthening of intermediate seg-
ments does not occur within a dendrite keeping all its branches, owing to
the packed intermingling of the cortical components.) The results, there-
fore, suggest that the increase in branching occurred predominantly at the
terminal segments. Terminal segments in the B group were found to be
considerably longer than the intermediate segments (Figs. 3, 5). If branch-
ing occurred at the termina1 tips, one would expect the mean length of the
674 UYLINGS ET AL.

intermediate segments in the EC and SC groups to increase in relation to

the mean length of the intermediate segments in the B group. This, how-
ever, was not observed, except for segments of the first order. These
findings strongly indicate that the new branches were mostly formed at a
considerable distance from the tips of the terminal segments. An increase
in the length of the first order intermediate segments due to branching at
the terminal tips of the first order terminal segments is unlikely in view of
the low number (N = 10) of these segments in the B group and the distri-
bution of the intermediate segment lengths in the EC and the SC rats.
Therefore, for adult rats of the EC and the SC groups it appeared that (i)
basal dendrites increase their branching predominantly at their terminal
segments, and at a considerable distance from the tips, and (ii) terminal
segments of basal dendrites increase in length.
The mode of growth of basal dendrites in these adult rats did not differ
from the mode of growth of dendrites during development. Berry and his
co-workers (3, 4, 14) reported that terminal growth occurred both in the
basal dendrites of layer V cortical pyramidal cells and in the developing
dendrites of Purkinje cells in the cerebellum. Furthermore, Ten Hoopen
and Reuver (28) deduced that during development basal dendrites grew
by branching at a considerable distance from the elongated terminal tip.

Eflects of Differential Experience im Adulthood

The present findings show a significant increase in thickness of frontal

and especially occipital cortex in the 142-day-old EC rats relative to both
the 112-day-old B rats and the 142-day-old SC rats, and an increase in the
branching frequency and length of terminal segments of area 17 pyramidal
neurons in both EC and SC rats relative to the B rats. In a normally
developing rat cortex the thickness increases until about Day 26 postnatal
life, decreases slightly from Day 26 until Day 106, and is approximately
constant afterward (9). The larger cortical thickness of the EC rats, there-
fore, can be considered to be induced by an environmental condition lasting
30 days and not by age, especially as the cortex thickness of the SC rats
is slightly smaller than that of the B rats that are 30 days younger.
Rats of the SC group showed an increase in dendritic length at Day
142 relative to the B group (Day 112). This could have been induced by
age only, although the possibility cannot be excluded that recaging and
increase in social exploration at Day 112 were the responsible factors. The
increase in the branching and length of the terminal segments of dendrites
of layers II to III pyramidal neurons in the “enriched condition” (EC)
group was higher than the SC group, significantly in orders 1, 2, and 3.
Given the finding of a radial extension of the basal dendrites, and given
the assumption that the diameter of already existing dendritic segments
 PLASTICITY OF C‘ORTICAL DENDRITES IN ADLILT RATS 675

did not decrease, it can be concluded that the EC produced an increase in

dendritic surface area. This could provide such neurons with additional
area for establishing synaptic contacts with other nerve cells. Growth cones
are considered to be indicative of growing neuronal processes (18). Such
growth cones were not observed in our Golgi-Cox material, which does
not exclude the possibility, however, that further outgrowth of dendrites
indeed occurred (21, 24).
In developing rats, Greenough and Volkmar (13) detected a relative
increase in branching of (basal and apical) dendrites and GIobus et al.
(11) reported an increase in numbers of dendritic spines induced by
environmental enrichment. In adult cats Rutledge et al. (24) observed a
further outgrowth in apical dendrites in layers II to III pyramidal cells in
the cortex contralateral to the stimulated suprasylvian gyrus. Furthermore,
some preliminary findings of increased branching in apical dendrites (but
not in basal dendrites) of layer V pyramidal cells in young adult rats were
also reported by Greenough (12). Effects of differential experience in
adulthood are reviewed in Uylings et al. (30), and include an increased
rate of glial multiplication in the rat neocortex (1) ; an increase in cortical
thickness [ (8) and this report] ; in cortical weight (2, 22) ; in the ratio of
RNA to DNA (Bennett, personal communication) ; and an improvement
of learning capacities (7). Thus, the effects of differential experience are
not limited to a short, “critical” period in the rat’s life. This implies that it
would be important to examine the possible therapeutic or recovery influ-
ences of enriched environments SWS~L Zatu upon animals that have lesions,
are deprived, or are malnourished. Some work along these lines has in fact
already appeared in the literature [see (30) for references].

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